Spectrochimica Acta Part A 78 (2011) 497–502

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Spectrochimica Acta Part A: Molecular and

Biomolecular Spectroscopy
journal homepage: www.elsevier.com/locate/saa

A simple spectrophotometric method for the determination of phosphate in soil,

detergents, water, bone and food samples through the formation of
phosphomolybdate complex followed by its reduction with thiourea
B. Shyla, Mahadevaiah, G. Nagendrappa ∗
Department of Chemistry, University of Mysore, Manasagangotri, Mysore 570006, India

a r t i c l e i n f o a b s t r a c t

Article history: A simple spectrophotometric method is developed here for the determination of phosphate present
Received 11 August 2010 in the samples of soil, detergents, water, bone and food based on the formation of phosphomolybdate
Received in revised form 26 October 2010 complex with the added molybdate followed by the reduction of the complex with thiourea in aqueous
Accepted 15 November 2010
sulfuric acid medium. The system obeys Beer’s law at 840 nm in the phosphate concentration range,
0.5–10.0 ␮g/ml. Molar absorptivity, correlation coefficient and Sandell’s sensitivity values are found to be
Keywords:
1.712 mol−1 cm−1 , 0.9769 and 0.0555 ␮g cm−2 respectively. For a comparison of the results determined
Phosphate determination
from the developed method, phosphate present in the same set of samples is determined separately
Spectrophotometry
Phosphomolybdate complex
following an official method. The results of the developed method are agreeing well with those of the
Molybdate official phosphomolybdate method.
Thiourea © 2010 Elsevier B.V. All rights reserved.
Soil
Detergents
Water
Bone and food samples

1. Introduction volumetry [14,15], flow injection spectrophotometry [16,17], HPLC

Phosphorous is the 11th abundant element in crustal rocks of
the earth, and all its known terrestrial minerals are orthophos-
phates [1]. The phosphate ion, PO4 3− is an inorganic polyatomic
ion sometimes found in lakes and streams as a result of agricul-
tural operations but it is originated either from animal wastes
or from fertilizers [1]. Phosphate is used in detergents and it is
also used in cola beverages in its acid form, H3 PO4 , to impart a
tart flavor [2]. Phosphate stimulates the growth of plankton and
aquatic plants which provide food for fish but excessive use of
fertilizers and detergents leads to over growth of aquatic plants
reducing dissolved oxygen level of water which is detrimental to
aquatic life [1,3]. As a consequence methods for determining phos-
phate are finding importance in the areas of medicinal, agricultural,
metallurgical and environmental sciences [4]. Since phosphate is
present in detergents [5], fertilizers [6], soil [7,8], meat [9] and
also in beverages [10] to monitor its level in such samples vari-
ous methods are found in the literature [11–19]. Such methods are
including atomic absorption spectrometry [11,12], titrimetry [13],
∗ Corresponding author. Tel.: +91 821 2419665.
E-mail address: gnagendrappa@yahoo.co.in (G. Nagendrappa).
[18], inductively coupled plasma emission spectrometry [19] and
spectrophotometry [20–23]. However, among such methods, it is
appearing from the literature [3,4,24] that the spectrophotometric
methods involving ammonium molybdate [3,4] and molybdovana-
date [24] are the most commonly practiced methods. Ammonium
molybdate spectrophotometric methods for phosphate are involv-
ing various reducing agents, for example, ascorbic acid [21],
stannous chloride [25], the mixture of hydrazine sulfate and ascor-
bic acid [26], and sodium sulfide [4]. A few of these methods are
requiring complicated and expensive equipments [18,19] and a
few of them are also involving extraction procedures [20,21] and
obviously limiting the practice of such methods in common labo-
ratories. In this background and like earlier phosphate method [4]
from here it is attempted again to develop a simple and straight
forward, spectrophotometric method for phosphate but with the
reducing agent thiourea. Since thiourea is commercially available,
economically cheaper, known for good shelf stability and already
used as a preservative in phosphate determination along with the
reducing agent, o-phenylenediamine dihydrochloride [27]. There
are literature methods [28,29] where thiourea is used as a reduc-
ing agent for iron(V) and iron(VI) and an official phosphomolybdate
method involving ascorbic acid is having a limitation associated
with ascorbic acid stability [30] that are prompted here to consider
thiourea as a reducing agent in the present method to reduce the

1386-1425/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.saa.2010.11.017
498 B. Shyla et al. / Spectrochimica Acta Part A 78 (2011) 497–502
phosphomolybdate formed in acidic medium from the test quan-
tity of phosphate and a known but excess quantity of molybdate
[4] and the level of reduced phosphomolybdate is monitored at
840 nm. The developed method is used effectively for the deter-
mination of phosphate present in the samples of soil, detergents,
water, bone and food. The results of phosphate determined from
the new method are in good agreement with the results of phos-
phate determined separately from an official phosphomolybdate
method for phosphate involving ascorbic acid as a reducing agent
[30].
2. Experimental
2.1. Apparatus
Elico spectrophotometer model SL 171 (Hyderabad, India) with
1 cm matched quartz cells and Sartorius digital balance readable
0.0001 g were used.
2.2. Reagents and solutions
All the reagents used here were of analytical grade and the water
used was distilled water.
2.2.1. Standard phosphate solution [4]
A weighed amount, 0.2077 g of sodium dihydrogen phosphate
dihydrate was transferred into a clean beaker. It was dissolved in
water and the solution was transferred into a 250 ml volumet-
ric flask and diluted to the mark with water [4]. The working
5.3 × 10−4 M phosphate solution was prepared by its further dilu-
tion.
2.2.2. Ammonium molybdate solution [31]
A weighed amount, 1.7081 g of ammonium molybdate was dis-
solved in about 150 ml warm water. A slightly milky solution
obtained was cooled to room temperature. It was transferred into
a 250 ml volumetric flask and diluted to the mark with water [31].
2.2.3. Thiourea [32]
A weighed amount, 2 g of thiourea [32] was transferred into a
clean 100 ml beaker. It was dissolved in about 50 ml water. It was
transferred into a 100 ml volumetric flask and diluted to the mark
with water.
2.2.4. Sulfuric acid
Sulfuric acid (0.25 N) was prepared by diluting the concentrated
sulfuric acid (≈36 N) with water.
2.2.5. Absorption spectrum of reduced phosphomolybdate
complex
The absorption spectrum (Fig. 1) of thiourea reduced phospho-
molybdate complex solution is having its maximum absorption at
840 nm.
2.2.6. Recommended procedure
Various volumes (0.1–2 ml) of the working phosphate solution
corresponding to 0.5–10 ␮g ml−1 , 0.5 ml of 5.54 × 10−3 M ammo-
nium molybdate solution, 0.5 ml of 0.25 N sulfuric acid and 1 ml of
2% thiourea solutions are to be added into a series of 10 ml vol-
umetric flasks. After about 10 min the solutions are to be diluted
to the volume with water and absorbance of the solutions are to
be measured at 840 nm against water. Calibration graph is to be
obtained by plotting absorbance values of the solutions against
their phosphate concentration.
0.55
0.45
Absorbance
0.35
0.25
0.15
0.05
500 600 700 800 900 1000
Wavelength, nm
Fig. 1. Absorption spectrum of thiourea reduced phosphomolybdate complex
formed from the solution containing 1 ml of 50 ␮g/ml phosphate + 0.5 ml of
5.54 × 10−4 M ammonium molybdate + 0.5 ml of 0.25 N sulfuric acid + 1 ml of 2%
thiourea diluted to 10 ml with water.
2.3. Applications
2.3.1. Soil sampling
The soil samples from an agricultural field were collected by dig-
ging small pits of 30 cm depth. The samples were thoroughly mixed
and dried under a shade. Grass and other external objects were
removed by hand picking. Then soil samples were sieved through
a 2.0 mm sieving apparatus. The sieved samples were preserved in
separate, labeled plastic bags.
2.3.2. Procedure for phosphate in a soil sample [30]
2.5 g of each of the soil sample was weighed and transferred to a
250 ml conical flask. It was shaken with a measured volume, 50 ml,
of sodium bicarbonate solution of pH 8.5 and continued the shaking
for another 30 min. Then it was kept aside for 20–30 min to attain
equilibration and filtered through a Whatmann filter paper. The
clear filtrate was collected in a 50 ml volumetric flask and diluted
the filtrate to the volume with water. A measured volume, 10 ml,
of the filtrate was transferred into a 50 ml beaker. It was acid-
ified by drop wise addition of about 1 ml of dilute sulfuric acid.
The acidification of the filtrate could be easily noticed by ceasing
of evolving gas bubbles. Then the solution was transferred into a
25 ml volumetric flask and diluted to volume with water. It was
used for the determination of its phosphate content following the
recommended procedure.
2.3.3. Procedure for the determination of phosphate in detergents
[4]
A weighed amount, 5 g of detergent washing powder of Indian
make was transferred into a silica crucible and the content was
ashed in an electric burner. The ash obtained was dissolved in water
and made acidic by adding dilute sulfuric acid. It was heated on a
water bath for about 10 min to expel hydrogen sulfide, nitrite, etc.
[4]. It was filtered through a Whatmann filter paper and collected
the filtrate in a 100 ml volumetric flask and diluted the solution to
the mark with water. Aliquots of the solution were used for the
phosphate analysis.
2.3.4. Procedure for water samples
Water sample from Kukkarahalli lake/Karanji lake/Lingabudi
lake/tap water was filtered through a Whatmann filter paper and
collected the filtered sample in a labeled sample bottle. Aliquots
of the sample were used for the determination of the phosphate
through the recommended procedure.
2.3.5. Procedure for the determination of phosphate in bone
2.3.5.1. Water soluble phosphate of bone sample. A selected bone
sample was cleaned, washed and dried in an electric oven at 100 ◦ C.
It was cooled and cut to small pieces and ground to powder in an
 B. Shyla et al. / Spectrochimica Acta Part A 78 (2011) 497–502 499

agate mortar. From that a weighed amount, 1 g, was transferred Table 1

Determination of phosphate in soil, detergent and water samples.
into a silica crucible. It was ashed in an electric furnace. The ash
obtained was mixed with distilled water and filtered it through a Sample Phosphate in RSD % Phosphate in RSD %
Whatmann filter paper and collected the filtrate in a 25 ml volu- ␮g ml−1 by ␮g ml−1 by official
developed method method
metric flask and made up to the volume with distilled water. It was
used for its phosphate determination following the recommended Soila 0.0037e 2.63 0.0035e 1.67
procedure. Soilb 0.0032e 3.10 0.0037e 2.30
Soilc 0.0030e 3.07 0.0028e 2.52
Soild 0.0040e 2.43 0.0041e 3.06
Detergent
2.3.5.2. Acid soluble phosphate of bone sample. The bone ash residue Company A 18.1 0.55 18.6 1.09
present on the filter paper was transferred into a 50 ml beaker. The Company B 15.5 1.69 15.5 1.57
residue was dissolved by treating it with 1 ml of concentrated sul- Company C 19.0 1.24 18.5 0.89
Tap water 02.0 3.71 2.0 3.38
furic acid and diluted to about 25 ml with water. It was boiled to Tap water 1.51 2.79 1.7 0.75
reduce the volume of the solution to about 10 ml. It was cooled and Kukkarahalli lake 2.90 1.73 3.4 2.24
transferred the solution into a 25 ml standard flask and diluted to water
the mark with water. It was used for its phosphate determination Lingabudi lake 2.40 2.50 2.9 1.29
water
following the recommended procedure.
Distilled water ND ND
a
Soil: cotton.
b
Soil: sunflower.
2.3.6. Determination of phosphate in wheat flour and ragi flour c
Soil: groundnut.
4 g of the wheat flour/6 g of the ragi flour was weighed and trans- d
Soil: banana.
ferred into a silica crucible and ashed the contents using an electric e
Values in percentage, value is an average of five replicates.
burner. The ash obtained was dissolved in about 1 ml of the con-
centrated sulfuric acid and diluted to about 20 ml with water. It Table 2
was boiled on a water bath for about 5 min and cooled. The clear Determination of phosphate present in bone, wheat and ragi flour samples.
solution obtained was transferred into a 50 ml standard flask and Sample Phosphate in RSD % Phosphate in RSD %
diluted to the volume with water. Aliquots of the solution were ␮g ml−1 by ␮g ml−1 by official
used for the phosphate determination through the recommended developed method method
procedure. Water soluble 1.73 3.54 1.8 3.38
phosphate (bone)
Acid soluble 9.56 1.98 9.20 1.29
3. Results and discussion phosphate (bone)
Wheat flour 9.60 2.03 9.95 1.28
Ragi flour 8.42 1.16 8.52 0.75
The developed method is based on the formation of phospho-
Value is an average of five replicates.
molybdate complex [3,4], due to the reaction between molybdate
and phosphate followed by its reduction with thiourea in aqueous
sulfuric acid medium. Under the optimized experimental condi- 4. Optimization
tion and with fixed concentrations of molybdate and the reducing
agent, the color intensity of the reduced phosphomolybdate solu- 4.1. Effect of concentration of ammonium molybdate solution
tion is found to be proportional to the amount of phosphate present
in sodium dihydrogen phosphate, soil, detergents, water, bone and The effect of ammonium molybdate on the color intensity of
food samples. The reaction conditions as well as the various exper- the reduced phosphomolybdate complex at constant volumes of
imental parameters affecting the development and stability of the 0.5 ml of 0.25 N sulfuric acid, 1 ml of 50 ppm phosphate, 0.5 ml of
colored complex were carefully investigated and optimized for 2% thiourea was examined with various volumes of (0.25–2 ml)
the quantitative determination of phosphate present in various of 5.54 × 10−3 M molybdate solution per 10 ml of total solution.
samples analyzed. The experimental variables such as concen- Absorbances of the solutions were measured at 840 nm. The results
tration of ammonium molybdate, concentration of sulfuric acid, obtained are shown in Fig. 2, that are accounting for the max-
concentration of thiourea, order of addition of the reagents and imum absorbance value for the solution containing 0.5 ml of
also color stability of the complex were carefully optimized for 5.54 × 10−3 M ammonium molybdate solution. Above that vol-
the effective determination of phosphate. The stoichiometry was
determined and found to be 1:1 with respect to phosphate and 0.06
molybdate [4]. For a comparison of phosphate results determined
from the new method, the phosphate content of the same set of 0.05
the samples was also determined separately following an official
Absorbance

0.04
phosphomolybdate method involving reducing agent ascorbic acid
[30]. The phosphate results so determined from both the meth- 0.03
ods are given in Tables 1 and 2 and the results are accounting
for their complementing nature. Therefore, the developed new 0.02
phosphomolybdate method with reducing agent thiourea for the 0.01
determination of phosphate can be used either as an independent
or an alternate method to the existing official phosphomolyb- 0
date method for phosphate involving reducing agent, ascorbic 0 0.5 1 1.5 2 2.5
acid [30]. Obviously, the advantages of the new method over the Volume of ammonium molybdate in ml
existing official method [30] are essentially attributable to nature
Fig. 2. Effect of different volumes, 0.25–2 ml 5.54 × 10−4 M ammonium molybdate
and stability of thiourea [27] over the nature of ascorbic acid
solution + 1 ml of 50 ␮g/ml phosphate + 0.5 ml of 0.25 N sulfuric acid + 1 ml of 2%
[30]. thiourea diluted to 10 ml with water.
500 B. Shyla et al. / Spectrochimica Acta Part A 78 (2011) 497–502
0.07
0.06
0.05
Absorbace
0.04
0.03
0.02
0.01
0 of 0.25 N hydrochloric acid for a total volume, 10 ml of the
0 0.5 1 1.5 2 2.5
Volume of 0.25N sulfuric acid in ml
Fig. 3. Effect of different volumes, 0.25–2 ml 0.25 N sulfuric acid + 1 ml of 50 ␮g/ml
phosphate + 0.5 ml of 5.54 × 10−4 M ammonium molybdate + 1 ml of 2% thiourea
diluted to 10 ml with water.
ume of molybdate solution, the absorbance of the solution was
decreasing; therefore, 0.5 ml of 5.54 × 10−3 M ammonium molyb-
date solution was used in the analysis of samples for phosphate.
4.2. Effect of concentration of sulfuric acid
The effect of concentration of sulfuric acid on absorbance was
investigated to achieve high absorbance value of reduced phospho-
molybdate solution by taking various volumes (0.25–2 ml) of 0.25 N
sulfuric acid, 0.5 ml of ammonium molybdate, 1 ml of phosphate
(50 ppm) and 1 ml of 2% thiourea per the total volume of 10 ml.
The solutions of the flasks were diluted to the volume with water.
Absorbance of the solutions was measured at 840 nm. The results
obtained are shown in Fig. 3 and are indicating that the solution
containing 0.5 ml of the 0.25 N sulfuric acid was more sensitive.
Therefore that volume of the sulfuric acid was used per 10 ml of the
solution throughout the experiment in determining the phosphate
concentration.
For the confirmation of the selecting sulfuric acid as the reac-
tion medium over other acid media, namely hydrochloric acid
and acetic acid, the following investigations were carried out. A
series of labeled 10 ml volumetric flasks were arranged. To each
flask the following quantities of the solutions, 1 ml of 50 ␮g ml−1
phosphate, 0.5 ml of 5.54 × 10−4 M ammonium molybdate, 0.5 ml
of 0.25 N sulfuric acid/0.5 ml of 0.25 N hydrochloric acid/0.5 ml of
0.25 N acetic acid followed by 1 ml of 2% thiourea were added. Then,
the solutions were diluted to the mark with water and shaken vig-
orously and kept aside for about 20 min. After that absorbance of
the solutions were measured at 840 nm against water. The results
obtained are given in Table 3. The results obtained for the sim-
ilar set of solutions but acidified with hydrochloric acid/acetic
acid were also determined and given in Table 3. Absorbance val-
ues of the solutions acidified either with sulfuric acid or with
hydrochloric acid are accounting for almost the same value, 0.06.
However, average absorbance value of the solutions that were
acidified with acetic acid indicates colorless nature. Therefore, the
calibration graph for the determination of phosphate could be
constructed using either 0.5 ml of 0.25 N sulfuric acid or 0.5 ml
Table 3
Effect of types of acid, 0.5 ml of 0.25 N sulfuric acid/0.25 N hydrochloric acid/0.25 N
acetic acid on the determination of phosphate.
Type of acid Absorbance
Sulfuric acid 0.0629
Hydrochloric acid 0.0666
Acetic acid 0.0004
Value is an average of five replicates.
Table 4
Effect of order of reagents addition.
Number Order of addition Absorbance
1 A+B+C+D 0.0920
2 B+C+D+A 0.0915
3 C+D+A+B 0.0893
4 D+A+B+C 0.0893
A, 1 ml of 50 ␮g ml−1 phosphate solution; B, 0.5 ml of 5.54 × 10−4 M ammonium
molybdate; C, 0.5 ml of 0.25 N sulfuric acid; D, 1 ml of 2% thiourea.
solution.
4.3. Effect of the concentration of the reducing agent
The influence of the concentration of the reducing agent,
thiourea on the phosphomolybdate complex was studied as above
but with different volumes of (0.5, 1.0, 1.5 and 2 ml) of 1% thiourea
solution per 10 ml of total volume. Absorbance values of the solu-
tions were found to be increasing with the increasing volume of
1% thiourea solution. When 1 ml of 1% thiourea solution was used
for constructing the calibration graph to determine the phosphate
content, the color development of the solutions was found to be
slow as a consequence it was opted to use 1 ml of 2% thiourea solu-
tion rather than 1 ml of 1% thiourea solution in constructing the
calibration graph for phosphate.
4.4. Effect of order of reagents addition
Order of the reagent addition was studied using optimized vol-
umes of the corresponding reagents through the recommended
procedure. The phosphate results obtained are shown in Table 4.
The results are indicating that the change in order of addition of
reagents is not affecting the absorbance values. However, for main-
taining the uniformity of the order of reagents addition, Series No. 1
of Table 4 was followed throughout the course of the determination
of phosphate.
4.5. Color stability of thiourea reduced phosphomolybdate
The color stability of the reduced phosphomolybdate of the
reaction system was studied carefully owing to the importance of
this parameter and its effect on reproducibility of the phosphate
results of the method. It was investigated by performing the exper-
iment using the recommended volumes and concentrations of the
reagents involved except phosphate that was 1 ml, 50 ␮g ml−1 per
10 ml of the reaction mixture. The phosphate results obtained are
shown in Fig. 4. And the results are indicating that there is a rapid
increase in the values of absorbance with time for about 20 min.
After 20 min there is a slow increase in the values of absorbance and
found to be stable for about 5 min. After that the absorbance values
are again increasing slowly with time. Therefore, the absorbance
measurement of the solutions were performed after 20 min but
completed before 30 min.
The same procedure was used for the phosphate determination
but by using one week old thiourea solution. The results obtained
are given in Fig. 5 and are indicating the rapid increasing trend of
the absorbance values with time for about 20 min. Later absorbance
values of the solutions are almost same for about another 40 min. In
addition, the color stabilities of thiourea reduced phosphomolyb-
date solutions as in Fig. 5 are more than those found from the freshly
prepared thiourea reduced phosphomolybdate solutions (Fig. 4).
These observations are complimenting the established preservative
nature of thiourea solution [27].
 B. Shyla et al. / Spectrochimica Acta Part A 78 (2011) 497–502 501

0.16 Table 5
Interference of foreign ions in the determination of 5 ␮g ml−1 of phosphate.
0.14
Ions Tolerance limit (ppm)
0.12
2+
Zn 90
Absorbance

0.1 K+ 209
Mg2+ 80
0.08
Fe3+ 4
0.06 V5+ 2.5
Cd2+ 48
0.04 Cu2+ 7
SO4 2− 638.4
0.02
Cl− 375.2
0
0 20 40 60 80 100
Time/min solutions. The precision of the method as expressed by the rela-
tive standard deviation is found to be less than 3.2% whereas its
Fig. 4. Effect of time on the color stability of thiourea reduced phosphomolybdate
solution having 1 ml of the 50 ␮g/ml phosphate solution, 0.5 ml of 5.54 × 10−4 M accuracy as expressed by the calculated relative error is 2.7%.
ammonium molybdate, 0.5 ml of 0.25 N sulfuric acid and 1 ml of 2% thiourea solution
diluted to 10 ml with water.
4.7. Interferences studies
0.12
In order to evaluate the selectivity of the new method for
0.1 phosphate, influence of select metal ions, sulfate and chloride
were examined over a range of their concentrations but for fixed,
0.08
Absorbance

5 ␮g ml−1 of phosphate solutions. The results obtained are given

0.06 in Table 5 and are indicating that all the ions except copper, iron
and vanadium are not interfering in the determination of phos-
0.04 phate. Copper(II), iron(III) and vanadium(V) could also be tolerated
0.02 respectively up to 7, 4, and 2.5 ␮g ml−1 for the effective determina-
tion of phosphate.
0
0 20 40 60 80 100
Time/min 5. Conclusion

Fig. 5. Effect of time on the color stability of thiourea reduced phosphomolybdate
solution having 1 ml of the 50 ␮g/ml phosphate solution, 0.5 ml of 5.54 × 10−4 M
ammonium molybdate, 0.5 ml of 0.25 N sulfuric acid and 1 ml of 2% one week old
thiourea solution diluted to 10 ml with water.
4.6. Calibration graph
Under the optimized experimental condition, the calibration
graph established for the phosphate determination in its concentra-
tion range 0.5–10 ␮g ml−1 is given in Fig. 6. It is having a straight
line with slope, 0.0126 and intercept, 0.0135. Molar absorptivity,
correlation coefficient and Sandell’s sensitivity values were cal-
culated and are found to be 1.712 × 10−3 mol−1 cm−1 , 0.9769 and
0.0555 ␮g cm−2 respectively. The precision and accuracy of the
method were studied by analyzing 1.0, 2.5 and 5.0 ppm phosphate
0.14
0.12
0.1
Absorbance
The developed new method for the spectrophotometric deter-
mination of phosphate is working on simple and straight forward
principles of reducing phosphomolybdate complex formed from
test quantity of phosphate and molybdate in excess followed by its
reduction with thiourea solution [28,29] leading to form molybde-
num blue which could be monitored at 840 nm. Chemicals involved
in the method are cheap and are considered to be normally avail-
able in common laboratories. The method is unlike other methods
since it is neither involving extraction [20,21] nor sophisticated
instruments like HPLC [18] nor flow injection spectrophotome-
ter [16] nor is requiring strict control of pH. The reducing agent,
thiourea employed here to reduce phosphomolybdate is a known
shelf stable and preservative [27] and preparation of its solution
is also simple and solution once prepared it is found to be very
stable when compared to the nature and stability of ascorbic acid
solution which is used as a reducing agent for phosphomolyb-
date to determine phosphate in an official method [30]. In that
official method [30] the reducing agent ascorbic acid solution is
to be prepared on daily basis. In addition, the developed new
method is working satisfactorily for the determination of phos-

phate present in various samples analyzed here. The phosphate

0.08
results obtained from the new method are also comparing well
0.06 with those of the phosphate results determined separately from
0.04 an official method [30]. Absorbance values of the thiourea reduced
phosphomolybdate solutions of the method are found to be not
0.02 completely independent of time; however, the phosphate results
0 obtained for the samples are reproducible when the determination
0 5 10 15 of phosphate was carried out through the recommended proce-
Concentration of phosphate in ppm dure. Therefore, the developed new phosphomolybdate method
with reducing agent thiourea for the determination of phosphate
Fig. 6. Calibration graph for the determination of phosphate, each solution is having can be used either as an independent or as an alternate method
0.5–10 ␮g/ml phosphate solution + 0.5 ml of 5.54 × 10−4 M ammonium molyb-
to the existing official phosphomolybdate method for phosphate
date + 0.5 ml of 0.25 N sulfuric acid + 1 ml of 2% thiourea solution diluted to 10 ml
with water.
involving reducing agent, ascorbic acid [30].
502 B. Shyla et al. / Spectrochimica Acta Part A 78 (2011) 497–502
Acknowledgments
Author, B. Shyla is grateful to the U.G.C and University of
Mysore for the U.G.C Meritorious Fellowship. Thanks are due to
Shashikumar, C., Assistant Professor (Agronomy), Krishi Vignyana
Kendra, Hiriyur, for his useful discussions and the interest taken
in providing the soil samples. Authors are also acknowledging the
suggestions of Samyuktha Moorthy since in the light of his sugges-
tions the manuscript is revised in the initial stage. Encouragements
of Jim R. Durig Editor of the journal and services of the Support Staff
of the journal have helped considerably in evolving the manuscript
to the present state.
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