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Holding Company for Drinking water and Waste water CWC/MOS/Test P/23
Cairo Water Company Issue No.: 01
Laboratories & Research Sector
Quality Control Administration Issue Date: 1/ 4/2018
Test Procedure
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Determination of Phosphate

TITLE: Determination of Phosphate

Name Signature

Prepared by Mai Abdullah Mai Abdullah

Saeed Ibrahim Saeed Ibrahim

Reviewed by
Warda Adel Warda Adel

Final reviewed by Mai Sadek Mai Sadek

Approved by Atef Dokhan Atef Dokhan

 Holding Company for Drinking water and Waste water CWC/MOS/Test P/23
Cairo Water Company Issue No.: 01
Laboratories & Research Sector
Quality Control Administration Issue Date: 1/ 4/2018
Test Procedure
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Determination of Phosphate

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 Holding Company for Drinking water and Waste water CWC/MOS/Test P/23
Cairo Water Company Issue No.: 01
Laboratories & Research Sector
Quality Control Administration Issue Date: 1/ 4/2018
Test Procedure
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Determination of Phosphate

CONTENTS

1. Purpose.
2. Scope and Application.
3. Principle.
4. Responsibility.
5. Environment conditions.
6. Equipment.
7. Reagents.
8. Precautions.
9. Sample handling and Storage.
10. Interference.
11. Procedure.
12. Calculation
13. Assuring the test results.
14. Criteria.
15. Test report.
16. References.
 Edited by Foxit Reader
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water and Waste water CWC/MOS/Test P/23
Cairo Water Company Issue No.: 01
Laboratories & Research Sector
Quality Control Administration Issue Date: 1/ 4/2018
Test Procedure
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Determination of Phosphate

1. PURPOSE
This procedure applied for determination of reactive phosphorus content using
Spectrophotometer according to SMWW Standard Methods for the Examination of Water
and Wastewater 23rd Edition, Method
The font No.4500-P
of number D. the
must be (Stannous
same Chloride Method).
size as writing and not bold
2. SCOPE AND APPLICATION
2.1 This procedure is applicable to all types of water sample such as raw water, water
during intermediate stages of the treatment, drinking water and ground water samples as
well as external or internal samples.

2.2 This procedure used a suitable Light path lengths for various concentration ranges are as
follows:
Approximate P Light Path
Range mg/L cm
0.3–2 0.5
0.1–1 2
0.007–0.2 10

2.3 So, Spectrophotometer with Light path length =1cm can’t used.

2.4 This procedure is suitable for range lies between 0.01 to 6 mg P/L.

2.5 The minimum detectable concentration is about 3µg P/L.

3. PRINCIPLE
Molybdophosphoric acid is formed and reduced by stannous chloride to intensely colored
molybdenum blue. This method is more sensitive and makes feasible measurements down to
7 µg P/L by use of increased light path length.

4. RESPONSIBILITY
All Chemists are responsible for carrying out and understanding the contents of this Test
Procedure.

5. ENVIRONMENT CONDITIONS
Rate of color development and intensity of color depend on temperature of the final solution,
each 1°C increase producing about 1% increase in color. Hence, hold samples, standards,
and reagents within 2°C of one another and in the temperature range between 20 and 30°C.
 Edited by Foxit Reader
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water and Waste water CWC/MOS/Test P/23
Cairo Water Company Issue No.: 01
Laboratories & Research Sector
Quality Control Administration Issue Date: 1/ 4/2018
Test Procedure
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Determination of Phosphate
6. EQUIPMENT JENWAY single beam, MODEL 6715
6.1 Spectrophotometer, for use at 690 nm.

6.2 Nessler tubes, 100ml, tall form, matched.

7. REAGENTS
7.1 Phenolphthalein indicator aqueous solution.

7.2 Strong acid solution: Slowly add 300ml conc. H2SO4 to about 600ml distilled water.
when cool, add 4ml conc. HNO3 ,and dilute to 1L.

7.3 Ammonium molybdate reagent I: Dissolve 25g (NH4)6Mo7O24.4H2O in 175ml distilled

water. Cautiously add 280ml conc. H2SO4 to 400ml distilled water, Cool, add
molybdate solution, and dilute to 1 L.

7.4 Stannous chloride reagent I: Dissolve 2.5g fresh SnCl2 . 2H2O in 100ml glycerol.
Heat in a water bath and stir with a glass rod to hasten dissolution. This reagent is
stable and requires neither preservatives nor special storage.

7.5 Stock phosphate solution: Dissolve in distilled water 219.5 mg anhydrous

KH2PO4 and dilute to 1L; (1ml = 50 µg PO43- - P)

7.6 Standard phosphate solution: Dilute 100ml from stock solution to 1L;
( 1ml = 0.005mg PO43- - P)

8. PRECAUTIONS
8.1 Rinse all glass containers with hot dilute HCl, then rinse several times in reagent water.

8.2 Preferably, reserve the glassware only for phosphate determination, and after use, wash
and keep filled with water until needed. If this is done, acid treatment is required only
occasionally.

8.3 Never use commercial detergents containing phosphate for cleaning glassware used in
phosphate analysis.

8.4 Use batches of chemicals low in phosphorus.

8.5 Use distilled reagent water in making reagents and dilutions.

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Determination of Phosphate
8.6 Volumetric measurement of sample and reagent is extremely important to analytical
accuracy.

9. SAMPLE HANDLING AND STORAGE

9.1 Collect samples in clean bottles, preferably glass rinsed with 1+ 1 HNO3, and examine
them as soon as possible after collection.

9.2 The minimum sample size is 100ml, in grab sample.

9.3 If dissolved phosphorus forms are to be differentiated, filter sample immediately after
collection within 1h through 0.45µm membrane filters.

9.4 Wash membrane filters by soaking in distilled water before use because they may
contribute significant amounts of phosphorus to samples containing low concentrations of
phosphate. Use one of two washing techniques: This procedure applied for determination
of reactive phosphorus content only not
9.4.1 soak 50 filters in 2L distilled water for 24h. dissolved or any form

9.4.2 soak 50 filters in 2L distilled water for 1h, change distilled water, and soak filters
an additional 3h.
9.4.3 Membrane filters also may be washed by running several 100ml portions of
distilled water through them. This procedure requires more frequent determination
of blank values to ensure consistency in washing and to evaluate different lots of
filters.

9.5 For long periods storage before analysis, preserve samples by freezing at or below -
10°C. In some cases 40mg HgCl2/L may be added. CAUTION: HgCl2 is a hazardous
substance; take appropriate precautions in disposal; use of HgCl2 is not encouraged.

9.6 Don’t add either acid or CHCl3 as a preservative when phosphorus forms are to be
determined.

9.7 For total phosphorus alone is to be determined, add H2SO4 or HCl to pH < 2 and cool to
4°C, or freeze without any additions. ( 28 day Maximum storage recommended).

9.8 Don’t store samples containing low concentrations of phosphorus in plastic bottles unless
kept in a frozen state because phosphates may be adsorbed onto the walls of plastic
bottles.
 Holding Company for Drinking water and Waste water CWC/MOS/Test P/23
Cairo Water Company Issue No.: 01
Laboratories & Research Sector
Quality Control Administration Issue Date: 1/ 4/2018
Test Procedure
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Determination of Phosphate

10. INTERFERENCE
10.1 Positive interference is caused by silica and arsenate only if the sample is heated.

10.2 Negative interferences are caused by arsenate, fluoride, thorium, bismuth, sulfide,
thiosulfate, thiocyanate, or excess molybdate.

10.3 Blue color is caused by ferrous iron but this doesn’t affect results if ferrous iron
concentration is less than 100mg/L.

10.4 Sulfide interference may be removed by oxidation with bromine water.

11. PROCEDURE
11.1 Preparation of calibration curve:
11.1.1 Prepare a calibration curve by using suitable volumes of standard phosphate
solution diluted to 50ml in Nessler tubes, ( note; range of standard according to
Light Path).

St. No. Volume of standard Volume of dist.water Conc. of standard

1 0.07 ml 49.93 ml 0.007 mg/l
2 0.09 ml 49.91 ml 0.009 mg/l
3 0.1 ml 49.9 ml 0.01 mg/l
4 0.25 ml 49.75 ml 0.025 mg/l
5 0.5 ml 49.5 ml 0.050 mg/l
6 0.8 ml 49.2 ml 0.080 mg/l
7 1 ml 49 ml 0.1 mg/l
8 1.5 ml 48.5 ml 0.15 mg/l
9 2 ml 48 ml 0.2 mg/l
10 4 ml 46 ml 0.4 mg/l
11 6 ml 44 ml 0.6 mg/l
12 8 ml 42 ml 0.8 mg/l
13 10 ml 40 ml 1.0 mg/l
14 13 ml 37 ml 1.3 mg/l
15 15 ml 35 ml 1.5 mg/l
16 20 ml 30 ml 2 mg/l

11.1.2 Add, with thorough mixing after each addition, 2ml molybdate reagent I and
0.25ml (5drops) stannous chloride reagent I.
 Holding Company for Drinking water and Waste water CWC/MOS/Test P/23
Cairo Water Company Issue No.: 01
Laboratories & Research Sector
Quality Control Administration Issue Date: 1/ 4/2018
Test Procedure
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Determination of Phosphate

11.1.3 Rate of color development and intensity of color depend on temperature of the
final solution.

11.1.4 Measure color After 10min, but before 12min, using the same specific interval
for all determinations

11.1.5 Read absorbance on a spectrophotometer, using a wavelength of 690 nm. Set

zero absorbance using distilled water blank.

11.1.6 Plot concentration of standard solution against absorbance. The calibration

curve may deviate from a straight line at the upper concentrations of the 0.3- to
2mg/L range.

11.2 Sample Measurement:

11.2.1 To 50ml sample containing not more than 200µg P and free from color
and turbidity, add 0.05ml (1 drop) phenolphthalein indicator.

11.2.2 If sample turns pink, add strong acid solution dropwise to discharge the color.

11.2.3 If strong acid more than 0.25ml (5 drops) is required, take a smaller sample
and dilute to 50mL with distilled water after first discharging the pink color
with acid.

11.2.4 Add, with thorough mixing after each addition, 2ml molybdate reagent I and
0.25ml (5 drops) stannous chloride reagent I.

11.2.5 Measure color after 10min, but before 12min, using the same specific interval
for all determinations.

11.2.6 Determine phosphate concentration from the calibration curve previously

prepared.

11.2.7 Always run a blank on reagents and distilled water.

11.2.8 Prepare at least one standard with each set of samples or once each day that
tests are made.
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Laboratories & Research Sector
Quality Control Administration Issue Date: 1/ 4/2018
Test Procedure
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Determination of Phosphate

12. CALCULATION

13. ASSURING THE TEST RESULTS

13.1 The laboratory apply internal quality control program that specifies the measures
required producing data with known precision and accuracy.

13.2 This program consists of control charts (X & R Charts)

13.3 The laboratory will construct the charts according to the work instruction CWC/MOS/
Work. I /01

14. CRITERIA
There are no acceptance and rejection criteria of the samples according to the Egyptian
Standards& Regulations (No. 458) for the year 2007.

15. TEST REPORT

The laboratory has different test reports according to required tests, for Phosphate test
Using form N4/lab & N5/lab& N 8-1/lab

16. REFERENCES
SMWW Standard Methods for the Examination of Water and Wastewater 23rdEdition,
2017, Method No. 4500-P D.