DETERMINATION OF PHOSPHATE

IN WATER SAMPLES

Fatma Basyoni Salem

Department of Analytical Chemistry Contents

Faculty ofPharmacy, Mansoura University
Mansoura, Egypt

CONTENTS

Page

Abstract 225
Introduction 226
Instruments, Materials and Methods 226
Results and Discussion 229
Conclusion 233
References 236

ABSTRACT

Orthophosphate and molybdate ions condense in sulphuric acid solution

to give molybdophosphoric acid, which upon selective reduction with
hydrazinium sulphate produces a blue color due to molybdenum blue. This
was readily conjugated with the cation dye Brilliant Blue, which exhibits
maximum absorption after its extraction in chloroform at 840 nm. Beer's
law is obeyed in the concentration range of 2.8 to 28.1 μg phosphate per 25
ml, with molar absorptivity 883700 L mol"1 cm"1. This procedure was
compared with the results obtained from titrimetric techniques. A
Potentiometrie method was used in the presence of a cupric ion selective
electrode to determine the excess copper ions with EDTA after phosphate
precipitation. The excess in copper ions was determined amperometrically
using thiomalic acid in acetic acid as a titrant at an applied voltage of +0.8V
versus a standard calomel electrode, with a rotating micro platinum
electrode.

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I. INTRODUCTION

The aim of this paper is to report:

a) A new spectrophotometric technique is used to determine phosphate in

potable waters using Brilliant Blue as a chromogenic agent to form a
conjugate system between the produced molybdenum blue and the dye. The
maximum absorbance is determined at 880 nm with a concentration range of
2.8 to 28.1 μg phosphate per 25 ml.
b) This is compared with the titrimetric study including Potentiometrie
and amperometric techniques. The excess copper is determined either by the
use of a copper ion selective electrode and EDTA as a titrant or by the use of
a rotating platinum electrode at an applied potential of +0.8V using
thiomalic acid in acetic acid as a titrant; it is used in the concentration range
of 1 χ 10"' to 1 χ 10"6 Μ solution of phosphate.
c) These techniques are applied for determination of some water samples
of Egypt, especially Dakahlia province, and some organophosphoric com-
pounds, e.g. malathion and parathion.

Ground water contains only very small amounts of phosphate, usually

less than 0.1 mg.l"1, with the exception of water from phosphate-containing
soil or polluted by organic matter. Condensed phosphates are used as
cleaning preparations in the treatment of some water supplies. Phosphorus
compounds are carried into natural water with waste water.
Several methods were used for the determination of phosphate including
spectrophotometric /1-5/, titrimetric 16,11, amperometric and Polarographie
/8,9/, atomic absorption /10/, thin layer /ll/, gas liquid chromatographic
/12/ and x-ray fluorescence /13/ techniques.

2. INSTRUMENTS, MATERIALS AND METHODS

An Orion Research microprocessor PH/millivolt meter 811 is used

equipped with the Orion model 94-29 cupric and Orion model 90-01 single
junction reference electrodes, while the apparatus for amperometiy is
reported in ref. UM, consisting of a rotating platinum microelectrode and a

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standard calomel electrode in addition to a Unicam SP 1800

spectrophotometer.
All solutions were prepared with analytical reagent grade chemicals
(BDH) using bidistilled water.

a) Phosphate stock solution was prepared as follows: 100 mg disodium

hydrogen phosphate (Na2HP04, 12 H 2 0) was dried at 60°C to constant
mass,then dissolved in 100 ml of water,
Phosphate working standard solution was prepared by diluting 10 ml of
the stock standard solution to 50 ml in a measuring flask, each 1 ml
containing 5 6.2 μg phosphate (P04)3". Standard solutions of malathion
and parathion are prepared by digesting 3.3 and 2.91 g of each organo-
phosphorus compound respectively in 10 ml of equal volume of 10%
nitric and sulphuric acid for 10 min in a boiling oil bath. After cooling,
the digest solution was diluted to one liter to obtain a 0.01 Μ solution.

b) Standard phosphate (Na2HP04, 12H20) 0.1 Μ solution was prepared by

dissolving 35.8 g in one liter of distilled water; serial dilutions were
done, down to 1 χ 10"6 Μ solution.

c) Standard EDTA solution 0.1 Μ was prepared by dissolving 37.2 g

reagent - grade EDTA in about 200 ml distilled water; this was then
diluted to one liter volume distilled water and used for Potentiometrie
standardization of cupric sulphate solution.

d) Standard 0.1 Μ cupric sulphate solution was prepared by dissolving

15.61 g of anhydrous cupric sulphate in acidified distilled water with
sulphuric acid and completing to one liter; serial dilutions were done.

e) Standard thiomalic acid was prepared in a concentration of 0.1 Μ

solution; serial dilutions were obtained and standardized ampero-
metrically.

f) Acetic acid solution 1:1 v/v.

g) Saturated solution of ammonium acetate.

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Reagents for the Spectrophotometric Method:

a) Brilliant Blue (0.1% w/v) in ethanol;

b) Chloroform for extraction;
c) Hydrazinium sulphate solution was prepared by dissolving 1.5 g of A.R.
hydrazinium sulphate in deionised water and completed to 1 L;
d) Molybdate solution was prepared by dissolving 12.5 g of A.R. sodium
molybdate (Na2Mo04, 2H 2 0) in 5 Μ sulphuric acid and diluting to 500 ml
with 5 Μ sulphuric acid.

Spectrophotometric Determination of Phosphate, Procedure (1)

A sample solution containing not more than 28.0 μg phosphate was

transferred into a 25 ml Pyrex tube. 5 ml of the molybdate solution, and 3 ml
of 10% sulphuric acid were added and warmed on a boiling water bath for
15 min, then 2.0 ml of hydrazinium sulphate solution was added. The
mixture in the tube was mixed well and immersed in a boiling water bath for
another 10 min, then removed and 0.5 ml Brilliant Blue solution were added
and heating continued for another 5 min. The mixture was cooled rapidly to
room temperature and extracted with chloroform three times (each time 5
ml) and the organic extract collected in a 25 ml volumetric flask and the
volume adjusted. The absoibance was measured at 840 nm against a reagent
blank within 2 hours.
The concentration can be obtained from a calibration curve previously
prepared in the range of 5.6 to 28.1 μg P043" per 25 ml.

Potentiometrie Determination of Phosphate, Procedure (2)

10.0 ml of standard phosphate solution in a concentration range of 1x10"'

to 1 χ 10"6 Μ solution was transferred into a 250 ml beaker followed by 5
ml of 1:1 acetic acid solution and 25 ml of cupric sulphate, then heated just
to boiling point and cooled. Cupric and reference electrodes were placed in
the solution to a depth of about 3 cm, and the function switch turned to the
mV position. The solution was thoroughly stirred, and a stable potential was
awaited and recorded. The solution was titrated with a suitable concentration
of standard EDTA according to the molarity of phosphate and cupric
solutions required.

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In each case the potential was plotted in mV versus mis titrant to

determine the end point graphically for the maximum slope AE/AV or from
the point where the second derivative A2E/AV2 becomes zero.

Amperometric Determination of Phosphate, Procedure (3)

The reaction mixture after boiling and then cooling as directed in

procedure (A) above was transferred into the cell. 10 ml of the supporting
electrolyte, consisting of 5 ml of (111) acetic acid and 5 ml saturated
ammonium acetate, was introduced. Meanwltile the applied voltage was
adjusted to +0.8V vs standard calomel electrode. The sensitivity control was
set to an appropriate value and the galvanometer spot to zero. The titrant
thiomalic acid solution was added from the burette in 0.5 ml portions until
within 1 ml of the equivalence point, then in 0.1 ml quantities for the
following 2 ml, subsequently in 0.5 ml portions. The gas stream was passed
through the solution for about 1 min after each addition (more dilute
solutions will require up to 3 min) to assist the precipitation of the copper
ions and then over the surface before reading the galvanometer deflection
(current). The current readings were corrected for the change in volume of
the solution due to the added reagent. The equivalence point was read off
from the amperometric titration curve drawn from the obtained result.

3. RESULTS AND DISCUSSION

In order to obtain suitable conditions for the determination of phosphate

in water samples, the effect of various factors was studied.

(a) Effect of Acidity

In the range of 0.5 - 3 ml of 10% acid, H 2 S0 4 provides the largest and

constant absorbance at 840 nm (Fig. 1), which is higher in molar
absorptivity than in the presence of hydrochloric acid where the absorbance
decreases more rapidly. At the same time condensed phosphate, probably
existing in water samples, is hydrolysed in this acid medium by heating
in a boiling water bath for 30 minutes.

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F.B. Salem Reviews in Analytical Chemistry

(b) Effect of Molybdate Concentration

Molybdate solution is used in the concentration range of 2 - 5 ml to

obtain the maximum absorbance at 840 nm.

(c) Effcct of Dye Concentration

Figure 2 shows the effect of change in concentration of 0.1% Brilliant

Blue solution. It is clear that in the range of 0.2 to 1.0 ml the maximum
absorbance is obtained at 840 nm, and can persist for 2 hours.

Vol. in mis.

Fig.(2): Effect of the volumes of Brilliant Blue.

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Inorganic phosphate reacts with a solution of molybdate in the range of 2

- 5 ml in the presence of 0.5 - 3 ml of 10% H 2 S0 4 acid to give an intense
yellow coloration, due probably to the complex molybdophosphoric acid,
which upon selective reduction with hydrazinium sulphate (0.2 ml),
produces a blue coloration due to molybdenum blue. This associates with the
oppositely charged cation dye Brilliant Blue in the range of 0.2 to 1.0 ml of
0.1%. The neutral extractable chelates species produced are extracted into
chloroform and measured spectrophotometrically at 840 nm within 45 min,
with molar absorptivity 88.37 χ 104 L mol"1 cm"1.
The spectrophotometry method has the following advantages: It is of
high sensitivity, used within the range of 0.095 μg to 0.095 mg per liter,
simple and less time consuming. It is of relatively low reagent blank
absorbance (about 0.1). However, the influence of additional ions that may
occur in natural waters, such as arsenate or silicate, have the ability to form
a gradual blue heteropolyacid complex. Arsenate interference can be
removed by reduction with potassium iodide in the presence of concentrated
hydrochloric acid and the produced arsenite is precipitated by H2S as As2S3.
Interference of sulphide can be removed by adding several mg of potassium
permanganate and shaking for 1-2 min; after that 1 ml of ascorbic acid
solution is added and the procedure is continued. Silicate ions are eliminated
by acidifying the sample solution, heating and filtering, or may be separated
by fuming with perchloric acid to dehydrate the silicic acid and render it
insoluble /15.16/.
It is well known that phosphate can be determined by precipitation as
MgNH4P04, 6H 2 0; the precipitate is then dissolved in dilute hydrochloric
acid, and excess EDTA is added to form the magnesium EDT-complex. The
excess EDTA is determined with standard magnesium chloride using
Solochrome Black (Eriochrome Black T) as indicator /14/. This method is
not sensitive: it needs filtration washing, so losses may occur and it is time
consuming. It is used for samples containing high concentrations of
phosphate ions. In the mentioned procedure filtration is not required and
direct determination of excess copper ions can be established. The Potentio-
metrie technique is used for determination of copper ions by titration with
EDTA in the presence of a copper ion selective electrode as indicator. The
equivalence point of the reaction will be revealed by a sudden change in
potential in the plot of e.m.f. readings against the volume of the titrating
solution; any method which will detect this abrupt change of potential may
be used. It can be used within the range of 1 χ 10"' to 1 χ 10"6 Μ solution.

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Excess copper can be amperometrically determined by complexation with

thiomalic acid in acetate buffer, using a rotating platinum micro electrode at
an applied voltage +0.8 vs standard calomel electrode; the sensitivity is
adjusted by using a galvanometer. The titration graph is plotted, the end
point evaluated and the concentration of the samples calculated. The
procedure can be used in a concentration range of 1 χ 10"3 to 1 χ ΙΟ"5 Μ
solution. The Potentiometrie method is superior to the amperometric method
for more concentrated solutions.
These methods are simple, easy, not time consuming and relatively free
from interferences, but arsenate ions have the same reaction with copper
solution. They interfere in concentrations as low as 0.1 m g . r ' , so it is
necessary that they be removed, as stated, before carrying out the
experiment.

4. CONCLUSION

These procedures were compared with each other using Student's t test,
considering the spectrophotometric method as a standard method. It was
found that no significant difference existed between them (t calculated is
between 1.59 to 1.83 and tabulated 2.03) at 95% confidence limit and 34
degrees of freedom as shown in Table 1.
These procedures are applied for the determination of phosphate ions in
water samples collected from different parts of Nile river in Egypt,
especially in Dakahlia province. It is necessary to obtain quantitatively the
total phosphorus present in the different samples. Phosphorus is released
from its combination with organic matter, e.g. malathion and parathion, by a
digestion or a wet oxidation technique. In most cases this can be performed
by boiling in a water bath for 30 min or an oil bath for 10 min after adding
10 ml of a mixture of sulphuric and nitric acid (each 10%), cooling, filtering
if necessary and almost neutralizing to Phenolphthalein with 5 Μ sodium
hydroxide /15,16/. The result indicates the sum of all inorganic phosphate,
i.e., the orthophosphate originally contained in addition to the acid
hydrolysable phosphate, present in the water samples.
It is clear from Table 2 that the concentration of phosphate in all water
samples is higher than the permissible limit of 10 - 50 μg phosphate /111.

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