SQ Prove Operating Manual en 2021 04

Operating Manual
Spectroquant® Prove
Spectroquant® Prove
Spectrophotometer 100 ● 300 ● 600
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Contents
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1 Spectrophotometers .................................................. 8
1.1 Photometry ............................................................................................................. 8
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1.2 The Spectrophotometers........................................................................................... 9
2 Photometric Test Kits............................................... 10

2.1 Basic Principle ....................................................................................................... 10
4
2.1.1 Spectroquant® Cell Tests.............................................................................. 10
2.1.2 Spectroquant® Reagent Tests ...................................................................... 11
2.2 Notes for Practical Use............................................................................................ 12
2.2.1 Measuring Range........................................................................................ 12
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2.2.2 Inuence o pH .......................................................................................... 13
2.2.3 Inuence o Temperature ............................................................................ 13
2.2.4 Time Stability ........................................................................................... 14
2.2.5 Inuence o Foreign Substances ................................................................... 14 6
2.2.6 Dosing the Reagents ................................................................................... 15
2.2.7 Shelf-life of the Reagents ........................................................................... 15
3 Sample Preparation ................................................. 16 7

3.1 Taking Samples .................................................................................................... 16
3.2 Preliminary Tests .................................................................................................. 16
3.3 Dilution ................................................................................................................ 17
3.4 Filtration .............................................................................................................. 18 8
3.5 Homogenization..................................................................................................... 19
3.6 Decomposition ...................................................................................................... 19
4 Pipetting System...................................................... 21 9
5 Analytical Quality Assurance (AQA) ......................... 22
5.1 Quality Control at the Manufacturer ......................................................................... 22
5.2 Quality Control for the User .................................................................................... 24 10
5.2.1 Checking the Spectrophotometer.................................................................. 25
5.2.2 Checking the Overall System ....................................................................... 26
5.2.3 Checking the Pipettes ................................................................................. 27
5.2.4 Checking Thermoreactors ........................................................................... 27 11
5.2.5 Testing or Handling Errors ......................................................................... 28
5.3 Determination o Sample Inuences (Matrix Eects) .................................................. 28
5.4 Denition o Errors................................................................................................. 28
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6 Overview ................................................................. 30
6.1 Scope of Delivery................................................................................................... 30
6.2 Overview of the Instrument..................................................................................... 30
6.3 Display and User Interace ...................................................................................... 31 13
7 Safety ...................................................................... 39
7.1 Intended Use ........................................................................................................ 40
7.2 General Safety Instructions ..................................................................................... 40 14
7.2.1 Function and Operational Saety .................................................................. 40
7.3 Target Group and User Qualication ......................................................................... 40
7.4 Handling Hazardous Substances............................................................................... 41
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8 Getting Started ........................................................ 42

8.1 General Notes on Handling ...................................................................................... 42
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8.2 Initial Setup .......................................................................................................... 42
8.2.1 Connecting the Power Supply ....................................................................... 42
8.2.2 First Power-on ........................................................................................... 43
8.2.3 Language Setup ......................................................................................... 44
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8.2.4 Date, Time and Country-specic Settings....................................................... 44
8.2.5 Self-test.................................................................................................... 45
8.3 Connecting Optional Peripheral Devices..................................................................... 46
5 8.3.1 Communication Ports .................................................................................. 46
8.3.2 Printer ...................................................................................................... 46
8.3.3 USB Memory Device ................................................................................... 47
8.3.4 Barcode Reader.......................................................................................... 47
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9 Operation................................................................. 48
9.1 Switching the Spectrophotometer On or O............................................................... 48
9.2 System Setup ....................................................................................................... 50
7 9.2.1 Information ............................................................................................... 51
9.2.2 Interace (Setup 1)..................................................................................... 52
9.2.3 Region (Setup 2)........................................................................................ 52
9.2.4 Quality (Setup 3) ....................................................................................... 55
8 9.2.5 Automation (Setup 4) ................................................................................. 57
9.2.6 User Management ...................................................................................... 59
9.2.7 Service ..................................................................................................... 60
9.2.8 Updates .................................................................................................... 62
9 9.2.9 Network and Prove Connect ......................................................................... 63
9.3 Measurements....................................................................................................... 64
9.3.1 Performing a Measurement .......................................................................... 65
9.4 Zero Adjustment ................................................................................................... 67
10 9.4.1 Notes on Zero Adjustment .......................................................................... 67
9.4.2 When to Repeat the Zero Adjustment? ......................................................... 68
9.4.3 Zero Adjustment or Concentration Measurement Methods ............................... 69
9.4.4 Zero Adjustment or Absorbance/Transmission Measurements (Ad hoc menu)..... 69
11 9.4.5 Zero Adjustment or Spectrum Measurements ............................................... 70
9.4.6 Zero Adjustment or Kinetic Measurements ................................................... 70
9.5 Method List ........................................................................................................... 70
9.5.1 Selecting a Method Manually ....................................................................... 70
12 9.5.2 Searching and Filtering the Method List ........................................................ 71
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9.6 Programming a User-dened Method ........................................................................ 72

9.6.1 User-dened Concentration Methods ............................................................. 73
9.6.2 Calibration Data and Calibration Function or Single-wavelength Methods........... 73 3
9.6.3 Programming/Modiying User-dened Methods (Single Wavelength) .................. 74
9.6.4 Calibration Data and Calibration Function or Special Methods
(e. g. Multi-wavelength)............................................................................... 84
9.6.5 Programming/Modiying User-dened Special Methods (e. g. Multi-wavelength)... 84 4
9.6.6 Programming a User-dened Spectrum Method .............................................. 88
9.6.7 Programming a User-dened Kinetic Method .................................................. 90
9.6.8 Copying/Duplicating a User-dened Method ................................................... 92
9.6.9 Modiying a User-dened Method rom the Method List .................................... 93 5
9.6.10 Deleting a User-dened Method rom the Method List ...................................... 93
9.6.11 Export User-dened Methods to a USB Memory Device .................................... 94
9.7 Measuring in Concentration Mode............................................................................. 95
9.7.1 Measuring Cell Tests with a Barcode.............................................................. 95
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9.7.2 Measuring Reagent Tests with AutoSelector.................................................... 97
9.7.3 Measuring Reagent-ree Tests and User-dened Methods ................................. 98
9.7.4 Exceeding the Upper or Lower Limits o the Measuring Range ........................... 99
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9.7.5 Method-specic Settings or Concentration Mode ...........................................100
9.7.6 Measuring Diluted Samples.........................................................................101
9.7.7 Sample Blank Value ...................................................................................102
9.7.8 Reagent Blank Value ..................................................................................104
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9.7.9 Automatic Turbidity Correction ....................................................................106
9.7.10 User Recalibration (Standard Adjustment) ....................................................107
9.7.11 MatrixCheck .............................................................................................111
9.7.12 User-dened Range ...................................................................................114 9
9.7.13 Dierentiation...........................................................................................116
9.7.14 Plausibility Check ......................................................................................117
9.8 Ad hoc Measurement (without selecting a specic method).........................................119
9.8.1 Ad hoc ABS/TRANS Measurement.................................................................. 120 10
9.8.2 Ad hoc Spectrum Measurement ...................................................................121
9.8.3 Ad hoc Kinetic Measurement .......................................................................123
9.9 Spectrum.............................................................................................................125
9.9.1 General Information ..................................................................................125 11
9.9.2 Recording the Spectrum .............................................................................125
9.9.3 Evaluating a Spectrum ...............................................................................127
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9.10 Kinetics ...............................................................................................................128

9.10.1 General Information ..................................................................................128
3 9.10.2 Recording Kinetics .....................................................................................128
9.10.3 Evaluating a Kinetic ...................................................................................130
9.11 AQA (Analytical Quality Assurance) .........................................................................131
9.11.1 Spectrophotometer Monitoring (AQA1) .........................................................131
4 9.11.2 Total System Monitoring (AQA2) ..................................................................133
9.11.3 AQA Overview...........................................................................................134
9.11.4 Perform AQA1 Status Check........................................................................135
9.11.5 Perform AQA2 Status Check........................................................................136
5 9.11.6 AQA1 Selection List ...................................................................................137
9.11.7 Activating and Deactivating an AQA1 Test .....................................................138
9.11.8 Editing an AQA1 Test .................................................................................138
9.11.9 Performing an AQA1 Test............................................................................141
6 9.11.10 Creating a User-dened AQA1 Test .............................................................143
9.11.11 AQA2 Selection List ...................................................................................144
9.11.12 Activating and Deactivating an AQA2 Test .....................................................145
9.11.13 Editing an AQA2 Test .................................................................................145
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9.11.14 Performing an AQA2 Test............................................................................148
9.11.15 Perform PipeCheck ....................................................................................150
9.12 Timer ..................................................................................................................153
9.13 Results and Measurement Datasets ...........................................................................154
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9.13.1 Displaying the Results ...............................................................................155
9.13.2 Show Details of One Result .........................................................................156
9.13.3 Filtering Results to Further Process Measurement Datasets .............................157
9 9.13.4 Panorama – Value Control Card ..................................................................158
9.13.5 Print Results and Measurement Datasets ......................................................159
9.13.6 Deleting results ........................................................................................160
9.13.7 Export Results and Measurement Datasets....................................................160
10 Data Transfer from Spectroquant® Prove with USB stick ..................................160
9.14 User Management .................................................................................................162
9.14.1 Activating/Deactivating the User Management Function ..................................163
9.14.2 Create an Administrator/User Account..........................................................164
11 9.14.3 Edit or Delete a User..................................................................................165
9.15 Login/Logout ........................................................................................................166
9.15.1 Change Password with User Rights Only .......................................................167
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10 Maintenance and Cleaning ................................... 168

10.1 Exchanging the Buer Battery ................................................................................168
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10.2 Exchanging the Halogen Lamp (Prove 100)...............................................................169
10.3 Cleaning ..............................................................................................................170
10.3.1 Cleaning the Housing and Display ................................................................170
10.3.2 Cleaning the Cell Compartment ...................................................................171
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10.3.3 Cleaning the Cell Compartment Cover and the Rear Cavity..............................173
10.3.4 Cleaning the Detector Lens .........................................................................174
11 Error Causes and Trouble-shooting ...................... 176 5

12 Technical Data ..................................................... 178
12.1 Spectroquant® Prove 100 .......................................................................................178
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13 Accessories and Test Media ................................. 184

13.1 Accessories ..........................................................................................................184
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13.2 Optional Equipment/Connection Cables ....................................................................184
13.3 Test Media ...........................................................................................................185
14 Appendix.............................................................. 186 8
15 List of Smart Icons on Display ............................. 188
16 Contents o log les ............................................. 195
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1
1 Spectrophotometers
1.1 Photometry
When a beam o light is transmitted through a ized using the transmittance T (or, respectively,
3
colored solution, then this beam loses its intensi- T in percent).
ty, in other words a part o the light is absorbed
by the solution. Depending on the substance T = I/I0
in question, this absorption occurs at specic
4
wavelengths. I0 = Initial intensity of the light
I = Intensity o the transmitted light
Monochromators (e. g. narrow-band intererence
5 lters, lattices) are used to select the wave- I the light is not absorbed at all by a solution,
length from the total spectrum of a tungsten- then this solution has a transmittance of 100 %;
halogen lamp (VIS spectrum), a deuterium lamp a complete absorption of the light in the solution
(UV spectrum) or, respectively, a xenon lamp. means 0 % transmittance.
6 The intensity of the absorption can be character-
The measure generally used or the absorption
o light is the absorbance (A), since this cor-
relates directly with the concentration o the
7 absorbing substance. The following connection
exists between absorbance and transmittance:
A = –log T
8
Experiments by BOUGUER (1698–1758) and
LAMBERT (1728–1777) showed that the ab-
sorbance is dependent on the thickness o the
9 absorbing layer o the cell used. The relationship
between the absorbance and the concentration
o the analyte in question was discovered by
BEER (1825–1863). The combination o these two
10 l natural laws led to the derivation o Lambert-
c Beer’s law, which can be described in the orm o
l0 the ollowing equation:
11 A = ελ · c · d
d ε λ = Molar absorptivity, in l/mol × cm

d = Path length of the cell, in cm
12 c = Concentration o the analyte, in mol/l
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8 Version 3.0 – 04/2021
1 Spectrophotometers – 1.2 The Spectrophotometers 1
1.2 The Spectrophotometers

The spectrophotometers that belong to the
3
Spectroquant® analysis system dier rom con-
ventional spectrophotometers in the following
important aspects:
4
• The calibration functions of all test kits are elec-
tronically stored
• The measurement value can be immediately
read o rom the display in the desired orm 5
• The method or the test kits (cell tests and
reagent tests)
belonging to the Spectroquant® analysis sys-
tem is automatically selected via the scanning 6
o the bar code
• All cells ormats used are automatically identi-
ed and the correct measuring range is select-
ed automatically 7
• Instrument-supported AQA ensures that mea-
surement results can be used as secure, repro-
ducible, and recognized analytical results
• New methods can be downloaded rom the in- 8
ternet site www.sigmaaldrich.com/photometry
and permanently stored in the instrument
For technical data and instructions or use please 9

refer to section 6 and the Analytical Procedures
and Appendices.
They can also be ound in the internet.
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Version 3.0 – 04/2021 9
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2 Photometric Test Kits
2.1 Basic Principle 2.1.1 Spectroquant® Cell Tests

By means of reagents, the component of a
3
sample to be analyzed is converted into a col-
1
ored compound in a specic reaction. The re-
6
agents or reagent mixtures contain – in addi-
tion to the reagent selective for a parameter
4
to be determined – a number o auxiliary sub-
stances that are essential for the course of the 2
reaction. These include, or example, buers 3 7
5 or adjusting the pH to the optimal value or the
reaction, and masking agents that suppress or 4
minimize the inuence o interering ions.

7
6 The color reactions are in most cases based

8
on standardized analytical methods specically
optimized in terms o ease o use, a low working
10
eort, and shorter reaction times. Furthermore,
7 methods cited in the literature or developed by
ourselves are also used. Details on the respec-
tive reerence procedures are stated in the pack-
age insert or else in the parameter overview.
8
5 9
9 1 Identication mark or the correct insertion

into the cell compartment of the spectropho-
tometer
2 Cat. No. of test kit
10 3 Designation of test kit
4 Risk phrases
5 Special cell in optical quality
6 Leakproof cap
11 7 Bar code or identication in the NOVA and
Pharo photometers
8 Details regarding contents
9 Highly precise dosage o the reagent
12 10 2D barcode or identication in the Prove
spectrophotometers
Additional reagent(s)
13
Certain cell tests, e. g. COD or nitrite, already
contain all necessary reagents in the cells, and
the sample must merely be added with a pi-
pette. In other tests, however, for reasons of
14
chemical compatibility it is necessary to sepa-
rate the test into two or three dierent reagent
mixtures. In such cases, besides the sample a
15 metered reagent must also be added.
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10 Version 3.0 – 04/2021
2 Photometric Test Kits – 2.1 Basic Principle 1
2.1.2 Spectroquant® Reagent Tests

3
The principle behind the reagent tests is that 6

the reagents necessary for the color reaction
are combined in the orm o liquid concentrates
or solid-substance mixtures. A ew drops o the
reagent concentrate are added to the sample. 7
This means that there is no need to dilute the
sample, which in turn enhances the sensitivity
o the detection. The procedure generally used
in classical photometry by which the sample is
8
made up to a dened volume in a volumetric
ask is dispensed with.
9
The method is selected automatically by means
o the scanning o the bar code by the
AutoSelector. All cells ormats used are auto-
matically identied and the correct measuring
10
range is selected automatically. Subsequently
the result is automatically shown on the display.
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Version 3.0 – 04/2021 11
1 2 Photometric Test Kits – 2.2 Notes for Practial Use
2.2 Notes for Practical Use

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2.2.1 Measuring Range
In photometry it is conventional practice to mea-

sure against the reagent blank value. Here the
4
analysis is carried out "blind", i.e. without any
analyte added. Instead o the sample volume,
the corresponding quantity o distilled or DI wa-
ter is used.
5
This reagent blank value is prestored in the
Absorbance
spectrophotometers belonging to the

Spectroquant® analysis system, which means
6 that – due to the high batch reproducibility – it
is possible to dispense with a separate measure-
ment of the reagent blank. At the lower limit
of the measuring range, the accuracy of the
7 determination can be enhanced by perorming
the measurement against a separately prepared
reagent blank. In some cases the intensity of
the color o the solution and thus the absorbance
8 can drop again when very high concentrations o
Concentration the analyte are present (see package insert o
test kit).
The intensity o the color o a solution, measured
9 as the absorbance, is proportional to the con-
centration of the respective analyte only within
a specic range. This measuring range (eective
range) is electronically stored in the spectropho-
10 tometers or each individual test kit.
Below the specied measuring range, either a

dierent cell or else another procedure must
11
be used. The lower limit of the measuring
range either takes the form of nonlinearity of
the calibration curve, as shown in the gure, or
else is given by the method detection limit. The
12
method detection limit of an analytical meth-
od is the lowest concentration o the analyte in
question that can be measured quantitatively
13 with a dened degree o probability (e. g. 99 %).
The upper limit of the measuring range is
the point at which the linear correlation between
the concentration and the absorbance ends. In
14 such a case the sample must be diluted accord-
ingly so that it lies ideally in the middle o the
eective range (least-error measurement).
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2 Photometric Test Kits – 2.2 Notes for Practial Use 1
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2.2.2 Inuence o pH 2.2.3 Inuence o Temperature
Chemical reactions follow an optimal course
only within a certain pH range. The reagents
contained in the test kits produce an adequate
4
buering o the sample solutions and ensure
that the pH optimal or the reaction in question
is obtained.
5
Absorbance
Strongly acidic (pH <2) and strongly alkaline (pH
>12) sample solutions can prevent the pH rom
being adjusted to an optimal range, since under
6
certain circumstances the buering capacity
o the test-kit reagents may not be sufcient.
Any necessary correction is made by the drop-
wise addition o diluted acid (reduces the pH) or 7
diluted lye (raises the pH), testing the pH with
suitable indicator strips ater each drop is added.
The addition o the acid or lye results in a dilu-
tion o the test solution. When up to ve drops 10 20 30 40
8
are added to 10ml o sample, the change in the Temperature (°C)
volume can be neglected, since the resultant er-
ror is lower than 2 %.
The temperature o the sample solution and the
The addition o larger quantities should be duly
reagents may have an eect on the color reac- 9
considered by adjusting the sample volume ac-
tion and thus on the measurement result. The
cordingly.
typical temperature course is illustrated in the
gure.
The specied pH values or the sample solution 10
and, wherever applicable, or the measurement
If the sample temperature is lower than 15 °C,
solution are dened in the respective package
alse-low results must be reckoned with. Tem-
inserts and in the Analytical Procedures and Ap-
peratures exceeding 30 °C generally inuence
pendices. 11
the stability o the compound that is ormed in
the reaction. The optimal temperature for the
color reaction is stated in the package inserts o
the respective Spectroquant® test kits.
12
CAUTION
Ater thermic decomposition procedures, the de-
termination of COD or total contents of nitrogen, 13
phosphorus, or metal, a sufficient waiting time
must be allowed or to permit the solution cool to
room temperature.
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1 2 Photometric Test Kits – 2.2 Notes for Practial Use
2.2.4 Time Stability 2.2.5 Inuence o Foreign Substances

3 Foreign substances in the sample solution can
• raise the measurement value as a result of an

amplication o the reaction
4 • lower the measurement value as a result of a
prevention of the reaction
Absorbance
A quantication o these eects is stated in tab-

5 ular form in the respective package inserts for
the most important foreign ions. The tolerance
limits have been determined or the individual
ions; they may not be evaluated cumulatively.
6
Suitability for use in seawater
A tabular survey (see appendix 1) provides inor-
mation on the suitability of the tests in connec-
7 tion with seawater and also on the tolerances or
30 60 salt concentrations.
Reaction time (minutes)
8
Most of the color reactions require a certain time
to reach the maximum color intensity. The solid
curve in the gure at the right gives a schematic
9 impression of a typical time course. The behav-
ior of relatively instable color reactions with time
is shown by the dotted curve. The reaction time
specied in the working instructions reers to the
10 period o time rom the addition o the last re-
agent until the actual measurement. In addition,
the package inserts or the individual test kits
also state the time interval in which the mea-
11 surement value does not change. The maximum
time interval is 60 minutes; this time should not
be exceeded, even in the case o stable color
reactions.
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2 Photometric Test Kits – 2.2 Notes for Practial Use 1
2.2.6 Dosing the Reagents

Small amounts o liquids are dosed by counting At the rst use replace the black screw cap o the 3
the number o drops rom a leakproo bottle. reagent bottle by the dose-metering cap. Hold
the reagent bottle vertically and, at each dosage,
press the slide all the way into the dose-metering
cap. Beore each dosage ensure that the slide is 4
completely retracted.
CAUTION
5
Reclose the reagent bottle with the black screw
cap at the end o the measurement series, since
the unction o the reagent is impaired by the
absorption of atmospheric moisture.
6
CAUTION
When using dropper bottles it is extremely im-
portant that the bottle be held vertically and 7
that the drops be added slowly (approx. 1 drop 2.2.7 Shelf-life of the Reagents
per second). I this is not observed, the correct
The Spectroquant® test kits are in most cases
drop size and thus the correct amount o reagent
stable or three years when stored in a cool, dry
are not achieved. 8
place. A few test kits have a lower shelf-life of 18
or 24 months or must else be stored in a rerig-
A positive-displacement pipette should be used erator.
or larger quantities o liquid or or the exact
dosage o smaller reagent quantities. In these COD Cell Tests must be stored protected rom 9
cases the reagent bottles are not tted light. The expiry date o the package unit is
with a dropper insert. printed on the outer label. The shel-lie may be-
come reduced when the reagent bottles are not
reclosed tightly ater use or when the test kit is
10
stored at temperatures higher than those speci-
ed.
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Solid substances are dosed either with the

dose-metering cap or with microspoons that are
integrated into the screw cap o the respective 13
reagent bottle. The dose-metering cap is used
or solid reagents or reagent mixtures that are
ree-owing. In all other cases the substances
are dosed with the microspoon. 14
In this case it is necessary to add only level

microspoonfuls.
To this end the spoon must be drawn over the
15
brim of the reagent bottle.
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Version 3.0 – 04/2021 15
1
3 Sample Preparation
Sample preparation covers all the steps nec-

essary before the actual analysis can be per-
3 ormed.
4 3.1 Taking Samples

The taking o samples is the rst and most Parameter Preservation
important step on the way to obtaining the
COD +2 to +5 °C max. 24 h
5 correct analysis result. Not even the most exact
or
method o analysis can correct any mistakes –18 °C max. 14 days
made in the taking o the sample. The objective
N compounds: analyze immediately, only in ex-
o the sampling procedure is to gain a sample
NH4-N, NO3-N, NO2-N ceptional cases
6 with a representative composition. The most im- +2 to +5 °C max. 6 h
portant precondition or gaining a representa-
P compounds: short-term storage, no preserva-
tive sample is the identication o the suitable
PO 4-P, P total tion;
sampling site. Here it must be borne in mind with nitric acid to pH 1, max. 4
7 that the solution to be investigated can display weeks
varying concentrations in dierent places at di-
Heavy metals short-term storage, no preserva-
ferent times. tion;
with nitric acid to pH 1, max. 4
8 In sampling, a distinction is made between weeks
manual and automatic methods. In many cases
a true picture of the average composition of the
sample can be obtained only once several indi-
9 vidual samples have been collected; this can be
done manually or with an automatic sampler. 3.2 Preliminary Tests
Correct measurement results can be obtained
Clean plastic or glass containers with a volume only within the measuring range specied or
10 of 500 or 1000 ml are generally suitable for each individual parameter. When dealing with
collecting samples. They should be rinsed sev- sample solutions of an unknown concentration,
eral times, under vigorously shaking, with the it is advisable to establish whether the sample
water to be investigated, and then lled ree o concentration is indeed within the specied
11 air bubbles and immediately closed tightly. The measuring range, ideally roughly in the middle o
containers must be protected against the eects the range. Preliminary tests enhance the ana-
o air and heat and then be orwarded or the lytical reliability and make the determination o
further analytical steps as soon as possible. In the necessary dilution ratios in the case o high
12 exceptional cases, preservation measures in the concentrations easier. MQuant® Test Strips are
form of short-term refrigeration at +2 to +5 °C very well suited or preliminary tests.
and chemical conservation can be taken.
13
Further useul tips on suitable sample vessels or
specic analytes and possible preservation and
storage conditions can be ound in EN ISO
5667-3, Water quality – Sampling – Part 3: Pres-
14
ervation and handling o water samples
(ISO 5667-3:2018).
15
16
16 Version 3.0 – 04/2021
3 Sample Preparation – 3.3 Dilution 1
3.3 Dilution
Dilution o samples is necessary or two reasons: Example
3
Step 1: Make up 2 ml o sample to 200 ml
• The concentration o the parameter under with distilled water;
investigation is too high, i. e. it lies outside the DF = 100, dilution number 1+99.
measuring range
4
• Other substances contained in the sample Step 2: Take 5 ml o the above solution
interere with the determination (matrix inter- and make up to 100 ml;
erence); alse-high or alse-low results may DF = 20, dilution number 1+19.
ensue 5
The dilution actor or the total dilution is calcu-
The ollowing auxiliaries are absolute prerequi- lated by multiplying the individual dilutions:
sites or the dilution o the sample:
DFtotal = DF1 × DF2 = 100 × 20 = 2000, 6
• Volumetric asks o varying sizes (e. g. 50, 100 dilution number 1+1999
and 200 ml)
• Positive-displacement pipette Simplied procedure
• Distilled or DI water Dilutions up to 1:10 can also be prepared without 7
volumetric asks in a glass beaker, measuring
Only dilutions carried out with these auxiliary the volumes o the sample and the dilution water
products are o sufcient reliability in the area o using a previously calibrated positive-displace-
trace analysis, to which photometry belongs (or ment pipette (see table or instructions). 8
the simplied procedure see page 17).
An important aspect here is that once the volu- Desired Volume Volume Dilution Dilution
metric ask has been lled up to the mark with dilution of of dis- factor number
distilled water the ask must be closed and the sample tilled 9
contents thoroughly mixed. (ml) water
The dilution factor (DF) resulting from the (ml)
dilution procedure is calculated as ollows: 1:2 5 5 2 1+1

10
1:3 5 10 3 1+2
Final volume (total volume)
DF = 1:4 2 6 4 1+3
Initial volume (sample volume)
1:5 2 8 5 1+4
The analytical result is subsequently multi- 11
1:10 1 9 10 1+9
plied by the dilution factor.
A calculation can be dispensed with when
the dilution is programmed into the spec-
trophotometer. The dilution number (see 12
table below) is entered and the measurement
value is subsequently calculated correctly and
immediately displayed.
All dilutions should be made in such a way that
13
the measurement value lies in the middle o the
measuring range. As a rule, the dilution actor
should never be higher than 100. In the event
14
that yet higher dilutions become necessary all
the same, then this must be done in two sepa-
rate steps.
15
16
Version 3.0 – 04/2021 17
1 3 Sample Preparation – 3.4 Filtration
3.4 Filtration
Strongly turbid samples require pretreatment Procedure or Microltration
3
beore they can be determined in a spectropho-
tometer, since the eect o turbidity can result 1 2
in considerable variations in the measurement
values and in alse-high readings. Care must be
4
taken here to ensure that the substance to be
determined is not contained in the suspended
material, in which case a sample decomposition
5 must be carried out.
Compounds that always occur in dissolved orm

(or example ammonium, nitrate, nitrite, chlo- 1 Draw out the liquid 2 Screw the syringe
6 rine, chloride, cyanide, uoride, orthophosphate,
to be ltered with tightly into the
and sulate) permit a previous ltration, even
the syringe. ront side o the
when the sample solution is strongly turbid.
membrane lter
attachment.
7 Weak turbidity is eliminated by the automatic
turbidity-correction feature built into the
3 4
spectrophotometer (see section 9.7.9); in such
cases it is not necessary to lter the sample
8 before analysis.
As a measure to distinguish between dissolved

and undissolved water-borne substances, the
9 water sample can be ltered through a simple
paper lter. Following the recommendations
stated in the reerence methods, membrane l-
ters with a pore size o 0.45 µm are required or 3 Hold the syringe 4 Filter the contents
10 ne ltration. upright and slowly of the syringe into
depress the piston the intended glass
upwards until the vessel.
membrane lter is
11
ully wetted and
free of air bubbles.
12
13
14
15
16
18 Version 3.0 – 04/2021
3 Sample Preparation – 3.5 Homogenization – 3.6 Decomposition 1
3.5 Homogenization
As a measure to ensure that a representative
3
sample can be taken in the presence of sus-
pended matter in the water sample in question,
or certain parameters – e. g. COD and the total
content of heavy metals – the sample must be
4
homogenized. This must be carried out using a
high-speed blender (2 minutes at 5000 – 20000
rpm and taking the sample while stirring.
5
3.6 Decomposition
Water-borne substances can be present in the The manner in which the sample is pretreated 6
sample or investigation in a variety o orms: as enables the three proportions to be distinguished
the ion, bound more or less solidly in a complex, rom each other. This can be illustrated using
or as a solid substance. a copper-containing wastewater sample as an
example. 7
Ion Complex
Example
8
Filtration
9
Decomposition Decomposition Filtration
Total content Dissolved proportion Dissolved proportion 10

Solid substance Solid substances
Cu(OH)2 Complexes Cu-
Complexes Cu- EDTA
EDTA Ions Cu2+ Ions Cu2+
Ions Cu2+ Result B Result C 11
Result A
Proportion:
Ionogenic =C
Complex = B–C 12
Solid
substances = A–B
Total content = A
13
Decomposition converts the substance to be
determined into an analyzable orm. In most
cases, decomposition agents take the orm o
acids in combination with oxidizing agents; in 14
exceptional cases (e. g. in the determination o
total nitrogen) an alkaline decomposition is more
eective. The type o decomposition procedure
used depends on the analyte to be determined 15
and the sample matrix.
16
Version 3.0 – 04/2021 19
1 3 Sample Preparation – 3.6 Decomposition
The ready-to-use sample-decomposition prod- The necessity or decomposition can be checked
3
ucts Spectroquant® Crack Set 10 and 20 are according to the ollowing diagram.
suited or the preparation o the sample materi-
als or the determinations stated in the table.
Decomposition
4
Determination of Sample preparation with Procedure Procedure
Total phosphorus * Crack Set 10/10C **
Measurement Measurement
5 Total chromium * Crack Set 10/10C
[= sum o chromate and
Result A Result B
chromium(III)]
Total metal Crack Set 10/10C

6 [= sum o ree and
complex-bound metal]
Total nitrogen * Crack Set 20 Decomposition No A and B Yes No decomposition

necessary identical? necessary
7 * The decomposition reagents are already contained in the
packs of the respective cell tests.
** Decomposition cells are included in the pack; empty cells For wastewater with a consistent composition,
are required or the decomposition or Crack Sets 10 and
20. this check as a rule need be carried out only
8 once. It is, however, advisable to check the re-
The decomposition processes are carried out in sult periodically.
the Spectroquant® thermoreactor (capacity: 12
or 24 decomposition cells) at 120 °C or, respec-
9 tively, 100 °C. Details regarding the heating
times and urther treatment can be ound in the
package inserts contained in the Spectroquant®
Crack Set packs.
10
In the event that the sample to be analyzed is a
highly contaminated material (high proportion o
organic substances) or water-insoluble samples,
11
decomposition using concentrated acids and
other agents is indispensible. Corresponding ex-
amples from the collection of applications for
12 real samples are available on request.
13
14
15
16
20 Version 3.0 – 04/2021
1
4 Pipetting System
Positive-displacement pipettes permit

3
• an exact dosage o the sample volume
• a precise measurement o sample and reagent
volumes and o the volumes o water or dilu-
4
tion purposes
Pipettes o varying volumes and also ones with a

xed volume are available. 5
Sources of error and hints on how to avoid
them:
Closely ollow the instructions or use contained 6
with the pipette in question.
• Check the pipetted volumes

a) by weighing using analytical scales (weighing 7
accuracy ±1 mg), 1 ml o water at 20 °C =
1000 g ±1 mg
b) using Spectroquant ® PipeCheck; this is a
photometric check o the pipette, and scales 8
are not necessary (see section 5.2.3)
• Avoidance o spread eects by rinsing the
pipette several times with the solution to be
pipetted 9
• Always exchange the pipette tip
• Draw up the liquid slowly and depress piston
completely to discharge the liquid
10
11
12
13
14
15
16
Version 3.0 – 04/2021 21
1
5 Analytical Quality Assurance (AQA)
3
The objective o analysis must always be to de-
termine the true content of the analyte in ques- Certificate of Final Inspection (full details)
tion as accurately and precisely as possible.
Device Name: Spectroquant® Prove 600
4 Serial no: 2005614705
Analytical Quality Assurance represents a suit- Software version: 1.4.5
able and indispensible method by which the Wavelength Accuracy ⃰
quality o the user's own work can be assessed, Equipment Nominal value Tolerance limit ⃰ ⃰ Actual value Result
errors in the measurement system diagnosed,

241.15 nm 240.0 - 242.4 nm 241.9 nm P
5 Holmium Oxide Liquid Filter

Hellma 667-UV5
361.30 nm 360.1 - 362.5 nm 361.4 nm P
and the comparability with the results obtained 640.55 nm 639.4 - 641.8 nm 641.1 nm P
using the respective reerence methods demon- Wavelength Precision / Reproducibility ⃰

strated. Equipment Wavelength Nominal value Actual value Result
6 Holmium Oxide Liquid Filter

241.15 nm
361.30 nm
≤0.10 nm
≤0.10 nm
0.01 nm
0.02 nm
P
P
Details regarding the necessity o AQA can be Hellma 667-UV5
640.55 nm ≤0.10 nm 0.06 nm P
ound in Memorandum A 704 o the German As-

Photometric Accuracy ⃰
sociation or the Water Sector, Wastewater, and Equipment Wavelength Nominal value Tolerance limit ⃰ ⃰ Actual value Result
7 Waste Materials (Deutsche Vereinigung ür Was- 440 nm 1.093 A 1.082 - 1.104 A 1.092 A P
serwirtschat, Abwasser und Aball e.V., DWA)

Neutral Density 1.0 Abs.
546 nm 1.003 A 0.996 - 1.011 A 1.004 A P
Hellma 666-F4
635 nm 1.024 A 1.016 - 1.031 A 1.023 A P
and in the corresponding sel-control/sel-mon- 440 nm 2.252 A 2.236 - 2.268 A 2.248 A P
itoring regulations o the German ederal states Neutral Density 2.0 Abs.
Hellma 666-F203
546 nm 1.994 A 1.982 - 2.005 A 1.995 A P
8 (available in English).
635 nm 1.928 A 1.916 - 1.940 A 1.931 A P
Photometric Precision / Reproducibility ⃰ @ 1.0 A
Causes or errors can include: Equipment Wavelength Nominal value Actual value Result
440 nm ≤0.003 A 0.000 A P

• the working materials used Neutral Density 1.0 Abs.
546 nm ≤0.003 A 0.000 A P
9
Hellma 666-F4
• the handling 635 nm ≤0.003 A 0.001 A P
• the sample under investigation
These errors have eects on both the accuracy

10 and precision o the results obtained.
11
5.1 Quality Control at the
Stray Light ⃰
Manufacturer Equipment Wavelength Nominal value Actual value Result
Spectrophotometers and photometric test kits Potassium Chloride

Hellma 667-UV1
198.00 nm ≤1.00 %T 1.00 %T P
12 possess specications that are adhered to and Sodium Nitrite 340.00 nm ≤0.10 %T 0.00 %T P
above all else also documented by the manuac-

Hellma 667-UV11
turer. Spectral Bandwidth ⃰ ⃰ ⃰

Equipment Nominal value Actual value Result
13 The certicate or the spectrophotometer Toluene in n-Hexane

Hellma 667-UV6
≤1.8 nm 1.1 nm P
enclosed with each device documents the quality Selftest Hardware P

o the measuring device. No visual flaws, no burrs, no loose parts and fastenings P
14 Date: 29/01/2020
Inspector: gberg
- This document has been generated using electronic data processing and is valid without signature. -
15 Merck KGaA, 64271 Darmstadt, Germany

www.analytical-test-kits.com
EMD Millipore Billerica. MA 01821 - USA
Merck KGaA is ISO 9001:2000 and ISO 14001 certified
16
22 Version 3.0 – 04/2021
5 Analytical Quality Assurance (AQA) – 5.1 Quality Control at the Manufacturer 1
3
The certicate or the test kit, available for Calibration function:
each lot produced, documents the quality o the The calculated unction must agree, within speci-
reagents contained in the test kit. ed tolerances, with the unction electronically
stored in the spectrophotometer. 4
Condence interval:
Lot Certificate Maximum deviation rom the desired value over
Chargenzertifikat / Certificado del lote
Spectroquant COD Cell Test
®
the entire measuring range; every measurement 5
®
Spectroquant CSB-Küvettentest value can be aected by this deviation; this pa-
®
Spectroquant Test en cubetas DQO
Cat.No. / Art.Nr. / Art. Nro. 1.14541.0001 n = 10 rameter is a measure for the accuracy.
Target value Sollwert Result Messergebnis /
Measuring Range / Messbereich / Intervalo de Valor nominal Resultado
25 - 1500 mg/l CSB / COD / DQO
6
medida (Standard / Patrón) (Standard / Patrón)
Standard deviation for the procedure:

mg/l COD/CSB/DQO mg/l COD/CSB/DQO
Lot no./ Charge-Nr. / Lote nro. HC613856 25 19
Expiry date / Verwendbarkeit / Fecha de caducidad 2019/11/30

190
350
184
351 Measurement or the dispersion o the measure-
Standard / Standard / Patrón Potassium hydrogen phthalate 1.02400 Lot 142400Z 510 508
Photometer / Photometer / Fotómetro
Wavelength / Wellenlänge / Longitud de onda
Reference / Referenz / Referencia
605 nm
675
840
674
838
ment values over the entire measuring range,
expressed in ± mg/l.
Cell / Küvette / Cubeta 16 mm (round / rund / redonda) 1.005 1.005
Tester / Prüfer / Verificador Fr. Brandner 1.170 1.163
Date / Datum / Fecha
File / Datei / Fichero
2016/11/17
1145410001_HC613856_EN
1.335
1.500
1.324
1.483 7
Target value Lot value
Calibration Function / Kalibrierfunktion / Función de calibración
Coefcient o variation or the procedure:

Sollwert Chargenwert
ISO 8466-1 / DIN 38402 A51
Valor nominal Valor del lote
Slope / Steigung / Pendiente Tolerance +/- / Tolerancia 1,00 ± 0,03 0,99 
Measurement or the dispersion o the measure-

Ordinate segment / Ordinatenabschnitt / Intersecto en ordenadas 0
Reagent blank / Reag.blindwert / Valor en blanco del react Tolerance +/- / Tolerancia 0,010 ± 0,010 A 0,010 A 
Confidential interval (P=95%)
± 25 mg/l ± 12 mg/l 
ment values over the entire measuring range,

Vertrauensbereich (95% Wahrscheinlichkeit) / Intervalo de confianza (95 % de probabilidad)
8
Standard Deviation of the Method
± 4,9 mg/l 
Verfahrensstandardabweichung / Desviación estándar del procedimiento
expressed in %. The smaller the standard devia-

Variation Coefficient of the Method
± 2,5 % ± 0,7% 
Verfahrensvariationskoeffizient / Coeficiente de variación del procedimiento
1500 tion/coefcient o variation or the procedure,

1200 the more pronounced the linearity o the calibra-
tion curve.
9
Result (mg/l)
900
600
300
10
0
0 300 600 900 1200 1500
Target value (mg/l)
Merck KGaA, Darmstadt, Germany
Quality control Head of Lab. / Laborleiter /

Qualitätskontrolle / Control de calidad Jefe de laboratorio
Merck KGaA, 64271 Darmstadt, Germany, Tel.: +49 (0)6151 72-2440

EMD Millipore Corporation, 400 Summit Drive, Burlington MA 01803, USA, Tel.: +1-978-715-4321
11
Sigma-Aldrich Canada Co. or Millipore (Canada) Ltd., 2149 Winston Park, Dr. Oakville,
Ontario, L6H 6J8, Phone: +1 800-565-1400
1/2
12
13
14
15
16
Version 3.0 – 04/2021 23
1 5 Analytical Quality Assurance (AQA) – 5.2 Quality Control for the User
5.2 Quality Control for the User

A complete check comprises the entire system,
3 Checking the Checking the handling Inuence
i. e. the working equipment and the mode o working equip- operations of the
ment sample
operation. The spectrophotometer oers an
optimum degree o support in this regard, in the
orm o the dierent quality mode. The instru-
4
ment, or the whole system (including reagents
and all accessories) are checked, depending
on which quality mode is selected. All checking
5 operations can thus be supported by the spec- Pipette Suspend the bottom sedi-
ment in the cell by swirling.
trophotometer and the check values accordingly
documented as per GLP (Good Laboratory Prac-
tice) recommendations (see section 9.11).
6
The ollowing diagram provides an overview re-
garding internal quality-assurance aspects:
Carefully pipette 3.0 ml of
the sample into a reaction
7 Test kit cell, close tightly with the
screw cap, and mix vigor-
ously. Caution, the cell
becomes very hot!
8
Test for
recovery
9 Heat the reaction cell in the

thermoreactor at 148 °C for
C2/25 CSB 1500
Me§bereich 100 1500

Chemischer Sauerstoffbedarf
mg/l CSB 14 mm
2 hours.
2 ml Probelösung
in ein Reaktions- Mischen
Küvette wird heiß, im Thermoreaktor
küvette geben erhitzen mind. 10 min
am Verschluss abkühlen
anfassen 148 C, 120 min
Mischen
Abkühlen auf
Raumtemperatur Messen
(mind. 30 min)
10 1 2 3
4 5 6
7 8 9
. 0 C
Photometers
11 Remove the reaction cell

rom the thermoreactor and
place in a test-tube rack to
cool.
12
13 Thermoreactor
Swirl the cell after

10 minutes.
14
=
Test for the
15 overall system
16
24 Version 3.0 – 04/2021
5 Analytical Quality Assurance (AQA) – 5.2 Quality Control for the User 1
3
5.2.1 Checking the Spectrophotometer
As soon as the spectrophotometer is activated
it runs a Sel-Check. This means the hardware
and the sotware o the spectrophotometer are 4
checked and compared with internal standards.
The spectrophotometer itsel is checked in the

AQA1 mode with the Spectroquant® 5
PhotoCheck: the pack includes round cells
containing stable test solutions (secondary
standards) for checking the spectrophotom-
eter at the 445, 525, and 690 nm wavelengths. 6
The test solutions are measured in a reference
spectrophotometer monitored with primary
standards, and the certicate stating the ab-
sorbance values is enclosed with the package 7
unit. These desired values with the permissible
tolerances are entered into the spectrophotom-
eter or else handwritten into the control chart.
For the measurement the cell is placed in the
8
compartment or the round cell and identied
by the spectrophotometer via the bar code, and
the measured absorbance is compared with the
9
desired value. The absorbance is shown on the
display and can be entered into the correspond-
ing control chart.
10
The measurement of four cells for a given wave-
length tests – in addition to the wavelength
accuracy – also the linearity of the absorbance
over the eective range. 11
The verication o the instrument, as required by
DIN/ISO 9000 or GLP, can be easily perormed
by using the Spectroquant® PhotoCheck. The 12
PhotoCheck hence oers the possibility to check
the instrument. All o the corresponding docu-
mentation required by these certication guide-
lines is done by the spectrophotometer automa- 13
tically.
14
15
16
Version 3.0 – 04/2021 25
1 5 Analytical Quality Assurance (AQA) – 5.2 Quality Control for the User
5.2.2 Checking the Overall System

The testing o the overall system includes check- They can be diluted to dierent nal concen-
3
ing the working equipment and checking the trations, which should preerably lie approxi-
handling operations. mately in the middle o the measuring range o
the respective test kit. The table presented in
The overall system can be checked using Appendix 2 provides an overview o the available
4
standard solutions o a known content, preer- CombiCheck and ready-to-use standard solutions.
ably with the Spectroquant® CombiCheck; this
corresponds with the AQA2 mode in the spec- Due to limited shel-lie characteristics, there are
5 trophotometer. no CombiCheck or ready-to-use standard solu-
tions available or certain parameters. Appendix
Spectroquant® CombiCheck are ready-to-use 3 is a compilation of standard working proce-
standard solutions that in terms o the ana- dures necessary to make your own solutions of
6 lyte concentration are nely adjusted to the a dened concentration. This allows the control
individual test kits. They contain a mixture o of parameters where there are no simple-to-
several analytes that do not interere with each prepare solutions available.
other. The standard solution (R-1) is used in the
7 same way as a sample. A double determination If the test for the overall system shows that all
is recommended as a measure to diagnose any requirements are ullled, the individual results
random errors. are agged as AQA2. I not, an error message
is given and the individual components o
8 Standard solutions for photometric applica- the instrument have to be checked in detail.
tions (CRM) are ready-to-use standard solu-
tions that in terms of the analyte concentration
are nely adjusted to the individual test kits. The
9 standard solution is used in the same way as a
sample. A double determination is recommended
as a measure to diagnose any random errors.
10 In addition to the CombiCheck and the standard

solutions for photometric applications, it is also
possible to use Certipur® standard solutions
or this checking procedure. These contain
11 1000 mg of the respective analyte per liter of
solution.
12
13
14
15
16
26 Version 3.0 – 04/2021
5 Analytical Quality Assurance (AQA) – 5.2 Quality Control for the User 1
5.2.3 Checking the Pipettes 5.2.4 Checking Thermoreactors

This is checked by means o the thermosensor.
3
The Spectroquant® PipeCheck is used to The thermoreactor is preheated as described in

check the pipettes. The pack contains cells lled the Instruction Manual. When the control lamp 7
with color-dye concentrates. Ater the addition goes out, the temperature is measured in any
o a predened volume o water using the pi- one of the bores of the thermoreactor. The fol-
pette in question, the cell is measured against lowing desired temperatures must be achieved:
8
a corresponding reerence cell also contained
in the pack. The dierence in the absorbance Block temperature 100 °C =
values o the measurement cell and reerence Desired temperature 100 ± 3 °C
cell must not exceed the tolerances given in the
9
package insert. I the tolerances are exceeded, Block temperature 120 °C =
the instructions given in the section 4 must be Desired temperature 120 ± 3 °C
ollowed accordingly.
Block temperature 148 °C = 10
Desired temperature 148 ± 3 °C
The even distribution o the temperature over all

bores can also be documented using the ther-
11
mosensor.
12
13
14
15
16
Version 3.0 – 04/2021 27
1 5 Analytical Quality Assurance (AQA) – 5.2 Quality Control for the User – 5.3 Determination of
Sample Influences (Matrix Effects) – 5.4 Definition of Errors
3 5.2.5 Testing or Handling Errors

The user’s own mode o operation must also be I the calculated dierence is equal to the
subjected to an exact analysis. concentration o analyte o the addition solution
that was added, the recovery rate is 100 %. In
4 The ollowing questions may serve as a guide: the case that the recovery rate lies outside a
• Is the test kit optimal for the measurement range o approx. 90 – 110%, a matrix interer-
assignment in question? ence is present.
• Is the test kit’s measuring range suitable?
5 • Were the operating instructions for the test
ollowed?
• Was the sample volume correct? 5. Denition o Errors
• Was the pipette handled properly? It is obvious that measurement results as a
6 • Was a new pipette tip used? rule may be associated with errors. This ap-
• Is the pH o the sample and measurement plies equally to standardized methods o analysis
solution correct? (reerence methods) and to routine analysis. The
• Was the reaction time adhered to? discovery and the minimization o errors must
7 • Does the sample and reagent temperature lie be the objective here.
within the correct range?
• Is the cell clean and ree rom scratches? A distinction is made between systematic errors
• Has the expiry date or the test kit been and random errors.
8 exceeded?
• Were suitable sample vessels used? Systematic errors are present when all the
• Were the vessels used or the samples results o an analysis deviate rom the true value
and preparation steps clean and ree rom with the same algebraic sign. Examples here
9 detergent residues? include: a wrong sample volume, a wrong pH, a
wrong reaction time, a sample-matrix inuence,
etc. Systematic errors thus aect the accuracy
10 o the method o analysis.
5.3 Determination of Sample
Inuences (Matrix Eects) Accuracy = Deviation o the measured concen-
tration from the true concentration
The inuence o other substances contained in
11 the sample may, under certain circumstances,
Random errors manifest themselves in the
be so great that their recovery rates lie in the
orm o a wide range o deviation o the results
region o several percent. It is recommended
of a given sample. These can be kept to a mini-
to check or any inuence by using the addition
12 mum by ensuring good operating techniques and
solution contained in the Spectroquant®
multiple determination with calculation o the
CombiCheck pack.
mean values. Random errors make the result o
the analysis unreliable; they inuence the preci-
A dened quantity o the addition solution
13 (R-2), which contains a known concentration o
sion.
the respective analyte, is added to the sample
Precision = Dispersion of the results among
and the recovery rate is determined. The ollow-
each other
ing dierence is then calculated:
14 Result (sample + addition solution) –
Result (sample)
15
16
28 Version 3.0 – 04/2021
5 Analytical Quality Assurance (AQA) – 5.4 Definition of Errors 1
3
The ollowing diagram illustrates the aspects o
accuracy and precision:
1 3
6
Accuracy: poor Accuracy: poor

Precision: poor Precision: good 7
Major errors have been made! The high degree o precision mistakenly indi-
2 4 10
Accuracy: good cates a correct value!

Precision: poor Accuracy: good
Calculation of the mean values from at Precision: good 11
least three – or better even more – parallel The ideal objective!
determinations yields an approximation o the
true value.
12
13
14
15
16
Version 3.0 – 04/2021 29
1
6 Overview
6.1 Scope of Delivery 6.2 Overview of the Instrument

• Spectrophotometer Packaging
3 The spectrophotometer is shipped in protective
• Power adapter
• Power connectors (3 pieces) transport packaging.
• Dust cover
• Zero cell CAUTION
4
• Quick Guide (A4 ormat) Retain the original packaging including the inner
• Safety instructions packaging to protect the instrument against hard
• Certicate o nal inspection knocks i it has to be transported. Please note that
5 damage caused by improper transport voids all
warranty claims.
Front of the instrument

6
1 Display and user 2 Flip-up cover
interface
7
10
11
12
13
14
3 Shaft for rectangular 4 Shat or round cells
cells
15
16
30 Version 3.0 – 04/2021
6 Overview – 6.2 Overview of the Instrument – 1
6.3 Display and User Interface
6.3 Display and User Interface

3
5 6 7 8
NOTE
The entire display is touch-sensitive. Make selec-
tions using a fingertip or special touch pen. Do
not touch the display with sharp objects (e. g. 4
the tip o a ballpoint pen).
• Do not place objects on the display, as doing 5

so may scratch it
• Touch buttons, words or symbols to select
them
• Scrollbars are provided to assist quick move- 6
ment through long lists
Ports at the rear of the instrument • Touch the arrow in the scrollbar to scroll up-
5 Socket for plug-in power supply unit wards or downwards through the list
6 LAN port • Following selection, the item is activated im- 7
7 USB Mini B port mediately
8 USB-A ports • Touching a main button outlines it in blue
• Selecting an item inverts it (with dark text be-
NOTE ing shown on a light background) 8
All connections comply with SELV. • Selecting a text inverts it (with dark text
being shown on a light background), e. g.
method-specic settings or concentration
mode "Show Absorbance" 9
• "0" is OFF, "I" is ON – the active selection is
displayed light grey with dark gure, in this
case the Show Absorbance is ON
10
11
12
13
14
15
16
Version 3.0 – 04/2021 31
1 6 Overview – 6.3 Display and User Interface
Main menu navigation

The main menu is always visible on the let: It "Methods" and "Results" are the most oten used
3 consist of two pages with four smart icons each. modes and they are at the top o the main menu
To switch between the two pages push at navigation.
the bottom on the left.
4 Methods Results List

List
5 System
Methods Setup
Settings
6 Ad hoc Login/Log-
out
AQA
7 Timer
NOTE
10 The menu selected is always outlined in
blue.
NOTE
11
Action buttons like "Start", "Save", "Print" give
the ollowing touch eedback:
12 Normal
Remains static
Active elds are always shown in bright color.

13
Pressed elds invert the color as long as the
chosen action is perormed.
14
Disabled
Draws 30 % of the normal state
Inactive, disabled elds show aint color.

15
16
32 Version 3.0 – 04/2021
6 Overview – 6.3 Display and User Interface 1
NOTE
The main menus "Settings (Method Settings)", 3
"Ad hoc", "AQA", "System (Instrument Settings)",
"Login/Logout", "Timer" open up a submenu.
Example "Settings":
4
To leave these, the submenu has to be closed
by touching the main menu button again, in this
case: 5
The main menu "Methods" comprises two main overview panels arranged as shown below: 8
the Concentration Measurement Overview and the Method List Overview.
Screen layout concentration measurement overview

9
2 Alerts
1 Menu 3 Time-
title stamp 10
4 Measure-
ment range
5 Results 11
12 Main
menu but-
tons 6 Citation
12
7 Action
buttons
13
11 Main
menu
selection
button: 10 Brief infor- 9 Notication symbols/ 8 Info bar with
14
Switch be- mation settings Information
tween the button
two main
menu 15
overviews
16
Version 3.0 – 04/2021 33
Screen layout method list overview
3 2 Dropdown 3 Dropdown
box box
(closed) (closed)
1 Menu
4 title
7 Focuses 4 Selection
selected
buttons
main menu
5 button
6 Name of
method 5 Scrollbar
7
10
Screen layout results overview
2 Filter menu
11 1 Menu
title
7 Focuses 3 Selection
selected buttons
12
main menu
button
6 Result 4 Scrollbar
13 list
14
5 Action
buttons
15
16
34 Version 3.0 – 04/2021
Main menu buttons
6
Method list Settings
List o all methods, irrespective o This button is used to activate meth-
mode od-specic settings (e. g. sample dilu-
tion, turbidity correction, zero adjust-
7
ment, sample blank, reagent blank)
10
11
Ad hoc AQA
For perorming measurements (ab- Overview and list o all Analytical
sorbance/transmission, spectrum, Quality Assurance (AQA) modes
kinetics) 12
Allows measurements to be per-
ormed without the need to create
methods
13
14
15
16
Version 3.0 – 04/2021 35
Main menu buttons
6
Results list System setup
List o all stored results This button is for optional instrument
settings (e. g. date, time, updates
7 etc.)
10
11 Login/logout Timer list

Check users in and out List of stopwatch functions
12
13
14
15
16
36 Version 3.0 – 04/2021
Overview of main buttons
3
Buttons Description
Method list
List o all methods, irrespective o mode
4
Settings
This button is used to activate method-specic settings
(e. g. sample dilution, turbidity correction, zero adjustment, sample blank, reagent blank)
Ad hoc
5
For perorming measurements (absorbance/transmission, spectrum, kinetics)
Allows measurements to be perormed without the need to create methods
Absorbance/Transmission Mode
Ad hoc submenu: perorm absorbance or transmission measurements
6
Spectrum Mode
Ad hoc submenu: record spectrum
Method list: create methods -> Spectrum Mode
7
Kinetic Mode
Ad hoc submenu: perorm kinetic measurement
Method list: create methods -> Kinetic Mode
8
AQA
Overview and list o all Analytical Quality Assurance (AQA) modes
AQA Status 1&2 9

AQA submenus: Status display o the period o validity and the outcome (passed/ailed)
AQA1
AQA submenu: List o AQA1 methods 10
AQA2
AQA submenu: List o AQA2 methods
11
Pipette check
AQA submenu: List o pipette-checking methods
Result list
12
List o all stored results
System setup
This button is or optional instrument settings (e. g. date, time, updates etc.)
13
Login/logout
Log users in and out
14
Timer list
List of stopwatch functions
15
16
Version 3.0 – 04/2021 37
Overview of action & selection buttons
3
Action & Selection Buttons Description
Start button
Start an action (e. g. measurement)
4
Start zero
Start zero adjustment or a method
5 Apply
Save
6
Stop
7
Close
8 Logout
User logout
Search method
9
Search/results list
Search unction, search criterion: method name, method number or item number
10
Filter cancellation button
Cancel all set lter options
11 Edit
For editing parameters
Create method
12
Print
Print to pd (USB device) or printer
13
Export button
All selected results are exported to an external memory device as .csv le
14 Import button
Updates/Methods are imported rom an external memory device into the instru-
ment
Delete
15 The selected items are deleted
16
38 Version 3.0 – 04/2021
1
7 Safety
1
2
2
This operating manual contains basic instruc- Safety instructions 3

tions that you must ollow during the commis- Safety instructions in this operating manual are 3
sioning, operation and maintenance o the spec- indicated by the warning symbol (triangle) in the
trophotometer. Consequently, all responsible let margin. The signal word (such as
personnel must read this operating manual care- "CAUTION") indicates the danger level. 4
ully beore working with the meter. Keep this The ollowing warning symbols are used: 4
operating manual in the vicinity of the meter.
This is a class A device. This equipment may

5
cause intererence in a residential installation. In 5
this case the user is encouraged to perorm ap-
propriate measures to correct the interference.
6
6
Symbols Description
WARNING
Hazardous area (general). The xenon lamp (UV/VIS) emits radiation in the ultraviolet 7
region, which may cause damage to the eyes. Never look directly in the radiation o this
light source without wearing proper eye protection. Protect your skin rom the direct ex-
posure to UV light.
WARNING
8
Dangerous electrical voltage.
8
WARNING
9
WARNING
Signies instructions that must be ollowed precisely in order to prevent serious dangers
9
to personnel.
10
CAUTION CAUTION
Signies instructions that must be ollowed precisely in order to avoid minor injuries to
10
personnel or damage to the instrument or the environment.
CAUTION
11
CAUTION
This is a cautionary notice with a warning symbol drawing your attention to the risk o
11
(limited) harm to personnel.
12
NOTE NOTE
Signies a notice drawing your attention to special characteristics. 12
REFERENCE
13
Used to indicate reerences to other documents. 13
14
Please pay attention to the separate saety instructions leaet (part o delivery scope) and read it 14
carefully.
15
15
16
Version 3.0 – 04/2021 39
1 7 Safety – 7.1 Intended Use – 7.2 General Safety Instructions –
7.3 Target Group and User Qualification
7.1 Intended Use

The intended use o the spectrophotometer con- Safe operation
3
sists exclusively o the carrying out o photomet- If safe operation is no longer possible, the spec-
ric measurements according to this operating trophotometer must be taken out o service and
manual. Observe the technical specications o secured against inadvertent use.
the cells in the operating manual. Any other use Safe operation is no longer possible if the spec-
4
is considered to be unauthorized. The spectro- trophotometer:
photometer was developed or perorming water • Has been damaged in transport
analyses in the laboratory. • Has been stored under adverse conditions over
5 a lengthy period o time
• Is visibly damaged
• No longer unctions as described in this manual
6 I you are in any doubt, contact the supplier o

your spectrophotometer.
7.2 General Safety Instructions
The spectrophotometer is built and inspected ac-
7 cording to the relevant guidelines and norms or
electronic instruments (see section 12). It left
the actory in a sae and secure technical condi-
tion.
8 7.3 Target Group and User
NOTE Qualication
The spectrophotometer must be opened, adjust- The spectrophotometer was developed or use
ed, or repaired only by specialist personnel au- in the laboratory. Perorming photometric deter-
9
thorized by the manuacturer. Noncompliance in- minations using test kits frequently involves the
validates any claim under warranty. handling o hazardous substances. We presume
that the operating personnel are familiar with
10 handling hazardous substances as a result o
their proessional training and experience. In
particular, the operating personnel must be able
7.2.1 Function and Operational Saety to understand and correctly ollow the saety
11 The smooth unctioning and operational saety o labels and saety instructions on the packages
the spectrophotometer can only be guaranteed and test kit inserts.
i the generally applicable saety measures and
the specic saety instructions in this operat-
12 ing manual are ollowed during operation. The
smooth unctioning and operational saety o the
spectrophotometer can only be guaranteed un-
der the environmental conditions that are speci-
13 ed in the operating manual. I the spectropho-
tometer is moved rom a cold environment to a
warm environment, the ormation o condensate
can cause it to function incorrectly. In this event,
14 wait until the meter reaches room temperature
before using it again.
15
16
40 Version 3.0 – 04/2021
7 Safety – 7.4 Handling Hazardous Substances 1
7.4 Handling Hazardous Substances 4

When developing Spectroquant test kits, the
®
manuacturer works diligently to ensure that the

tests can be carried out as saely as possible.
Some dangers posed by hazardous substances 5
cannot always be avoided, however.
WARNING
6
Improper handling o certain reagents can ad-
versely affect your health. Always follow the
saety labels on the packaging and the saety in-
structions on the package insert. The protective 7
measures speciied there must be careully ol-
lowed.
Safety data sheets 8

The chemical data sheets contain all the instruc-
tions needed to handle them saely, and give
details o potential risks as well as actions to
be taken preventively and i a danger is posed.
9
Work safely by following these instructions.
10
11
12
13
14
15
16
Version 3.0 – 04/2021 41
1
8 Getting Started
8.1 General Notes on Handling 8.2 Initial Setup

The Spectroquant Prove spectrophotometer is
® Proceed as follows:
3 • Connect the power adapter (see section 8.2.1)
an optical precision instrument. It should always
be handled with care, especially when in mobile • Switch on the spectrophotometer (see section
use. Always protect the instrument rom condi- 8.2.2)
tions that could damage the mechanical, optical • Set the language (see section 8.2.3)
4
and electrical components. Please note the ol- • Set the date and time (see section 8.2.4)
lowing in particular: • Run the self-test (see section 8.2.5)
• The temperature and humidity during opera-
5 tion and storage must be within the limits NOTE
specied in the "Technical Data" section (see
For our operating manual please visit:
section 12)
The instrument must never be exposed to www.sigmaaldrich.com/spectroquant
the following: For more inormation about the technical videos
6 please visit:
• Extreme dust, humidity and moisture
• Intense light and heat www.sigmaaldrich.com/photometry
• Fumes that are corrosive or contain high con-
7 centrations of solvents
In addition take care of the following:
• For measuring, the instrument must be placed 8.2.1 Connecting the Power Supply
on a at surace Power is supplied through the power adapter
8 • Spilled liquid or other material should be re- provided. The power adapter supplies the spec-
moved immediately (see section 10.3) trophotometer with the required voltage and
• I a cell has broken in the cell holder, the cell type o current (24 V DC).
holder should be cleaned immediately (see
9 section 10.3)
• The cover should always be closed when the CAUTION
spectrophotometer is not in use The line voltage at the user location must fulfil
• When the spectrophotometer is being trans- the speciications stated on the power adapter
10 ported, the cell compartment must be empty (the speciications are also indicated in the oper-
ating manual). Only ever use the 24 V power
adapter provided. Please note that damage
caused by using a dierent power adapter than
11 the one supplied voids all warranty claims.
12
13
14
15
16
42 Version 3.0 – 04/2021
8 Getting Started – 8.2 Initial Setup 1
8.2.2 First Power-on

After switching on the spectrophotometer for the 3
rst time you are automatically guided through
the language, date and time setup procedures.
1 5
7
1. Press the ON/OFF button 1 . The spectro-
photometer gives an audible signal (beep)
2
and starts booting or approximately 2 min-
utes. You will see the ollowing display: 8
9
1
10
Connecting the power adapter:
1. Connect the miniplug 1 o the power adapter
to the socket 2 of the spectrophotometer. 11
2. Connect the power adapter to a wall socket.
2. The display switches to language setup (see

12
section 8.2.3).
13
14
15
16
Version 3.0 – 04/2021 43
1 8 Getting Started – 8.2 Initial Setup
8.2.3 Language Setup 8.2.4 Date, Time and Country-specic

3 The software supports several languages. When Settings
you switch on the spectrophotometer for the During initial setup, having set the language op-
rst time, a list o language options is automati- tion you are automatically guided through the
cally displayed ater the boot procedure. date and time setup procedure.
4
5 1
6
2
1. Select the desired language 1 . 1. Tap on the Date format button 1 .

2. Tap on the Save button 2 to conrm. 2. The calendar view pops up 2 . You can now
8 enter the date.
NOTE
The saving process of changing the language re-
9 quires some seconds.
2
10 4
3
11
12 3. Tap on OK 3 to conrm.
4. Tap on the Arrow button 4 to choose the
country-specic basic date setting. The date
ormat can be set and displayed or EU and
13 US.
14
15
16
44 Version 3.0 – 04/2021
8 Getting Started – 8.2 Initial Setup 1
8.2.5 Self-test
Following language, date and time setup the
3
spectrophotometer performs a self-test.
6 4
8 5
10
9
5
7
1
6
5. Tap on the Time format button 5 . The nu-
meric key panel 6 pops up. Now you can
enter the time.
6. Tap on OK 7 to conrm. 1. Remove all cells and close the cell compart- 7
7. Tap on the Arrow button 8 to choose the ment cover.
country-specic basic time setting. The time 2. Start the self-test with the Start button 1 .
ormat can be set and displayed or EU and 3. The spectrophotometer performs the self-
US. test. 8
8. Tap on the Arrow button 9 to choose the
decimal separator "."/"," used in your coun- Self-test
try. The sel-test covers:
9. Tap on the Save button 10 to conrm. • Checks on memory, processor, internal inter- 9
aces, lter and lamp
After initial setup is complete, you can change • A calibration of the wavelengths
the date and time at any time in the setup/date/
time menu (see section 9.2.3). When the sel-test has ended, the display shows 10
the main menu.
11
12
13
14
15
16
Version 3.0 – 04/2021 45
1 8 Getting Started – 8.3 Connecting Optional Peripheral Devices
8.3 Connecting Optional

NOTE Peripheral Devices
3
The spectrophotometer also automatically runs a
calibration of the wavelengths after every 100th 8.3.1 Communication Ports
measurement. A corresponding message pops
4 up in the display as long as the calibration
operation is in progress.
7
You can connect the ollowing peripheral devices
to the spectrophotometer:
• Printer (see section 8.3.2)
8 • USB mass storage device (see section 8.3.3)
• Barcode reader (see section 8.3.4)
NOTE
9 I you want to connect several USB devices such
as a USB PC keyboard and a USB memory de-
vice to the instrument, you can increase the
number of USB-A sockets by attaching a com-
10 mercially available USB-2 hub with separate
power supply.
11
8.3.2 Printer
Printers can be connected to the spectrophoto-
12 meter as ollows:
Connecting the Spectroquant® Prove using a

commercially available USB-A to USB Mini B
13 cable enables its data to be printed out.
NOTE
14 All postscript printers can be used with
Spectroquant® Prove.
15
16
46 Version 3.0 – 04/2021
8 Getting Started – 8.3 Connecting Optional Peripheral Devices 1
3
8.3.3 USB Memory Device
Using a USB memory device (e. g. a USB mass
storage device), you can 4
• Update the instrument rmware and method
data (see section 9.2.8)
• Transer data to the USB memory device
(see section 9.13.7) 5
USB memory devices are connected to the

USB-A port.
6
NOTE
Please follow the instructions on using USB
memory devices 7
(see section 9.13.7).
8
8.3.4 Barcode Reader
The barcode reader enables the simplied ac-
quisition o alphanumerical character strings and 9
can be used in all operating situations that re-
quire the entry o text or numerals. The barcode
reader is connected to the USB-A port.
10
NOTE
The barcode reader must support USB/HID.
11
12
13
14
15
16
Version 3.0 – 04/2021 47
1
9 Operation
9.1 Switching the Spectrophoto-

meter On or O
3
2
6
Switching on 4. Start the self-test with the Start button 2 .

1. Press the ON/OFF button 1 . The spectro- 5. The spectrophotometer performs the self-
7
photometer gives an audible signal (beep) test.
and starts booting or approximately 2 min-
utes. You will see the ollowing display:
Self-test
8 The sel-test covers:
• Checks on memory, processor, internal inter-
aces, lter and lamp
• A calibration of the wavelengths
9
9
NOTE
The spectrophotometer also automatically runs a
10 calibration of the wavelengths after every 100th
measurement. A corresponding message pops
up in the display as long as the calibration
operation is in progress (see section 8.2.5).
11
When the sel-test has ended, the display shows
2. After the booting process the screen shows the main menu.
the sel-test dialog.
12
Starting the self-test
3. Remove all cells and close the cell compart-
13 ment cover.
14
15
16
48 Version 3.0 – 04/2021
9 Operation – 9.1 Switching the Spectrophotometer On or Off 1
3
NOTE NOTE
After the self-test, the system automatically You can set a user-deined time or this unction
checks the status o activated AQA tests. An (see section 9.2.5). 4
overview o any ailed and/or expired AQA tests
automatically opens (see section 9.11).
Switching o
5
6
4
Energy-saving mode – display

Press the ON/OFF button 4 to switch the spectro-
photometer o.
9
9
NOTE
The instrument has an Auto-Power-Off function,
which switches it automatically off after a user- 10
3
deined time. This unction is not active out o
the box, but you can turn it on in "System
(Instrument settings)".
11
12
13
The spectrophotometer automatically switches

o the backlight o the display 3 when no but-
ton has been tapped within a period o 10 min- 14
utes. The backlight is switched on again with the
next tap.
The button unctions are activated only ollowing
a further tap. 15
16
Version 3.0 – 04/2021 49
1 9 Operation – 9.2 System Setup
9.2 System Setup

3 General instrument setup is carried
out in the "System" menu.
6
Buttons Description
Information
This submenu displays the ollowing inormation about the device:
7 Sotware/method versions, device class, lamp counter and serial number
Interface
This submenu displays the ollowing settings options and standard settings:
8 Audible signals – ON, Backlight – 100 %, Print to pd – ON
Region
9 This submenu displays the ollowing settings options and standard settings:
Language, date, time and country zone EU/US, decimal separator – "."/"," (dot or comma)
Quality
Quick zero – OFF, AQA1 and AQA2 lock – OFF, Zero Adjustment expiry – ON (interval:
7 days), Use expired reagents – OFF, Service reminder – ON
Automation (Setup 4)
Energy saving mode – ON (10 minutes), Auto Power o – OFF, Auto log o – OFF,
Auto store – ON, Auto print – OFF, Sample ID popup – OFF
User management
Activation o user management and administrator settings, User login required – OFF
13 Service
This submenu displays the ollowing settings options:
Various service unctions such as backup, restore, export o log or system data and
import o methods
14 Update
This submenu displays the option or perorming sotware and method updates
15
16
50 Version 3.0 – 04/2021
9 Operation – 9.2 System Setup 1
Buttons Description 3
Network
This submenu displays the setting options or connecting the Prove device with a network
4
Prove Connect
This submenu displays the settings options or connecting the Prove device with the Prove
Connect sotware (the Prove Connect sotware is optionally available, order No. Prove Connect
to LIMS Y110860001) 5
9.2.1 Information 6
This submenu displays the ollowing
inormation and settings options:
7
2 8
8
3
5
9
6
7
8 Reset button for lamp counter in
Prove 100 10
Press this button after the change of the halogen
lamp to reset the lamp counter to zero.
1 Update Version
Instrument version number
11
2 MMI Version
Man-machine interfaces version number

3 MCS Version
Measurement and control sotware version 12

number
4 Methods Version
Method version currently in use

5 Device
13
Class o device being used
(Prove 100 | 300 | 600)
6 Lamp counter
Lietime/service perormance o the lamp 14

7 MCS Serial
Instrument serial number
15
16
Version 3.0 – 04/2021 51
9.2.2 Interface 9.2.3 Region

This submenu displays the ollowing This submenu displays the ollowing
3
inormation and settings options: inormation and settings options.
4
9
1
11
5 10 3 2
3 2
4
6
7 9 Audible signals 1 Language

Ticking/unticking the box switches the audible 2 Date/time
alert on or o. 3 Country zone EU/US
10 Print to pd le 4 Decimal separator
8
Activating the pd printer transers all print jobs
to the external memory device (e. g. USB mass Language
storage device) in pd ormat. To change the user language, proceed as ollows:
11 Backlight
9
This setting enables you to adjust the backlight
brightness to the ambient light conditions.
10 1 2
12
11
12 13
1. Select System 1 .
2. Select Region (Setup 2) 2 .
13
1. Select System.
2. Select Interace (Setup 1).
3. Set the backlight contrast 11 to the desired
14 level using the drop-down menu 12 .
4. Tap on the Save button 13 to store and close
the settings.
15
16
52 Version 3.0 – 04/2021
Country zone EU/US, decimal separator

3
Opening the drop-down menu allows the country
settings to be changed.
3
4
5
3
5
3. Select the desired language in the drop- 6
down menu 3 .
4. Tap on the Save button 4 to store your
changes and close the settings by touching
the System button again. 7
• Date display US/EU 3
• Time display US/EU 4
Date/time
• Decimal separator "."/"," (dot or comma) 5
Depending on country, the date ormat is pre-
8
sented in the sequence e. g. Day.Month.Year
(DD.MM.YY) or Month/Day/Year (MM/DD/YY or NOTE
MM.DD.YY). To reset or change the gures, pro- Make sure that the decimal separator used is the
ceed as ollows: same which is used in your Excel sotware to
9
avoid any problems with the ormat o your csv
files.
10
1 2
7
11
6
8 12
1. Select System 1 .
2. Select Region (Setup 2) 2 . 13
3. Tapping on the Date eld 6 pops up a calen-
dar 7 . Here you can set the date and tap on
OK 8 to conrm your selection.
14
15
16
Version 3.0 – 04/2021 53
10
5 11
6 4. Tapping on the Time eld 9 pops up a selec-

tion eld 10 . Here you can set the time and
tap on OK 11 to conrm your selection.
7
9 12
10
5. Tap on the Save button 12 to store all of the
changed settings.
11
12
13
14
15
16
54 Version 3.0 – 04/2021
9.2.4 Quality
This submenu displays the ollowing
3
To activate a unction, tap on the input eld 1 .

A tick 2 appears when the setting option is ac-
4
1
5 11
tivated. To cancel settings, tap on the C button
6 12 3 . This works only for changes that have been
made but not yet stored. To accept the settings,

7 5
2 13 tap on the Save button 4 .
8
9 4 3
10 6
7
Possible settings (items 5 to 12 ):
Pos. Name Function – active Function – inactive
5 QuickZero Perormance and direct saving o the zero Perormance and saving o the zero adjust- 8
(Possible only or adjustment or: ment only or the currently selected con-
concentration meth- • all 12 wavelengths that are used or mea- centration method.
ods) suring Spectroquant® test kits
• the wavelength currently being used in 9
the Concentration mode
6 AQA1 – Lock device I an invalid AQA1 test is present, a warning I an invalid AQA1 test is present, a warning
message appears. The instrument is locked message appears. Measurements can still
or all measurements except AQA1 tests. be perormed. The instrument is not locked. 10
7 AQA2 – Lock I an invalid AQA2 test is present or a I an invalid AQA2 test is present or a
method concentration method, a warning message concentration method, a warning message
appears when this method is selected. The appears when this method is selected.
performance of a concentration measure- Measurements can still be perormed. The
11
ment using this method is locked. instrument is not locked.
8 Zero adjustment When the pre-set expiry date in days 13 An already perormed zero adjustment
EXP is reached, the zero adjustment must be does not have to be repeated.
(Possible only or repeated.
12
concentration meth-
ods)
9 Use expired re- The use of Spectroquant® test kits beyond The use of Spectroquant® test kits beyond
agents the date o expiry is allowed. A warn- the date o expiry is not allowed. A warn-
13
ing message appears when the barcoded ing message appears when the barcoded
Spectroquant® cell or the AutoSelector Spectroquant® cell or the AutoSelector is
is inserted into the instrument; ater the inserted into the instrument. No measure-
warning message is conrmed, however, ment can be made.
the measurement is still carried out. 14
10 Service reminder The instrument automatically signals the The instrument does not signal the need to
need to service the instrument. service the instrument.
15
16
Version 3.0 – 04/2021 55
3 Pos. Name Function – active Function – inactive
11 Deactivate message When results are higher than the measuring Results beyond the measuring range o
indicating results range o the selected method, the display the selected method are also shown as a
beyond measuring o the device shows “HI”. numerical value. Negative measurement
4 range When results are lower than the measuring results may also be shown here.
range o the selected method, the display (This unction can be o help in calculating
o the device shows “LO”. statistical parameters, e. g. in determining
limits o detection.)
5 12 Allow deletion o When the user management is activated, No results in the result list can be deleted.
results in the result results in the result list can be deleted.
list (This unction can be activated only when
the user management is activated by a user
with administrator status.)
6
10
11
12
13
14
15
16
56 Version 3.0 – 04/2021
9.2.5 Automation
This submenu displays the ollowing 3
7 Auto logo
The instrument logs the active user out after the

4
1 1
5 10
user-dened time has elapsed and no entry has
2
6 11 been made. The screen changes to the login/
logout screen.
7 12 5
When the unction is activated, a eld pops
8
up or setting the time. Here you can enter
9 4 3 your individual time in minutes and conrm by
tapping the OK button. Accept the time setting 6
by tapping on the Save button.
8 Prex S-ID
This unction enables you to prex a sample ID

with a recurrent character string (e. g. Fountain), 7
which is then saved together with the sample ID.
1 Energy-saving mode
When the unction is activated, a eld pops up
To activate a unction, tap on the input eld 1 .
to enter data. Here you can enter the individual
A tick 2 appears when the setting option is acti-
string o characters or description, conrming 8
vated. To cancel settings, tap on the C button
your entry by tapping the OK button. Accept the
3 . This works only for changes that have been
entry by tapping on the Save button.
made but not yet stored. To accept the settings,
9 SampleID popup
tap on the Save button 4 .
5 Energy-saving mode Ater each measurement, an input window 9
automatically pops up to enter the sampleID.
Ater a user-dened time has elapsed, the back-
The data can be entered via the virtual keyboard
light o the display switches o automatically.
o the display, a keyboard connected via the USB
The backlight is switched on again with the next
port, or a hand-held scanner connected via the 10
tap.
USB port.
your individual time in minutes and conrm by
11
tapping the OK button. Accept the time setting
6 Auto power o
Ater a user-dened time has elapsed, the 12

instrument switches o automatically.
your individual time in minutes and conrm by 13
tapping the OK button. Accept the time setting
14
15
16
Version 3.0 – 04/2021 57
3 10 Auto store 13
The instrument automatically saves the

measurement results in the Concentration mode
in the result list.
4
NOTE
The instrument is capable o storing 2,000 indi-
5 vidual measurements o the measurement
modes concentration, absorbance/transmission,
and/or multiple wavelengths as well as 20 data
sets with results of the spectrum or kinetics
6 methods or each o these modes. It operates on
the FIFO storage principle (irst in – irst out), When the barcode scanner is deactivated, a
meaning that when all storage locations are corresponding symbol 13 is constantly shown in
occupied, the next time a result is to be stored the upper status bar o the display.
7 the oldest result on the list is automatically
overwritten. Accordingly it is advisable to
regularly back up stored data sets on external
media (see section 9.13.7).
8 The results o the measurement modi AQA1,
AQA2, MatrixCheck, and PipeCheck are managed
separately. A total o 500 results are stored.
Results that determine a system or method
9 status are not overwritten, even when all
storage locations are already occupied.
11 Auto print
10
The instrument automatically starts a print
operation after the measurement has been
completed. Precondition: a USB memory device
(print as pd) or PostScript printer (print on
11
paper) is connected.
12 Barcode scanner
This unction enables the barcode scanner

12 or decoding round cells and AutoSelectors to
be deactivated. When the barcode scanner is
deactivated, the automatic method recognition
o barcoded round cells and AutoSelectors is
13 switched o and the method must be selected
manually.
This unction comes in handy when e. g. you
wish to run your own methods in round cells.
14
15
16
58 Version 3.0 – 04/2021
9.2.6 User Management

This submenu oers the ollowing 3
setting options and is only acces-
sible to administrators once "Login
required" is active:
4
4. Enter the user password 4 .
5. Conrm the user password 5 .
6. Tap on OK to conrm 6 .
7. The newly created user appears in the pick- 5
list 7 .
1
6
1
7
3
Activate user management:
4
When the "Login required" box 1 is ticked, us-
2 5
ers have to log in in the "Login/Logout" menu
8
with their name and password to get certain ac-
cess user rights (see section 9.14.1).
NOTE 9
Edit user, e.g. change password:
Once "Login required" is ticked, it can only be 1. Select the user 1 whose password is to be
deactivated by an administrator. changed.
2. Tap on edit 2 to edit the user.
10
3. Enter the new user password 3 .
7 2 4. Conrm the password 4 .
5. Tap on OK 5 to conrm.
3
11
4
5
1 6
12
Create user: 13
1. Tap on the Add new user eld 1 .
2. Assign the user rights by tapping on the but-
ton 2 : ull rights = administrator; restricted
rights = user (see section 9.14). 14
3. Enter the user name 3 .
15
16
Version 3.0 – 04/2021 59
9.2.7 Service
3
This submenu oers the ollowing
setting options:
4
4 6
1 1
7 7 4
2
5 3
6
5
3
2
6
7 Delete user: To export or import data you need a commer-

1. Select the user to be deleted 1 . cially available USB mass storage device.
2. Tap on the Delete button 2 .
3. A window pops up asking "Do you really 1 Backup/restore
8 want to delete user?". Create a backup on the USB device.
Tap on OK 3 to conrm. 1. Attach a blank (no data present) USB mass
4. The user is deleted rom the pick-list 1 . storage device to the spectrophotometer.
2. Tapping on the Export button 4 automati-
9
cally stores the spectrophotometer data on
the USB mass storage device.
3. A message appears in the ino window 5
10 when the data have been successully trans-
erred.
Importing a backup rom the memory device.

1. Attach the USB mass storage device contain-
11
ing the backed up data to the spectropho-
tometer.
2. Tapping on the Import button 6 automati-
12 cally imports the spectrophotometer data to
the instrument and stores them there.
3. A message appears in the ino window 5
when the data have been successully trans-
13 erred.
14
15
16
60 Version 3.0 – 04/2021
2 Exporting log and/or system les 3 Importing user-dened methods 4

1. Tick the Log or SysData box 7 . 1. Attach the USB mass storage device contain-
2. Attach a blank (no data present) USB mass ing the method list with the user-dened
storage device to the spectrophotometer. methods to the spectrophotometer.
The memory capacity of USB mass storage 2. Tapping on the Import button 6 automati- 5
device should be at least 512 MB. cally imports the spectrophotometer data to
3. Tapping on the Export button 4 automati- the instrument and stores them there.
cally stores the spectrophotometer data on 3. A message appears in the ino window 5
the USB mass storage device. when the data have been successully trans- 6
4. A message appears in the ino window 5 erred.
when the data have been successully trans-
erred. NOTE
7
Only users rom the Administrator group have
NOTE the right to perform the setting options in the
When log iles are exported, three separate log Service submenu.
iles are created and exported: an error log ile, 8
a user log ile, and a service log ile. The iles
are saved on the USB mass storage medium in
the ollowing older structure:
Parent older: PROVE 9
Subolder: Log
Subolder: Serial number o the device,
underscore, “Date”, underscore YYMMDD,
underscore hhmm 10
Example: “PROVE\Log\SN1529610052_
Date_201208_1001”
11
12
The error and service log iles contain

inormation that can be o use in dealing with 13
service issues. The user log file contains
inormation on activities carried out by the user,
e. g. changes in the system settings. For urther
details on the contents o the log iles and their 14
interpretation please refer to section 16.
15
16
Version 3.0 – 04/2021 61
9.2.8 Updates
3 Firmware and method updates
ensure your spectrophotometer is
always up-to-date.
4 NOTE 1. Download the update le rom the website

Only users rom the Administrator user group onto a USB memory device.
may perorm irmware and method updates. 2. Unzip the le with the entire older structure
into the main directory o the USB mem-
5 The update comprises: ory device. Ater unzipping, a older titled
• The latest rmware “PROVE” with a subolder titled “Update”
• New or modied method data appears in the main directory o the USB
memory device.
6 3. Select System 1 .
NOTE
4. Tap on the Update button 2 .
A irmware and method update does not alter 5. Attach the USB memory device with the up-
user-deined data (such as settings, custom date to the spectrophotometer.
7 methods or measurement data).
The update is transferred to the spectro-

photometer by using a USB mass storage 3
8 device for temporary storage.
NOTE
To keep the instrument always up-to-date, we
9
recommend always installing the latest update.
The relevant updates are available on
www.sigmaaldrich.com/photometry.
10
Firmware and method update spectro-
photometer 6. Tap on the Import eld 3 starts the search
11 or update les on USB device. Search needs
some time (approx. 1 minute).
7. You will be asked whether you want to install
the update version on the Prove. Conrm
12 with OK.
1
8. The installation process is reported in the in-
ormation window and conrmed by the nal
2 message that the data have been success-
13 ully transerred.
9. Conrm with OK.
14
15
16
62 Version 3.0 – 04/2021
9.2.9 Network and

Prove Connect
3
Connect the Prove spectrophotometer
with a network and the Prove
Connect sotware (optionally
available, order No. Prove Connect to 4
LIMS Y110860001)
The broad range o settings options are

described in a separate manual.
5
7
10. The instrument then shuts down and re-
boots. The boot screen appears in the dis-
play. Depending on the data volume, this
procedure may take several minutes. 8
10
4
11
11. If the import was unsuccessful, a corre-

sponding message 4 appears. Try again.
12
Beore this, check whether the older struc-
ture described above in item 2 is present on
the USB memory device.
13
14
15
16
Version 3.0 – 04/2021 63
1 9 Operation – 9.3 Measurements
9.3 Measurements
The spectrophotometer can be used to perorm the measurements listed below.
3
Type of Measurement Description
Concentration • Pre-programmed methods that can be executed using Spectroquant® test kits or self-
4 prepared reagents
• User-programmed methods
Absorbance/transmission • Single-wavelength measurements for establishing the absorbance or transmission of

solutions
5 • Multiple-wavelength measurements for establishing the absorbance or transmission of
solutions
Spectrum • Programmed methods or establishing the absorbance or transmission o solutions over a
dened wavelength range
6
Kinetics • Programmed methods or establishing the absorbance or transmission o solutions over a
dened period
Quality checks Instrument-supported analytical quality assurance:

• Instrument check (AQA1)
7 • Method-specic system check – pre-programmed or all Spectroquant® standards (AQA2)
• Pipette volume control (PipeCheck)
• Check intererences rom oreign substances (MatrixCheck)
10
11
12
13
14
15
16
64 Version 3.0 – 04/2021
9 Operation – 9.3 Measurements 1
9.3.1 Performing a Measurement 3

Measurements can be perormed using rect-
angular cells o various path lengths (10, 20,
50 mm/100 mm Prove 600) and Spectroquant®
round cells. Insert cells as ollows to start the 4
Measuring with rectangular cells with open
measurement:
lid:
Measuring with a round cell with closed lid insert AutoSelector
5
6
6
2
4
7
10
5
7
1 11
3
• Open the ip-up cover 4 by pushing it back

with your ngers
• Insert the barcoded Spectroquant® round cell 12
• Insert the AutoSelector vertically into the cell
through the opening 1 , ensuring that the
compartment 5 , ensuring that the white posi-
white position mark 2 on the cell is aligned
tion mark 6 on the AutoSelector is aligned with
with the positioning mark on the spectrophoto-
the positioning mark on the spectrophotometer 13
meter 3
7
• Measurement starts automatically, and the
• The photometer is ready to measure
measurement result is displayed in the concen-
tration measurement overview (see page 33)
NOTE 14
I the barcode cannot be read, please see section
9.7.1.
15
16
Version 3.0 – 04/2021 65
1 9 Operation – 9.3 Measurements
3
Measuring with rectangular cells with open Measuring with rectangular cells with open
lid: lid:
Insert rectangular cells (10, 20, 50 mm) Insert 100 mm rectangular cells (Prove 600)
4
5
10
9
11
10
9 • Remove the top o the round cell compartment
8 including the AutoSelector 10
• Insert the 100 mm rectangular cell vertically
11 into the cell holder 11 . Make sure that you hold
it with both hands on the small edges while
• Insert the rectangular cell vertically into the inserting it carefully
cell compartment 8 , ensuring that the cell is • Measurement starts automatically, and the
12 ush against the let side o the cell holder 9 measurement result is displayed in the concen-
at all times tration measurement overview (see page 33)
• Measurement starts automatically, and the
measurement result is displayed in the concen- NOTE
13 tration measurement overview (see page 33)
Please see Analytical Procedures and Appendices
or detailed measurement procedures.
14
15
16
66 Version 3.0 – 04/2021
9 Operation – 9.3 Measurements – 9.4 Zero Adjustment 1
9.4 Zero Adjustment

Minimum lling volumes or the cells used A valid zero adjustment is required or calcula-
3
tion of measurement values in Concentration,
Absorbance/% Transmission, Special/Multi-
Cell Filling
volume wavelength and Kinetic modes. During zero
(minimum adjustment, the absorbance o a cell lled with
4
distilled water ("zero cell") is measured and
10 mm Rectangular Standard 2 ml
stored.
10 mm Rectangular Semimicro 1 ml A zero adjustment must be perormed or each
cell type. The zero adjustment or concentration 5
20 mm Rectangular Standard 4 ml
methods is stored within the spectrophotom-
20 mm Rectangular Semimicro 2 ml eter separately or each cell type. The period o
50 mm Rectangular Standard 8 ml validity o the zero adjustment or concentration
methods can be edited in the System settings 6
50 mm Rectangular Semimicro 4 ml
(see section 9.2.4). When a zero adjustment has
100 mm Rectangular Standard 16 ml already been perormed or the inserted cell
round 4 ml type and the selected method, the date o the
most recent zero adjustment 1 is displayed in 7
the info line 2 .
10
2
1
9.4.1 Notes on Zero Adjustment 11

Zero adjustment with round cells
• Only use clean, scratch-ree round cells and
distilled water. 12
The minimum lling level is 20 mm. A ready
zero cell is contained within the scope o deliv-
ery of the spectrophotometer
• A ready zero cell can, in principle, be used or 13
an indenite period o time. We recommend,
however, that you regularly check the zero cell
or visible contamination and scratches and
rell or exchange it i necessary (at least every 14
24 months)
• Insert the round cell until it touches the bot-
tom o the round cell compartment
15
16
Version 3.0 – 04/2021 67
1 9 Operation – 9.4 Zero Adjustment
3
Zero adjustment with rectangular cells 9.4.2 When to Repeat the Zero
• With rectangular cells, zero adjustment must Adjustment?
be carried out using the same cell type (manu- We recommend that you repeat the zero
acturer and cell material [e. g. optical glass, adjustment in the following cases:
4 quartz glass, plastic]) as the one that will be • I the spectrophotometer was subject to me-
used or measurement. This is important be- chanical stress such as strong shock or trans-
cause cells o dierent manuacturers have di- port
ferent absorption characteristics. When chang- • I the ambient temperature has changed by
5 ing the cell type, repeat the zero adjustment more than 5 °C since the last zero adjustment
with the new type • At least once a week. The interval to repeat a
• Prior to zero adjustment, clean the rectangular zero adjustment is set in the instrument to 7
cell and ll it with distilled water. The minimum days. You can change this under "System (In-
6 lling level is 20 mm strument Settings)"
• Rectangular cells always have to be inserted in • I a new cell type (dierent manuacturer, di-
the cell compartment with the same orienta- erent glass type) is used
tion or measurement and zero adjustment • Basically, each time you want to measure with
7 (e. g. cell inscription always on the let side) the highest possible accuracy
• Insert the rectangular cell until it touches the
bottom and let edge o the holder. The opaque
NOTE
sides o the rectangular cell must point to the
8 ront and rear. The spectrophotometer detects I an interval to repeat a zero adjustment is set
stray light. If there is too much stray light, a you will be prompted to repeat it ater the inter-
message prompts you to close the cell com- val has passed. You can also repeat a zero ad-
partment cover justment by selecting a method, then touching
9 the "Settings" icon. Choose "Zero adjustment"
and insert a zero cell to start the measurement.
NOTE
Ordering inormation or cells is provided in sec-
10 tion 13. The cells listed in section 13 are special-
ly intended or use with the Spectroquant® test
kit system. Note that the spectral transparency
o the cell must be suitable or the intended ap-
11 plication (example, quartz cell or UV range).
12
13
14
15
16
68 Version 3.0 – 04/2021
9 Operation – 9.4 Zero Adjustment 1
9.4.3 Zero Adjustment or Concen- 3

tration Measurement Methods
A concentration method must be selected to
start the zero adjustment. There are two ways
to select the concentration method: 4
• By inserting a barcoded cell or the barcoded 5 4
AutoSelector
• By manually selecting the concentration meth-
od via the method list (see section 9.5) 5
6
1
2
4. Insert the zero cell according to cell type.
Zero adjustment starts automatically and, 7
i the zero adjustment is passed, a tick 4
appears in the status display eld or the
Zero adjustment 3 .
In the case o a method which only mea- 8
sures the sample at a single wavelength
the absorbance of the Zero value 5 is also
1. Once the concentration method is selected displayed.
touch Settings 1 . 5. With a cell inserted, the zero adjustment 9
2. Tap on the Zero adjustment button 2 . can be repeated manually by tapping on the
Start zero button 6 .
6. Tapping on the OK button accepts the zero
adjustment value or the method.
10
3 7. The screen changes to show the concentra-
tion measurement screen.
8. The instrument is ready to start measuring
11
the sample.
9.4.4 Zero Adjustment or Absorbance/

12
Transmission Measurements
(Ad hoc menu)
Zero adjustment must always be done
beore starting a measurement series and is 13
3. The Zero Adjustment screen opens. The
automatically prompted by the instrument (see
status display eld or the Zero adjustment
section 9.8.1).
is blank 3 .
NOTE 14
Cells must be absolutely clean and scratch-ree.
For zero adjustment, always use a cell o the
same type to measure the sample.
15
16
Version 3.0 – 04/2021 69
1 9 Operation – 9.4 Zero Adjustment – 9.5 Method List
9.5 Method List

3
9.4.5 Zero Adjustment or 9.5.1 Selecting a Method
Spectrum Measurements Manually
Zero adjustment must always be done Select a method rom the method list.
4 beore starting a measurement series and is
automatically prompted by the instrument (see 2
sections 9.8.2 and 9.9.2). 1
3
5 NOTE 4 5 6 7 8
Cells must be absolutely clean and scratch-ree.

For zero adjustment, always use a cell o the
same type to measure the sample.
6
9.4.6 Zero Adjustment or Kinetic

7 Measurements
Zero adjustment must always be done
beore starting a measurement series and is 1. In the main menu tap on the Method button
automatically prompted by the instrument (see 1 .
8 sections 9.8.3 and 9.10.2). 2. The screen 2 changes and the list o all
methods is displayed. The methods are listed
NOTE alphabetically. The arrow bar 3 on the right
9 Cells must be absolutely clean and scratch-ree. edge indicates that the list contains more
For zero adjustment, always use a cell o the methods urther up or down.
same type to measure the sample. 3. Select the required method.
4. The screen changes to show the method.
10 5. The instrument is ready to start measuring.
NOTE
The method designation contains the ollowing
11 information
4 Method number
5 Method name
6 Measuring range + unit (i the method is
12 suited or several cell ormats, the measuring
range over all cell ormats is displayed)
7 Citation (convertible)
8 Cat. No. (6 digits) or reerence to application
13 (“App” or test-kit-ree methods)
14
15
16
70 Version 3.0 – 04/2021
9 Operation – 9.5 Method List 1
3
9.5.2 Searching and Filtering the
Method List
You can search and lter the method list in order
to make it easier to nd the method you are 10
4
looking or:
5
9
7
2. Set lter by criteria 10 :
• All methods
• Last used methods: last six used in alphabetic
order 8
1. Filter by method type 9 : • Frequently used methods: six most requently
• All used in alphabetic order
• Concentration • Factory pre-programmed methods only
• Kinetic • User-specic methods only 9
• Spectrum • Area o application (e. g. brewery, color, oils,
sugar)
10
11
12
13
14
15
16
Version 3.0 – 04/2021 71
1 9 Operation – 9.5 Method List – 9.6 Programming a User-defined Method
9.6 Programming a User-dened

Method
3
11 14 1 2
4
12
13
3. Search by character string 11 . Proceed 1. Select Methods 1 in the main menu.

7 as follows: 2. Tap on Add New Method 2 in the method
1. Tap on the button 11 . list.
2. The key pad 12 pops up.
3. Enter search criteria: method name, method
8 number, item number (rst six digits without
any decimal point). I you type in ewer than
4 5 6
three characters, the search runs only at the
start o all search criteria (e. g. “ch” yields
9 hits such as “Chlorine”, “Chloride” etc.).
Typing in three or more characters starts a
search over the entire character string of the
search criteria (e. g. “nitr” yields hits such as
10 “Nitrate”, “Nitrite”, “Free amino nitrogen”
etc.).
4. Tap on OK 13 to activate the search lter.
5. The method list shows all methods matching
11 the search criteria. 3. The input window opens.
4. Select the type o method.
• Concentration method 4
NOTE
• Spectrum method 5
12 Searching with subscripted characters is also • Kinetic method 6
possible. To search or a method with subscript- 5. The screen changes.
ed characters you need to place an underscore 6. To continue with the programming turn to
in ront o the character to be subscripted. Tap the corresponding Sections 9.6.1 to 9.6.7.
13 on the C button 14 to deactivate the search il-
ter.
14
15
16
72 Version 3.0 – 04/2021
9 Operation – 9.6 Programming a User-defined Method 1
9.6.1 User-dened Concentration 3

Methods
Overview Line types
For Concentration Mode you can develop and The dependency between the nominal value and
store your own user-dened methods under absorbance is oten linear in a wide range as 4
method numbers 1001 to 1100. shown in 1 or non-linear as in 2 .
The spectrophotometer software supports you • Example o a linear calibration unction ater a
in creating the methods. Two dierent method 10-point calibration 1 . In the case of a linear
types can be programmed. dependency, the calibration unction is deter- 5
mined by means o linear regression. The slope
• Single-wavelength methods and axis intercept (E0) are the characteristics
• Multi-wavelength methods (max. 5 wave- o the calibration line:
lengths)
6
1
9.6.2 Calibration Data and Calibration 7

Absorbance
Function or Single-wavelength

Methods
In photometry, the calibration unction describes
the dependency between the measured param- 8
eter (e. g. concentration) and the photometric
measurement result (e. g. absorbance) o a
sample.
Nominal value (e. g. concentration) 9
The knowledge o this dependency is a pre-
requisite or the development o a photometric
method. The calibration unction is usually de- • Example o a non-linear calibration unction
termined by means o a series o measurements after 10-point calibration 2 . In the case of a
with standard solutions o known concentrations non-linear dependency, the calibration unc- 10
(nominal value), e. g. as a 10-point calibration. tion is calculated by a polynomial unction:
2
11
Absorbance
12
13
Nominal value (e. g. concentration)
NOTE 14
In measuring operation, the reverse calibration
unction is used to output a measured absor-
bance as a concentration value. 15
16
Version 3.0 – 04/2021 73
1 9 Operation – 9.6 Programming a User-defined Method
3 9.6.3 Programming/Modiying User-dened Methods (Single

Wavelength)
To program a user-dened single-wavelength method, proceed as ollows:
1. Select the method type "Concentration" (see section 9.6).
4 2. The screen changes.
5
1
5
3
4
6
2
6
9
7
7 8
9 3. Fill out input eld items 1 – 8 .

Item Input eld Possible input
1 Name Any name

NOTE
10 2 Wavelength Freely selectable (in nm) It is also possible to display characters in
subscript and superscript. To put a character in
16 (round), 10, 20, 50 or
3 Cell subscript, an underscore “_” must be placed in
100 mm
ront o the character in question (e. g. entry or
11 4 Citation form * e. g. PO4-P H2O = H_2O). To put a character in superscript,
a circumlex “^” must be placed in ront o the
5 Unit * e. g. mg/l character in question (e. g. entry or m2 = m^2).
0.001, 0.01, 0.1, 0.25, 0.5
12 6 Resolution
or 1 4. Tapping on the calibration eld 9 changes
Lower and upper Any value between zero and the screen.
7
limit of the mea- the highest concentration of
suring range the standard solutions used
13
8 User-dened Any value between zero and
range * the highest concentration of
the standard solutions used
14 9
Calibration func- (See examples on the ol-
tion lowing pages)
* Optional
15
16
74 Version 3.0 – 04/2021
4
11
10
5
5. You now have the following options for plot-

7
ting a calibration curve:
• Entry or measurement o value pairs 10
• Enter a unction directly into the elds E0,
A0 – A5 11
8
10
11
12
13
14
15
16
Version 3.0 – 04/2021 75
Enter value pairs
4 7
6
8 1
2
3
5 9
5
10
11
6
6
7
4 15 14 13 12
Enter the value pairs, nominal value (concentra- 3. "Fit E0" 5 can be activated as an additional
8 tion)/measured absorbance einer o an already option. With "Fit E0" activated, concentra-
available test series with the following value tion 0 (= reagent blank value) intercepts the
pairs: absorbance axis at the associated E0 value.
• E0 2 = reagent blank (see section 9.7.8) 4. Once all values are available, tapping on the
9 • At least one, max. eleven value pairs in various eld “Function” 9 calls up an overview of
concentrations the calculated coefcients. Tapping on the
eld “Graph” 10 shows the calibration curve.
1. Enter E0 2 , concentration o the standard
10 solution 3 and associated absorbance 4 us-
ing the keypad 1 . Tapping on the + button NOTE
5 allows urther (up to eleven) value pairs The calculated unction maps the calculation o a
to be entered. The UP&DOWN buttons 6 are result (e. g. concentration) via a measured ab-
11 activated i more than our value pairs are sorbance in the form of a polynomial equation of
entered. the ollowing type:
2. Activate the eld “Linear” 7 to calculate a
linear unction. When “Linear” is not activat- C = A0 + A1 x (Abs – E0) + A2 x (Abs – E0)2
12
ed, a non-linear unction o the second order
is automatically calculated (square unction). where:
C = measurement result (e. g. concentration)
NOTE A0, A1, A2 = coeicients (polynomial)
13
If you wish to calculate a linear function, you Abs = measured absorbance
require at least the E0 value and 2 value pairs. I E0 = absorbance o the reagent blank value
you wish to calculate a non-linear function, you
14 require at least the E0 value and 3 value pairs. 5. You can now enter an ID or a specic batch
code or the calibration. Tapping on the eld
“Batch ID” 11 opens a virtual keyboard.
Enter the code and conrm by tapping on
15 the OK button.
16
76 Version 3.0 – 04/2021
6. To close the determination o the You now have the ollowing options: 3
coefcients, conrm the data by tapping on
the OK button 12 . • Close the programming/processing o the
7. Tap on the Export button 14 to transfer method by tapping on the Save button 17 .
the data in the CSV ormat to an external All entered data are accepted. A method 4
storage medium. number appears 18 .
8. Tap on the Print button 15 to print out the Close the screen by tapping on the X
data. button 19 . The screen changes to the
9. To close the operation without accepting the method list.
5
data, tap on the X button 13 . All entered • To call up coefcients, value pairs, or the
data are deleted. graph again, tap on the eld “Calibration”
16 .
6
• Cancel the programming/processing o the
method by tapping on the X button 19 .
All entered data are deleted. The screen
changes to the method list. 7
16
8
17 19
18
10
11
12
10. Tapping on the OK button 12 conrms the

entered data, and the screen changes to the 13
method screen. A check appears in the eld
“Calibration” 16 .
14
15
16
Version 3.0 – 04/2021 77
Measure value pairs (see section 9.7.10)
4 7
6
8 1
2
3
5 9
5
10
11
6
6
7
4 15 14 13 12
1. Activate the Absorbance button E0 2 (a blue

8 rame appears).
2. Insert the cell with E0 (reagent blank value).
NOTE
9 16
I no zero adjustment has been made or the set 18

measurement conditions (wavelength and optical 19 20
path length), a zero adjustment is automatically
10 prompted. Please ollow the instructions that 17 22 21
appear.
11 3. The screen changes. Measurement starts

automatically.
The measured absorbance is shown 16 .
You have the option to perform multiple or
12 repeated measurements.
These can be perormed by reinserting the
cell or, i the cell has already been inserted,
by tapping on the Start button 17 .
13 The median value 18 ,and also the number
19 o measurements that have already been
perormed are shown.

Tapping on the arrow button 20 shows the
14 individual results o the measurements that
have been made.
Tap on the OK button 21 to conrm the
median value.
15 Tapping on the X button 22 closes the
operation.
The screen changes.
16
78 Version 3.0 – 04/2021
10. You can now enter an ID or a specic batch 3

4. Use the numerical keypad 1 to enter the
next concentration 3 and activate the code or the calibration. Tapping on the eld
corresponding absorbance eld 4 (a blue “Batch ID” 11 opens a virtual keyboard.
rame appears). Enter the code and conrm by tapping on the
OK button. 4
5. Insert the cell with the measurement
solution o the corresponding concentration. 11. To close the determination o the
Follow the procedure described above in coefcients, conrm the data by tapping on
step 3. the OK button 12 .
12. Tap on the Export button 14 to transfer
5
6. Perorm steps 4 and 5 or all values that are
required. the data in the CSV ormat to an external
7. Activate the eld “Linear” 7 to calculate storage medium.
a linear unction. When “Linear” is not 13. Tap on the Print button 15 to print out the
6
activated, a non-linear unction o the data.
second order is automatically calculated 14. To close the operation without accepting the
(square unction). data, tap on the X button 13 . All entered
data are deleted. 7
NOTE
If you wish to calculate a linear function, you
require at least the E0 value and 2 value pairs. I
you wish to calculate a non-linear function, you 8
require at least the E0 value and 3 value pairs.
8. „Fit E0“ 8 can also be selected as a urther 23

9
option. With "Fit E0" activated, concentration
0 (= reagent blank value) intercepts the
absorbance axis at the associated E0 value. 24 26
9. Once all values are available, tapping on the 10
eld “Function” 9 calls up an overview of
the calculated coefcients. Tapping on the
eld “Graph” 10 shows the calibration curve.
25
11
NOTE
The calculated unction maps the calculation o a
result (e. g. concentration) via a measured
absorbance in the form of a polynomial equation 12
o the ollowing type:
C = A0 + A1 x (Abs – E0) + A2 x (Abs – E0)2

13
where:
A0, A1, A2 = coefcients (polynomial)
Abs = measured absorbance Tapping on the OK button 12 conrms the 14
E0 = absorbance o the reagent blank value entered data, and the screen changes to the
method screen. A check appears in the eld
15
16
Version 3.0 – 04/2021 79
Enter a function:
3 You now have the ollowing options: This option can be used when an evaluation
unction is already available or has already been
• Close the programming/processing o the calculated previously by a calculation pro-
method by tapping on the Save button 24 . All gramme on the basis o available data.
4 entered data are accepted. A method number Enter a unction to calculate the concentration
appears 25 . rom the absorbance (reverse calibration unc-
Close the screen by tapping on the X button tion). You can enter on the spectrophotometer
26 . The screen changes to the method list. the coefcients o a polynomial equation
5 • To call up coefcients, value pairs, or the o the ollowing type:
graph again, tap on the eld “Calibration” 23 .
• Cancel the programming/processing o the C = A0 + A1 x (Abs – E0) + A2 x (Abs – E0)2 +
method by tapping on the X button 26 . A3 x (Abs – E0)3 + A4 x (Abs – E0)4 + A5 x
6
All entered data are deleted. The screen (Abs – E0)5
changes to the method list.
where:
7 C = measurement result (e. g. concentration)
A0, A1, A2, A3, A4, A5 = coefcients
(polynomial)
Abs = measured absorbance
8 E0 = absorbance o the reagent blank value
10
11
12
13
14
15
16
80 Version 3.0 – 04/2021
Enter coefcients:
1 4
3
5
2
5
4
6
7
9 8 7 6
1. Enter E0 1 and the required coefcients

A0 – A5 2 on the keypad 3 . At least one
8
coefcient (A1) must be entered.
2. You can now enter an ID or a specic batch
9
“Batch ID” 4 opens a virtual keyboard. 10
Enter the code and conrm by tapping on
the OK button.
3. Once all coefcients have been entered, 10
tapping on the eld “Graph” 5 shows the 11 13
calibration curve.
4. To close the determination o the coefcients,
conrm the data by tapping on the OK button 11
6 . 12
the data in the CSV ormat to an external
storage medium. 12
6. Tap on the Print button 9 to print out the
data.
7. To close the operation without accepting the
data, tap on the X button 7 . All entered 13
data are deleted.
14
8. Tapping on the OK button 6 conrms the
entered data, and the screen changes to the
method screen. A check appears in the eld 15
16
Version 3.0 – 04/2021 81
3 You now have the ollowing options: Example 2 (non-linear calibration function)
The coefcients o the reverse calibration unc-
• Close the programming/processing o the tion are determined by multiple regression.
method by tapping on the Save button 11 . All When doing so, the concentration has to be on
4 entered data are accepted. A method number the Y axis and the absorbance on the X axis. The
appears 12 . absorbance o the individual value pairs must
Close the screen by tapping on the X button always be corrected by the reagent blank.
13 . The screen changes to the method list.
5 • To call up coefcients, value pairs, or the X value Y value
graph again, tap on the eld “Calibration” 11 .
Absorbance Absorbance - RB* Concentration
• Cancel the programming/processing o the
method by tapping on the X button 13 . 0.010 0.000 0.0 mg/l
6
All entered data are deleted. The screen 0.020 0.010 0.1 mg/l
changes to the method list.
0.070 0.060 0.2 mg/l
Example 1 (linear calibration function) 0.150 0.140 1.0 mg/l

7
For entry as a ormula, you can determine the
0.325 0.315 2.0 mg/l
coefcients o the reverse calibration unction by
linear regression. When doing so, the concentra- 0.490 0.480 3.0 mg/l
8 tion has to be on the Y axis and the absorbance 0.655 0.645 4.0 mg/l
on the X axis. The absorbance o the individual
0.825 0.815 5.0 mg/l
value pairs must always be corrected by the
reagent blank. * = reagent blank
9
X value Y value Calculated calibration function (third order
polynomial):
Absorbance Absorbance - RB* Concentration
C = -0.044983 + 7.4807 × A -4.5229 × A2 +
10 0.050 0.000 0.0 mg/l * 3.8305 × A3
0.250 0.200 1.0 mg/l
or
0.451 0.401 2.0 mg/l
11 0.648 0.598 3.0 mg/l Calculated calibration unction (th order

0.850 0.800 4.0 mg/l polynomial):
C = -0.093083 + 9.9988 × A -27.549 × A2 +
1.053 1.003 5.0 mg/l
78.315 × A3 -99.226 × A4 + 46.604 × A5
12 * = reagent blank
Calculated calibration function:

C = 0.0027 + 4.9914 × A
13
14
15
16
82 Version 3.0 – 04/2021
Graphic presentation of the calibration function
As already described in the sections above, ater 3

entering/measuring value pairs or entering a unc-
tion, the calibration curve can be graphically called
up by tapping on the “Graph” button .
4
5
1
2
6 5 4 7 3 9 8 10
9
1. The calibration function 1 appears in the 2. The X axis shows the absorbance values,
graph. When the function has been calculat- the Y axis the corresponding results (e. g.
ed using value pairs, the coefcient o deter- concentration). Tapping on the X/Y button 10
3 changes the presentation o the axes
mination “R2” 2 is also shown.
around.
NOTE The formula of the calibration function 1
The unction determined in this way is used or
that is shown remains unchanged. 11
3. When the unction has been calculated
calculating a result (e. g. concentration) via a
via value pairs, these pairs are shown in a
measured absorbance in the orm o a polynomi-
separate eld 4 . The orward and back keys
al equation o the ollowing type:
5 , 6 can be used to call up the next value 12
pair. Tap on the C button 7 to reset the
C = Polynomial (Abs)
view.
where:
the data in the CSV ormat to an external 13
storage medium.
Abs = [measured absorbance o the sample or
standard] minus [absorbance of the reagent
data.
blank value (E0)] 14
6. Tap on the X button 10 to close the view
of the graphic presentation. The screen
changes.
15
16
Version 3.0 – 04/2021 83
3 9.6.4 Calibration Data and Calibration 9.6.5 Programming/Modiying User-

Function or Special Methods dened Special Methods
(e. g. Multi-wavelength) (e. g. Multi-wavelength)
In this mode you can carry out measurements Proceed in the ollowing manner to programme a
4 with special methods and unctions. You can use special method:
the ollowing unctions or these methods:
• Measurements at dierent wavelengths

5 • Multiple measurements at one wavelength
(e. g. beore and ater adding a reagent)
• Use o procedure variables. Procedure vari- 1
ables provide a value that has to be en-

6 tered prior to each measurement on the
spectrophotometer (e. g. volume, pH value or
temperature)
• Check whether a value meets a condition. With
7 a condition you can check a value or validity
(e. g. absorbance value, procedure variable or
the result o a ormula)
1. Select the method type "Concentration" 1
8
Formula editor or convenient programming o (see section 9.6).
any user-dened methods (see section 9.6.5).
10
3
11
12 2. Tapping on arrow 2 beside Concentration

opens the selection list.
3. Select Special Method 3 rom the displayed
selection list.
13
14
15
16
84 Version 3.0 – 04/2021
5. Fill out input eld items 4 – 13 . Tapping on

the input elds 5 , 11 , 12 , 13 changes the
4
4
screen. These can be programmed as de-
8 7 6
scribed in steps on the ollowing pages.
5 9
5
NOTE
11 12
It is also possible to display characters in
10 13 subscript and superscript. To put a character in
subscript, an underscore “_” must be placed in 6
ront o the character in question (e. g. entry or
H2O = H_2O). To put a character in superscript,
a circumlex “^” must be placed in ront o the
4. The screen changes. character in question (e. g. entry or m2 = m^2). 7
4 Name Any name 8
5 Wavelength Up to 5 wavelengths denable
6 Cell 16 (round), 10, 20, 50 or 100 mm 9

7 Citation form * e. g. PO4-P
8 Unit * e. g. mg/l
10
9 Resolution 0.001, 0.01, 0.1, 0.25, 0.5 or 1
Lower and upper limit o the mea- Any value between zero and the highest concentration o the standard
10
suring range solutions used
11
11 Procedure variable * Procedure variables provide a value that has to be entered prior to each
measurement on the spectrophotometer (e. g. volume, pH value or tem-
perature)
12 Formula unction Formula editor or convenient programming o any user-dened methods 12
13 Condition * With a condition you can check a value or validity (e. g. absorbance value,
procedure variable or the result o a ormula).
* Optional 13
14
15
16
Version 3.0 – 04/2021 85
4 19
15
5 16 14 17 18 21 20 22 23
6
6. Input eld 5 – wavelength: Up to ve Additional entry elds 19 are added by tapping
wavelengths can be set. Add urther entry on the + button 20 .
elds or wavelengths 15 by tapping on the Tapping the Delete button 21 removes the most
7 + button 14 . Tapping the Delete button 16 recently programmed input row 19 . Tapping on
removes the most recently programmed the Save button 22 accepts the input. Tapping on
entry eld 15 . Tapping on the Save button 17 the X button 23 closes the screen and opens the
accepts the input. Tapping on the X button previous input mask.
8 18 closes the screen and opens the previous
input mask.
10 5
11 11 24
11
12 7. The number o wavelengths created appears 9. The number o variables created appears in
in the display eld 5 . the display eld 11 .
8. Tapping on the Procedure variable button 11 10. Tapping on the Formula button 24 changes
changes the screen. Here you can program the screen. Using the ormula editor, you
13 up to ve dierent procedure variables 19 . can now create a reely denable unction
Dene the ollowing values. rom the variables and wavelengths you have
• Name = name of the variable dened.
(e. g. temperature)
14
• Min = lower limit of the variable value
• Max = upper limit o the variable value
• RES = decimal places in the variable value
(e. g. 0.1)
15
• Unit * (optional) = unit o the variable value (°C)
16
86 Version 3.0 – 04/2021
30
29
4
25 26 31 32 5
6
11. Tapping on the OK button 25 accepts the 13. Optional tapping on the Condition button 28
input. Tapping on the X button 26 closes the changes the screen. Here you can dene a
screen and opens the previous input mask. condition or valid measurements (e.g. absor-
bance wavelength 1 ≤ 2.50). Tapping on the 7
Variable and/or wavelength button 29 accepts
the selection and displays it in the pre-select-
ed output eld 30 .
8
NOTE
When methods are used that are subject to a
27
condition, the measurement result is calculated
only when the condition is met. 9
28
14. Tapping on the OK button 31 accepts the

input. Tapping on the X button 32 closes the 10
screen and opens the previous input mask.
12. An excerpt o the ormula you have dened
appears in the display eld 27 .
11
12
13
14
15
16
Version 3.0 – 04/2021 87
9.6.6 Programming a User-dened

Spectrum Method
3
For Spectrum Mode you can develop and store
your own userdened methods under method
34
numbers 1101 to 1120.
4 The spectrophotometer software supports you in
creating the methods. To create a user-dened
spectrum method, proceed
as ollows:
5
33 35
1
15. Tap on the Save button 33 . The method is

automatically given a number generated by
7 the system 34 . All values are stored.
16. Tap on the X button 35 to leave the edit
method screen.
8
36
1. Dene the method type (see section 9.6).
2. Enter a name or the method 1 . This name
9 is displayed in the method list.
10
11
17. The screen changes to show the method list
36 .
18. The method is now created and stored in the

12
instrument.
NOTE
13 To ind the method more quickly, use the ilter
function (see section 9.5.2).
14
15
16
88 Version 3.0 – 04/2021
3
8
6
4
2
4
5 5
9 7 9 7
Item Input Possible input 7. Tap on the Save button 7 . The method is
eld automatically given a number generated by
the system 8 . All values are stored. 7
1 Name Any name
8. Tap on the X button 9 to leave the edit
2 Start & Wavelength range Prove 300 | 600: method screen.
3 Stop 190 – 1100 nm
Wavelength range Prove 100: 320 – 8
1100 nm
11 10
Peak
4 Threshold value or peak detection
detection
Sampling rate for the wavelength

9
5 Interval
range
6 Selection Choice between absorbance or trans-

button mission
10
3. Dene the wavelength range or the method.

Start 2 & Stop 3 .
4. Determine the sensitivity 4 of the 11
method. 9. The screen changes to show the method list
5. Set the interval 5 . Here you can choose 10 .
between 0.1 nm, 1 nm, 5 nm. 10. The method is now created and stored in the
6. Set the choice between absorbance or trans- instrument.
12
mission 6 .
NOTE
To ind the method more quickly, use the ilter 13
function 11 (see section 9.5.2).
NOTE
A spectrum can comprise up to a maximum o 14
1000 measurement points. I the entry is invalid,
it is shown in red and cannot be accepted.
15
16
Version 3.0 – 04/2021 89
9.6.7 Programming a User-dened

Kinetic Method
3
10
4
1
2 5
5
6
3
7
6
8
7 9
4 11
8
For Kinetic Mode you can develop and store 4. Create the user-dened parameters
your own user-dened methods under method • Wavelength 3
9 numbers 1201 to 1220. The spectrophotometer • Unit 4
sotware supports you in creating the methods. • Interval 5
To create a user-dened kinetic method, proceed Prove 100 Minimum: 00:00:10 (hh:mm:ss)
as ollows: Prove 300/600 Minimum: 00:00:05 (hh:mm:ss)
10 • Delay 6
1. Dene the method type (see section 9.6). • Duration 7
2. Enter a name or the method 1 . This name • Slope factor 8
is displayed in the method list. 5. Tap on the Save button 9 . The method is
11 3. Select type o measurement: absorbance or automatically given a number generated by
transmission measurement 2 by tapping on the system 10 . All values are stored.
the button or the required measurement 6. Tap on the X button 11 to leave the edit
(the activated selection is light grey). method screen.
12
NOTE
Invalid entries are shown in red and cannot be
accepted.
13
14
15
16
90 Version 3.0 – 04/2021
3
1 Name Any name
4
2 Selection button Choice between absorbance or transmission
3 Wavelength Wavelength range Prove 300 | 600: 190 – 1100 nm

Wavelength range Prove 100: 320 – 1100 nm
5
4 Unit * User-dened input when an end result is to be calculated (e. g. enzyme activity U/ml)
5 Interval Time intervals between the measuring points (hh:mm:ss)
6 Wait Lead time to start o the series o measurements (hh:mm:ss) 6

Dierence in time between the rst and last measuring point in the series o measurements
7 Duration
(hh:mm:ss)
8 Slope factor * User-dened input or calculating a result = "slope actor" × "∆ A/min" (instrument automati- 7
cally calculates the dierence in absorbance/minute = ∆ A/min)
* = Optional (only useul or absorbance)
8
13 12
10
11
7. The screen changes to show the method list
12 .
8. The kinetic method is now created and 12

stored in the instrument.
NOTE
13
To ind the method more quickly, use the ilter
function 13 (see section 9.5.2).
14
15
16
Version 3.0 – 04/2021 91
9.6.8 Copying/Duplicating a
User-dened Method
3
1. Search and select the method (see section
9.5.2).
5 4
7
5. The method is created under the method
name with the addition "name-copy" 4 and
appears in the method list.
8 1 6. Modiy the duplicated method as needed
3
2
(see section 9.6.3).
10
2. Activate the method by tapping on the arrow
in the right column o the method list 1 .
11 3. A selection o various method editing possi-
bilities shows up 2 .
4. To copy the method, tap on the Copy button
3 .
12
13
14
15
16
92 Version 3.0 – 04/2021
9.6.9 Modiying a User-dened Method 9.6.10 Deleting a User-dened Method

rom the Method List rom the Method List
3
1. Search and select the method (see section 1. Search and select the method (see section
9.5.2). 9.5.2).
1
3
2
1
5
3
2
7
2. Activate the method by tapping on the arrow 2. Activate the method by tapping on the arrow
in the right column o the method list 1 . in the right column o the method list 1 .
3. A selection o various method modication 3. A selection o various method modication
possibilities shows up 2 . possibilities shows up 2 . 8
4. To modiy the method, tap on the Edit but- 4. To delete the method, tap on the Delete but-
ton 3 . ton 3 .
5. Now ollow the description in the sections on
programming a: 9
• User-dened concentration method (see sec-
tion 9.6.1)
• User-dened spectrum method (see section
9.6.6) 10
• User-dened kinetic method (see section 9.6.7)
5 4
11
12
5. The conrmation box asking i you really
want to delete the selected method pops up.
6. To delete the method, tap on the OK button
4 to conrm or on the X button 5 to cancel 13
the deletion process.
CAUTION
14
Following your conirmation the method is per-
manently deleted. Beore deleting any method,
we recommend you export a backup copy o it to
an external memory device. 15
16
Version 3.0 – 04/2021 93
3
9.6.11 Export User-dened Methods to
a USB Memory Device
4
1
5 2
7. Ater the OK button 4 has been pressed, 1. Filter the user-dened method in the method
the screen changes to Method List. The list, e. g. by using the search lter User 1 .
8 deleted method no longer appears in the 2. Activate the method by tapping on the arrow
2 .
method list.
10
3
11 4
12 3. A selection o various method modication

possibilities shows up 3 .
4. Make sure that an USB memory device is
connected to the spectrophotometer.
13 5. To export the method, tap on the Export
button 4 .
14
15
16
94 Version 3.0 – 04/2021
9 Operation – 9.7 Measuring in Concentration Mode 1
9.7 Measuring in Concentration

Mode
3
4
2
3 5
1 4
5 7
9.7.1 Measuring Cell Tests with a

Barcode 8
Inserting a cell with a barcode starts a measure- 2. The measurement result 1 is displayed. The
ment (see section 9.3). position of the measurement result in the
1. Insert the barcoded round cell through the measuring range is shown on the measuring
opening in the cover. The white position line range display bar 2 with a blue line. 9
on the cell has to be aligned with the posi-
tion mark on the spectrophotometer. Insert NOTE
the cell until it touches the bottom of the
round cell compartment. The spectropho-
Measurement results beyond the measuring 10
range are marked separately in the display (see
tometer selects the method based on the
section 9.7.4).
barcode and automatically starts measuring.
3. Further options: 11
NOTE
• Select a dierent citation orm by tapping on
I the barcode cannot be read, a message the citation orm display eld 4 (e. g. NH4 <–>
appears. In this case you can make a new NH4-N)
attempt by using the Spectroquant® round cell • Select a dierent measuring unit by tapping on 12
bearing the barcode or the AutoSelector as the unit display eld 3 (e. g. mg/l <–> mmol/l)
described. Care must be taken to ensure that • Make urther settings such as dilution or blank
the positioning mark on the round cell/ value measurements with "Settings" (see sec-
AutoSelector is aligned with the position mark on tion 9.7.5)
13
the spectrophotometer.
Alternatively, you can close the message and
then select the required method manually rom
the method list. 14
15
16
Version 3.0 – 04/2021 95
1 9 Operation – 9.7 Measuring in Concentration Mode
3
4. Content of the information bar in the
concentration mode:
7
5
8
Tapping on the Information bar button 5
opens the extended inormation bar. In the
concentration mode the ollowing inormation is
9 displayed:
100049 Calcium S-ID 0063 1+9

10 Article number
(rst 6 digits o order Method Name Sample ID with prex "S-ID" Sample Dilution
no.)
HC123456 EXP 12/31/2015 AQA2 EXP 08/19/2015 10 mm

11 Lot number test kit Expiration date test kit AQA2 status (valid until date/no. Path length o inserted cell
with Prex "EXP" o measurements) with Prex "AQA2
EXP"
ZA 08/20/2015 U-CAL 08/20/2015 RB 0,050 A 08/20/2015 SB 0,010 A

12 Date o Zero Adjust- Date user calibration Date + value user reagent blank with Value sample blank with
ment with Prex "ZA" with Prex "U-CAL" Prex "RB" Prex "SB"
13
14
15
16
96 Version 3.0 – 04/2021
9.7.2 Measuring Reagent Tests with

AutoSelector
3
Select the method by inserting the AutoSelector NOTE
and the spectrophotometer is ready to start Measurement results beyond the measuring
measuring. range are marked separately in the display (see
section 9.7.4). 4
1. Open the cell compartment cover.
2. Insert the AutoSelector in the cell compart-
4. Further options:
ment or round cell. The white position line
• Select a dierent citation orm by tapping on
has to be aligned with the position mark on 5
the citation orm display eld 4 (e. g. NH4 <–>
the spectrophotometer. Insert the
NH4-N)
AutoSelector until it touches the bottom.
• Select a dierent measuring unit by tapping on
the unit display eld 3 (e. g. mg/l <–> mmol/l)
NOTE 6
• Perorm urther settings such as dilution or
I the barcode cannot be read, a message blank value measurements through Settings
appears. In this case you can make a new (see section 9.7.5)
attempt by using the Spectroquant® round cell 7
bearing the barcode or the AutoSelector as
described. Care must be taken to ensure that
the positioning mark on the round cell/
AutoSelector is aligned with the position mark on 8
the spectrophotometer.
Alternatively, you can close the message and
then select the required method manually rom
the method list. 9
3. Insert the rectangular cell until it touches

the bottom and let edge o the holder. The
opaque sides o the rectangular cell must 10
point to the ront and rear. Inserting the
rectangular cell (1, 2, 5, [10 cm Prove 600
only]) automatically selects the correct mea-
suring range. The spectrophotometer starts 11
measuring automatically. As the spectropho-
tometer has an inbuilt ambient light protec-
tion there is no need to close the cell com-
partment cover. 12
13
14
15
16
Version 3.0 – 04/2021 97
9.7.3 Measuring Reagent-ree Tests and

User-dened Methods
3
User-dened methods and reagent-ree methods 4. Further options:
generally have no barcode and hence no auto- • Select a dierent citation orm by tapping on
matic method recognition. In such a case, select the citation orm display eld 4 (e. g. NH4 <–>
4 the method manually. NH4-N)
• Select a dierent measuring unit by tapping on
1. Select the method manually (see section the unit display eld 3 (e. g. mg/l <–> mmol/l)
9.5.1). • Perorm urther settings such as dilution or
5 2. The spectrophotometer is ready to start blank value
measuring. measurements through Settings (see section
3. Depending on the type, insert the cell as ol- 9.7.5)
lows: • Detailed inormation on the individual mea-
6 surement is shown in the information bar 5
Round cell: (see section 9.7.1)
Insert the round cell in the cell compartment or
round cells until it touches the bottom. NOTE
7 For some methods, e. g. chlorophyll, the options
Rectangular cell: listed under item 4 are not possible.
Insert the rectangular cell vertically so it touches
the bottom and let edge o the cell compart-
8 ment. The opaque sides o the rectangular cell
must point to the ront and rear. As the spectro-
photometer has an inbuilt ambient light protec-
tion there is no need to close the cell compart-
9 ment cover.
NOTE
10 Measurement results beyond the measuring
range are marked separately in the display (see
section 9.7.4).
11
12
13
14
15
16
98 Version 3.0 – 04/2021
9.7.4 Exceeding the Upper or

Lower Limits of the Measuring
3
Range
Measurement results outside the measuring

range are shown separately in the display.
2 4
If a result is beneath the lower limit of the
measuring range, a blue arrow 3 appears at the
lower measuring range limit, while a result above
the higher limit of the measuring range prompts
1
5
a blue arrow at the higher measuring range limit
5 .
A measurement result lying ar outside the

measuring range is not shown, and instead 6
either the text “Lo” (or very low results) or
respectively “Hi” 4 (or very high results)
appears.
Depending on the method, the measurement
7
result is displayed 1 as long as it remains inside NOTE
the measuring range between the upper and
In the event that the display o “HI” or “LO” or
lower limit. The position of the measurement
results lying beyond the measurement range is
result in display bar 2 . 8
deactivated in the “Quality” menu o the system
settings (see section 9.2.4), under certain cir-
cumstances - e. g. the measurement of analyte-
free samples - measurement results with a mi-
nus sign may also appear. This is entirely inten- 9
tional and is not a deect o the Prove spectro-
photometer. Experienced users know that every
measurement result is subject to a so-called
measurement uncertainty (actual result = dis- 10
played result ± measurement uncertainty). Many
3
users also need to determine measurement re-
sults for analyte-free samples, e. g. in connection
with method-validation operations. This is why 11
the display o measurement results with a minus
sign, which in certain conditions may be due to
the measurement uncertainty of the measure-
ment system, is allowed when the HI/LO display
12
5
is deactivated.
4
NOTE 13
For methods involving a special measurement
procedure, e.g. the chlorophyll analysis, results
beyond the measuring range are shown by “ --- ”.
14
15
16
Version 3.0 – 04/2021 99
9.7.5 Method-specic Settings or

Concentration Mode
3
1 9
5
2 10
3 11
6
4
5
7
6
8 7
10 When a method is selected, the Settings menu

can be activated. Depending on the method se-
lected, the settings available or the method are
as ollows:
11 6 Reagent blank
1 Dilution 7 Recalibration
2 Turbidity correction (global setting) 8 MatrixCheck
3 Show absorbance (global setting) 9 User-dened range
12 4 Zero adjustment (see section 9.4) 10 Dierentiation
5 Sample blank 11 Plausibility (general setting)
13 NOTE
The individual settings are greyed out i they are
not available or the selected method. Switching
a gobal setting (turbidity correction, show absor-
14 bance) ON activates it or ALL methods where it
is applicable.
15
16
100 Version 3.0 – 04/2021
9.7.6 Measuring Diluted Samples

I the concentration o a sample exceeds the
3
measuring range o a method, you can speci-
3
cally dilute the sample so that the concentration
o the diluted sample is within the measuring 4
range o the method. A valid measurement can

4
be perormed by this means. Once the actor or
the dilution has been entered, the meter con-
verts the concentration to that o the undiluted
sample. 5
5
NOTE
Optimum measurement results are achieved i
the concentration o the diluted sample is in the 6
middle o the measuring range o the method a- 2. Select Dilution 2 and conrm. The input
ter diluting. screen 3 for Dilution appears.
3. Tap on the dilution value in the display elds
4 , enter the actor on the keypad and tap 7
Having selected the method, enter the dilution
on OK button 5 .
as ollows:
8
2
7
1
9
10
11
1. Open the Settings menu 1 . 4. The spectrophotometer is ready to start
measuring.
The display 6 changes to measuring mode.
12
The dilution that was just entered is used or the
next measurement.
13
14
15
16
Version 3.0 – 04/2021 101
9.7.7 Sample Blank Value

The value entered or the dilution is only valid By measuring and using a sample blank value,
3
or the selected method. The dilution actor is measurement errors due to coloring and turbid-
deleted: ity o the sample matrix can be largely elimi-
• When the spectrophotometer is switched o nated. The sample blank value is a characteristic
• When dilution value 0 (= no dilution) is entered o the sample (coloration) to be currently deter-
4
in the Dilution screen. mined. The sample blank is diluted depending on
the method being used, but does not contain any
NOTE color reagents. The pH is the same as that o the
5 test sample.
I a dilution actor is active, it is indicated on the
display during measurement in the orm [1 + x]
7 . The dilution actor is also shown in the inor- NOTE
mation bar at the bottom (see section 9.7.1). The Adding reagents dilutes the sample. This can
6 maximum dilution is 1 + 999. change the pH o the sample. For this reason the
sample blank also has to be diluted and the pH
value adjusted accordingly. The sample blank
value is valid or the next measurement only.
7 The sample blank value can be determined by
single or multiple determination. With multiple
determination, the sample blank value is calcu-
lated as the median rom the single measured
8 values. Having selected
the method, measure the sample blank value as
ollows:
9
10 1
11
12
1. Open the Settings menu 1 .
13
14
15
16
102 Version 3.0 – 04/2021
NOTE
3
The use o the sample blank value is indicated by
the symbol 8 in the display being lit. The sam-
4
ple blank value is also shown with preix "SB" in
the information bar at the bottom (see section 4
5 9.7.1).
6 5
6
2. Select Sample Blank Value 2 .
3. Insert the cell with a suitable sample blank.
The sample blank measurement starts auto-
matically. The value is used only or the next 7
measurement.
4. The rst single measurement or the sample
blank value is perormed. The ollowing data
is displayed as the result: 8
• The measured absorbance rom the (last)
single measurement 4
• The median rom all single measurements
carried out so ar 5 9
5. If necessary, carry out further single mea-
surements to calculate the median.
10
8
11
12
13
6. Tap on OK 6 to accept the measurement.
7. The screen changes to the measuring mode
7 .
14
8. The spectrophotometer is ready to start
measuring.
15
16
Version 3.0 – 04/2021 103
9.7.8 Reagent Blank Value

The evaluation of the photometric measurement Having selected the method, measure the re-
3
always refers to the comparison value of a test agent blank value as ollows:
solution without the substance to be determined
(reagent blank value). Thus the inuence o the
basic absorbance of the reagents on photometric
4
measurement is compensated or. In practice,
the reagent blank value is measured with the
1
same amount o distilled or DI water instead o
5 sample. With photometric concentration deter-
mination, the reagent blank value is a constant.
The method data or all measurements with 2
Spectroquant® test kits (Concentration Mode)
6 include an exactly determined reagent blank
value. This value is overwritten if you measure
the reagent blank value yourself.
7 NOTE 1. Open the Settings menu 1 .

You can increase precision i you determine the 2. Select Reagent Blank Value 2 .
reagent blank value with a test from a new lot
and use the reagent blank value or all urther
8 measurements with that lot. This is especially
recommended or measurements in the vicinity
of the lower limit of the measuring range. To en-
4
able the assignment in the result documentation
9 at a later time, the batch code saved in the bar- 5
3
code o the inserted round cell or o the AutoSe-
lector is also documented; alternatively, you can 6
also enter the Lot Number on the reagent pack-
10 aging (Lot ID) when determining the blank value.
7
The actory blank values always remain stored in

11 the instrument and can be activated at any time.
3. Insert the cell with a suitable reagent blank.
The reagent blank values you measured yoursel
The reagent blank measurement starts auto-
also remain stored in the spectrophotometer
matically.
until they are overwritten by a new blank value
4. The Lot ID 3 is read rom the barcode. It
12 measurement.
can be edited manually, however.
5. The rst single measurement or the reagent
The reagent blank value can be determined by
blank value is perormed. The ollowing data
single or multiple determination. With multiple
is displayed as the result:
13 determination, the reagent blank value is calcu-
• The measured absorbance rom the (last)
lated as the median rom the single measured
single measurement 4
values.
• The median rom all single measurements car-
ried out so ar 5
14 6. The unction “User RB” is activated 6 .
7. If necessary, carry out further single mea-
surements to calculate the median.
8. Tap on OK 7 to accept the measurement.
15
16
104 Version 3.0 – 04/2021
NOTE
3
In the event that a Lot ID dierent rom the one
9
used to measure the reagent blank value is used
in a subsequent measurement, this is recognized
by the barcode o the inserted round cell or o 4
the AutoSelector. The activated user reagent
8 blank value is then automatically deactivated
and a corresponding message appears in the
display. Tapping “OK” clears the message and 5
the measurement is perormed automatically
without taking a user reagent blank value into
account.
6
8 .
10. The spectrophotometer is ready to start

measuring. 7
NOTE
The use o the reagent blank value is indicated
by the symbol 9 in the display being lit. The re- 8
agent blank value and its date o measurement
are also shown with preix "RB" in the inorma-
tion bar at the bottom (see section 9.7.1).
9
10
11
12
13
14
15
16
Version 3.0 – 04/2021 105
9.7.9 Automatic Turbidity Correction

The turbidity correction unction activates the
3
automatic recognition and compensation o the
light absorption caused by turbid substances.
Following activation, the unction remains per-

4
manently switched on. Measured values that 3
were measured with turbidity correction are la-
beled with the turbidity correction button on the
5 display 6 and in the documentation (printout
4
and memory).
NOTE
6 The turbidity correction unction is not active
when the spectrophotometer leaves the factory. 3. Select turbidity correction 3 with 0 = deac-
The setting or automatic turbidity correction is tivated and 1 = activated (in light grey).
possible with all methods where the turbidity 4. Accept the settings with OK 4 .
7 correction makes sense. Where a method does
not permit turbidity correction, the button 2 is
greyed out.
8 Having selected the method, activate the turbid-

6
ity correction unction as ollows:
9 5
2
1
10

5 .
11 6. The spectrophotometer is ready to start
measuring.
NOTE
12
The use o the turbidity correction is indicated
1. Open settings 1 .
by the symbol 6 in the display.
2. Select turbidity correction 2 .
13
14
15
16
106 Version 3.0 – 04/2021
9.7.10 User Recalibration (Standard

Adjustment)
3
A ew pre-programmed methods and the user- Calibration or user-dened methods
dened methods or concentration measurement
provide the option to optimize the original cali- 1. Select the method manually (see section
bration stored with the method by means o a 9.5.1).
4
user recalibration. When creating a user-dened
method you can also allow a user recalibration NOTE
(see section 9.6). I a zero adjustment has not been perormed,
When a method is called up or which a user the spectrophotometer informs you that you 5
recalibration is required, measurement is only need to perorm a zero adjustment.
possible with a valid user calibration. The use o
the user recalibration is documented along with
the measured value and indicated in by the cor- 6
responding symbol in the display. Activation
o user recalibration and its date are also shown
with prex "U-CAL" in the inormation bar at the
bottom (see section 9.7.1). 7
NOTE
A user calibration is always stored or the meth- 1
od presently called up. A user calibration is 8
erased only i a new user recalibration is carried
out.
9
2. Tap on "Recalibration" 1 .
3. The screen changes.
10
11
12
13
14
15
16
Version 3.0 – 04/2021 107
Calibration for Spectroquant® methods

3
NOTE
2
This option is applicable only for very few
4 Spectroquant® methods.
3
4
1. Select the method manually (see section
9.5.1), or by inserting a barcoded cell/
5 AutoSelector.
6 5
NOTE
6 I a zero adjustment has not been perormed,
4. The data 2 of the current calibration are the spectrophotometer informs you that you
shown. need to perorm a zero adjustment.
5. Tap on the “Value pair” button 3 to switch
7 to the view of the value pairs.
6. Tap on the “Graph” button 4 to switch to
the graphic presentation of the calibration
curve. 1
8
the data in the CSV ormat to an external
storage medium.
9 8. Tap on the Print button 6 to print out the 2
data.
A user recalibration can be perormed by any

10 of the means below. User recalibration by
• Entry o a unction (see section 9.6.3)
2. Tap on the Settings button 1 . The screen
• Entry as a value pair (see section 9.6.3)
changes.
• Measurement of value pairs (see section 9.6.3)
3. Tap on the Recalibration button 2 . The
11
screen changes.
12
13
14
15
16
108 Version 3.0 – 04/2021
10
4
7
4
3 5 5
12
13
11 6 8 16 15 17 14 7
4. Enter the user-dened nominal values (min NOTE

1/max 11) in the input elds provided 3 via To calibrate urther user-deined nominal values
the keyboard 4 . Tap on the “+” button 5 8
(max. 11), repeat steps 11 – 14 o the
o the numerical keyboard to add lines or procedure. Be sure to activate each input ield
further value pairs. Use the cursor buttons for the measurement values.
6 to navigate the screen up and down or
the lines of the value pairs.

9
11. Activate the eld “Linear” 9 to calculate
5. Activate the Absorbance button E0 7 (a blue
a linear unction. When “Linear” is not
rame appears).
activated, a non-linear unction o the
6. Insert the cell with E0 (reagent blank value).
second order is automatically calculated 10
Measurement starts automatically.
(square unction).
7. The measurement result appears in the
activated input eld.
NOTE
8. Activate the input eld 8 of the absorbance 11
o the next concentration. If you wish to calculate a linear function, you
9. Insert the cell with the measurement require at least the E0 value and 2 value pairs. I
solution (= standard + reagents acc. to the you wish to calculate a non-linear function, you
method description o the selected method) require at least the E0 value and 3 value pairs.
12
o the activated concentration.
10. Insert cell with Standard 1 measuring 12. "Fit E0" 10 can be activated as an additional
solution. Measurement starts automatically. option. With "Fit E0" activated, concentration
0 (= reagent blank value) intercepts the 13
absorbance axis at the associated E0 value.
14
15
16
Version 3.0 – 04/2021 109
3 13. Activating the UserCalOn button 11 calculates

the measurement results or the method
based on the user-dened calibration 18
that was perormed. To restore the actory-
4 set calibration or the method, deactivate the
UserCalOn button 11 .
14. Once all values are available, you can tap on
the eld “Graph” 12 to view the calibration
5 curve.
NOTE
The calculated unction maps the calculation o a
6 result (e. g. concentration) via a measured 16. To close the determination o the
absorbance in the form of a polynomial equation coefcients, conrm the data by tapping on
o the ollowing type: the OK button 14 . I the eld “U-CAL on” has
been activated, an icon 18 pops up in the
7 C = A0 + A1 x (Abs – E0) + A2 x (Abs – E0)2 measurement display.
17. To call up the calibration data again, select
where: Settings 1 and Recalibration 2 again.
C = measurement result (e. g. concentration) 18. Tap on the Export button 15 to transfer
8
A0, A1, A2 = coeicients (polynomial) the data in the CSV ormat to an external
Abs = measured absorbance storage medium.
E0 = absorbance o the reagent blank value
9 data.
15. You can now enter an ID or a specic batch
20. To close the operation without accepting the
data, tap on the X button 17 . All entered
“Batch ID” 13 öopens a virtual keyboard.
data are deleted.
10 Enter the code and conrm by tapping on the
OK button 14 .
11
12
13
14
15
16
110 Version 3.0 – 04/2021
9.7.11 MatrixCheck
The MatrixCheck unction is used to check i Practical instructions
3
the photometric determination is disturbed by • Ater evaluating the measured value o the
other substances present in the sample (sample sample, the
matrix). The MatrixCheck can be carried out by spectrophotometer suggests or the Matrix-
spiking or by diluting. The spectrophotometer Check to spike
4
enables a simplied MatrixCheck with the aid or dilute the sample and standard with suitable
of the Spectroquant® CombiCheck R-2 addition volumes.
solution or a pre-programmed ready-to-use CRM For each spiking or dilution, the relevant nomi-
standard. The MatrixCheck can be carried out nal concentration value is displayed 5
immediately. The volumes required or the sam- • To be able to reliably recognize matrix eects
ple and standards are displayed on the screen. by spiking, the increase in volume after spiking
The MatrixCheck is then carried out with a single should be small
spiking. For the MatrixCheck with a standard o • To be able to reliably recognize matrix eects 6
your own, however, you can enter the number of by diluting, the dilution actor should be high
spikings or dilutions yoursel (max. 3). • You can carry out the MatrixCheck as a series
of measurements, consisting of up to three
MatrixCheck by spiking determinations with dierent spiking volumes 7
For the MatrixCheck by spiking, the photomet- or dilutions respectively
ric determination is repeated ater a dened • Prepare all test sample solutions simultane-
amount o analyte has been added to the test ously at the
sample in the orm o standard solutions. Recov- beginning of the series of measurements 8
ery o the addition is automatically calculated as
ollows: NOTE
The spectrophotometer suggests the optimal
Recovery o addition [%] = 100 x 9
version or the MatrixCheck. Depending on the
{measurement value (sample + standard solution) – concentration of the sample in relation to the
measurement value (sample)}/{nominal value measuring range the spectrophotometer sets
(sample + standard solution) – measurement spiking or dilution. I both are possible you can
value (sample)} 10
make your own choice.
A matrix disturbance is likely i recovery is less

than 90 % or more than 110 %.
11
MatrixCheck by diluting
For the MatrixCheck by dilution, the photometric
determination is repeated ater the test sample
12
has been diluted with distilled water.
The nominal value or the determination is calcu-

lated rom the dilution, provided that there is no 13
disturbance due to the sample matrix. Ater the
photometric determination the measured value
is compared with the nominal value and the re-
covery rate is calculated. A matrix disturbance is 14
likely if recovery is less than 90 % or more than
110 %.
15
16
Version 3.0 – 04/2021 111
Performing the MatrixCheck

3
1. Measure the original sample without spiking
or diluting it (see section 9.7).
4 2. The measured value is displayed.
3. Tap on the Settings button 1 .
1
5. Tap on MatrixCheck 2 .
5 6. The screen changes. The ollowing elds ap-
pear:
7. Perorm the required settings i you wish to
2 make your own choice 3 , 4 , 5 , 9 , 10 .
6 8. Tap on the Start button 11 .
5 9
7
8 4 3
10
10
11
11
6 7 8
12
13
14
15
16
112 Version 3.0 – 04/2021
Item Field name Description
3 Toggle switch Dilution/Spiking Dilution/Spiking option. (Acceptance o the instrument pre-sets is recommend-
ed. Switching the option is possible only as long as this does not cause values 4
to be outside the measuring range.)
4 max. dierence Permitted deviation rom the nominal value in %
5 Standard ID Selection menu or actory-programmed standards or user-dened standard 5

(concentration denable). This option is only active or spiking
6 Sample (ml) Figure or the sample volume
Figure or the standard volume, in case o dilution the volume o distilled water 6
7 Standard (ml)
is displayed
8 Target value (mg/l) Expected measurement value
7
9 Concentration Only active or user-dened standard (concentration denable).
10 Delete button Remove rows that are not required
8
If Spiking is Active:
9
1 2
3 4
10
11
1. Mix the sample with the dened standard 1 5. The display shows the actual value 2 , re-
12
and perorm the test procedure as described covery in % 3 and the assessment o the
in the package insert. MatrixCheck 4 (ok/ail).
2. Insert the prepared cell.
13
3. The instrument starts measuring automati-
cally.
4. Repeat this procedure or each standard ad-
dition.
14
15
16
Version 3.0 – 04/2021 113
9.7.12 User-dened Range

If Diluting is Active: The unction “User-dened range” can be
3
used to set acceptance ranges (limits) or
measurement results. These acceptance
ranges can be oriented to legal and/or other
specications.
4 1 5 With the unction activated and the lower and
6 7 upper limits for measurement results set, the
setting “User-dened range” is also shown
5 in the measurement-range bar of the result
screen. Ater the measurement has been made,
the position of the measurement result on the
8 9 10
measurement-range screen can be used to
6 check whether it is within the dened limits.
Ater selecting the method, the user-dened
range can be activated as ollows:
1. Dilute the sample as required 1 and per-
7 orm the test procedure as described in the
package insert. 2
2. Insert the prepared cell.
3. The instrument starts measuring automati- 1
8 cally.
4. The display shows the actual value 5 , re-
covery in % 6 and the assessment o the
MatrixCheck 7 (ok/ail).
9
NOTE
If the AutoStore function is active the results are
10 stored automatically and can be recalled rom
the result list. If AutoStore is not active, the re- 1. Open the menu “Method settings” 1 .
sults are lost when the Close button 10 is 2. Select “User-dened range” 2 .
touched. In this case touch Printer 8 or
11 Save button 9 to store or print the results be-
ore closing the MatrixCheck.
12
1 2
13
14
3. Activate the input eld 3 for the lower limit.
15
16
114 Version 3.0 – 04/2021
9. The spectrophotometer is now ready to start

3
the measurement.
4
5
5
8
6
4. A virtual numerical keypad 4 pops up.
Now enter the numerical value for the lower
limit and tap the OK button 5 to conrm.
5. Repeat this procedure to enter the upper 10. After the measurement, the position of 7
limit. the measurement result is shown on the
measurement-range screen 8 .
11. To deactivate the user-dened range, open
the “Settings” menu 1 and select “User- 8
dened range” 2 .
6 10
9
11
12. Tap on the “C” button 9 . The user-dened 12

range is reset. The upper and lower limits
are shown as zero values.
7
13
14
6. Tap the OK button 6 to accept the settings.

7. The screen changes to the Measurement mode.
8. The “User-dened range” setting is shown in 15
the measurement-range bar 7 of the result
screen.
16
Version 3.0 – 04/2021 115
9.7.13 Dierentiation
For certain methods there is a special
3
“Dierentiation” unction that enables the
user to dierentiate between varying chemical
formulas of the analyte when interpreting
measurement results. In some methods or the
4
determination o chlorine, or example, total
chlorine can be distinguished rom ree chlorine
and the proportion o bound chlorine can be
5 10 automatically calculated and displayed.
Ater selecting the method, you can activate the
dierentiation mode in the ollowing manner:
6
13. Accept settings by tapping on the OK button
10 .
2
14. The screen changes to the Measurement
1
7 mode.
the measurement.
9
1. Open the menu “Method settings” 1 .
2. Select “Dierentiation” 2 .
10
11
3
12 4
13
3. Activate the dierentiation unction by
tapping on the shift key 3 . The function is
activated when “I” appears against a light
14 gray background 3 .
4. Accept the settings by tapping on the OK
button 4 .
5. The screen changes to the mode “Measure-
15 ment with dierentiation”.
the measurement.
16
116 Version 3.0 – 04/2021
9.7.14 Plausibility
NOTE The unction “Plausibility” is available or the
3
For those methods or which the dierentiation ammonium methods.
option is available, the procedures or activation Once it has been activated, the unction remains
and the measurement are individually described permanently switched on or all ammonium
in detail in the manual section “Analyical methods.
4
Procedures and Appendices”.
NOTE
In the practical environment it has emerged that
when the sample contains very high ammonium 5
concentrations a dierent color occurs and the
measurement signal in the photometric
5 6 measurement is no longer proportional to the
ammonium concentration. 6
In these cases, the reaction solution is no longer
yellow-green to green in color, the yellow
component becomes diminished, and the
solution turns turquoise to blue in color. 7
In this case a reliable statement on the
ammonium content is no longer possible, in the
worst case resulting in a substantial
7. To deactivate the dierentiation unction, misinterpretation of the ammonium content with 8
open the menu “Method settings” 5 . serious consequences for the environment.
The submenu shows the available settings When the optional plausibility check in the Prove
options. spectrophotometer is activated, the
8. Select “Dierentiation” 6 . measurement is carried out at several 9
9. Deactivate the dierentiation unction by wavelengths and the yellow component o the
tapping on the shift key 3 . The function is reaction solution is also measured. The Prove
deactivated when “0” appears against a light spectrophotometers are thus capable of
gray background. recognizing this deviation in the color and giving 10
10. Accept the settings by tapping on the OK a corresponding warning signal.
button 4 . This plausibility check helps the user to avoid
11. The screen changes to the Measurement misinterpreting the results of the ammonium
mode. concentration.
11
the measurement.
12
13
14
15
16
Version 3.0 – 04/2021 117
Ater selecting the method, you can activate the

3
plausibility check in the ollowing manner:
1
2
6
NOTE
The active status of the plausibility check func-
tion is shown on the display by the symbol 5 .
7 1. Open the menu “Method settings” 1 .
2. Select “Plausibility” 2 .
9
3
10 4
11
3. Activate/deactivate 3 the plausibility
check unction with 0 = deactivated or 1 =
activated (light gray).
12 4. Accept the settings by tapping on the OK 7
button 4 .
13 the measurement.
NOTE
14 I the plausibility check unc¬tion is active and a
very high ammonium concentration is present in
the sample (concentration considerably beyond
the measuring range o the selected method), a
15 window with a warning message 6 pops up.
Ater conrming by tapping on the OK button,
the measurement result is shown as “--- ” 7 .
16
118 Version 3.0 – 04/2021
9 Operation – 9.7 Measuring in Concentration Mode – 9.8 Ad hoc Measurement 1
(without selecting a specific method)
NOTE
3
I a zero cell (cell with distilled water) is
measured as the sample when the plausibility
check func-tion is active, the warning message
and the result “---” 7 also appear. 4
The reason or this is that the zero cell does not
show the yellow color described above.
9.8 Ad hoc Measurement (without selecting

a specic method) 6
Tapping on the ad hoc button in the main menu changes the
screen and the older that allows the type o ad hoc measure-
ment to be selected.
7
Absorbance/
Transmission Spectrum Kinetic
10
11
1 2 3
12
13
14
15
16
Version 3.0 – 04/2021 119
1 9 Operation – 9.8 Ad hoc Measurement (without selecting a specific method)
9.8.1 Ad hoc ABS/TRANS

Measurement
3
Proceed as ollows to perorm an
ad hoc ABS/TRANS measurement.
4
8
1
2 5 3
5
6 4 7 6 9
7
1. Select type o measurement: absorbance or 4. The screen changes 8 .
transmission measurement by tapping on 5. Perorm zero adjustment by inserting a cell
the button 1 or the required measurement with distilled water or tapping on the Start
8 (the activated selection is light grey). zero button 9 .
2. Dene the wavelength(s) or the measure-
ment(s). Do this by tapping on the button 2 .
A blue frame appears. Now enter the re- 10
9 quired wavelength by tapping on the keypad
3 .
NOTE
10 Tapping on the + button 4 brings up a new input
ield. Dierent wavelengths can be programmed
11
or the measurement. The selection can be delet-
ed by tapping on the Delete symbol 5 .
11
NOTE
Invalid entries are shown in red and cannot be 6. The Start zero button changes to the Start
12 accepted. button. The instrument is ready to start mea-
suring.
7. Start the measurement by inserting a cell
3. Conrm your input by tapping on the Start
with sample or tapping on the Start button
button 6 .
13 To cancel, tap on the X button 7 .
11 . The measurement results are displayed
in the absorbance column 10 .
14
15
16
120 Version 3.0 – 04/2021
9 Operation – 9.8 Ad hoc Measurement (without selecting a specific method) 1
9.8.2 Ad hoc Spectrum

Content of the information bar in ABS/ Measurement
TRANS measurements
3
Proceed as ollows to perorm a
spectrum measurement.
4
1
2
5
2
3
4
5 6
7
16 mm S-ID 0125 NOTE
Path length o inserted cell Sample ID with prex "S-ID" A spectrum can comprise up to a maximum o
empty
1000 measurement points. I the entry is invalid,
it is shown in red and cannot be accepted. 8
empty
1. Select type o measurement: absorbance or

transmission measurement by tapping on
the button 1 or the required measurement 9
(the activated selection is light grey).
2. Dene the wavelength range or the method.
Start & Stop 2 .
3. Determine the sensitivity of the measure-
10
ment 3 .
4. Set the interval 4 . Here you can choose
between 0.1 nm, 1 nm, and 5 nm 4 .
11
12
13
14
15
16
Version 3.0 – 04/2021 121
Content of the information bar for Ad hoc

3
Spectrum Measurements
4
6
6 7
7
5. Tap on the Start button 5 . ADHOC SPECTRUM S-ID 0033
6. The screen changes 6 . Measurement mode Sample ID
7. Perorm zero adjustment by inserting a cell with prex
8 with distilled water or tapping on the Start "S-ID"
zero button 7 (baseline determined). 10 mm 400 - 900 nm 1 nm

Path length of in- Scan range Scan
serted cell interval
9 empty
10
8
11
12 8. Start the measurement by inserting a cell

with sample or tapping on the Start button
8 .
9. The screen changes, the instrument is ready

13 to start measuring.
14
15
16
122 Version 3.0 – 04/2021
9 Operation – 9.8 Ad hoc Measurement (without selecting a specific method) 1
9.8.3 Ad hoc Kinetic

Measurement
3
Proceed as ollows to perorm a ki-
netic measurement.
4
9
1
3 4
2 5 5
6
8
10 6
7
4. The screen changes 9 .
5. Perorm zero adjustment by inserting a cell
with distilled water or tapping on the Start
zero button 10 . 8
10
1. Select type o measurement: absorbance or
11
transmission measurement by tapping on
the button 1 or the required measurement 11
(the activated selection is light grey).
2. Set the measuring range and the duration.
• Wavelength 2 6. Start the measurement by inserting a cell
• Unit 3 with sample or tapping on the Start button 12
• Interval 4 11 .
• Delay 5 7. The screen changes, the instrument is ready
• Duration 6 to start measuring.
• Slope factor 7 13
NOTE
Invalid entries are shown in red and cannot be
accepted. 14
3. Tap on the Start button 8 .

15
16
Version 3.0 – 04/2021 123
Content of the information bar for Ad hoc

3
Kinetics Measurements
7
ADHOC KINETIC S-ID 0034
Measurement mode Sample ID
with prex
8 "S-ID"
10 mm 00:00:30 00:00:05
Path length of Duration Time
inserted cell interval
9 empty
10
11
12
13
14
15
16
124 Version 3.0 – 04/2021
9 Operation – 9.9 Spectrum 1
9.9 Spectrum
9.9.2 Recording the Spectrum 3

9.9.1 General Information
With the Spectrum function, the absorbance or 1. Select the method rom the method list.
transmission in dependency on the wavelength is
measured and recorded. Record the baseline: 4
The wavelength range can be reely selected
within the measuring range of the spectropho-
tometer. The interval is selectable (0.1 nm,
1 nm, 5 nm). 5
A spectrum can be recorded in ad hoc mode (see

section 9.8), or loaded as a stored method (see
section 9.6.6). 6
Baseline
A baseline has to be recorded beore a spectrum
can be recorded. The baseline has to cover at 7
least the wavelength range of the spectrum be-
ing recorded. Once the baseline is measured, it
remains stored in the spectrophotometer until
• A new baseline is recorded 8
• Ad hoc Spectrum Mode is exited
• The loaded Spectrum method is exited
• The spectrophotometer is switched o
9
1
10
2. Zeroing against air: Tap on the Start zero 11

button 1 .
The spectrophotometer records the baseline.
Or
Zeroing against reerence solution: 12
Insert cell with reference solution. The
spectrophotometer records the baseline
automatically.
3. Wait until the baseline has been ully record- 13
ed. Once the baseline has been recorded,
the spectrophotometer is ready to start
measuring.
14
15
16
Version 3.0 – 04/2021 125
1 9 Operation – 9.9 Spectrum
3 Content of the information bar in spectrum

measurements
6
4. Insert the cell with the sample vertically
until it touches the bottom (rectangular cells
should touch the let edge o the cell com-
7
partment; the opaque sides o the rectangu- 1103 user spectrum S-ID 0187 S1
lar cell must point to the ront and rear). Sample ID with prex
Method number Method name
"S-ID"
8 16 mm 400 – 500 nm 1 nm
Path length of Scan range Scan interval
inserted cell
empty
9
10 2
11
5. Recording o the spectrum starts automati-
cally.
6. Once the spectrum of your sample has been
12 recorded you have the ollowing options:
• Evaluate the spectrum on the display immedi-
ately (see section 9.9.3)
• Print the spectrum by tapping on the Printer
13 button 2 either as a graph on a connected
printer – or as a pd le, i Print to pd is acti-
vated and a USB device is connected
• Save the spectrum to the result list. If
14 AutoStore is on, this is done automatically
15
16
126 Version 3.0 – 04/2021
9 Operation – 9.9 Spectrum 1
9.9.3 Evaluating a Spectrum

3
4
15 9
14 8
5
13 7
12 6 6
3
11 10
2 4 1 5 2
9
A spectrum can be evaluated immediately ater 5. The following mathematical functions for
measurement. Stored spectra can be loaded and various evaluating and calculating operations
evaluated rom the results list as well. The ol- are available or selection: 10
lowing tools are available or editing: • Derivate 12 : calculates the derivative o the
total spectrum. To calculate the second and
1. Moving to individual measurement points: third derivative, the unction can be carried
• Activate action icon 1 and use the next let/ out several times 11
next right icons 2 to move to all individual • Compare spectrum 13 : loads a second spec-
measurement points. The coordinates (wave- trum into the same diagram or direct com-
length and absorbance) o the respective mea- parison
surement point appear in the Ino box 3 • Subtract spectrum 14 : subtracts a stored 12
• Activate icon 4 to move to maximum values spectrum from the current spectrum
and icon 5 to move to minimum values • Add spectrum 15 : adds a stored spectrum to
2. Switch between graph view 6 and table the current
view 7 . spectrum 13
3. In the graph view use icons 8 to zoom out
and 9 to zoom in. Use navigation icon 10 to NOTE
optimize the position o sections o the graph The addition and subtraction o two spectra can
in the display. be applied only to the common wavelength
14
4. Tap icon 11 to return to the original spectrum. range of both spectra.
15
16
Version 3.0 – 04/2021 127
1 9 Operation – 9.10 Kinetics
9.10 Kinetics
3
9.10.1 General Information
The kinetics function enables the temporal trac-
ing of the absorbance or transmission of a sam-
4 ple at a certain wavelength.
The spectrophotometer automatically calcu-
lates the slope between two adjacent measuring
points rom the available measurement data.
5 The catalytic activity can also be determined and
displayed i required. To record the kinetics, the
spectrophotometer carries out single measure- 2
ments at regular intervals (measuring interval)
6 and stores the measured values as a time unc-
tion.
Kinetics can be recorded in ad hoc mode (see 2. Once the kinetic method has been selected,
7 section 9.8), or loaded as a stored method (see the screen changes rom the method list to
section 9.6.7). the Kinetics screen. The Start zero button 2
is active.
3. Insert the zero cell according to cell type.
8 9.10.2 Recording Kinetics Zero adjustment starts automatically.
NOTE
9 1 The zero adjustment can also be perormed
without a cell being inserted (measurement
against air). Tap on the Start zero button to start
the procedure.
10
11
12 1. Select the method rom the method list by

using the lter option 1 .
13
14
15
16
128 Version 3.0 – 04/2021
9 Operation – 9.10 Kinetics 1
Content of the information bar in kinetics

3
measurements
1201 Kinetik 1 S-ID Demo 4

Sample ID with pre-
Method number Method name
x "S-ID"
10 mm 00:01:00 00:00:05
Path length of Duration Time interval 5
inserted cell
empty
3 6
7
4. Following successul zero adjustment the
Start zero button 2 becomes the Start but-
ton 3 .
5. The instrument is ready to start measuring 8
the sample.
6. Insert the cell vertically until it touches
the bottom (rectangular cells should touch
the let edge o the cell compartment; the 9
opaque sides o the rectangular cell must
point to the ront and rear).
7. Recording o kinetics starts automatically.
10
11
12
13
14
15
16
Version 3.0 – 04/2021 129
1 9 Operation – 9.10 Kinetics
9.10.3 Evaluating a Kinetic

3
4
5
5 6
6
8
10 9
7
4 4
8
A kinetic can be evaluated immediately ater • Change to tabular view 7

10
measurement. Stored kinetics can be loaded and • Change to graphic display 8
evaluated rom the results list as well. The ol- • Cursor function 9 for incremental moves to
lowing tools are available or editing: ranges
• Let/Right tab unction 4 to gradually scan the • Call up original screen 10
11
kinetics with the indication o x and y values
• Zoom function to scale a section up 5 or
down 6
12
13
14
15
16
130 Version 3.0 – 04/2021
9 Operation – 9.11 AQA (Analytical Quality Assurance) 1
9.11 AQA (Analytical

Quality Assurance)
3
General information 9.11.1 Spectrophotometer Monitoring

The objective o analytical quality assurance (AQA1) 4
(AQA) is to secure correct and precise measure-
ment results (see section 5). At least one set o test standards such as
Spectroquant® PhotoCheck or Certipur® is re-
NOTE quired or spectrophotometer monitoring. The 5
administrator species which test standard has
Only persons in the administrator user group
to be used as the minimum requirement or
have access to the settings or AQA checks. Ev-
AQA1 monitoring. The extent o the monitoring
ery registered user can carry out the AQA check
can be expanded with urther test standards. 6
(see section 9.14).
The ollowing can be tested:
• Photometric accuracy
Analytical quality assurance (AQA) can be car-
• Wavelength accuracy
ried out in two steps independent o each other:
• Stray light test 7
• AQA1: Monitoring o the spectrophotometer
• Spectral resolution (only Prove 600)
• AQA2: Total system monitoring.
AQA2 covers the spectrophotometer, the test
NOTE
that is used, the accessories and the user's way 8
o working. Monitoring includes a check proce- Only persons in the administrator user group
dure that has to be successully repeated by the have access to the settings or AQA checks. Ev-
user within a certain period (AQA interval). ery registered user can carry out the AQA check
(see section 9.14).
9
NOTE
AQA monitoring is not active in the delivery Photometric accuracy
state. Normally test items of known absorbance val-
ues at dened wavelengths are used to monitor 10
photometric accuracy. The instrument is pre-
programmed with standard AQA1 tests that can
be perormed using Spectroquant® test items.
Such test items are, or instance: Spectroquant® 11
PhotoCheck, Certipur® UV/VIS-Standard 1A,
Certipur® UV/VIS-Standard 1.
Each pack is provided with a lot-dependent test
certicate with all nominal values (absorbances) 12
and tolerances o the test standards. These
nominal values and tolerances are already pre-
programmed in the spectrophotometer. Compare
them with the lot-dependent values and adjust 13
them i needed (see section 9.11.8).
14
15
16
Version 3.0 – 04/2021 131
1 9 Operation – 9.11 AQA (Analytical Quality Assurance)
NOTE Stray light test

3
The value of the tolerance consists of the toler- Normally test items with edge lter properties
ance o the standard (listed in the lot-speciic are used to monitor the eects o stray light.
certiicate) and the speciied tolerance o the The instrument is pre-programmed with stan-
spectrophotometer (see Technical Data Section dard AQA1 tests that can be perormed using
4
12). Spectroquant® test items. Such test items are,
or instance: Certipur® UV/VIS-Standard 2. Each
NOTE pack is provided with a lot-dependent test cer-
5 ticate with all nominal values and tolerances
Pay attention to the stability of the test stan-
o the test standards. These nominal values are
dard. A check on the values in the spectropho-
already pre-programmed in the spectrophotom-
tometer is required whenever a new pack o test
eter. Compare them with the technical data o
standards is used. The values must be adjusted
6 the spectrophotometer (see section 12).
if necessary.
NOTE
Wavelength accuracy
The value of the tolerance consists of the toler-
Normally test items o known absorbance max-
7 ance o the standard (listed in the lot-speciic
ima at dened wavelengths are used to monitor
certiicate) and the speciied tolerance o the
wavelength accuracy. The instrument is pre-pro-
spectrophotometer (see section 12).
grammed with standard AQA1 tests that can be
8 perormed using Spectroquant® test items. Such
test items are, or instance: Certipur® UV/VIS- Spectral resolution
Standard 6. Spectral resolution can be monitored using the
Each pack is provided with a lot-dependent test spectrum o a 0.02 % solution o toluene in hex-
certicate with all nominal values (wavelengths ane. The minimum ratio between the absorbance
9
with absorbance maxima) and tolerances o in the maximum at 269 nm and the absorbance
the test standards. These nominal values and in the minimum at 266 nm is a measure for
tolerances are already pre-programmed in the the spectral resolution. The instrument is pre-
10 spectrophotometer. Compare them with the programmed with standard AQA1 tests that can
lot-dependent values and adjust them i needed be perormed using Spectroquant® test items.
(see section 9.11.8). The test item used here is: Certipur® UV/VIS-
Standard 5.
11 NOTE
The value of the tolerance consists of the toler-
ance o the standard (listed in the lot-speciic
certiicate) and the speciied tolerance o the
12 spectrophotometer (see section 12).
13
14
15
16
132 Version 3.0 – 04/2021
9.11.2 Total System Monitoring (AQA2)

For total system monitoring, standard solutions
3
with a dened analyte content are required.
NOTE
Only persons in the administrator user group 4
have access to the settings for AQA checks. Any
registered user can perorm the AQA2 test.
Spectroquant® CombiCheck standards are ready-
to-use multiparameter standards, i.e. they can 5
be used or several test kits (methods). Standard
solutions (CRM) are ready-to-use single-parame-
ter standards, i.e. they can be used or single
test kits (methods). In addition to the above so- 6
lutions, single-parameter standard solutions
(z.B. Certipur®) can also be used. They are ad-
justed by dilution to the respective end concen-
tration. The end concentration should be approx- 7
imately in the middle o the measuring range.
NOTE 8
Suitable CombiCheck standards and single-pa-
rameter standards are listed in the catalog
"Water, Food and Enviromental Analytics" or on
the Internet. 9
10
11
12
13
14
15
16
Version 3.0 – 04/2021 133
9.11.3 AQA Overview

This main menu oers the ollow-
3
ing submenus:
6
4
7 1 2 3 4 5
Item Designation Description

9
1 AQA1 status Summary o the status o all activated AQA1 tests. (OK, ail, expired, next test in xx days)
2 AQA2 status Summary o the status o all activated AQA2 tests. (OK, ail, expired, next test in xx days)
10
3 AQA1 Activate, edit, perorm and create AQA1 tests
4 AQA2 Activate, edit, perorm and create AQA2 tests
11 5 PipeCheck Perform pipette check tests
Activated AQA1 test(s) and/or AQS2 test(s) are ailed or expired. Tapping the symbol
6 Attention
changes the display and opens an overview o the AQA tests involved.
12
13
14
15
16
134 Version 3.0 – 04/2021
9.11.4 Perform AQA1 Status

Check
3
To check the current AQA1 status of
the instrument, proceed as ollows.
1
6
7
1. Tap on the AQA1 status button . 1
2. The screen changes and a status overview o

the activated AQA1 tests is shown.
8
9
4
5
10
2
11
12
3
3. The screen shows the ollowing inormation: • Three dierent status symbols 5 : 13
• AQA1 method number 2 ! = expired/invalid; ü = test OK; - = test
• Name of the AQA1 test 3 ailed
• Number o days or which the AQA1 test is still 4. For quality or documentation purposes we
valid beore a new test has to be perormed 4 recommend printing the list.
14
15
16
Version 3.0 – 04/2021 135
9.11.5 Perform AQA2 Status

Check
3
To check the current AQA2 status of
the instrument, proceed as ollows.
6 1
7
1. Tap on the AQA2 status button 1 .
2. The screen changes and a status overview o
the activated AQA2 tests is shown.
8
7
10
11
12
3 4 5 6
13 3. The screen shows the ollowing inormation: • Three dierent status symbols 7 :
• AQA2 method number 2 ! = expired/invalid; ü = test OK; - = test
• Name of the AQA2 test 3 ailed
• Measuring range of the AQA2 check function 4 4. For quality or documentation purposes we
14 • Number o days or which the AQA2 test is still recommend printing the list.
valid beore a new test has to be perormed
5 , or number of measurements before a new
test has to be perormed 6

15
16
136 Version 3.0 – 04/2021
9.11.6 AQA1 Selection List

3
1
6
7
1. Tap on the AQA1 button 1 .
2. The screen changes and a list o all AQA1
tests stored in the instrument is shown.
8
4 9
2 6
10
11
12
3 5
3. The screen shows the ollowing inormation: • Three dierent status symbols 5 to remind 13
• AQA1 method number 2 you that a test is due. ! = expired/invalid;
• Name of the AQA1 test 3 ü = test OK; - = test ailed
• Number o days or which the AQA1 test is still • An empty circle means that the AQA1 test is
valid beore a new test has to be perormed 4 not activated 14
• Input buttons or editing 6 and creating 7
AQA1 tests
15
16
Version 3.0 – 04/2021 137
9.11.7 Activating and Deactivating an 9.11.8 Editing an AQA1 Test

AQA1 Test
3
Depending on the test type selected
• Photometric accuracy
4
6 • Wavelength accuracy
• Stray light test
• Spectral resolution (Prove 600 only)
5
the particular display screen changes. The input
values should be taken rom the lot-specic cer-
ticates or the test items and adjusted to suit
6 as ollows:
1. Tap on the Edit button 6 .

7
1
8
8

10 2. The screen changes. Depending on the AQA1
test selected a type-specic screen appears.
2. The screen changes. Make the AQA1 type specic changes in the
3. Tapping on the ON/OFF button 8 activates/ corresponding screen (see ollowing screens
11 deactivates the AQA1 test. (I = on, 0= o, and table).
the light grey background shows which state
is active).
12 NOTE
Before activating the lot-specific values for the
current test item, compare with the existing en-
13 tries or the input ields and make changes ac-
cordingly (see section 9.11.8).
14
15
16
138 Version 3.0 – 04/2021
Photometric accuracy (P) Stray light test (S)

3
1 1
6 7 8
2 3 4 2 3 4
4
5 5
13
12 5
12
9 10 11 9 10 11
Wavelength accuracy (W) Spectral resolution (Prove 600 only) (R)

7
1 1
6 7 8
2 3 4 2 3 4 8
5 5
14
12 9
12
9 10 11 9 10 11
10
11
12
13
14
15
16
Version 3.0 – 04/2021 139
3
Item Display eld Description Test
type
P, W,
1 Name Name of the test item
S, R
4 P, W,
2 EXP Expiry o the test item as shown in the certicate
S, R
P, W,
3 Lot number Lot number o the test item as shown in the certicate
S, R
P, W,
5 4 "0"/"I" (OFF/ON) Deactivate/activate the test
S, R
Test interval (input in weeks). For an active test the instrument reminds you when an P, W,
5 Interval
AQA1 test is due. S, R
6 Cell Pre-set name of the cell P, W
6 7 Target values Lot-specic nominal values P, W
8 Tolerance Tolerance range for the measurement value. P, W
(Tolerance = measurement uncertainty o the test item + specication o the spectro-
photometer)
7 9 Save Adopt values into the instrument
P, W,
S, R
10 Delete Delete user-dened AQA1 tests P, S
P, W,
11 Close Close display eld
S, R
8 P, W,
12 Numeric eld Touch the individual eld to enter values
S, R
13 Transmission Pre-set instrument-specic value (in % transmission) S
Nominal (Amax/ Pre-set instrument-specic value (minimum ratio between absorbance in the maximum
9 14
Amin) and absorbance in the minimum)
R
3. Make your individual changes in the type- NOTE

specic screens and store them by the Save AQA1 tests pre-programmed by the manuactur-
10
button 9 . To close the editing screen touch er (e. g. PhotoCheck) cannot be deleted.
the Close button 11 .
11
12
13
14
15
16
140 Version 3.0 – 04/2021
9.11.9 Performing an AQA1 Test

3
3
1
5
6
NOTE 4. The screen changes. A type-specic screen
In order to perorm an AQA1 test it must be ac- appears.
tivated (see section 9.11.7). 5. For urther actions ollow the on-screen
7
commands 3 .
6. Insert the corresponding reerence cell (e. g.
the zero cell as reerence cell in the Photo-
Check). 8
7. Insert ollowing test cell as prompted in the
command line.
9
5
2 4
10
11
12
2. The screen changes and you are presented 8. A tick appears following a successful test 4 .
with a list o all AQA1 tests stored in the 9. I all test steps are successul and the AQA1
instrument. test is passed, a tick appears in the com-
3. Select an AQA1 test by tapping on its name mand line 5 . 13
2 .
14
15
16
Version 3.0 – 04/2021 141
Content of the information bar in AQA1

3 NOTE measurements
I the test has ailed, a - symbol appears in the
display instead o the tick 4 symbol for a suc-
4 cessful test. If it is necessary to insert multiple
cells in succession during the test and one o the
individual test steps does not pass, check wheth-
er you have used the correct cell as prompted in
5 the command line. Repeat the measurement
with the correct cell. I one o the individual test
steps in a multiple series is not passed, the
AQA1 is not passed and the procedure is termi-
6 nated.
7 114693 PhotoCheck
Article number Method name
(rst 6 digits o order
no.)
8 HC123456 EXP 11/28/2016 12

week(s)
Lot number item Expiration date item Interval
with Prex "EXP"
9 Absorbance check
12
week(s)
Description of the
Interval
checked unction
10
11
12
13
14
15
16
142 Version 3.0 – 04/2021
9.11.10 Creating a User-dened AQA1

Test
3
3 4 4
5
1
It is possible to create two dierent user-dened 4. The screen changes. In addition, selection
types of AQA1 tests. options appear or the two types listed above 7
• Photometric accuracy (photometric accuracy 3 ; stray light test 4 ).
• Stray light test 5. Tapping on the particular Option buttons 3
1. Tap on the AQA1 button 1 . or 4 changes the screen. The entry mask
for that type appears. 8
10
11
2. The screen changes and you are presented 6. For editing (see section 9.11.8).
with a list o all AQA1 tests stored in the 12
instrument. NOTE
3. Tap on the Add button 2 . In the case o the user-deined AQA1 test or
photometric accuracy the input ields or the test
conditions (cell designation, 13
testing wavelength, nominal values for absor-
bance and absorbance tolerance) are added to
the screen by tapping on the
+ button 5 . 14
15
16
Version 3.0 – 04/2021 143
9.11.11 AQA2 Selection List

3

4 2. The screen changes and a list o all AQA2
tests stored in the instrument is shown.
1
6
7 5 6
9 3 7
4 8
10
11
3. The screen shows the ollowing inormation: • Number of measurements before a new AQA2
12 • Number o the method being tested 2 test has to be perormed 6
• Name o the method being tested 3 • Four dierent status symbols 7 : ! = test
• Display o the measuring range o the method expired/invalid; ü = test OK; - = test ailed;
being tested 4 = not activated
13 • Number o days or which the AQA2 test is still • Buttons to edit 8 AQA2 tests
valid beore a new test has to be perormed 5
14
15
16
144 Version 3.0 – 04/2021
9.11.12 Activating and Deactivating an 9.11.13 Editing an AQA2 Test

AQA2 Test
3
2
4
2
1
5
3 4
1. Tap on the Edit button 1 . The input values should be taken rom the lot-
specic certicates or the AQA2 test items and
2. The screen changes. 7
3. Tapping on the ON/OFF button 2 activates/ adjusted to suit as ollows.
deactivates the AQA2 test. (I = on, 0 = o, 1. Tap on the Edit button 1 .
the light grey background shows which state 2. The screen changes. A method-specic
is active). screen appears. 8
See example screen on the next page.
NOTE 3. This submenu oers the ollowing setting
options:
Before activating the lot-specific values for the
See description in the table below the ex- 9
current test item, compare with the existing en-
ample screens on the next page.
tries or the input ields and make changes ac-
4. Make your individual changes in the type-
cordingly (see section 9.11.13).
specic screens and store them by the Save
button 3 . 10
5. To close the editing screen touch the Close
button 4 .
11
12
13
14
15
16
Version 3.0 – 04/2021 145
1
4
3 4
8 12
5
9
6 11
7
13
2 5
8
9 7
10
10
11
12
13
14
15
16
146 Version 3.0 – 04/2021
3
Item Display eld Description
1 Name Name o the method being tested
Selection of Selection o the AQA2 standard (a choice can be made between Spectroquant®
2 4
standards pre-programmed standards such as
CombiCheck and a reely denable standard)
3 EXP Expiry o the standard as shown in the certicate or the standard
4 Lot number Lot number o the standard as shown in the certicate or the standard 5
5 "0"/"I" (ON/OFF) Deactivate/activate the test
6 Interval mode Test interval choice between weeks and and number o measurements
6
7 Interval (values) Enter test interval. For an active test the instrument reminds you when an AQA2 test is due.
8 Target value Lot-specic nominal value
9 Tolerance Tolerance range for the nominal value* 7

10 Save Adopt values into the instrument
11 Close Close display eld

8
12 Numeric eld Touch the individual eld to enter values
First 6 digits o the order number o the pre-programmed test kit corresponding to the se-
13 Article number
lected method
* The user must edit the tolerance range according to his/her requirements. The tolerance range should include typical errors 9
(measurement uncertainty) o the test medium used and o the method being checked. The typical error o the test medium
used can be taken rom the batch-specic certicate o the test medium. The typical error o the method being checked
must be calculated by the user under his/her conditions.
10
11
12
13
14
15
16
Version 3.0 – 04/2021 147
9.11.14 Performing an AQA2 Test

3
4 3
5 1
6
NOTE 5. Perorm an analysis as per the method de-
In order to perorm an AQA2 test it must be ac- scription using the selected AQA2 standard
tivated (see section 9.11.12). as sample and insert the cell.
7 6. Measurement starts automatically.
7. The measurement result appears in the dis-
play.
8. A tick appears following a successful test 4 .
8
9
2
4
10
11
2. The screen changes and a list o all AQA2 NOTE

tests stored in the instrument is shown. I the test has ailed, a - symbol appears in the
12 3. Select an AQA2 test by tapping on its name display instead o the tick 4 symbol for a suc-
2 .
cessful test.
4. The screen changes and shows the nominal
values and the tolerances 3 for the mea-
13 surement with the selected AQA2 standard.
14
15
16
148 Version 3.0 – 04/2021
Content of the information bar in AQA2

3
measurements
7
114942 Nitrate CombiCheck 20
Article number Method name AQA2 Standard Name
(rst 6 digits o order
no.)
8
HC123456 EXP 12/31/2015 AQA2 EXP 08/28/2015 10 mm
Lot number test kit Expiration date Interval Path length
test kit with Prex o inserted
"EXP" cell 9
ZA 08/21/2015 RB 0,100 A 08/21/2015
Date o Zero Adjust- Date + value user reagent
ment with Prex "ZA" blank with Prex "RB"
10
11
12
13
14
15
16
Version 3.0 – 04/2021 149
9.11.15 Perform PipeCheck

3
4. The screen changes 3 .
7 7
4
5 5
6
1
6
7
1. Tap on the PipeCheck button 1 . 5. Insert the reference cell.
2. The screen changes and a list o all Pipe- 6. After the reference cell has been successful-
Check tests stored in the instrument is ly measured, tick 4 appears on the display.
8 shown.
9
2
10
11
3. Select a PipeCheck by tapping on the name 7. A window pops up in which a name or code
12 2 . can be entered or the pipet being tested
(e. g. manuacturer, serial number or similar).
Tap on the input eld 8 and enter the name
or code. Tap the OK button to accept the
3
13 entry.
14
15
16
150 Version 3.0 – 04/2021
3
9. Insert the "Check" cell.
10. Ater the "Check" cell has been successully
measured, tick 5 appears on the display.
11. The dierence between reerence and Check 4
cell is calculated automatically. I this value
6 is within the pre-programmed tolerance,
the PipeCheck is passed shown by text and

tick 7 . 5
6
9
NOTE
9
I this value o the dierence is outside the tol-
erance, the test has ailed and a - 9 appears.
10
11
12
13
14
15
16
Version 3.0 – 04/2021 151
Content of the information bar in PipeCheck

3 measurements
7 114692 PipeCheck 2.0 S-ID SN123

Article number Method name Pipet designation/
(rst 6 digits o order no.) (incl. pipette volume) identication
HC123456 EXP 31.12.2017

8 Lot number item Expiration date item
with Prex "EXP"
empty
10
11
12
13
14
15
16
152 Version 3.0 – 04/2021
9 Operation – 9.12 Timer 1
9.12 Timer
You can use the timers to remind you
3
by an acoustic signal of a time inter-
val that has expired.
The spectrophotometer has two types o timers: 5

4
• The user-dened timer is a timer that can be
reely assigned
• Pre-programmed timer is a timer which oers
preset reaction times (<= 15 min) or pre-pro- 5
grammed actory methods
6
2
4. The screen changes back to the timer over-

view.
5. Tap on the timer name 5 and the screen 7
changes 6 .
8
6
The spectrophotometer manages all timers in 9

the timer list. The timer list is opened with the
Timer list button 1 .
If you want to manually enter time intervals, use
the user-dened timer unction. 7 8 9 10
3 11
6. Tap on the Start button 7 to start the timer.
Tap on the Stop button 8 to stop the timer.
The count down is interrupted.
Tap on Start button 7 again will continue 12
4
the count down. When the count down is
nished an acoustic signal sounds. The
insertion o a barcoded round cell enables
a measurement to be started immediately. 13
The insertion of an AutoSelector enables a
method to be selected immediately. Tapping
on the Start button 7 is active again and the
timer can be started once more. 14
2. The input window 3 opens. 7. Tap on the X button 9 to close the timer.
3. Enter the required time by tapping on the The display changes to timer list.
corresponding eld and conrm with OK 4 .
15
16
Version 3.0 – 04/2021 153
1 9 Operation – 9.13 Results and Measurement Datasets
9.13 Results and Measurement

Datasets
3
All results and measurement datasets in all mea-
surement modes are stored automatically in the
result list, as the instruments are delivered with
4 a factory pre-set AutoStore function (see section
9.2.5).
All stored results and measurement datasets can 2
be retrieved, exported and printed out.
5
NOTE 1
The instrument is capable o storing 2000 indi-
vidual measurements o the measurement
6
modes concentration, absorbance/transmission,
and/or multiple wavelengths as well as 20 data
I the AutoStore unction is deactivated, results
sets with results of the spectrum or kinetics
and datasets have to be stored manually ater
7 methods or each o these modes. It operates on
each measurement.
the FIFO storage principle (irst in – irst out),
1. Tap on the Save button 1 .
meaning that when all storage locations are
2. The system suggests a sample ID 2 .
occupied, the next time a result is to be stored
3. Tapping on Save 1 again stores the result
8 the oldest result on the list is automatically
under the suggested sample ID.
overwritten. Accordingly it is advisable to
regularly back up stored data sets on external
media (see section 9.13.7).
9 The results o the measurement modi AQA1,
AQA2, MatrixCheck, and PipeCheck are managed
separately. A total o 500 results are stored. 3
Results that determine a system or method

10 status are not overwritten, even when all
storage locations are already occupied.
4
11
4. To replace the sample ID by an individual

12 one, tap on the sample ID eld 2 .
5. Enter a new sample ID with the key pad 3 .
6. Conrm with the OK button 4 .
13
14
15
16
154 Version 3.0 – 04/2021
9 Operation – 9.13 Results and Measurement Datasets 1
9.13.1 Displaying the Results

3
1 3 3 3 3 3 4 5
5
8
2 2 2
10
6
8
7 6 9
10
By tapping on the Results button 1 you can ac- • Export selected results 6 . To export results
cess the result list via the main menu. they have to be selected by ticking the box 10
• Print selected results 7 . To print results they
The menu oers you the ollowing choices: have to be selected by ticking the box 10 11
• Sort ascending/descending 2 • Scrollbar 8 . Scroll through the result list by
• Filter result list 3 – (see section 9.13.3) tapping on the arrows of the scrollbar
• Clear lter 4 • Panoramic view o selected results rom one
• Select/deselect all results 5 . specic method 9 (see section 9.13.4) 12
• Results are selected by ticking the box 10 .
Remove the selection by tapping on the tick
symbol and the box will be emptied
13
14
15
16
Version 3.0 – 04/2021 155
7
1 2 3 4 5 6 7
The individual result line shows the ollowing • Result (e. g. in units or passed/ailed, depend-
8 inormation: ing on measurement mode) 4
• Measurement mode (e. g. concentration, spec- • Analyte tested 5
trum, kinetic, AQA) 1 • Sample ID 6
• Method number 2 • Tick box to select a result line to print or ex-
9 • Date and time o measurement 3 port the results 7
9.13.2 Show Details of One Result

10
11
12 1
13
1. Tap on an individual result line 1 2. A screen showing all details o one result or
dataset opens.
14 3. Tap on the print button 2 to print the indi-
vidual dataset on a printer or as a pd.
15
16
156 Version 3.0 – 04/2021
9.13.3 Filtering Results to Further

Process Measurement Datasets
3
1 2 3 4 5
Specied lter criteria can be set to select cer- • Filter by date. Tapping on the Date button
tain results and datasets to be exported, print- 3 brings up an input mask or the date range
7
ed, displayed or deleted. from … to ...
The ollowing criteria can be set: A calendar screen pops up.
• Filter using the individual measuring
mode 1 . The list shows the ollowing modes: 8
8 8
concentration, kinetic, spectrum, ad hoc,
AQA1, AQA2, PipeCheck and MatrixCheck
9
2
9
10
11
• Use the orward and back buttons 8 to scroll
through the calendar. Select the required date
and conrm by tapping on the OK button 9 .
The calendar closes. 12
• Filter using character strings. Tapping on • Tap on the OK button 6 to adopt the selected
the Search icon 2 brings up the keypad dis- period or ltering.
play eld. Enter search criteria such as method
name, method number or article number (rst NOTE 13
six digits o order number) no decimal point.
Keeping your nger on the “Forward“ and “Back“
Tap on OK to activate the search lter
buttons 8 scrolls through the calendar orwards
and backwards year by year.
14
15
16
Version 3.0 – 04/2021 157
9.13.4 Panorama – Value Control Card

The panorama function allows you to get a
3
graphical view o selected results rom one
10 single method (e. g. ammonium). The graphical
view corresponds to a value control card.
4
5
1
6
• Filter by sample ID 4 . Tapping on the but-
2
ton brings up a selection bar 10 which you can
use to lter according to sample ID
7
11
1. Use the lter option to create a result list or
5
one single method (see section 9.13.3).
8 2. Select the results to be included into the
panorama value control card by ticking the
boxes 1 .
3. The Panorama button is activated 2 .
9
10 12
• Filter by user name 5 . Tapping on the button 5
brings up a selection bar showing you the us-

11
ers created or the instrument. When you se-
lect a user, the measurement data list displays 4 4
all measured results or that user
12
NOTE
Tapping on the C button 11 cancels all of the fil-
4. Tapping on the Panorama button 2 opens
ters and the complete measurement data list ap-
the graphical view of the panorama value
13 pears. You can also search in iltered lists or set
control card or the selected values.
more filters, the filter status is shown in
5. Tap on the Forward/Backward buttons 4 to
the Infobar 12 .
get the individual datasets 5 for each value
displayed.
14
15
16
158 Version 3.0 – 04/2021
9.13.5 Print Results and Measurement

Datasets
3
I a postscript printer is connected to the spec-
trophotometer, results and measurement datas-
ets can be printed on paper by touching the Print
button (see section 8.3.2). 2 4
In addition, you can print the results and mea-
surement datasets as a pd. Connect an USB
memory device and activate "Print to pd" in the
submenu "Interace" in the "System" menu (see 5
section 9.2.2). To print to pd touch the Print but-
3
ton.
Print to pd will create the older PROVE on your 6

USB memory device. This older has several
subolders. You will nd the pd print les in the 2. Select the results to be included into the
older "Print". printout by ticking the boxes 2 .
3. Tap on the Print button 3 to print the re- 7
NOTE sults.
4. The in-progress icon appears during the
While printing more than one result, only results
data-export procedure.
rom a single measuring mode can be printed at
one time as each measuring mode has a dier-
5. I you print as pd to the USB memory de- 8
vice, wait for some more time before remov-
ent printing format.
ing the USB device to make sure all data are
transerred.
NOTE
9
In the Spectrum and Kinetics measurement
modes the results must be selected and printed
out separately.
10
11
1
12
13
1. Select the measuring mode or the results

you want to print, e. g. concentration 1 .
14
15
16
Version 3.0 – 04/2021 159
9.13.6 Deleting Results 9.13.7 Export Results and Measurement

The unction “Delete Results” is available only Datasets
3
when the user administration is activated (see In many cases it is advisable to export results
section 9.2.6) and the unction “Delete Results” and measurement data sets, e. g. to archive
is activated in the “Quality” menu o the system them or to analyze them in greater detail using
settings (see section 9.2.4). a suitable sotware tool. Besides oering data
4
transmission to a suitable memory device via the
USB port, the Spectroquant® Prove spectropho-
tometer also supports the automatic transfer of
5 measurement data via a local network
(Spectroquant® Prove Connect to LIMS, Y11086).
Spectroquant® Prove Connect is optionally avail-
able at www.proveconnect.com.
6
Data Transfer from Spectroquant®
1
Prove with USB Memory Device
7 NOTE
The reliability o data stored on USB memory
devices depends on the quality o the memory
When the unction is activated, the symbol or
device and the data transmission.
8 the unction “Delete” 1 is shown in the screen
of the results list.
Data are stored partly or not at all i or example:
• The power supply o the external memory de-
9 vice is interrupted during the write process, or
• The external memory device is prematurely
disconnected rom the spectrophotometer dur-
2
ing the data backup
10
To prevent a data loss we recommend the ol-
lowing:
• Store all data internally in the spectrophotom-
11 3 eter rst
• After performing a backup leave the USB
memory device connected to the spectropho-
tometer for some time
12 1. Click on the results that you wish to delete • Check whether the stored data is complete,
2 . e. g. on a PC
2. Tap “Delete” 3 to delete these results. • Use the USB memory device or data transport
but not or permanent data storage
13
NOTE
The deletion o results is documented in the user
log ile (or urther details on log iles please see
14 section 9.2.7 and section 16).
15
16
160 Version 3.0 – 04/2021
3
NOTE
The data are exported as CSV iles. To open
them in a spreadsheet make sure that your PC
calculation program is set to the same decimal 4
separator as the spectrophotometer (see section
8.2.4).
1
6
8
1. Select the results you want to export by
ticking the box 1 .
9
10
11
2
12
2. Export the selected results by tapping on the
Export button 2 .
3. The in-progress icon appears during the 13
data-export procedure.
4. When the in-progress icon disappears, wait
for some more time before removing the
USB device to make sure all data are trans- 14
erred.
15
16
Version 3.0 – 04/2021 161
1 9 Operation – 9.14 User Management
9.14 User Management

The Spectroquant® Prove allows the manage- Administrators and users log in to the spectro-
3
ment o up to 100 users. Every user is member photometer with their user name and password.
o a user group with dened user rights. Thus, documented measured values can later be
assigned to the user.
4 User groups
There are three hierarchical user groups: NOTE
• Administrator (top level) The user management function is not active in
• User (user account registered by the adminis- the Spectroquant® Prove when it leaves the fac-
5 trator) tory. Every user can carry out all unctions. Acti-
• Guest (user without user account) vating the user management creates an admin-
istrator user account.
6 User rights in detail
Action Administrator User Guest
7 Change system date/time X
Change GUI language X X
Firmware update X
8 Language update X
Method update X
Export error log X X

9 Run AQA1/AQA2 X X
Create/edit AQA1/AQA2 X
Display AQA status X X

10 Display AQA results X X
User management On/O X
Run measurements X X X
11
Evaluate measurements X X X
Export results and measurement datasets X X X
Handle user-dened methods X X

12
Recalibrate actory pre-programmed methods X X
Backup/restore X
13
14
15
16
162 Version 3.0 – 04/2021
9 Operation – 9.14 User Management 1
9.14.1 Activating/Deactivating the User

Management Function
3
I the user management unction is activated by
ticking 1 , each user has to log in to the spec-
trophotometer. After the login, the user has
certain rights depending on the user group. Only 4
an administrator can deactivate the user man-
agement unction. I the unction is deactivated, 1
every user has full instrument rights.
2
5
2. Tap on the box 1 . A tick appears.

3. Tap on the Save button 2 and the user
management is active. 7
4. An administrator can deactivate the User
management unction by tapping on the box
1 to remove the tick.
5. Tap on the Save button 2 to store the 8

changes. The User management is deacti-
1. Tapping on "System" and "User" opens the vated.
screen to activate the user management.
9
10
11
12
13
14
15
16
Version 3.0 – 04/2021 163
1 9 Operation – 9.14 User Management
9.14.2 Create an Administrator/

User Account
3
1
6
3
4
4
4
5 2
On the right part 1 o the screen "User Manage- 5. I the new user has been created success-
ment" administrators or users can be created. fully, the user name appears on the left 6 .
7 1. Tap on the Add button 2 to start to create
an administrator/user.
2. Select whether you want to create an admin- 7
istrator or user by tapping on eld 3 .
8
NOTE
The irst person who is created is automatically
assigned administrator rights. Here it is not
9
possible to select “Administrator” or “User”.
3. Enter name, password and conrm password

10 in the ollowing elds 4 .
7
11
12
5
13
4. The administrator resp. user is created by NOTE

14 tapping on the OK button 5 . Error messages 7 appear if the user name al-
ready exists or the password is not conirmed
correctly.
15
16
164 Version 3.0 – 04/2021
9 Operation – 9.14 User Management 1
9.14.3 Edit or Delete a User

NOTE 3
Edit or delete a user is possible only or
4
administrators. A regular user cannot access the
User management submenu and can change the
password in the Login/Logout menu only (see 4
section 9.15.1). 5
7 6
5
1
2
6
4. You can edit the access rights (Administrator
3
or User) 4 and change the password 5 .
5. Conrm your changes with the OK button 6 . 7
6. To delete a user proceed as beore and tap
on the Delete button 7 .
1. Tap on the System menu button and select 8

the submenu User.
2. Select the user from the list on the left 1 by
tapping on the corresponding name 2 .
9
9 8
10
11
7. Conrm your changes with the OK button 8 .
You can quit your decision with the X button
9 .
12
13
14
15
16
Version 3.0 – 04/2021 165
1 9 Operation – 9.15 Login/Logout
9.15 Login/Logout
I the user management is activated (see
3
section 9.14.1), login is mandatory to receive
user or administrator rights. Without login only
restricted guest rights are granted.
To login into the spectrophotometer proceed
4
as ollows:
5
4
7
6
6
1
2 3
7 8
8 1. Select Login/Logout 1 from the main menu. 5. Enter the user password by using the key
2. Tap on the arrow to open the user list 4 . pad 7 and conrm it with OK 8 .
NOTE 9
9 I a guest selects Login/Logout 1 from the main
menu the Logout button 2 is inactive. To contin-
ue as a guest press the Start button 3 .
10
11
5
12 6. The screen changes to the start screen. The

user name o the logged in user appears in
the upper status bar 9 .
13
3. Select the user name from the user list 5 .
4. Tap on the entry eld 6 .
14
15
16
166 Version 3.0 – 04/2021
9 Operation – 9.15 Login/Logout 1
9.15.1 Change Password with User 2

Rights Only
I you are registered with user rights only, you
have to change your password in the Login/ 3
10 Logout mode.
1
4
3
5
2
7. Wrong entry o the password creates the

warning "Invalid login" 10 . Repeat the proce-
6
dure above with the correct password.
To Logout as user or administrator

1. To change your password you have to be 7
logged in (see section 9.15). After successful
login your user name appears in the upper
status bar 1 .
8
4
1
9
2 3
4
10
1. Select Login/Logout 1 from the main menu.

2. Tap on the Logout button 2 . 11
3. The screen changes to the Guest mode.
4. To continue as a guest touch the Start
2. Select Login/Logout 2 from the main menu.
button 3 .
3. Tap on the lock icon 3 . 12
4. Enter the new password and conrm it in the
NOTE
entry elds 4 .
To continue as a dierent administrator or user, 5. Store your changes with the Save button 5 .
open the user list by tapping on the arrow 4 6. Login with your new password (see section 13
and select the user rom the list. Enter the cor- 9.15).
responding password as described above. 7. The screen changes back to the Login/
Logout mode.
Press Start again to continue. 14
15
16
Version 3.0 – 04/2021 167
1
10 Maintenance and Cleaning
10.1 Exchanging the Buer Battery

3 3. Insert the new batteries in the battery
compartment, making sure the polarity is
correct. Use only lithium ion batteries of the
type CR 2032. Insert the battery with the
4
writing acing upwards. The ± signs on the
batteries must match the ± signs in the bat-
tery compartment.
5
1
7 1
Battery service life
The power consumption of the clock is very low.
The service life of high-quality batteries is at
8
least ve years.
4. Close the battery compartment cover 1 .
Disposal of batteries
Dispose of the batteries at a suitable facility
9 NOTE
according to local legal requirements. Please
do not dispose o the batteries with household I you leave the spectrophotometer switched on
refuse. during the
Within the European Union, the batteries are exchange or insert the new batteries within a
10 minute ater taking out the old ones, the date
removed at specialized treatment centers at the
end o the instrument's lie. Instruments are tak- and time are retained in the spectrophotometer.
en to one o these specialized treatment centers
11 via the recycling system set up for this purpose.
12
13
2
14
1. Open the battery compartment cover 1 .

15 2. Remove the old batteries 2 from the bat-
tery compartment using a pair o tweezers.
16
168 Version 3.0 – 04/2021
10 Maintenance and Cleaning – 10.2 Exchanging the Lamp (Prove 100) 1
10.2 Exchanging the Halogen Lamp

(Prove 100)
3
WARNING
Beore exchanging the lamp, switch the instru-
ment o and disconnect the power plug rom the 4
power supply! I the lamp has snapped o or is
broken, it must be exchanged by the Customer
Service Department, as there is a risk of signifi-
cant injury! 5
CAUTION
There is a risk o explosion i unsuitable lamps
6
are used.
Use only the lamp that is intended or use with
your instrument.
7
Disposal of the lamp
Dispose o the lamp at a suitable acility accord-
ing to local legal requirements. Please do not
dispose o the lamp along with household reuse. 8
Within the European Union, the lamp is removed
at specialized treatment centers at the end o
the instrument's life. Instruments are taken to
one o these specialized treatment centers via 9
the recycling system set up for this purpose.
3. Open the lamp compartment cover and re-

Procedure 10
move it.
11
12
1
13
1. Place the spectrophotometer bottom-side up 4. Careully remove the lamp module rom the
14
on a soft surface. lamp compartment. Do not touch the lamp.
2. Remove the screws of the lamp compartment Use the mount 1 to take it o.
cover with an appropriate screw driver.
15
16
Version 3.0 – 04/2021 169
1 10 Maintenance and Cleaning – 10.2 Exchanging the Lamp (Prove 100) – 10.3 Cleaning
10.3 Cleaning
If a cell has broken or in the event of a reagent
3
accident, the spectrophotometer must be cleaned
immediately.
4 WARNING
Cells can contain dangerous substances. I the
contents are released, ollow the saety instruc-
tions o the Material Saety Data Sheet (MSDS).
5 If necessary, apply appropriate protective mea-
1 sures (protective goggles, protective gloves
etc.).
6 CAUTION
5. Install the new lamp module in the lamp
compartment. Use the mount 1 to insert Do not turn the spectrophotometer upside down
the lamp module. to remove the liquid. Doing so can cause the liq-
7 Do not touch the lamp to ensure that the uid could come into contact with electronic com-
service lie o the lamp is not aected! ponents and damage the spectrophotometer.
6. Close the lamp compartment cover with an
appropriate screw driver. CAUTION
8 The spectrophotometer has two drains at the
NOTE bottom through which the contents of broken
Upon renewed use o the spectrophotometer, re- cells or spilled liquid can drain o without caus-
boot the instrument and reset the lamp counter ing any damage to the instrument.
9 to zero (see section 9.2.1). If the self-test is
passed the instrument is ready or urther mea-
surements.
10.3.1 Cleaning the Housing and Display
10
CAUTION
The housing components are made o synthetic
materials. Avoid any contact with acetone, simi-
11
lar solvents and detergents containing such sol-
vents. Wipe o any splashes immediately.
Displays: Avoid contact with concentrated miner-
al acids, concentrated caustic solutions, benzyl
12
alcohol and methylene chloride. Wipe o any
splashes immediately.
13 Clean the spectrophotometer housing as ollows:

• I the housing surace is dirty, wipe it with a
sot cloth and mild soapy water
• Remove any chemicals splashes as soon as
14 possible
• For disinection, brie use o isopropanol is per-
mitted
Clean the displays with a sot cloth, i needed
15 moistened with clean water and a mild deter-
gent.
16
170 Version 3.0 – 04/2021
10 Maintenance and Cleaning – 10.3 Cleaning 1
10.3.2 Cleaning the Cell Compartment
CAUTION Prove 600: 3

2. Take the round cell holder 3 out. Take the
The cell compartment components are made o
cell compartment 4 with both hands, put
synthetic materials. Avoid any contact with ace-
the let and right orengers against the
tone, similar solvents and detergents containing
interior let and right o the compartment. 4
such solvents. Wipe o any splashes immediately.
3. Remove the cell compartment by pulling
evenly with both hands and hold it in a hori-
Routine cleaning of the cell compartment is zontal position.
normally unnecessary. Remove dust and slight 5
contamination with a moist, lint-free cloth. In
NOTE
case o spilled reagents switch the instrument
o and take the cell compartment out. Rinse it Put the cell compartment back in place by the
with clean water. same procedure. 6
Use isopropanol briey to remove persistent It is important that the compartment is com-
contamination (e. g. reagent residues). pletely pushed down to avoid alse measurement
results.
CAUTION 7
Clean the cell compartment only while wearing Prove 100
proper gloves to protect your hands! Prove 300 2
8
Make sure that you wear appropriate gloves and
clean the cell compartment as ollows:
1. Open the cover 1 . Prove 600
3
9
Prove 100 and 300:
2. Take the cell compartment 2 with both
Prove 600
hands, put the let orenger against the let 4
side o the interior o the compartment and 10

take the round cell holder with your right
hand.
3. Remove the cell compartment by pulling
evenly with both hands and hold it in a hori- 11
zontal position.
1
12
13
14
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Version 3.0 – 04/2021 171
1 10 Maintenance and Cleaning – 10.3 Cleaning
3 In case a cell is broken in the cell compartment

proceed as ollows:
1. Switch o the spectrophotometer 1 and dis- 1
connect the power plug 2 .
4 2. Liquid might have drained onto the lab bench
through the drains on the bottom o the in-
strument. Remove the instrument and clean
the bench.
5 3. Wipe the bottom of the instrument clean
without turning it upside down. 2
4. Remove the cell compartment as described
above.
6 5. Carefully remove all broken glass, e. g. with
a pair o tweezers.
6. Rinse the cell compartment thoroughly with
clean water. Dry it with a lint-free cloth. Use
7 isopropanol briey to remove persistent con-
tamination.
7. In case the non-removable part of the com-
partment is dirty clean it with a clean cloth.
8 8. Insert the cell compartment as described
above.
NOTE
9
Upon renewed use o the spectrophotometer,
reboot the instrument. I the sel-test is passed
the instrument is ready or urther measure-
10 ments. If the self-test fails, check whether the
detector lens is dirty and clean it (see section
10.3.4).
11
12
13
14
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172 Version 3.0 – 04/2021
10 Maintenance and Cleaning – 10.3 Cleaning 1
10.3.3 Cleaning the Cell Compartment

Cover and the Rear Cavity
3
CAUTION 3. Clean the cover and the rear cavity with
clean water and
The cell compartment cover is made o synthetic
dry it with a sot lint-ree cloth.
materials.
4. Insert the cover: Put the round ttings at
4
Avoid any contact with acetone, similar solvents
both sides 3 into the sliding cavities let and
and detergents containing such solvents. Wipe
right by pressing the ttings 2 again and
o any splashes immediately.
moving it slowly and careully back and orth
5
until the round ttings sit and the cover
Clean the cell compartment cover as ollows: slides perectly again.
• If the surface of the cell compartment cover is
dirty, wipe it with a sot cloth and mild soapy
water 6
• Remove any chemicals splashes as soon as
possible 2 Press
• For disinection, brie use o isopropanol is per-
mitted
2 Press 7
You can remove the cell compartment cover in

case anything is spilled or allen into the rear 3
8
cavity. Proceed as ollows:
Rotate
10
1
11
12
13
1. Open the cell compartment cover 1 .
2. Press the two ttings inside o the cover 2
and rotate the cover slightly. It will turn and
you can take it out. 14
15
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Version 3.0 – 04/2021 173
1 10 Maintenance and Cleaning – 10.3 Cleaning
10.3.4 Cleaning the Detector Lens

Routine cleaning o the detector lens is normally
3 3
unnecessary. Cleaning the detector lens can be
necessary in the ollowing cases: 4
• I the lens is visibly smudged, e. g. ater a cell
has broken or ater a reagent accident (see
4
section 10.3.2)
• If the self-tests fails
5 NOTE
I the lens is oten smudged make sure to pro-
tect the instrument rom dirt, dust and evapo-
rates o chemicals. Check operation conditions o
6 temperature and humidity. They have to be in
accordance with the values speciied in the tech-
nical data sheet (see section 12).
9
2
10
Proceed as ollows to clean the detector lens: 2. Cut o the end (approx. 2 cm) o a Dacron®
The detector lens is situated on the ront let swab, e. g. HY-LiTE® sampling pen, Cat. No.
side o the 1.30102.0021.
11 rectangular cell compartment 3 . 3. Grasp the cut-o end with the tip o a pair
o tweezers or small pliers. Clean the lens 4
1. Switch o the spectrophotometer 1 and with the dry head o the swab. To do
disconnect the power plug 2 . so, move the head rom the center o the
12 lens outward in
circles. If there is persistent contamination,
moisten the swab with a little deionized wa-
ter or isopropanol.
13
NOTE
Upon renewed use o the spectrophotometer,
14 reboot the instrument. I the sel-test is passed
the instrument is ready or urther measure-
ments.
15
16
174 Version 3.0 – 04/2021
1
10
11
12
13
14
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Version 3.0 – 04/2021 175
1
11 Error Causes and Trouble-shooting
Error Cause Remedy
3 Sel-test does not start. • A cell is inserted in one o the cell • Remove the cell
Start button is inactive compartments • Then tap on the Start button
• A oreign object is inserted in one o the • Remove the oreign object
two cell compartment • Then tap on the Start button
4 • The cell compartment is dirty • Clean the cell compartment (see section
10.3.2)
• Restart the instrument
• Instrument deective v Contact service department
5
Sel-test ailed • System check: Instrument deective • Contact service department
• Lamp check: Lamp deective • Prove 100: Change the lamp
• Prove 300 | 600: Contact service depart-
ment
6
• Wavelength check: • Remove the oreign object
• Foreign bodies in the cell compartment • Clean the lens (see section 10.3.4)
• Lens dirty I this happens repeatedly, check the oper-
• Instrument deective ating conditions (see section 8.1)
7 • Contact service department
The message "system • System has rozen • Switch the instrument o, wait 1 minute
error" shows up and turn it on again. I error message re-
8 mains contact service department
Instrument does not • Operating condition undened or EMC • Disconnect instrument from the electricity
react to touchscreen load unallowed supply, wait 1 minute and re-connect
operation
9
Measuring range under- • Selected method's measuring range • Select method with suitable measuring
cut or exceeded not suitable to sample concentration range
• Dilute the sample
10
Obviously incorrect • Cell dirty • Clean the cell
measured values • Dilution set incorrectly • Set the dilution
• Selected method not suitable • Select dierent method
11 • Zero measurement incorrect • Perorm zero measurement
• Blank value incorrect • Re-measure the blank value
12
13
14
15
16
176 Version 3.0 – 04/2021
11 Error Causes and Trouble-shooting 1
Error Cause Remedy
Data transfer to USB • Power supply o USB is interrupted • Connect power supply 3
not working • USB has been disconnected while • Wait for one more minute before
data transer was still running disconnecting the USB device rom the
instrument
Connected printer does • Print to pd is activated • Deactivate print to pd

4
not print • Printer is not a PostScript printer • Connect a printer that can interpret Post-
Script
Data format in PC cal- • Decimal separator not adjusted to PC • Use the same decimal separator in 5
culation program not calculation program the instrument which is used in the PC
correct (see section 8.2.4)
10
11
12
13
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Version 3.0 – 04/2021 177
1
12 Technical Data
The serial number o the spectrophotometer is printed on the type plate at the rear o the instru-
ment starting with "SN". The serial number is also stored in the instrument and can be looked up
3 under "System" and submenu "Inormation", the last line MCS serial contains the serial number o the
instrument (see section 9.2.1).
12.1 Spectroquant® Prove 100

4
Spectroquant® Prove 100
Measuring technology Spectrophotometer with reference beam technology

5
Wavelength range 320 – 1,100 nm
Lamp type Tungsten halogen lamp
Measuring modes Concentration, absorbance, transmission, multi wavelengths, spectra and kinetics in ab-
6 sorbance and transmission mode
Spectral bandwidth 4 nm
Wavelength resolution 1 nm (scan 0.1 nm)

7 Wavelength reproducibility ± 0.2 nm
Wavelength accuracy ± 1 nm
Stray light ≤ 0.1 % transmission at 340 nm

8 Photometric range ± 3.0 Abs
Absorbance resolution 0.001 Abs
Absorbance reproducibility ± 0.003 absorbance at 1 absorbance between 320 nm and 900 nm

9
Absorbance accuracy at 340 – 900 nm
1 absorbance: ± 0.005 absorbance
2.5 absorbance: ± 0.010 absorbance
10
Scan Limits freely selectable within the wavelength range
Increment: 0.1/1/5 nm
Scan speed: up to 170 nm/min (depending on the increment)
Smart Screen Display Resistive touch screen

11
Live ID barcode Automatic 2-D barcode reading system or all Spectroquant® cell and reagent tests
Barcode contains lot, expiry, and calibration data. Data stored with each measurement
Cell size 16 mm round cells, 10, 20 and 50 mm rectangular cells with automatic recognition
12
Minimum lling volumes 16-mm round cells: 4 ml
10-mm rectangular cells (Standard): 2 ml (Semimicro): 1 ml
13
Cell holder Removable for easy cleaning
Methods Programmed methods o all Spectroquant® cell and reagent tests, additional user-dened
methods:
14 99 concentration mode, 20 kinetic mode, 20 wavelength scans
15
16
178 Version 3.0 – 04/2021
12 Technical Data – 12.1 Spectroquant® Prove 100 1
4
Applications Free pre-programmed applications: bromate, brewery packages (MEBAK/EBC methods),

sugar (ICUMSA), oil (DOBI, olive oil) 5
Ambient light protection Measurement with open shat possible due to proprietary solution (patent pending)
AQA prime Individual settings or all methods in

AQA1 mode: instrument check using PhotoCheck and/or Certipur ® standards
6
AQA2 mode: system check using CombiCheck or CRM standard solutions
Monitoring functions Instrument-supported pipette check and sample matrix check
Ad hoc measurement Direct access to absorbance/transmission, kinetic and spectrum measurement

7
Sotware and method up- Free updates on our website (www.sigmaaldrich.com/photometry) via internet and USB
date stick
Communication interfaces USB: 2 × USB-A (or printer, USB memory devices, keyboard or bar code reader),
1 × USB-mini-B 8
Ethernet: LAN connection
Data storage 2,000 single measured values rom the measuring modes concentration, absorbance/%
transmission and multi wavelengths. 20 measurement result records o spectra and
kinetics methods each 9
Languages English, German, Spanish, French, Italian, Brazilian-Portuguese, Chinese (simplied and
traditional), Japanese, Russian, Bulgarian, Czech, Danish, Dutch, Greek, Hungarian,
Indonesian, Malay, Macedonian, Norwegian, Polish, Romanian, Serbian, Slovene, Swed-
ish, Thai, Turkish, Vietnamese, Korean 10
Protection class IP 31 or optics and electronics
Power supply Power supply with 3 cables (1.2 m long) tting US, EU and UK plugs
Total cable length 3 m (1.8 and 1.2 m)
11
Power requirements 100 V – 230 V, 50 – 60 Hz
Power consumption Standard working condition: 15 W; power save mode: 8.4 W
Temperature Operation: 10 – 35 °C; storage: -20 °C to +60 °C or 24 hours

12
Allowable relative humidity Operation: 20 – 80 % rH, storage in ambient relative humidity conditions o 20 % to 95
%
Non-condensing
Dimensions 416 × 276 × 237 mm (width × depth × height) 13

Weight approx. 6.8 kg
Warranty 12 months
EMC Directive 2004/108/EC, EN IEC61326-1, IEC61326-1 14

Instrument safety EN 61010-1, UL IEC61010-1
15
16
Version 3.0 – 04/2021 179
1 12 Technical Data – 12.2 Spectroquant® Prove 300

3

4
Lamp type Xenon ash lamp
sorbance and transmission mode
5 Spectral bandwidth 4 nm
Wavelength resolution 1 nm (scan 0.1 nm)
Wavelength reproducibility ± 0.2 nm

6
Stray light ≤ 0.1 % transmission at 340 nm; ≤ 1 % transmission at 198 nm
Photometric range ± 3.0 Abs

7
Absorbance reproducibility ± 0.003 absorbance at 1 absorbance between 200 nm and 900 nm

8 1 absorbance: ± 0.005 absorbance
2.5 absorbance: ± 0.008 absorbance

9 Increment: 0.1/1/5 nm
Smart Screen Display Resistive touch screen
10 Live ID barcode Automatic 2-D barcode reading system or all Spectroquant® cell and reagent tests
Cell size 16 mm round cells, 10, 20 and 50 mm rectangular cells with automatic recognition

11 10-mm rectangular cells (Standard): 2 ml (Semimicro): 1 ml

12
Methods Programmed methods o all Spectroquant® cell and reagent tests, additional user-dened
methods:
99 concentration mode, 20 kinetic mode, 20 wavelength scans
13
14
15
16
180 Version 3.0 – 04/2021
3

sugar (ICUMSA), oil (DOBI, olive oil)
Ambient light protection Measurement with open shat possible due to proprietary solution (patent pending) 4
AQA1 mode: instrument check using PhotoCheck and/or Certipur® standards
5
date stick
6
1 × USB-mini-B
7
kinetics methods each
Languages English, German, Spanish, French, Italian, Brazilian-Portuguese, Chinese (simplied and 8
ish, Thai, Turkish, Vietnamese, Korean
Protection class IP 31 or optics and electronics 9


10
Allowable relative humidity Operation: 20 – 80 % rH, storage in ambient relative humidity conditions o 20 % to 95 11
%
Non-condensing
Dimensions 416 × 276 × 237 mm (width × depth × height)
Weight approx. 6.8 kg 12

Warranty 12 months
EMC Directive 2004/108/EC, EN IEC61326-1, IEC61326-1
Instrument safety EN 61010-1, UL IEC61010-1

13
14
15
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Version 3.0 – 04/2021 181
1 12 Technical Data – 12.3 Spectroquant® Prove 600

3

4
Lamp type Xenon ash lamp
sorbance and transmission mode
5 Spectral bandwidth 1.8 nm
Toluene/hexane ratio > 1.4 – the correlation o spectral bandwidth to resolution or a toluene in hexane solu-
tion standard measured at ambient temperature 25 °C
6 Wavelength resolution 1 nm (scan 0.1 nm)
Wavelength reproducibility ± 0.1 nm
7 Stray light ≤ 0.1 % transmission at 340 nm; ≤ 1 % transmission at 198 nm
Photometric range ± 3.3 Abs

8 Absorbance reproducibility ± 0.003 absorbance at 1 absorbance between 200 nm and 900 nm

9 2.5 absorbance: ± 0.006 absorbance

Increment: 0.1/1/5 nm
10
Smart Screen Display p-cap glass touch screen
Live ID barcode Automatic 2-D barcode reading system or all Spectroquant® cell and reagent tests
11 Cell size 16 mm round cells, 10, 20, 50 and 100 mm rectangular cells with automatic recognition

12 50-mm rectangular cells (Standard): 8 ml (Semimicro): 4 ml
100-mm rectangular cells (Standard): 16 ml
13 Methods Programmed methods o all Spectroquant® cell and reagent tests, additional user-dened
methods:
99 concentration mode, 20 kinetic mode, 20 wavelength scans
14
15
16
182 Version 3.0 – 04/2021
3

sugar (ICUMSA), oil (DOBI, olive oil)
Ambient light protection Measurement with open shat possible due to proprietary solution (patent pending) 4
AQA1 mode: instrument check using PhotoCheck and/or Certipur® standards
5
date stick
6
1 × USB-mini-B
7
kinetics methods each
Languages English, German, Spanish, French, Italian, Brazilian-Portuguese, Chinese (simplied and 8
ish, Thai, Turkish, Vietnamese, Korean
Protection class IP 31 or optics and electronics 9


10
Allowable relative humidity Operation: 20 – 80 % rH, storage in ambient relative humidity conditions o 20 % to 95
11
%
Non-condensing
Dimensions 416 × 276 × 237 mm (width × depth × height)
Weight approx. 6.8 kg 12

Warranty 12 months
EMC Directive 2004/108/EC, EN IEC61326-1, IEC61326-1
Instrument safety EN 61010-1, UL IEC61010-1 13
14
15
16
Version 3.0 – 04/2021 183
1
13 Accessories and Test Media
13.1 Accessories
3
Description Order No.
Halogen lamp module or Spectroquant® Prove 100 1.74010.0001
Case for Spectroquant spectrophotometer Prove 100 | 300 and 600

®
1.73020.0001
4
Rectangular cells 10 mm (1 pack = 2 pcs) 1.14946.0001

5
Semi-microcells 50 mm (1 pack = 2 pcs) 1.73502.0001
Rectangular cells quartz 10 mm (1 pack = 2 pcs) 1.00784.0001
6 Empty cells 16 mm ∅ (1 pack = 25 pcs) with screw cap 1.14724.0001
Zero Cell (1 pack = 1 pc) 1.73503.0001
Rectangular cell 100 mm 1.74011.0001
7 Prove Connect to LIMS Y110860001
13.2 Optional Equipment/Connection Cables

8
Optional equipment Description Order No.
USB barcode reader (hand-held scanner) Trade

9
USB PC keyboard Trade
Connection cables Cable with USB-mini-B and USB-A plug Trade
10
11
12
13
14
15
16
184 Version 3.0 – 04/2021
13 Accessories and Test Media – 13.3 Test Media 1
13.3 Test Media

3
Description Order No.
Test media or instrument check Spectroquant® PhotoCheck 1.14693.0001

(AQA1)
Certipur® UV/VIS Standard 1 potassium dichromate solution 1.08160.0001 4

to check absorbance according to DAB and Ph.Eur.
Certipur® UV/VIS Standard 1A potassium dichromate solution 1.04660.0001

to check absorbance at 430 nm according to DAB and Ph.Eur.
5
Certipur® UV/VIS Standard 2 – sodium nitrite solution 1.08161.0001
to check scattered light according to DAB and Ph.Eur.
Certipur® UV/VIS Standard 3 – sodium iodide solution 1.08163.0001

to check scattered light according to DAB and Ph.Eur. 6
Certipur® UV/VIS Standard 5 – toluene solution in n-hexane 1.08165.0001
to check resolution power according to Ph.Eur.
Certipur® UV/VIS Standard 6 – holmium oxide solution 1.08166.0001

7
reerence material or wavelength according to DAB and Ph.Eur.
Test media or system check Spectroquant® CombiCheck, CRM and Certipur® standard solutions
(AQA2) and MatrixCheck (AQA3) are listed in the catalog "Water, Food and Enviromental Analytics" and on the Internet
under www.sigmaaldrich.com
8
Test equipment for pipet volume Spectroquant® PipeCheck 1.14962.0001
10
11
12
13
14
15
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Version 3.0 – 04/2021 185
1
14 Appendix
Absorbance Logarithmic dimension or the absorption o the sample; negative decadal logarithm o the
transmission.
3
Analysis instructions The exact workow or carrying out the detection procedure is described in the analysis
instructions.
AQA Analytical Quality Assurance.

4 AQA1 1st step o analytical quality assurance: monitoring o the instrument.
AQA2 2nd step o analytical quality assurance: monitoring o the total system.
AQA2 labeling In the documentation, measured values are given an AQA2 labeling i the measurement was
5 carried out with AQA2.
AutoSelector Plastic cylinder with barcode. It transmits the code or a reagent test kit to the spectropho-
tometer. To insert it into the spectrophotometer open the lid and place it into the round cell
compartment.
6 Barcode 2-D barcode containing inormation on method number, expiry and lot number. I needed, it
also contains data or a calibration update. Barcode is read by the inbuilt barcode reader.
Baseline Reference value for the spectrum of reference absorbances or reference transmissions.
7 Cell Vessel to take a liquid sample or measurement in a spectrophotometer. The cell material
(mostly glass) must have certain optical eatures to be suitable or photometry.
Citation forms Dierent orms o representing a measured concentration value that can be derived rom
each other. The method or the determination o phosphate, or example, provides a mea-
8 sured value or phosphorus P. This measured value can alternatively be given in the citation
forms PO 4, PO 4-P or P2O5.
CombiCheck Multiparameter standards used to check the total system or a method and or the Matrix-
Check.
9 Concentration Mass or amount o a dissolved substance per volume, e. g. in g/l or mol/l.
Detection procedure The detection procedure designates the general principle o how a sample is brought into
a orm suitable or measurement. Dierent methods can be based on the same detection
procedure.
10
Kinetics Measurement over a period o time.
Log les These contain the automatically recorded log o all or specic actions o processes in the
device.
11
MatrixCheck Check on whether the photometric determination is disturbed by other sample ingredients
(sample matrix). The MatrixCheck can be carried out by spiking or diluting.
Measuring solution Name or the sample ready to be measured. A measurement sample is obtained rom the
analytical sample (primary sample), usually through work-up. Measuring solution and analyti-
12
cal sample are identical when there has been no work-up.
Measured value The measured value is the special value o a measured parameter to be determined. It is
expressed as
a combination o the numerical value and unit (e. g. 3 m; 0.5 s; 5.2 A; 373.15 K).
13
Method A method comprises a chemical detection procedure and special method data (calibration
line) that is required to evaluate the measurement results. How to carry out the method up
to measuring with the spectrophotometer is described in the analysis instructions. All
Spectroquant® Prove spectrophotometers contain a database with methods. Furthermore,
14 user-dened methods can be entered in the database as well.
PhotoCheck standard Stable color solution with dened absorbance values or the check o AQA1 on the spectropho-
tometer.
15
16
186 Version 3.0 – 04/2021
14 Appendix 1
Reagent blank The evaluation of the photometric measurement always refers to the comparison value of a
test solution without the substance to be determined (reagent blank value). Thus the inu-
ence o the basic absorbance o the reagents on photometric measurement is compensated 3
or. For all measurements with Spectroquant® test kits (concentration mode) there is an
exactly determined reagent blank value stored in the spectrophotometer. This value can,
however, be overwritten by a reagent blank value measured by yoursel. I needed the 2-D
barcode on the cell tests and the AutoSelector may also contain an updated reagent blank
4
which overwrites the pre-programmed reagent blank in the instrument.
Recovery Recovery is the ound measured value divided by the deault value (percentage).
Example: Deault value 20 mg/l; Found 19.7 mg/l => recovery 98.5 %.
Recovery in MatrixCheck Recovery in MatrixCheck means recovery o the addition. Example o calculation: value with- 5
out addition = 100; addition o 20 = 120 theoretical value. Measured value = 115, only 15
rom the addition o 20 were ound, recovery = (115-100)/(120-100) = 75 %
Sample blank The sample blank value is a characteristic o the sample (coloration) to be currently deter-
mined. 6
The blank is diluted depending on the method being used, but does not contain any color
reagents. The pH is the same as that o the test sample.
Spectrum Distribution o the intensity, transmission or absorbance depending on the wavelength.

7
Standard Sample with a dened concentration o the analyte to be determined.
Test kit (test) A test kit contains all reagents that are required or the photometric determination o the
sample according to the analysis instructions.
Transmission Part of the light that passes through the sample. 8

Turbidity Light attenuation caused by diuse scattering at undissolved substances.
Zero adjustment Adjusting a spectrophotometer with a water-lled cell.

9
10
11
12
13
14
15
16
Version 3.0 – 04/2021 187
1
15 List of Smart Icons on Display
Main menu Buttons Description
3 Method list
List o all methods, irrespective o mode
Settings
4 This button is used to activate method-specic settings
(e. g. sample dilution, turbidity correction, zero adjustment, sample blank, reagent blank)
Ad hoc
For perorming measurements (absorbance/transmission, spectrum, kinetics)
5 Allows measurements to be perormed without the need to create methods
AQA
Overview and list o all Analytical Quality Assurance (AQA) modes
6 Results list
List o all stored results
System – instrument setup

7 This button is or optional instrument settings (e. g. date, time, updates etc)
Login/logout
Check on users in and out
8
Timer list
List of stopwatch functions
9
Info Buttons Description
Methods information
10
Main menu selection button – switches between 2 main menu overviews
Switch between dierent citations (NH4, NH3 etc)

11
Switch between dierent units (mg/l, ppm etc)
Sub-menu Buttons Description

12
Absorbance/Transmission Mode
Ad hoc submenu: perorm absorbance or transmission measurements
Result list: lter concentration mode
13 Concentration
Method list: create methods -> Concentration Mode
Result list: lter Ad Hoc ABS/Trans measurements
Spectrum Mode
14 Ad hoc submenu: record spectrum
Method list: create methods -> Spectrum Mode
Result list: lter spectrum mode
15
16
188 Version 3.0 – 04/2021
15 List of Smart Icons on Display 1
Kinetic Mode 3
Ad hoc submenu: perorm kinetic measurement
Method list: create methods -> Kinetic Mode
Result list: lter kinetic mode
AQA Status 1&2 4

AQA submenus: Status display o the period o validity and the outcome (passed / ailed)
AQA1
AQA submenu: List o AQA1 methods 5
AQA2
AQA submenu: List o AQA2 methods
6
Pipette check
AQA submenu: List o pipette checking methods
Information 7
System submenu displays the ollowing inormation about the device:
Sotware/method versions, device class, lamp counter and serial number
Interface
System submenu displays the ollowing settings options – and standard settings: 8
Audible signals – ON, Backlight – 100 %, Print to pd – ON
Region
System submenu displays the ollowing settings options – and standard settings:
Language, date, time and country zone EU/US, decimal separator – "." (dot) 9
Quality
Quick zero – OFF, AQA1 and AQA2 lock – OFF, Zero Adjustment expiry – ON (interval:
7 days), 10
Use expired reagents – OFF, Service reminder – ON
Automation
Energy saving mode – ON (10 minutes), Auto Power o – OFF, Auto log o – OFF, 11
Auto store – ON, Auto print – OFF, Sample ID popup – OFF
User management
Activation o user management and administrator settings, User login required – OFF 12
Service
System submenu displays the ollowing settings options:
Various service unctions such as backup, restore, export o log or system data and
import o methods 13
Update
System submenu displays the option or perorming sotware and method updates
14
15
16
Version 3.0 – 04/2021 189
1 15 List of Smart Icons on Display
3 Network
This submenu o the “Instrument settings” menu displays the setting options or
connecting the Prove device with a network
Prove Connect
4 This submenu o the “Instrument settings” menu displays the setting options or
connecting the Prove device with Prove Connect
Selection & Description

Action Icons
5 Start
Start zero
6 Start zero adjustment or a method
Apply
7
Save
8
Stop
9 Close
Logout
10 User logout
Search method
11
Reset resp. clear lter options
12 Edit
For editing parameters
Create method
13
Duplicate/copy
The selected method is duplicated/copied
14
Print
15
16
190 Version 3.0 – 04/2021

Action Icons
3
Export button
All selected methods are exported to an external memory device
Import button 4
Updates/methods are imported rom an external memory device into the instrument
Delete
The selected items are deleted 5
Notication Icons in Description

Menu "Settings"
Dilution
6
Activate and notiy predilution
Turbidity on
Activate and notiy the turbidity correction
7
Show absorbance
Activate and notiy display o absorbance value in the result screen
8
Zero adjustment
Perorm zero adjustment
9
Sample blank value
Activate and notiy Sample blank value
Reagent blank 10
Activate and notiy user-dened Reagent blank
Recalibrate
Activate and notiy user-dened recalibration 11
MatrixCheck
Activate MatrixCheck
12
User dened measurement range
Activate user-dened lower and upper limit o the measuring range
13
14
15
16
Version 3.0 – 04/2021 191
2
Toggle Button Description
OFF/ON Button
0 = O, I = On – the part displayed in light grey is active – here: 0 = OFF
3
Date/measurement
Switch between date or measurement interval (AQA2); active here: measurement interval
4 Absorbance/transmission
Switch between absorbance or transmission mode; active here: transmission mode
Spiking/dilution
5 Switch between spiking and dilution (MatrixCheck); active here: dilution
Action Icons on Description

Datepicker/Keyboard/
6 Calculator
Back
7
Close
Clear
8
Delete
9
Apply
10
Add
11 Radio Buttons/ Description

Checkboxes
Warning
Warnig symbol check ino box
12
Barcode scanner deactivated
The barcode scanner or reading out the Live ID barcode on round cells and AutoSelectors
has been deactivated
13
Locked
Change password
14 Choosen
Check mark
15
16
192 Version 3.0 – 04/2021
2
Action Icons,
e.g. Result List
Search list
Search unction, search criterion: method number, method name or article number (rst 6
3
digits)
Set date/date lter
4
Sample ID
Search/results list. Search unction, search criterion: sample ID
5
Select all/select none
Panorama view 6
Graphic representation o measurement series (control card, control chart, or trend
analyses)
Selection & Action Icons Description

e.g. User-dened Meth- 7
ods
Set value pair(s) for method calibration
8
Set formula for method calibration
9
Show graphic view

10
e.g. Spectrum Kinetic
Table view of values
11
Return to the recorded spectrum
Next left/Step/next right 12
Zoom out
13
Zoom in
14
View of peak max of a spectrum
15
16
Version 3.0 – 04/2021 193

e.g. Spectrum Kinetic
3
View of peak min of a spectrum
4 Calculate and show sum of spectra
Calculate and show dierence o spectra

5
Overlay of spectra
6
First-order derivative of a spectrum
7 Navigation in graphic view
Status Icons Description

8
Attention
Warning symbol check ino box
9 Time
Time Stamp
Passed
10 Status of a check; ü = passed
O
Status of a check; = inactive
11
Failed
Status of a check; - = ailed
12 Expired
Status of a check; ! = overdue
Progress
13 The instrument is in progress
Progress
The instrument is in progress
14
15
16
194 Version 3.0 – 04/2021
1
16 Contents of log files
As already mentioned in section 9.2.7, the

procedure or the export o log les involves the
creation and export o three separate les. 3
16.1 Error log le 16.2 User log le

The error log le contains the documented col- The user log le contains the documented col- 4
lection o error messages generated by the lection o relevant operations and changes in the
operating system. These error messages are settings initiated by the user.
presented in a coded ormat. The operations are saved in a coded ormat.
Each entry contains details on the date and time o Each entry contains details on the date and time, 5
the error, the error type, the process and the com- the user, and the action.
ponents involved, and the severity o the error.
The content o the error log le supports the
processing o service issues and remote diagno- 6
ses.
The inormation documented in the user log le are coded in the ollowing way:
7
Group Action code Additional information
0 = General 1 = Power On -
2 = Power O (by User) - 8
3 = Auto Power O -
10 = Sel-Test passed -
11 = Sel-Test ailed 1 = System-, 2 = Lamp-, 3 = WLCheck
20 = Login -
9
21 = Logo (by User) -
1 = AQA1 Management 0 = AQA1 Locking switched O -

1 = AQA1 Locking Switched On - 10
10 = De-Activated AQA1 Check Check ID No
11 = Activated AQA1 Check Check ID No; Interval
12 = Changed AQA1 Interval Check ID No; Interval
20 = AQA1 Check ailed Check ID No
11
22 = AQA1 Check passed Check ID No
2 = AQA2 Management 0 = AQA2 Locking switched O -

1 = AQA2 Locking Switched On - 12
10 = De-Activated AQA2 Check Method No;
11 = Activated AQA2 Check Method No; Interval; Mode
12 = Changed AQA2 Interval Method No; Interval
13 = Changed AQA2 Mode (M/D) Method No; Mode
13
20 = AQA2 Check ailed Method No
22 = AQA2 Check passed Method No
3 = General AQA 0 = Allow expired reagents - 14

1 = Prohibit expired reagents -
10 = Override AQA2 overdue Method No
11 = Override pass on reagents Method No
20 = Result deleted Method No
15
4 = Zero Management 1 = Zero Adjustment Wavelength; Pathlength
16
Version 3.0 – 04/2021 195
1 16 Contents of log files
Group Action code Additional information
3 5 = General Settings 0 = User Management O -

1 = User Management On -
10 = Change Date New Date (Format?)
11 = Change Time New Time (Format?)
4 12 = Change Language New Language (Enumeration)
13 = Change Zero Expiration New Interval (Days)
6 = System 1 = Instrument Sotware Up-date Version (n;m;r)
5 10 = Backup
11 = Restore
Example o entries in a user log le: 16.3 Service log le

6 Devicename;Prove 600 The service log le contains the documented col-
Serialnumber;1529610052 lection o operations carried out in the course o
Software Version;ISW 1.5.0 service operations by service engineers.
Timestamp;UserId;Actiongroup;Actioncode;Ino1 The error messages are presented in a coded
7
;Info2;Info3;Info4 format.
201208_0959;logout;0;10;0;0;0;0
201208_0959;anonymous;0;20;0;0;0;0
201208_1000;pm;0;20;0;0;0;0
8
201208_1000;pm;1;0;0;0;0;0
201208_1000;pm;2;0;0;0;0;0
201208_1000;pm;3;0;0;0;0;0
9 201208_1000;pm;3;20;9001;0;0;0
201208_1001;pm;3;20;9002;0;0;0
Explanation of an example:
10 201208_1001;pm;3;20;9002;0;0;0
Timestamp = 201208_1001
UserId = pm
11 Actiongroup = 3 = General AQA
Actioncode = 20 = Result deleted
Ino1 = 9002 = Method No
Ino2 = 0 = no urther details
12 Ino3 = 0 = no urther details
Ino4 = 0 = no urther details
Interpretation = on 8 Dec 2020 at 10:01am,

13 user “pm” made a change in the group “Qual-
ity (General AQA)”. A result o method 9002 was
deleted. No urther details.
14
15
16
196 Version 3.0 – 04/2021
We provide inormation and advice to our customers on application technologies and regulatory matters to the best o our knowledge and ability,
but without obligation or liability. Existing laws and regulations are to be observed in all cases by our customers. This also applies in respect to any
rights of third parties. Our inormation and advice do not relieve our customers of their own responsibility for checking the suitability o our prod-
ucts or the envisaged purpose.
The lie science business o Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada.
Manuactured by Merck KGaA, Frankurter Straße 250, 64293 Darmstadt, Germany
Distributed by EMD Millipore Corporation, 400 Summit Drive, Burlington MA 01803, USA
Sigma-Aldrich Canada Co. or Millipore (Canada) Ltd., 2149 Winston Park, Dr. Oakville, Ontario, L6H 6J8
The vibrant M, Supelco, Certipur, and Spectroquant are trademarks o Merck KGaA, Darm-
stadt, Germany or its afliates. All other trademarks are the property o their
respective owners. Detailed inormation on trademarks is available via publicly accessible
resources.
© 2021 Merck KGaA, Darmstadt, Germany and/or its afliates. All Rights Reserved.

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Operating Manual

2 Photometric Test Kits............................................... 10

3 Sample Preparation ................................................. 16 7

8 Getting Started ........................................................ 42

9.6 Programming a User-dened Method ........................................................................ 72

9.10 Kinetics ...............................................................................................................128

10 Maintenance and Cleaning ................................... 168

11 Error Causes and Trouble-shooting ...................... 176 5

13 Accessories and Test Media ................................. 184

d ε λ = Molar absorptivity, in l/mol × cm

1.2 The Spectrophotometers

For technical data and instructions or use please 9

2.1 Basic Principle 2.1.1 Spectroquant® Cell Tests

minimize the inuence o interering ions.

6 The color reactions are in most cases based

9 1 Identication mark or the correct insertion

2.1.2 Spectroquant® Reagent Tests

The principle behind the reagent tests is that 6

2.2 Notes for Practical Use

In photometry it is conventional practice to mea-

spectrophotometers belonging to the

Below the specied measuring range, either a

2.2.4 Time Stability 2.2.5 Inuence o Foreign Substances

• raise the measurement value as a result of an

A quantication o these eects is stated in tab-

2.2.6 Dosing the Reagents

Solid substances are dosed either with the

In this case it is necessary to add only level

Sample preparation covers all the steps nec-

4 3.1 Taking Samples

dilution procedure is calculated as ollows: 1:2 5 5 2 1+1

Compounds that always occur in dissolved orm

As a measure to distinguish between dissolved

Decomposition Decomposition Filtration

Total content Dissolved proportion Dissolved proportion 10

Total metal Crack Set 10/10C

Total nitrogen * Crack Set 20 Decomposition No A and B Yes No decomposition

Positive-displacement pipettes permit

Pipettes o varying volumes and also ones with a

• Check the pipetted volumes

4 Serial no: 2005614705

Analytical Quality Assurance represents a suit- Software version: 1.4.5

able and indispensible method by which the Wavelength Accuracy ⃰

errors in the measurement system diagnosed,

5 Holmium Oxide Liquid Filter

using the respective reerence methods demon- Wavelength Precision / Reproducibility ⃰

6 Holmium Oxide Liquid Filter

ound in Memorandum A 704 o the German As-

serwirtschat, Abwasser und Aball e.V., DWA)

Photometric Precision / Reproducibility ⃰ @ 1.0 A

440 nm ≤0.003 A 0.000 A P

• the handling 635 nm ≤0.003 A 0.001 A P

• the sample under investigation

These errors have eects on both the accuracy

Spectrophotometers and photometric test kits Potassium Chloride

above all else also documented by the manuac-

turer. Spectral Bandwidth ⃰ ⃰ ⃰

13 The certicate or the spectrophotometer Toluene in n-Hexane

enclosed with each device documents the quality Selftest Hardware P