Phosphorus, Reactive

(Orthophosphate)

DOC316.53.01119

USEPA1 PhosVer 3 (Ascorbic Acid) Method2

Method 8048

0.02 to 2.50 mg/L PO43

Powder Pillows or AccuVac Ampuls

Scope and application: For water, wastewater and seawater.

1
2

USEPA Accepted for reporting for wastewater analyses. Procedure is equivalent to USEPA and Standard Method 4500-P-E for
wastewater.
Adapted from Standard Methods for the Examination of Water and Wastewater.

Test preparation
Instrument-specific information
Table 1 shows sample cell and orientation requirements for reagent addition tests, such
as powder pillow or bulk reagent tests. Table 2 shows sample cell and adapter
requirements for AccuVac Ampul tests. The tables also show all of the instruments that
have the program for this test.
To use the table, select an instrument, then read across to find the applicable information
for this test.
Table 1 Instrument-specific information for reagent addition
Instrument

Sample cell orientation

Sample cell

DR 6000

The fill line is to the right.

2495402

DR 3800
DR 2800
DR 2700
DR 1900
DR 5000

The fill line is toward the user.

DR 3900
DR 900

The orientation mark is toward the user.

2401906

Table 2 Instrument-specific information for AccuVac Ampuls

Instrument
DR 6000

Adapter

Sample cell

2427606

DR 5000
DR 900
DR 3900

LZV846 (A)

DR 1900

9609900 or 9609800 (C)

DR 3800

LZV584 (C)

2122800

DR 2800
DR 2700

Before starting
Install the instrument cap on the DR 900 cell holder before ZERO or READ is pushed.
For the best results, measure the reagent blank value for each new lot of reagent. Replace the sample with deionized water
in the test procedure to determine the reagent blank value. Subtract the reagent blank value from the sample results
automatically with the reagent blank adjust option.
Review the Safety Data Sheets (MSDS/SDS) for the chemicals that are used. Use the recommended personal protective
equipment.
Dispose of reacted solutions according to local, state and federal regulations. Refer to the Safety Data Sheets for disposal
information for unused reagents. Refer to the environmental, health and safety staff for your facility and/or local regulatory
agencies for further disposal information.

Items to collect
Powder pillows
Description

Quantity

PhosVer 3 Phosphate Reagent powder pillow, 10-mL

Sample cells. (For information about sample cells, adapters or light shields, refer to Instrumentspecific information on page 1.)

Refer to Consumables and replacement items on page 6 for order information.

AccuVac Ampuls
Description

Quantity

PhosVer 3 Phosphate Reagent AccuVac Ampul

Beaker, 50-mL

Sample cells (For information about sample cells, adapters or light shields, refer to Instrumentspecific information on page 1.)

Stopper for 18-mm tubes and AccuVac Ampuls

Refer to Consumables and replacement items on page 6 for order information.

Sample collection and storage

Collect samples in clean glass or plastic bottles that have been cleaned with 6 N (1:1)
hydrochloric acid and rinsed with deionized water.
Do not use a detergent that contains phosphate to clean the sample bottles. The
phosphate in the detergent will contaminate the sample.
Analyze the samples as soon as possible for best results.
If immediate analysis is not possible, immediately filter and keep the samples at or
below 6 C (43 F) for a maximum of 48 hours.
Let the sample temperature increase to room temperature before analysis.

Phosphorus, Reactive, PhosVer 3 Method (2.50 mg/L)

Powder pillow procedure

Start

1. Start program 490 P

React. PP. For information
about sample cells,
adapters or light shields,
refer to Instrument-specific
information on page 1.
Note: Although the program
name can be different
between instruments, the
program number does not
change.

2. Prepare the sample: Fill

a sample cell with 10 mL of
sample.

3. Add the contents of one

PhosVer 3 Phosphate
Reagent Powder Pillow to
the cell. A blue color
develops if phosphorus is in
the sample.

4. Immediately close the

sample cell. Shake
vigorously for
2030 seconds.

5. Start the instrument

timer. A 2-minute reaction
time starts.

6. Prepare the blank: Fill a

second sample cell with
10 mL of sample.

7. When the timer expires,

clean the blank sample cell.

8. Insert the blank into the

cell holder.

If the sample was digested

using the Acid Persulfate
digestion, a 10-minute
reaction period is
necessary.

Zero

9. Push ZERO. The display

shows 0.00 mg/L PO43.

Read

10. Clean the prepared

sample cell.

Phosphorus, Reactive, PhosVer 3 Method (2.50 mg/L)

11. Insert the prepared

sample into the cell holder.

12. Push READ. Results

show in mg/L PO43.

AccuVac Ampul procedure

Start

1. Start program 492 P

React. PV AV. For
information about sample
cells, adapters or light
shields, refer to Instrumentspecific information
on page 1.
Note: Although the program
name can be different
between instruments, the
program number does not
change.

2. Prepare the blank: Fill

the sample cell with 10 mL
of sample.

3. Prepare the sample:

Collect at least 40 mL of
sample in a 50-mL beaker.
Fill the AccuVac Ampul with
sample. Keep the tip
immersed while the
AccuVac Ampul fills
completely.

4. Close the AccuVac

Ampul. Shake for
approximately 30 seconds.
Accuracy is not affected by
undissolved powder.

Zero

5. Start the instrument

timer. A 2-minute reaction
time starts.

6. When the timer expires,

clean the blank sample cell.

7. Insert the blank into the

cell holder.

8. Push ZERO. The display

shows 0.00 mg/L PO43.

If the sample was digested

using the Acid Persulfate
digestion, a 10-minute
reaction period is
necessary.

Read

9. Clean the AccuVac

Ampul.

10. Insert the prepared

sample AccuVac Ampul into
the cell holder.

11. Push READ. Results

show in mg/L PO43.

Phosphorus, Reactive, PhosVer 3 Method (2.50 mg/L)

Interferences
Interfering substance

Interference level

Aluminum

More than 200 mg/L

Arsenate

Interferes at any level

Chromium

More than 100 mg/L

Copper

More than 10 mg/L

Hydrogen Sulfide

Interferes at any level

Iron

More than 100 mg/L

Nickel

More than 300 mg/L

Highly buffered samples or

extreme sample pH

Can prevent the correct pH adjustment of the sample by the reagents. Sample pretreatment may be necessary. A pH range of 210 is recommended.

Silica

More than 50 mg/L

Silicate

More than 10 mg/L

Turbidity or color

Samples with a high amount of turbidity can give inconsistent results. The acid in the
reagents can dissolve some of the suspended particles and variable desorption of
orthophosphate from the particles can occur.

Zinc

More than 80 mg/L

Accuracy check
Standard additions method (sample spike)
Use the standard additions method (for applicable instruments) to validate the test
procedure, reagents and instrument and to find if there is an interference in the sample.
Items to collect:

Phosphate standard solution, 50 mg/L PO43 ampule

Ampule breaker
Pipet, TenSette, 0.11.0 mL and tips
Mixing cylinders, 25-mL (3)

1. Use the test procedure to measure the concentration of the sample, then keep the
(unspiked) sample in the instrument.
2. Go to the Standard Additions option in the instrument menu.
3. Select the values for standard concentration, sample volume and spike volumes.
4. Open the standard solution.
5. Prepare three spiked samples: use the TenSette pipet to add 0.1 mL, 0.2 mL and
0.3 mL of the standard solution, respectively, to three 10-mL portions of fresh sample.
Mix well.

Note: For AccuVac Ampuls, add 0.2 mL, 0.4 mL and 0.6 mL of the standard solution to three
50-mL portions of fresh sample.

6. Use the test procedure to measure the concentration of each of the spiked samples.
Start with the smallest sample spike. Measure each of the spiked samples in the
instrument.
7. Select Graph to compare the expected results to the actual results.
Note: If the actual results are significantly different from the expected results, make sure that
the sample volumes and sample spikes are measured accurately. The sample volumes and
sample spikes that are used should agree with the selections in the standard additions menu. If
the results are not within acceptable limits, the sample may contain an interference.

Standard solution method

Use the standard solution method to validate the test procedure, the reagents and the
instrument.
Phosphorus, Reactive, PhosVer 3 Method (2.50 mg/L)

Items to collect:

50 mg/L phosphate standard solution

100-mL volumetric flask, Class A
4-mL volumetric pipet, Class A and pipet filler safety bulb
Deionized water

1. Prepare a 2.00-mg/L phosphate standard solution as follows:

a. Use a pipet to add 4.00 mL of a 50-mg/L phosphate standard solution into the
volumetric flask. (Alternately, use one of the available mixed parameter
standards. These standards contain 2.0 mg/L phosphate.)
b. Dilute to the mark with deionized water. Mix well. Prepare this solution daily.
2. Use the test procedure to measure the concentration of the prepared standard
solution.
3. Compare the expected result to the actual result.
Note: The factory calibration can be adjusted slightly with the standard adjust option so that the
instrument shows the expected value of the standard solution. The adjusted calibration is then
used for all test results. This adjustment can increase the test accuracy when there are slight
variations in the reagents or instruments.

Method performance
The method performance data that follows was derived from laboratory tests that were
measured on a spectrophotometer during ideal test conditions. Users can get different
results under different test conditions.
Program

Standard

Precision (95% Confidence Interval)

Sensitivity
Concentration change per 0.010 Abs change

490

2.00 mg/L PO43

1.982.02 mg/L PO43

0.02 mg/L PO43

492

2.00 mg/L PO43

1.982.02 mg/L PO43

0.02 mg/L PO43

Summary of method
Orthophosphate reacts with molybdate in an acid medium to produce a mixed
phosphate/molybdate complex. Ascorbic acid then reduces the complex, which gives an
intense molybdenum blue color. The measurement wavelength is 880 nm (DR 1900:
710 nm) for spectrophotometers or 610 nm for colorimeters.

Consumables and replacement items

Required reagents
Description

Quantity/Test

Unit

Item no.

100/pkg

2106069

25/pkg

2508025

Quantity/Test

Unit

Item no.

Beaker, 50-mL

each

50041H

Stoppers for 18-mm tubes and AccuVac Ampuls

6/pkg

173106

PhosVer 3 Phosphate Reagent Powder Pillow1, 10-mL

OR

PhosVer 3 Phosphate Reagent AccuVac Ampul

1

PhosVer is a registered trademark of Hach Company.

Required apparatus
Description

Phosphorus, Reactive, PhosVer 3 Method (2.50 mg/L)

Recommended standards
Description

Unit

Item no.

16/pkg

17110

500 mL

17149

500 mL

256949

Drinking Water Standard, Mixed Parameter, Inorganic for F-, NO3N, PO43, SO42

500 mL

2833049

Wastewater Effluent Standard Solution, Mixed Parameter, for NH3-N, NO3-N, PO43,
COD, SO42, TOC

500 mL

2833249

4L

27256

Phosphate Standard Solution, 10-mL Voluette Ampule, 50-mg/L as PO4

Phosphate Standard Solution, 50-mg/L as PO4
Phosphate Standard Solution, 1-mg/L as PO4

Water, deionized

Optional reagents and apparatus

Description

Unit

Item no.

each

4103600

each

2405200

25/pkg

2677925

each

2196800

12/pkg

2087076

Mixing cylinder, graduated, 50-mL

each

189641

Flask, volumetric, Class A, 100-mL glass

each

1457442

Hydrochloric Acid Solution, 6.0 N (1:1)

500 mL

88449

Paper, pH, 014 pH range

100/pkg

2601300

Phosphate Treatment Powder Pillows

100/pkg

1450199

Phosphate Standard Solution, 10-mg/L as PO4

946 mL

1420416

Phosphate Standard Solution, 15-mg/L as PO43

100 mL

1424342

Phosphate Standard Solution, 100-mg/L as PO4

100 mL

1436832

Phosphate Standard Solution, 10-mL ampule, 500 mg/L as PO4

16/pkg

1424210

Phosphate Standard Solution, 500-mg/L as PO4

100 mL

1424232

each

1970001

AccuVac Drainer
AccuVac Ampul Snapper

AccuVac Ampul vials for sample blanks

Ampule Breaker, 10-mL Voluette Ampules

Bottle, sampling, with cap, low density polyethylene, 250-mL

Pipet, TenSette , 0.11.0 mL

Pipet, TenSette , 1.010.0 mL

each

1970010

50/pkg

2185696

1000/pkg

2185628

50/pkg

2199796

250/pkg

2199725

each

1451504

Pipet tips for TenSette Pipet, 0.11.0 mL

Pipet tips for TenSette Pipet, 0.11.0 mL
Pipet tips for TenSette Pipet, 1.010.0 mL
Pipet tips for TenSette Pipet, 1.010.0 mL
Pipet, volumetric, Class A, 4.00-mL

Phosphorus, Reactive, PhosVer 3 Method (2.50 mg/L)

FOR TECHNICAL ASSISTANCE, PRICE INFORMATION AND ORDERING:

In the U.S.A. Call toll-free 800-227-4224
Outside the U.S.A. Contact the HACH office or distributor serving you.
On the Worldwide Web www.hach.com; E-mail [email protected]

Hach Company/Hach Lange GmbH, 19892014. All rights reserved.

HACH COMPANY
WORLD HEADQUARTERS
Telephone: (970) 669-3050
FAX: (970) 669-2932

01/2014, Edition 9