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IMPORTANT NOTE

In order to avoid any malfunctioning of the instrument, follow these specifications before
normal operations:

A.
As Photometer is unpacked, let it functioning for about one hour before
starting to work with it.

B.
Check Flow cell if is dirty, clogged or creates air bubbles and eliminate the
trouble as indicated in maintenance section.

C.
Check peristaltic pump tubing if they are clogged or smashed or unshaped
and if necessary replace them.

D.
Check peristaltic pump's aspiration (minimum 0.5 ml).

IMPORTANT NOTE

In Case of malfunctioning of the instrument or softwares lock up, its possible to

restore the content of the EEPROM to the original factory settings. Note that
restoring all the settings will also overwrite the modified methods, therefore is
suggested to have a printout of your own methods settings to avoid to loose
them. Follow these steps to restore factory settings on the EEPROM.

1. Switch off the instrument

2. Switch the unit on keeping pressed the ESC key
3. The LCD display will ask for confirmation to restore EEPROM data with
factory settings
4. Confirm the restore process
5. Switch off the unit as prompted on the screen
6. Switch it on again
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PRIME EVOLUTION SEMI AUTOMATIC ANALYZER

TABLE OF CONTENTS

IMPORTANT NOTE..................................................................................................................................................................1

SECTION 1. INTRODUCTION...........................................................................................................................................3
1.1 BASIC CHARACTERISTICS....................................................................................................................................... 3
1.2 TECHNICAL SPECIFICATIONS................................................................................................................................. 6
1.3 INSTALLATION............................................................................................................................................................ 7
1.4 LANGUAGES SELECTION AND INIZIALIZATION PROCEDURE ..................................................................... 7
1.5 CONNECTION WITH THE DILUTER SAMPLER.................................................................................................... 8
1.6 STARTING OPERATIONS AND MAIN MENU ...................................................................................................... 9
SECTION 2. ROUTINE OPERATION............................................................................................................................10
2.1 AUTOZERO.................................................................................................................................................................. 10
2.2 START ANALYSIS..................................................................................................................................................... 12
2.3 CALIBRATION............................................................................................................................................................ 15
2.4 MULTI-STANDARD CALIBRATION..................................................................................................................... 16
2.5 SAMPLES ANALYSIS................................................................................................................................................ 17
2.6 PROFILE FUNCTION.................................................................................................................................................. 18
2.7 EXECUTING TESTS USING DISPOSABLE CUVETTES ........................................................................................ 20
2.8 DIRECT ABSORBANCE READINGS ....................................................................................................................... 22
SECTION 3. METHODS PROGRAMMING.....................................................................................................................22
3.1 METHODS PARAMETERS....................................................................................................................................... 23
3.2 CREATING AND MODIFYING METHODS............................................................................................................ 24
3.3 MULTI-STANDARD METHODS............................................................................................................................. 25
3.4 PRINTING METHODS STORED IN MEMORY...................................................................................................... 26
SECTION 4. MAINTENANCE AND TROUBLE-SHOOTING......................................................................................27
4.1 ADJUST PERISTALTIC PUMP STEPS.................................................................................................................... 27
4.2 SELECT PRINTER........................................................................................................................................................ 28
4.3 QUALITY CONTROL.................................................................................................................................................. 29
4.4 MENU' TEST ................................................................................................................................................................ 30
4.5 PERIODIC MAINTENANCE...................................................................................................................................... 31
4.6 TROUBLE-SHOOTING............................................................................................................................................... 32
SECTION 5. NON ROUTINE MAINTENANCE..............................................................................................................33
5.1 LAMP ASS'Y REPLACEMENT................................................................................................................................. 33
5.3 PERISTALTIC PUMP'S TUBES REPLACEMENT.................................................................................................. 34
PRIME EVOLUTION'S CALCULATION MODES ............................................................................................................35

ON LINE INTERFACING........................................................................................................................................................37

MATHEMATICAL MODELS FOR MULTI STANDARD CURVES ...............................................................................38

PRINTABLE ASCII CHARACTERS TABLE.....................................................................................................................39

ON SCREEN FLAG CODES LIST........................................................................................................................................41

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SECTION 1. INTRODUCTION

1.1 BASIC CHARACTERISTICS

The PRIME EVOLUTION is an integrated semi-automatic photometer,all functions of the

instrument, storage of tests parameters and calculation of calibration curves and end results are
microprocessor controlled.The keyboard allows the operator to recall directly up to 22 methods
commonly used in standard routines, as well is provided a key to recall tests by numerical keys, to
perform and modify tests up to 150.

Main parts of PRIME EVOLUTION are:

Photometer, composed of a light source placed at the center of a filterwheel,a 50 ml flow cell and
a solid state detecting system. The filterwheel contains an interference filter set.The light source is a
low consumption, high intensity quartz-jodine lamp, and interference filters are narrow band type
in the range of 340 nm to 620 nm.

Aspiration system, based on an electronic controlled peristaltic pump that operates aspiration of
reaction mixture as well as washing solutions in the flow cell.Instrument asks confirmation to
operator before doing any aspiration in the flow cell.

User Interface, composed of an LCD Graphic Dot Matrix Display, to visualize all menus,
operative messages and all the graph, such as the Levy-Jennings graph and the multistandard
curve for the immuno chemistry tests. A 24 keys, membrane type keypad is provided to access
directly 22 tests , all operative levels and the instrument's functions. Furthermore a 40 columns
High Speed thermal graphic printer is built in , to produce hard copies of test results and
parameters.

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3
4
5

1- DISPLAY LCD 4- ASPIRATION PROBE

2- ASPIRATION SYSTEM 5- ASPIRATION LEVER
3- MEMBRANE TYPE KEYPAD 6- MAIN POWER SWITCH

7 8 9 11 10

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7- WASTE OUT 10- PRINTER PARALLEL PORT
8- AC POWER INPUT 11- SERIAL RS 232C PORT
9- FAN

3 2
7
4

1- THERMAL PRINTER
2- PRINTER RESET KEY
3- PAPER ADVANCE KEY
4- DISPLAY LCD
5- MULTIFUNCTIONS KEYBOARD
6- ASPIRATION LEVER
7- PHOTOMETER COMPARTMENT

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1.2 TECHNICAL SPECIFICATIONS

PHOTOMETER
6 Interference Filters 340,405,505,546,570,620 nm and 1 avalaible free position for an optional
filter, tolerance of +/- 2 nm max.

Quartz-Jodide Lamp 12V 10 W - Long Life 2000 hours

50 l micro flow cell

Temperature control in flow cell effectued by Peltier elements tolerance of 0.1 C - Selectable
temperatures: Off - 25C - 30C - 37C

DIGITAL PROCESSOR
16 BIT Linear AD conversion

Based on 8088 ? P

Display LCD Graphic Dot Matrix

24 keys membrane type keypad

40 columns built-in Graphic thermal printer - parallel port for external printer

RS 232C serial port for on line data transfer

PHYSICAL CHARACTERISTICS
Dimensions: 400(W) x 350(D) x 160(H) mm

Weight: 8 Kg

Power Supply: 220V 50Hz - Opzionale 110V 60Hz

Power Consumption: 80 W

TYPES OF MEASUREMENTS
End Point
Kinetics Vs Factor
Kinetics Vs Standard
Bichromatics

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Serum Blank
Immunoturbidimetrics
Multi-Standard
Initial Rate
Direct Absorbance

1.3 INSTALLATION

After Unpacking check following :

Check the presence of power cord and waste discharge tubing.

Check if instrument's power supply matches with your country power outlet (USA-Japan 110V
60Hz-Europe 220V 50Hz-UK 240V 50Hz).

Check for the presence of ground line of good quality.

Check for the presence of clean power supply line, free from spikes and fluctuations. If necessary
use an AC stabilizer.

Avoid placing the instrument exposed to direct sun light.

Connect waste tubing to waste bottle, position the bottle below the equipment, connect power cord
to AC outlet and PRIME EVOLUTION is ready to work.

1.4 LANGUAGES SELECTION AND INIZIALIZATION PROCEDURE

The PRIME EVOLUTION can be set to operate in different languages. At the moment only 3
languages are available: Italian, English and Chinese. To select the operating language for the
instrument proceed as follows:

a. Switch on the instrument by keeping pressed the ESC key, on the LCD display will appear
the following screen:

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Press the key corresponding to the language desired and therefore press Enter to confirm the
selection. Note that for the Chinese language only its suggested to reboot the system to
make changes effective.

After having selected the language and pressed the Enter key to confirm such a selection, the system
will visualize the initialisation screen, which could be useful to restore the original instruments factory
settings. This function can be used whenever the instrument shows some malfunctioning due to
erroneous data writing on the internal EEPROM.

The display LCD will show the following screen:

To restore the original factory settings and initialise the instrument press the 1 key and confirm with
Enter.

IMPORTANT NOTE

Initializing the instrument the operator will loose all the user modified methods
and the calibrations data.

1.5 CONNECTION WITH THE DILUTER SAMPLER

In case you have purchased the optional Diluter/Sampler unit, refer to the following instructions to
connect it to the PRIME EVOLUTION photometer:

1. Connect the flat cable provided with the Diluter/Sampler to the parallel port located on the rear
panel of the PRIME EVOLUTION and to the apposite connector located on the rear part of the
Diluter/Sampler.
2. Switch on the PRIME EVOLUTION, then switch on the Diluter/Sampler and perform one
priming cycle by pressing the SINGLE PRIME lever on the top front of the Diluter.
3. Always keep the Diluter/Sampler switched on while working with the PRIME EVOLUTION
connected to it, this in order to avoid any malfunctioning in operation of PRIME EVOLUTION.

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4. To access to the functions of the Diluter/Sampler, symply press the DIL key on PRIME
EVOLUTION and select desired function for the Diluter.

On the LCD panel, once DIL key is pressed, will appear the following selection screen:

** REV 4.6/Profili Date: 24/08/04 **

SETTING ON DILUTER

At this prompt perform with the Diluter a SINGLE PRIME cycle and the display will show the
following:

** REV 4.6/Profili Date: 24/08/04 **

DILUTER
Diluter Sampler = 1 Dispenser = 2 [ ]

By entering 1 or 2 and pressing the ENTER key, it's possible to select the desired operation mode of
the Diluter, as a Sampler in a SIP and DIP mode, or as a diluter with the pipette used only for picking
up samples and reagent vial connected directly to the Diluter.
Once the function is selected, the LCD goes back to the main menu, then select the test to be
performed and the screen will demand you the operations required to be performed with the Diluter's
pipette.

Refer to the Diluter/Sampler Operation Manual for more detailed informations about the operative
procedures to be performed.

1.6 STARTING OPERATIONS AND MAIN MENU

Switch on the equipment using the main switch, located on the front of the instrument.At the moment
the instrument is switched on, the DATE request appears on the LCD panel:

REV 4.6/Profili*

Date (ddmmyy) : ( )

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Once the DATE is entered by the numerical keys, the Main menu will be displayed :

** REV 4.6/Profili Date: 24/08/04 **

F1=Autozero F2=Methods
F3=Maintenance F4=Start

By pressing the corresponding function key the sub-menues related to selected function,
F1,F2,F3,F4 are accessed.

SECTION 2. ROUTINE OPERATION

2.1 AUTOZERO

Option F1 on Main Men offers the possibility to zero the instrument completely before the start of
routine operation.This way the flow-through cell and the filters can be checked at the selected
wavelenghts.
Note: it's not suggested to start the routine with this, as every analysis will commence with an
Autozero. It's important to switch on the instrument at least 30-40 minutes before
operations, this in order to let the lamp and the electronics to stabilize and reach operative
temperature.

Before starting make sure the cuvet is filled with clean distilled water, the option asks operator to
select one of the avalaible filters:

** REV 4.6/Profili Date: 24/08/04 **

AUTOZERO
Filter 340 405 505 546 570 620 7 8 : [ ]

Note: Number 7 corresponds to a free position for an optional filter, and 8 corresponds to BLIND
position (0% transmission).

Selected filter is being checked and "Auto-zeroed".This means that the 0 Abs. (100% Transmission)
is being stored. The electronic values (Voltages) of these settings are stored and applied during
reading.If voltages found exceed factory set limits, a warning is displayed on LCD display for the
filter in question. Once the desired filter is entered, press the Enter key, on the display will appears:
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** REV 4.6/Profili Date: 24/08/04 **

AUTOZERO
WASH

Then immediately:

** REV 4.6/Profili Date: 24/08/04 **

AUTOZERO
ASPIRATION

Dip the aspirator probe in a test tube filled with clean distilled water and press the aspiration lever
once: the water will be aspirated into the flow cell and the screen will display in sequence:

** REV 4.6/Profili Date: 24/08/04 **

AUTOZERO
Wait for incubation

** REV 4.6/Profili Date: 24/08/04 **

AUTOZERO
Wait for reading

** REV 4.6/Profili Date: 24/08/04 **

FILTER Temperature O.D. AUTOZERO

340 37 0.0000 (ENTER=OK)

At this point press the ENTER key,in case everything works fine on the screen will appear:

TEMPERATURE OK

Then press the Enter key to return to the Main Menu.

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In case there are problems with the aspiration or with the filter or with the lamp, the following error
message will be displayed:

** REV 4.6/Profili Date: 24/08/04 **

FILTER Temperature O.D. AUTOZERO

340 37 OUT OF LIMIT (ENTER=OK)

Press Enter to return to the Main Menu and check for the fault ( aspiration or other cause ).

2.2 START ANALYSIS

Selecting the option F4 of the main men, the operation mode is entered, PRIME EVOLUTION
asks for the the method to be done, it's possible to press one of the keys with indicated a chemistry,
i.e. GOT, ALP etc., to recall directly the method to be performed:

** REV 4.6/Profili Date: 24/08/04 **

START OPERATION

TEST?_

or depress the key F4 / # TEST, in this case the screen will display:

** REV 4.6/Profili Date: 24/08/04 **

Test number ( 1 - xxx) : [ ]

Where the xxx means the last test available in the memory.

After entering number of desired method , followed by ENTER, or after pressing the key related to
the desired method, if printer is activated (see maintenance section) the date and the selected test will
be printed and the instrument will jump to the Autozero function automatically:
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** REV 4.6/Profili Date: 24/08/04 **

AUTOZERO
WASH

** REV 4.6/Profili Date: 24/08/04 **

AUTOZERO
ASPIRATION

Insert aspiration probe into a vessel filled with distilled water and push the aspiration lever,placed
behind of the aspirating probe.
Water is aspirated into the flow cell and the photometer is auto-zeroed.

In case everithing is right, the display will visualize:

TEMPERATURE OK

Now press the Enter key to continue,the system asks to aspirate reagent blank (if programmed for
the method), executes a reading and displays the measured value:

** REV 4.6/Profili Date: 24/08/04 **

14 GOT/AST R1274 BLANK

WASH

** REV 4.6/Profili Date: 24/08/04 **

14 GOT/AST R1274 BLANK

ASPIRATION

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Dip the aspirator probe into a test tube containing blank reagent and press the aspirator lever, after
the preincubation and the reading time.

On the LCD panel will be displayed the O.D. of the reagent blank:

** REV 4.6/Profili Date: 24/08/04 **

14 GOT/AST
O.D. BLANK : 1.5220

after which operator may select out of:

ESC : goes back to main men

ASP. REPEAT : repeats aspiration (before aspirating, empty flow cell by aspirating air)
O.D. REPEAT : repeats reading on the solution in the flow cell
ENTER : accepts the reading and steps to next display (Ex. GOT).

At this point we can continue by testing a sample or we can calibrate, if in the method specified we
have entered the standard concentration value:

** REV 4.6/Profili Date: 24/08/04 **

14 GOT/AST R1274 F1=CALIBRATE F2=NO

STD. CONC.=46 FACTOR=1768

The STD. CONC. is the value programmed for the test in question.The factor may be the
programmed factor OR the factor resulting from the last calibration.

By selecting F1 a calibration of the method may be executed.

F2 goes straight to the determination using the displayed factor.However if the method selected has
not been calibrated before, bypassing with F2 will not be possible and the operator will be urged to
calibrate first.

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2.3 CALIBRATION

** REV 4.6/Profili Date: 24/08/04 **

14 GOT/AST R1274 F1=CALIBRATE F2=NO

STD. CONC.=46 FACTOR=1768

After pushing F1, the cursor jumps to the STD. CONC. item.In case a different standard is being
used, it's value can be entered right away.By pressing ENTER key,the current value is accepted,

** REV 4.6/Profili Date: 24/08/04 **

WASH

The Flow cell is emptied

** REV 4.6/Profili Date: 24/08/04 **

ASPIRATE

The operator dips aspiration probe into the reaction mixture (Standard + Reagent) and pushes the
ASPIRATION LEVER.

The liquid is aspirated and sample is measured:

** REV 4.6/Profili Date: 24/08/04 **

14 GOT/AST R1274 CALIBRATION

1st O.D. STD.: 1.0056 FACTOR : 1782

After this,the operator has the option to:

ENTER : Confirms and accepts displayed value. Accepted values are printed.
O.D. REPEAT : Repeats the reading on the same liquid.
ASP. REPEAT : Aspirates again and repeats reading.

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2.4 MULTI-STANDARD CALIBRATION

In case a Multi-Standard test has been selected for calibration, the operator will be prompted 6 times
to aspirate each one of the calibrators:

** REV 4.6/Profili Date: 24/08/04 **

Aspirate Calibrator 1

** REV 4.6/Profili Date: 24/08/04 **

Aspirate Calibrator 2

** REV 4.6/Profili Date: 24/08/04 **

Aspirate Calibrator 3

** REV 4.6/Profili Date: 24/08/04 **

Aspirate Calibrator 4

** REV 4.6/Profili Date: 24/08/04 **

Aspirate Calibrator 5

** REV 4.6/Profili Date: 24/08/04 **

Aspirate Calibrator 6

After having aspirated the last calibrator,the Curve is calculated and the result is printed.

On the LCD display will appear the curve calculated and by pressing the key ESC the operator will
have access to the functions to accept or modify the curve of calibration obtained:

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** REV 4.6/Profili Date: 24/08/04 **

F1=Store F2=Change model

F3=Eliminate point F4=Restore point

F1=Store :Accepts the curve and stores it in non volatile memory.

F2=Change model :Change programmed mathematical model (see App. 6-C) and
recalculates curve.
F3=Eliminate :With this option 1 calibrator can be excluded from the curve, a new
curve is calculated.
F4=Restore :This function restores a calibrator previously eliminated. A new
curve is calculated each time

In case of a bad Curve-FIT, the display visualizes the error message:

< Regress. error>

By trying another model or eliminating one or more points a better curve may be found.

IMPORTANT NOTE

By pressing twice the ESC key when the LCD display shows the calibration curve, the
program is exited and therefore the operator looses all the calibration data and the curve
itself. Once the curve is recalculated and it results satisfactory to the operator, always store
it before performing any operation.

2.5 SAMPLES ANALYSIS

After calibration display prompts for the first patient:

** REV 4.6/Profili Date: 24/08/04 **

PAT. I.D. 1st O.D. FIT RES. GOT/AST

1 (F1=Input I.D. F2=not)

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F1 offers the possibility to enter a numeric patient ID, to be confirmed with ENTER. If no ID is
wanted push F2.

The instrument prompts the operator to aspirate the first sample.First sample will be read, the result
printed, the cuvet emptied and the system continues to next patient:

** REV 4.6/Profili Date: 24/08/04 **

PAT. I.D. 1st O.D. FIT RES. GOT/AST

1 0.9856 1.00 35

By pressing the Enter key, the value is accepted and the display visualizes the next patient's request:
** REV 4.6/Profili Date: 24/08/04 **

PAT. I.D. 1st O.D. FIT RES. GOT/AST

2 (F1=Input I.D. F2=not)

and so on. Pushing ESC key, interrupts the sequence and operator is prompted to wash the flow cell:

** REV 4.6/Profili Date: 24/08/04 **

WASH

** REV 4.6/Profili Date: 24/08/04 **

ASPIRATE

Once water has been aspirated into the flow cell, the system returns to the main menu.

2.6 PROFILE FUNCTION

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With this software revision ( Rev. 4.6 ) its possible to have the printout of the results of all tests
performed for a single patient on a sort of patients report.

To proceed its necessary to enter the data at the beginning of the operations, and to enter the patient
ID when required.

At the moment that the LCD displays asks for the first sample for the first test to be aspirated, it
shows :

** REV 4.6/Profili Date: 24/08/04 **

PAT. I.D. GLUCOSE

1 (F1=Input I.D. F2=not)

at this stage press the F1 key :

** REV 4.6/Profili Date: 24/08/04 **

PAT. I.D. GLUCOSE

1 ___

And type in the patient ID, note that only numerical ID can be used.

** REV 4.6/Profili Date: 24/08/04 **

PAT. I.D. GLUCOSE

1 02

then proceed as described.

When operations are completed and all tests have been performed, press the F3 key to enter in
Maintenance menu :

** REV 4.6/Profili Date: 24/08/04 **

F1=Adj. perist.pump F2=Select printer

F3=QC / PROFILE F4=Test

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then select F3 to enter the QC / PROFILE :

** REV 4.6/Profili Date: 24/08/04 **

F1= QC F2=PROFILE

At this stage press the F2 key to enter the PROFILE MENU :

** REV 4.6/Profili Date: 24/08/04 **

Patient ID : _______

the system requires the ID of the patient, once this is entered, the report will be printed, collecting all
tests results for that patient.
Once the printing has been completed repeat the operations with anoth ID to get another Report.

The use of the Profile function on the PRIME EVOLUTION reduces the number of methods which
can be stored onto the instrument. With this software revision the number of methods which can be
stored either for clinical chemistry or immunochemistry is 99 test.

Profiles will remain stored into the memory until a new date is entered and however the max number
of profiles which can be stored is 700 profiles.

If either the date or the ID are not entered, the system will not work correctly.
By performing the immediate repetition of a sample, the value which is stored for the profile function
is the last one obtained.

2.7 EXECUTING TESTS USING DISPOSABLE CUVETTES

It's possible to use either flow cell and disposable cuvettes to execute readings with PRIME
EVOLUTION photometer.
The instrument is factory provided with a flow cell installed in the photometer compartment, but is
supplied a metal adapter to be used with disposable cuvettes.

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To operate with disposable, press the O.C. key , the LCD will demand:

** REV 4.6/Profili Date: 24/08/04 **

SETTING
READING ONLY (Y)

Then after a few seconds the LCD will be back to the main menu, now the
instrument is set to operate with disposable cuvettes, the flow cell's
temperature and the peristaltic pump are unoperative.

Extract the flow cell from its compartment, having care of not damaging the
tubings, and insert it into the housing located on the left part of the
photometer block.

Effectued this operation, instrument is ready to operate with disposable

cuvettes.

Follow the figure to insert adapter into the photometer's housing:

To return to normal operations with the flow cell, extract the cuvette adapter from the
photometer housing and insert again the flow cell, with particular care to all tubings, adjust
the aspiration tubing making it passing through the metallic tubing located on the front of
the instrument, then simply press again the O.C. key, the LCD display will show the
following:

** REV 4.6/Profili Date: 24/08/04 **

SETTING
READING ONLY (N)

Now the setting for disposable Cuvettes operation is deactivated and the operation mode
with flow cell is active.

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2.8 DIRECT ABSORBANCE READINGS

To perform with this instrument a direct absorbance reading, follow these simple steps:

1. Press the O.D. key to enter the Direct Absorbance Reading function, the screen will show you the
Autozero request:

** REV 4.6/Profili Date: 24/08/04 **

AUTOZERO
Filter 340 405 505 546 570 620 7 8 [ ]

Enter a wavelenght and perform autozero,ref to section 2.1 , until the LCD will show :

** REV 4.6/Profili Date: 24/08/04 **

Filter O.D. READING ADC ( F1 = PRINT)

340 0.0002

2. Then press Asp. Repeat key to aspirate solution to be read, after preincubation time the LCD will
display the Absorbance of the sample.

3. Press F1 key to print absorbance read

4. To execute other solutions readings repeat items from 2.

5. Pressing O.D. Repeat key it's possible to read again the absorbance of solution in flow cell

6. Press ESC key to return in Main men

NOTE: The instrument always makes a first air aspiration in the flow cell, to wash any residuals of
previous solutions.

NOTE: When a wavelenght it's selected after another one, leave some time , about 5-6 minutes, to
stabilize the lamp as the voltage of the lamp changes according to the filter selected, this to make
longer the lamp's life.

SECTION 3. METHODS PROGRAMMING

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PRIME EVOLUTION is an " Open " instrument, as the operator can modify or create his own
methods of clinical chemistry, or immunochemistry according to reagents used, operative conditions
or specific needs.

3.1 METHODS PARAMETERS

Heart of programming methods in PRIME EVOLUTION Photometer, are the parameters, items to
be modified to create a new program either in immunochemistry or clinical chemistry.In PRIME
EVOLUTION programming you have these end user modifyable parameters:

1= Endpoint
2= Kinetic-Enzymatic tests
Type of Reaction (1-5) : 3= Initial Rate
4= Bichromatic Endpoint
5= Serum Blank
Name of test (15 char.) : See Appendix D
Lower limit (units) : Lower pathological limit
Upper limit (units) : Upper pathological limit
Temperature Cuvette : 0-25-30-37C 0= Temp. Off
Blank (Y=1/N=0) : Execute a reading of reagent blank
Reagent Blank subtraction : Correct samples for reagent blank
Lower limit Reag. O.D. : Warning level Reagent O.D.
Upper limit Reag. O.D. : Warning level Reagent O.D.
Linearity limit : Warning level for linearity
Reagent Volume (l) : These values will be read by the external Diluter
Sample Volume (l) : These values will be read by the external Diluter
Standard Concentration : Value of standard to be used
Factor for calculating end results; in case any
Factor : standard is used, the calculated factor is stored
Incubation Time : Time liquid should stay in the flow cell before
measurement. *
Duration of reading : Timespan during which reaction is monitored (typ.
20 sec.) **
Reaction Incr.(0)/Decr.(1) : Direction of reaction (e.g. CPK=0/GOT=1)
O.D. level substrate Depl. : Warning for substrate depletion.
Save calibration factor : Factor can be saved (1) to avoid too many
calibrations.
Number of controls (0-9) : Number of controls done at beginning of routine.
Number of decimals(0-4) : No. of digit after floating Point
Filter 1 : Enter 340-405-505-546-570-620 or 7 (optional)
Filter 2 : As above in case of bichromatic

(*) A minimum of 5 sec is reccomended to stabilize liquid in cuvet

(**)Only in case of kinetic reactions and Initial Rate.

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3.2 CREATING AND MODIFYING METHODS

Selecting option F2 (Methods) on Main men, the LCD panel displays a sub-men with 4 options:

** REV 4.6/Profili Date: 24/08/04 **

F1=Autozero F2=Methods
F3=Maintenance F4=Start Operation

Press F2:

** REV 4.6/Profili Date: 24/08/04 **

F1=Modify Method F2=Immunochemistry

F3=Print TEST F4=Print all TEST

Each sub-item can be selected by pressing the corresponding function key.With option F1, a test can
be configurated by assigning parameters described in chapter 3.1.Up to 150 methods can be
programmed and stored in the non-volatile RAM of the instrument.

Internally tests are being numbered from 1 to 150, but 28 methods are preprogrammed on leaving
the factory.

The option starts with asking the operator the test number wanted, the default is the first free (non
programmed method) number. However the operator can press a key with a chemistry code printed
on it, i.e. GGT, to modify that method or can press the F4 key in order to enter the number of the
test to be modified:

** REV 4.6/Profili Date: 24/08/04 **

Test number (1-xxx) : (_ )

where 'xxx' represents the highest test number + 1 programmed up to this moment.By hitting ENTER
key,the next free number is selected for creation. Otherwise a pre-programmed method's number can
be entered.

If selected method's number already has been selected in another class (immunochemistry), the
program warns the operator and ask for confirmation.

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Subsequently parameters described in chapter 3.1 can be entered by depressing enter key.

NOTE* As PRIME EVOLUTION is provided only with numerical keypad, all alfabetical characters
are entered by a 3 number ASCII code, described in Appendix-D.

3.3 MULTI-STANDARD METHODS

With the sub-option F2, tests with multiple standards (Immunochemistry) may be configurated:

** REV 4.6/Profili Date: 24/08/04 **

F1=Modify Method F2=Immunochemistry

F3=Print TEST F4=Print all TEST

Press F2:

** REV 4.6/Profili Date: 24/08/04 **

Test number (1-xxx) : (_ )

Then enter the desired or the last available test number, in case you want to create a completely new
method, and press enter to confirm.

Option asks for the same parameters as option F1, but in addition it asks for the following
parameters:

Model (1-6) : One of 6 mathematical model (see Appendix 6-

C)
Maximum Variance % : Max permitted deviation in % from calculated
standard curve.
Lot. No. : Lot number of reagent used.
CALIBRATOR 1 : Value of cal. 1
CALIBRATOR 2 : Value of cal. 2
CALIBRATOR 3 : Value of cal. 3
CALIBRATOR 4 : Value of cal. 4
CALIBRATOR 5 : Value of cal. 5
CALIBRATOR 6 : Value of cal. 6

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3.4 PRINTING METHODS STORED IN MEMORY

With option F3 it's possible to have a printout of all parameters of individual method:

** REV 4.6/Profili Date: 24/08/04 **

F1=Modify Method F2=Immunochemistry

F3=Print TEST F4=Print all TEST

Press F3:

** REV 4.6/Profili Date: 24/08/04 **

Test number (1-xxx) : (_ )

On entering a Test number, actual parameters of that test are being listed by the printer.

With option F4 it's possible to obtain a printout of all parameters of all tests stored in the RAM of the
instrument:

** REV 4.6/Profili Date: 24/08/04 **

F1=Modify Method F2=Immunochemistry

F3=Print TEST F4=Print all TEST

Press F4:

** REV 4.6/Profili Date: 24/08/04 **

PRINT ALL TESTS

F1=Only Full Name F2=With Parameters

Pressing F1 you can print only the names of tests associated with test numbers, pressing F2 it's
possible to obtain tests printouts with all parameters associated.

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SECTION 4. MAINTENANCE AND TROUBLE-SHOOTING

The option F3 of the Main men offers 4 sub-options:

** REV 4.6/Profili Date: 24/08/04 **

F1=Autozero F2=Methods
F3=Maintenance F4=Start Operation

Press F3:

** REV 4.6/Profili Date: 24/08/04 **

F1=Adj. perist.pump F2=Select printer

F3=QC/Profile F4=Test

4.1 ADJUST PERISTALTIC PUMP STEPS

Adjustment of peristaltic pump has 3 functions:

To switch off pump completely in case no flow cell is used (reading only of disposable cuvettes);
To adjust volume aspirated into the flow cell.The volume must be enough to rinse the flow cell
sufficiently, but not higher than the total reaction mixture;
To adjust volume aspirated during a Wash Cycle;

If volume is too high, air will be sucked into the cell.If volume is too low, more liquid than normal
stays behind in the reaction wells, and the cell may not be sufficiently cleaned.

As the tubing of peristaltic pump gets older, it looses some of its elasticity and the volume may get
lower too.

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** REV 4.6/Profili Date: 24/08/04 **

F1=Adj. perist.pump F2=Select printer

F3=QC/Profile F4=Test

Press F1, LCD will visualize following message:

** REV 4.6/Profili Date: 24/08/04 **

No. of steps for Aspiration: xxx

Now it's possible to have the following choices:

The current number of steps is displayed
The number of steps can be increased or decreased by entering a new number between 100 and
999 and confirming with ENTER:
Current number is not to be modifyed so press simply ENTER for confirmation to jump to next
screen:

** REV 4.6/Profili Date: 24/08/04 **

No. of steps for wash: xxx

Repeat the choices above described and press ENTER again.Upon Enter the aspiration or wash
cycle is being executed.

4.2 SELECT PRINTER

Standard the PRIME EVOLUTION has got a standard thermal printer "on board".It's however
possible to replace the on-board printer by a faster external printer.

** REV 4.6/Profili Date: 24/08/04 **

F1=Adj. perist.pump F2=Select printer

F3=QC/Profile F4=Test

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Press F2:
** REV 4.6/Profili Date: 24/08/04 **

MAINTENANCE
On Board=0 External=1 No=2

With this option of Maintenance programs, can be selected one of these three situations:

0. The On Board Printer is activated.

1. Results are printed by an external printer connected to parallel 25 pin (male) connector on the
rear of instrument.
2. No Printer, only to be used in exceptional cases.

The PRIME EVOLUTION model is available with a graphic high speed thermal printer.

4.3 QUALITY CONTROL

This option permits calculation of Mean, S.D. and C.V. of a series of results. When in the
maintenance menu:

** REV 4.6/Profili Date: 24/08/04 **

F1=Adj. perist.pump F2=Select printer

F3=QC/Profile F4=Test

Press F3:

** REV 4.6/Profili Date: 24/08/04 **

F1= QC F2=PROFILE

Then press F1:

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** REV 4.6/Profili Date: 24/08/04 **

QUALITY CONTROL
Value:

Enter one value after another.After last value is entered, push ESC key and the results are calculated
and printed.

By pressing then the F3 key corresponding to the option to visualize the graph, on the LCD will
appear the Levy-Jennings graph related to the data entered:

The graph obtained can be also printed on the internal thermal printer by pressing the F3 key.

4.4 MENU' TEST

This option permits to perform several mechanical and electronic functions:

** REV 4.6/Profili Date: 24/08/04 **

F1=Adj. Perist.pump F2=Select printer

F3=QC/Profile F4=Test

Press F4:

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** REV 4.6/Profili Date: 24/08/04 **

F1=Wash perist.pump F2=Aspirating

F3=Positioning filter F4=Temperature

F1: To aspirate water or cleaning solution into the flow cell.

F2: This option is used to check aspirated volume, to check for air bubbles in the flow cell.

F3:
** REV 4.6/Profili Date: 24/08/04 **

TEST
Filter: 340-405-505-546-570-620-7-8 [ ]

This option offers the possibility to make a pure absorbance reading of a solution.
Select a wavelenght and the filter will be positioned: a reading is made and the result printed.

Note* Position 8 represents "Dark" (0% transmission).

F4:

** REV 4.6/Profili Date: 24/08/04 **

TEST
Temperature of cuvet: 0-25-30-37: [ ]

Selecting a temperature will change the temperature into the flow cell.Time to change temperature
from (i.e.) 25 to 37C is typically about 30 secs , but the thermal stabilization could require more
time, typically 2 minutes.

4.5 PERIODIC MAINTENANCE

Follow this scheduled maintenance periodically to have your PRIME EVOLUTION always at its
best performance:

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Flow cell should be rinsed thoroughly with
distilled water at the end of te routine.
Before switching off instrument, flow cell
and tubings are to be filled with distilled
DAILY MAINTENANCE water.
If the instrument is not going to be used
for a long period of time, wash the flow
cell with BPC washing solutions
Extracleaning or Hypocleaning, then rinse
with distilled water.
Wash the flow cell with BPC washing
solution Hypocleaning, a deproteinizing
WEEKLY MAINTENANCE solution,then rinse.It's possible to use
BPC Extracleaning solution after the
Hypocleaning and according to working
time of the instrument
Replace tubing of peristaltic pump once
QUARTERLY MAINTENANCE every three months, as referred in section
5.3.

4.6 TROUBLE-SHOOTING

Before calling our Service Department, please check following items to find a faster solution of
occurring troubles;

SYMPTOMS POSSIBLE CAUSES AND ACTIONS

Too tight or too loose rotor of peristaltic
pump
Liquid not being aspirated Silicon tubings of peristaltic pump
damaged and to be replaced
Leaky motor not functioning
Lamp of Photometer burned out
Autozero of filterwheel gives 0V on all filters Connector of lamp ass'y disconnected or
failed
Filterwheel blocked or not in selected
position

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Flow cell not properly filled with water
Flow cell dirty(to be cleaned)
Little air bubbles building up into the
Erroneous Autozero cuvet(to be cleaned)
Tubing disconnected,leaky or clogged
Peristaltic pump malfunction
Not correct position of filterwheel
Check for little air bubbles into the
Erroneous non reproducible results cuvet(to be cleaned)
Check aspirating system
Check if the lamp flickers, try to replace
it.

SECTION 5. NON ROUTINE MAINTENANCE

5.1 LAMP ASS'Y REPLACEMENT

Lamp ass'y consists in a lamp, complete of its fittings, wires and connector.
The ass'y is fixed to photometer with two screws.To replace the lamp ass'y follow these steps:

1. Switch off the instrument

2. Remove the cover of the photometer
3. Remove the two screws
4. Pull connector from its fitting
5. Pull out burned lamp ass'y
6. Install the new one operating in reversed order

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WARNING:
Always avoid to touch the bulb with your fingers, as the human skin excretions will be burned into the
quartz and destroy the U.V. emission characteristics.
A lamp accidentally touched by human skin may be carefully cleaned with a piece of soft cloth.

After having installed a new lamp ass'y, the system has to be recalibrated in the following way:
Switch on instrument and let the system stabilize for at least 10 minutes
Aspirate clean distilled water into the flow cell, position the filterwheel at 340 nm and make a
reading(as indicated in section 4.4), the system makes a reading and prints the result:

** REV 4.6/Profili Date: 24/08/04 **

Filter 340 Volt x.xxxx D.O. y.yyyy

Adjust Gain potentiometer on Photometer PCB to read Volt=4.2000

Select filter 8 (Offset) and push enter
The offset value is displayed and should be between 0.0020 and 0.0030.It can be adjusted with
potentiometer marked Offset, near the GAIN potentiometer. After it's adjusted, the reading can
be repeated by pushing Enter again.

NOTE* Potentiometer GAIN is the screw next to cuvet

NOTE**Care that Flow cell is clean and distilled water used is clean and not contaminated.

5.3 PERISTALTIC PUMP'S TUBES REPLACEMENT

Tubing of peristaltic pump is pressed to the rotor of the pump by a semi circular leader.This leader is
mounted on a plate, which in turn is kept in position by an hand tightened screw.

To replace the tubing:

Untighten the screw

Remove the old tubing
Put in the new tubes
Press the leader by hand against the tubing
Tighten the screw
Use the option Aspirate of the TEST program (4.4) to check the
functioning

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Step 2
After replacement of peristaltic pump tubing, it's necessary to perform the adjustment of the steps for
aspirating and washing.
To perform these adjustments refer to section 4.1 and proceed as follows:

** REV 4.6/Profili Date: 24/08/04 **

F1=Adj. perist.pump F2=Select printer

F3=QC/Profile F4=Test

In this Maintenance men press F1 key:

** REV 4.6/Profili Date: 24/08/04 **

No. of steps for Aspiration: xxx

Here you can enter by pressing numerical keys, the value necessary to meet the following
specifications:

Levels of liquid ingoing and outgoing to and from the

flow cell, must be almost equals
There should not be air bubbles into tubings connected
to flow cell

Proceed in the same way in washing step adjustment

option.

APPENDIX 6-A

PRIME EVOLUTION'S CALCULATION MODES

All these calculation types can be used either with factor or with Standard.

Calculation 1 ENDPOINT

Conc. Standard
Conc. Sample = Abs. Sample x
Abs. Standard

Conc. Standard
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The Calibration Factor can be stored as F.
Abs. Standard

In case the calculation is made using factor or stored calibration, the formula becomes:

Conc. Sample = Abs. Sample X F

Calculation 2 KINETIC

During the reading interval programmed for the test, every 80 ms the absorbance is measured.
At the end of time span, a straight line is calculated through the points of measurement, using the
linear regression method.
The coefficient of correlation (FIT) is used as a check on the linearity (must be > 0.98).
The calculated slope, expressed in Abs./min. is used to calculate the activity:

U/l = Abs./min. X F

or:
Conc. Standard
Conc. = Abs./min. X
Abs./min. (Standard)

Results are FLAGGED:

In case of increasing (+) reactions:
If the first reading > of start-Absorb. as programmed for method in question
If the first reading < (RB+SE)
RB= Abs. Reagent Blank as measured at start of batch
SE= O.D. for substrate Expletion as programmed for method

In case of decreasing (-) reactions:

If the first reading < (RB-SE)

APPENDIX 6-A
Continued

Calculation 3 INITIAL RATE

This corresponds to fixed time kinetics.

Calculation is based on the Delta absorbance over the reading interval : one reading at the start (end
of incubation time) and the second at the end.

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Conc. Sample = Abs. Sample X F

Can be used also a standard insteadof F as in EndPoint calculation mode.

Calculation 4 BICHROMATIC

Calculation based on the Delta Absorbance between two wavelenghts.The formula is like in
EndPoint.

Conc. Sample = Abs. Sample X F

Calculation 5 SERUM BLANK

This type of calculation is based on the Delta abs. of a sample with reagent and a serum blank
(Sample + Diluent).
To this end, two reaction wells are being used for each sample.
Calculation as for EndPoint.

APPENDIX 6-B

ON LINE INTERFACING

INTERFACE

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Serial Interface used is of the type RS-232 C.

The serial port for connection to a Lab Computer is the upper 25 pins, female, on the back of
PRIME EVOLUTION.
Connections are as follows:

Pin 2 - TX (transmit)
Pin 3 - RX (receive)
Pin 7 - GND (signal ground)

WORD FORMAT

1200 Baud
8 Data bit
1 Stop bit
NO Parity

APPENDIX 6-C

MATHEMATICAL MODELS FOR MULTI STANDARD CURVES

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For the calculation of the non-linear standard curves the operator may choose out of 6 mathematical
models.

MODEL 1: 4 Parameters Log-Logit

1
R=Ro + Kc
1 + exp [-(a+b.lnC)]

MODEL 2: 5 Parameters Log-Logit

1
R=Ro + Kc
1 + exp [-(a+b.lnC+c.C)]

MODEL 3: 5 Parameters Exponential

R= Ro + K exp [ a.lnC + b.(lnC)2 + c.(lnC)3]

MODEL 4: 5 Parameters Polynomial

(R-Ro) (R-Ro)2 (R-Ro)3

ln C = a + b + c + d
100 (100)2 (100)3

MODEL 5: 4 Parameters Logit

1
R= Ro + Kc
1+exp [-(a+b.C)]

MODEL 6: Linear with offset

C = a + b.R
APPENDIX 6-D

PRINTABLE ASCII CHARACTERS TABLE

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As indicated in the Methods programming section, PRIME EVOLUTION is not provided with an
alfabetical keyboard, so to enter alfabetical characters it's necessary to enter them in numerical
ASCII format.
When PRIME EVOLUTION asks for an alfabetical entering, just press sequentially number keys
corresponding to letter wanted, then when you've entered complete word, press enter key to
confirm.

No. ASCII No. ASCII No. ASCII

032 Space 064 @ 096 '
033 ! 065 A 097 a
034 " 066 B 098 b
035 # 067 C 099 c
036 $ 068 D 100 d
037 % 069 E 101 e
038 & 070 F 102 f
039 ' 071 G 103 g
040 ( 072 H 104 h
041 ) 073 I 105 i
042 * 074 J 106 j
043 + 075 K 107 k
044 , 076 L 108 l
045 - 077 M 109 m
046 . 078 N 110 n
047 / 079 O 111 o
048 0 080 P 112 p
049 1 081 Q 113 q
050 2 082 R 114 r
051 3 083 S 115 s
052 4 084 T 116 t
053 5 085 U 117 u
054 6 086 V 118 v
055 7 087 W 119 w
056 8 088 X 120 x
057 9 089 Y 121 y
058 : 090 Z 122 z
059 ; 091 [ 123 {
060 < 092 \ 124 |
061 = 093 ] 125 }
062 > 094 ^ 126
063 ? 095 _ 127

APPENDIX 6-E

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ON SCREEN FLAG CODES LIST

L Beyond linearity limit fixed in methods

Dilution is needed

P Out of normal values range fixed in method

If first O.D. (in kinetics reaction as GOT and GPT) is minor than blank reagent O.D. - Substrate
expl. limit O.D. as programmed in method. It means reagent not working properly or activity of
sample is extremely high-try dilution-

FC In immunochemistry means the sample is out of calibration curve.

CK Near reagent Blank O.D. it means O.D. out of range programmed in method.
Check reagent.

CK Near C1,C2,C3 it means control serum value is out of the range programmed in
Control Sera Assay
Valid only if control sera value are programmed(not=0) see 3.8

ERROR MESSAGES ON SCREEN

WARNING RESET PROBLEM SWITCH OFF AND ON AGAIN

Tells operator to switch instrument off, wait 10 seconds and then switch it on again to reset the
CPU.In case there are still problems on reset, contact service.

WARNING INCORRECT FILTERWHEEL RESET

Problems of motor or photocoupler for home position, clean photocoupler or contact service.

FILTER OUT OF RANGE - INCORRECT AUTOZERO

Check if lamp is burned out, flow cells dirty or clogged, in case this message does not disappear,
contact your dealer.

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