MAYFEB Journal of Environmental Science

Vol 1 (2016) - Pages 20-29

The Spectrophotometric Evaluation of

Phosphate in Soil Samples
F.E. Adelowo
1
Department of Pure and Applied Chemistry, Ladoke Akintola University of Technology, P.M.B

4000, Ogbomoso, Nigeria, feadelowo@lautech.edu.ng

S.O. Oladeji
1
Department of Pure and Applied Chemistry, Ladoke Akintola University of Technology, P.M.B

4000, Ogbomoso, Nigeria, oladejioluwole@gmail.com

K.A. Odelade
2
Department of Pure and Applied Biology, Ladoke Akintola University of Technology, P.M.B 4000,

Ogbomoso, Nigeria, kennybramm@gmail.com

Abstract

The chemistry of soil indicate that it contain several chemical components ranging from organic to inorganic

compounds. Some of these include phosphates, nitrates and sulphates amongst others. Phosphorus has a strong

affinity for soil thereby increasing the rate of dissolved phosphorus that will be transported in runoff. The

phosphate levels could increase in the soil due to natural and human factors such as dumping of refuse from nearby

areas, use of fertilizers and human excretes. Therefore, it is important to analyze the levels of orthophosphate

(expressed in part per million) in soil using spectrophotometric methods. This was analyzed by molybdenum blue

phosphorus method using UV-Visible spectrophotometric method and the phosphates level was measured at 880 nm.

The calibration curve was formed using concentrations of 0.0 – 16.0 ppm. The samples analyzed showed high levels

of phosphates which could be due to natural and agricultural factors.

Keywords: calibration curve, chemical components, molybdenum blue phosphorus method, phosphates levels,

spectrophotometric methods.

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1. INTRODUCTION

Phosphorus is one of the essential elements present in the soil and it is classified as a macronutrient because of

the large amounts that are required by plants. They serve many functions in plants; one of the major roles of

phosphorus in living organisms especially in plants is the transfer of energy. In soil that contains adequate

amount of phosphorus, normal growth and development are inevitable [1]. Despite phosphorus been widely

distributed in nature, it do not occur in free state because of they are too reactive in soil and exist as phosphate.

The simplest of this is orthophosphate, has the chemical formula of PO43-.

The chemistry of soil indicate that it contain several chemical components ranging from organic to inorganic

compounds. Scientifically, soil is defined as the outermost or weathered superficial layer of the earth's crust.

They consist mainly of decomposed parent rock material (organic matter), mineral (inorganic) matter, water, air

and organisms [2]. From recent researches, scientists discovered that soil contains chemical constituents or

compounds such as phosphates, nitrates and sulphates. Plants depends on soil and water, they absorbs nutrients

from the soil and also moisture (water) which aids proper growth and development [3]. The interest in the

chemistry of soil phosphate indicates the influence and applications of phosphate in the soil. There have been

many methods proposed and developed for the qualitative and quantitative analysis of phosphate in the soil.

Some of these methods include titrimetric, complex-gravimetric, colorimetric, atomic absorption spectroscopic

and spectrophotometric methods [4, 5]. One of the simplest and common methods is spectrophotometric

method; the advantages they have over other methods include: no extraction, sophisticated instruments and

easily accessible. Despite the approach in determination of phosphate in soil, different methods have been

employed such as ammonium molybdate and molybdovanadate [5]. In soil digestion, several approaches have

been employed, concentrated and diluted acids such as HCl, H2SO4, HNO3 and H3PO4. The amount of

phosphate digested depends on the concentration of the acid, amount of acids or combinations, the amount of

soil phosphate and time of digestion of the soil samples [3 – 5].

The use of fertilizers (phosphates) by farmers increases on daily basis due to the increasing demands for

agricultural produce. The positive outcomes of the application of these fertilizers, such as, increasing in food

produce, nutrients of the soil have always been focus, thereby neglecting the negative impacts. Therefore, it is

important to analyze the levels of orthophosphate in soil using spectrophotometric methods using concentrated

solutions of HCl and H2SO4 for acid digestion.

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 MAYFEB Journal of Environmental Science
Vol 1 (2016) - Pages 20-29

II. METHODOLOGY

A. Chemicals and Reagents

All the chemicals and reagents used were analytical reagent grade unless otherwise stated and the solutions were

prepared in double distilled water.

B. Preparation of Standard Solutions (Stock Solution)

About 0.717 g of potassium dihydrogenphosphate (KH2PO4) was accurately weighed and dissolved in distilled

water and the volume was made 500 ml in a volumetric flask. The phosphate solutions of different

concentrations (0.5 to 3.5 ppm) were prepared by serial dilution of the stock solution.

C. Preparation of Working Solution of Ammonium Molybdate

About 1.7081 g of ammonium molybdate was accurately weighed and dissolved in 100 ml of warm

distilled water; a milky colouration was observed, cooled and transferred into 250 ml volumetric flask.

D. Preparation of Dilute Hydrochloric Acid

A measured amount of 40 ml of concentrated hydrochloric acid solution was diluted and made up to mark in a

100 ml of volumetric flask.

E. Preparation of Dilute Trioxonitrate (V) Acid

A measured amount of 40 ml of concentrated trioxonitrate (V) acid solution was diluted and made up to mark in

a 100 ml of volumetric flask.

F. Preparation of Hydrazine Hydrate

About 0.125 g of hydrazine sulphate was accurately weighed and dissolved with double distilled water in a

clean beaker in a 100 ml standard flask and made up to calibrated mark.

G. Preparation of Samples

H. Soil Sample Collection and Preparation

Soil sample was taken from LAUTECH farm, Ogbomoso, Nigeria on 15th April, 2016. The sample and

was dried for 15 days, sieved and stored for further analysis. The soil samples were collected systematically

from eight points in the afternoon and air dried for 15 days after which it was subjected into an oven for barely

one hour at about 30-40 °c to obtain total dryness. These samples were sieved using a sieve mesh size of 20 mm.

The sieved sample obtained was kept inside a clean, air-tight sample bottles inside an oven and this was stored

at 4 0C for further analysis.

I. Soil Sample Analysis for Phosphates

About 2 g of the sieved soil sample was accurately weighed and transferred to a 250 ml conical flask, 80 ml of

HCl and H2SO4 (ratio of 3 to 1) was added and shaken in a mechanical shaker for 45 minutes. The digestion was

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Vol 1 (2016) - Pages 20-29

done for 7 hours. This was then filtered using filter paper whatmann No. 42. About 20 ml of the filtered digested

sample was taken, 4ml of ammonium molybdate, 3 ml of hydrazine sulphate was added and kept in a water bath

for 30 minutes. The blue colour observed was measured spectrophotometrically.

III. RESULTS AND DISCUSSION

A. Determination of Maximum Absorbance

The absorption spectral data of the Potassium dihydrogenphosphate used as standard against the reagent

blank (water) exhibit the wavelength of maximum absorbance around 880 nm using CECIL CE 7200, 7000

series UV- Visible Spectrophotometer.

B. Calibration Curve for the Standard (KH2PO4)

In molybdenum blue method, at optimum analytical conditions, the amount of phosphates in the soil samples is

proportional to the intensity of colour (in KH2PO4). The calibration curve of the absorbance from the UV-

Visible Spectrophotometer of concentrations 0.5 – 4.0 ppm respectively is given below in Fig 1 from table 1.

The process involves the formation of molybdophosphoric acid from ortho phosphate and an excess of

molybdate in acidic solution followed by reduction to give molybdenum blue. The equation of the reaction

involve include:

PO43-+ 12(NH4)2MoO4 + 24H+ → (NH4)3PO4.12MoO3 + 21NH4+ + 12H2O [(NH4)3PO4.12MoO3+ (N2H4.H2O) →

molybdenum blue]

C. Phosphate Analysis in (Soil) Samples

The analytical results of phosphate analysis in soil samples are shown in table 2. The absorbance of the

phosphate level in these samples was determined at 880 nm. The corresponding phosphate concentration was

obtained from the calibration curve (figure 1) by interpolation. The concentrations were expressed in part per

million (ppm) of the respective soil samples. From table 2, different phosphate concentrations were obtained for

samples at different locations. The highest concentration of phosphate was obtained at L1 with a concentration

of 16.000 ppm; the lowest concentration was obtained at L5 with concentration of 1.480 ppm. The uses of

fertilizers, herbicides, fungicides and even manure have been discovered to be the major pollutants in the farm.

There is no health-based guideline proposed for phosphate consumption, thou, medically high intake of

phosphate can cause health problems like cancer, ulcers, kidney problems, brain damage and gastrointestinal

effects.

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The use of fertilizers, herbicides, fungicides in farm help improve the quality of farm produce. The

environmental protection agency has a recommended limit for phosphates levels in soil and water bodies. For

streams and rivers the Environmental Protection Agency recommended limit is 0.1 part per million (0.1 ppm).

The following recommendations are also given for phosphate consumptions: total phosphorus; no more than

0.1 ppm for streams which do not empty into reservoir, 0.05 ppm for streams discharging into

reservoirs and 0.025 ppm for reservoirs [2]. Manmade sources of phosphate include human sewage,

agricultural run-off from crops, sewage from animal feedlots, pulp and paper industry, vegetable and

fruit processing, chemical and fertilizer manufacturing, and detergents [8].

TABLE 1: The Absorbance, at obtained from different concentration (ppm) of potassium dihydrogenphosphate.

C AT

0.0 0.00

2.0 0.23

4.0 0.38

6.0 0.58

8.0 0.76

10.0 0.83

12.0 1.17

14.0 1.38

16.0 1.59

C = Concentration (ppm); AT= absorbance

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 MAYFEB Journal of Environmental Science
Vol 1 (2016) - Pages 20-29

1.8

1.6 y = 0.0964x
R² = 0.9901
1.4

1.2

1
absorbance

0.8

0.6

0.4

0.2

0
0 5 10 15 20
concentration (ppm)

Fig 1 Calibration curve for the standard solution (KH2PO4)

The highest concentration of phosphates at L1 could be attributed to excessive application of phosphate

fertilizers and manure. Through investigations, there were inventions of the plants grown in this farm years back

which prompt the practise of bush burning, use of fungicides and insecticides. The farm was fumigated

immediately after planting of the seed to prevent diseases, insect attack and weed growth. There was an

indication that the use of manure (cow dung) and inorganic fertilizers (phosphate fertilizers) was the usual

practice in this farm. The use of fertilizers in agriculture is paramount when compare with herbicides,

fungicides and nematocides uses. The high level of phosphates in soil samples taken from maize fields (L1, L3,

L6 and L2) could be due to the use of phosphates fertilizers and have higher level when compared to soil samples

taken from lands sprayed with phosphates fungicides. The lowest phosphate concentration recorded in L5 could

be due to the reduction in the use of phosphate fertilizers, crop rotation and human activities such as weeding

with hoes rather than applications of herbicides. Though, the farmland was previously a waste disposal site for

livestock and human wastes. Human being was discovered to excrete 1.3 to 1.5 g of phosphorus per day [6].

These could be responsible for the phosphate level in the soil.

The phosphate in fertilizers and manure is initially quite soluble and available. Most phosphate fertilizers have

been manufactured by treating rock phosphate with acids to make it soluble. Manure contains soluble, organic

and inorganic phosphates compounds. When these are applied to the soil, various reactions occur that make the

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phosphate less soluble and available. Gradually reactions occurs in which the adsorbed phosphate and the easily

dissolved compounds of phosphate form more insoluble compounds that results in decreases in soil phosphate.

TABLE 2 The Absorbance of the water samples and their corresponding concentrations obtained from the calibration curve

Sampling Area C (ppm)

L-1 16.00

L-2 14.80

L-3 15.10

L-4 12.80

L-5 11.65

L-6 14.97

L-7 12.05

L-8 13.05

L-1 – L8 = Sampling lands

Plants take up phosphorus from the soil solution as orthophosphate ion: either as HPO42- or H2PO4-. This

proportion in which these two forms are absorbed is determined by the soil pH. The mobility of phosphates in

soil is very limited and therefore, plant roots can take up phosphorus only from their immediate surroundings.

There have been many symptoms of phosphorus deficiency in plants, theses are scale down to; stunted growth,

dark purple colour of older leaves (figure 2), inhibition of flowering and root system development. In most

plants, these symptoms will appear when phosphate in the leaves is below 0.2%. High phosphates levels in soil

scientifically are not advisable. This can leads to excessive weed growth, stunted plant growth, harms beneficial

root fungi, which helps plants absorb water and nutrients, decreases plant ability to utilize zinc (leading to

bleaching of plant tissue) and decreases the plant availability of iron (leading to yellowing between leaf veins)

[7-9].

Soil phosphate level varies, depending on field location, past management and time of year. The

spectrophotometric method of phosphate analysis is an easy, efficient and a suitable method for the qualitative

and quantitative analysis of phosphate in soil samples. In soil analysis, acid digestion method is quite a suitable

method of extracting phosphate in soil; the acid digestion has some limitation and these can impose health or

environmental effects. There are simple methods or tests in determining the phosphate levels in soil samples;

these are quick hand test and phosphate colour test. They involves no extraction, no chemicals or capital-

intensive, no environmental or health issues, and simple qualitative and quantitative method of phosphate

analysis.

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TABLE 3 Phosphorus test results and recommendations for corn in Nebraska soils based on standard extractable P tests and water soluble

po4test for a 1:1 soil: water mixture

Soil phosphorus test relational values (ppm) by P-test PO4 Categories Recommended Fertilizer to apply to corn* Ib

method P2O5/acre & (P/ac)

1 2 3
Water Soluble Olsen P Bray 1-P Relative PO4 Broadcast Band

PO4 level

0-5 0-3 0-5 Very low 35 (80) 17 (40)

5-10 4-10 6-15 Low 17 (40) 9 (20)

11-15 11-16 16-24 Medium 0 0

20-20 17-20 25-30 High 0 0

˃20 ˃20 ˃30 Very High 0 0

1
Water soluble P (PO4) test (Hach trademark) for 1:1 soil: water mixture based on comparison with Bray-1 and Olsen P tests for 19

benchmark soils (Bray 1 for soils with pH <7.2; Olsen for soils pH >7.2). Water soluble Aquachek based P recommendations agreed for

twelve soils (63%), were borderline for three (total 79%). Four 1:1 soil water mixture/water soluble PO4 tests indicated higher available P

than standard Bray 1 & Olsen P tests.

2
Based on Nebguide “Fertilizer Suggestions for Corn” G74-174-A, Revised Sept. 2001.

3
Recommendations are based on use of synthetic P fertilizer but can be used for *organic sources of P such as rock phosphate or soft

phosphate which can supply equivalent levels of available P over time.

There is no specific “available” fraction of phosphorus in soils. The available phosphorus is what is in solution

plus what can be expected to become soluble from minerals and organic matter over the growing season.

Therefore, soil tests cannot extract the exact available amount from the soil, but rather an amount that reflects

what might become available. From table 3, the acceptable concentration of phosphorus in nutrient solutions is 5

– 10 ppm, even though it was found to be reduced to 0 - 5 ppm, some scientist believe the recommended value

is 10 ppm. In nutrient solutions that flow continuously, the concentration can be low as 1 – 2 ppm [10, 11].

The soil samples all exceeded the recommended phosphate concentration level for soil. The high

concentration of phosphates is due to agricultural practices, natural and human factors such as erosion,

leaching and weathering and also, runoff from nearby farm and nearby residential areas. In order to reduce the

level or concentration of phosphate in soil sample, the use of phosphate fertilizer should be reduced and use of

starter phosphate fertilizers should be encouraged.

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Fig 2 Maize corn showing deficiency of phosphate in the soil in L-1 farm

Phosphate availability can be managed by liming acid soils, using measures that increase organic matter, and

proper placement of Phosphate fertilizer affecting how efficiently phosphate is used by crops. Phosphate losses

can be reduced by applying appropriate measures to reduce erosion and runoff. Adequate Phosphate levels

encourage vigorous root and shoot growth, promote early maturity, increase water use efficiency and grain

yield. Thus, Phosphate deficiency stunts vegetative growth and grain yield. Soil phosphorus is relatively stable

in soil, and moves very little compared to nitrogen.

IV. CONCLUSION

The spectrophotometric analysis using ammonium molybdate as reductants proved effective analytical method

for the determination of phosphate level in soil samples. The phosphate concentration in the samples was

measured at 880 nm. The phosphates levels are high in all the samples when compared to the Environmental

Protection Agency. This could be due to natural such as human factors such as fertilizers, fungicides or

herbicides application and natural factors such as erosion and weathering. In the attempt to prevent the effects

of high phosphates, the use of phosphate containing agricultural materials such as fertilizers, herbicides,

fungicides should be reduce and the farmers should be enlighten on the negative impact of over dependence of

phosphate materials can cause.

Contributions to Knowledge

The following contributions to knowledge can be drawn from the research:

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(i) Phosphates in the soil are factors dependent such as pH, moisture content, and temperature.

All these factors contribute to the amount of phosphorus levels and lime and fertilizer requirements.

(ii) The amount of phosphate uptake in soil depends mainly on the plant or crop uptake or

requirement. Application of fertilizer depends on the purpose of the fertilizer.

(iii) Use a starter fertilizer when planting in cold, wet soils—particularly when soil tests are not

high.

(iv) Use best management practices to reduce phosphate levels such practices include crop

rotation, erosion and runoff prevention to avoid phosphorus losses and protect soil-water quality.

Conflict of Interest

The authors declare that there is no conflict of interest.

REFERENCES

[1] M.A. Chaube and V.K. Gupta, ‟Spectrophotometric determination of phosphate in polluted waters by Solvent Extraction of

Molybdenum Blue”, Analyst, vol. 108, pp. 1141-1144, 1983

[2]. T.E. Weier, C.R. Stocking and M.G. Barbour, "An introduction to plant Biology", 5thedition, University of California, Davis

California, 1973.

[3]. S.O. Oladeji, F.E. Adelowo and K.A. Odelade, ‟Evaluation of Phosphate Level in Water Samples (Ogbomoso Rivers) using UV-

Visible Spectrophotometric Method", Int J Sci Res Env Sci, vol. 4, issue 4, pp. 0102-0108, 2016

[4]. E.B. Hans, H.K. Hans, H. Per and T.N. Torsten, ‟Phosphate in Soil", Journal of Molecular and Cellular Cardiology, vol. 26, issue

1, page 41, 1994

[5]. V.C. Verma, “System of technical control for cane sugar factories in India”, Published by STAI , vol. 33, 1988.

[6]. A. Skoog, J. Holler and A. Nieman ‟Principles of Instrumental Analysis", Fifth ed. Thomson Learning, Inc: pp. 329-353. 1998.

[7]. A.I. Vogel’s ‟A Text Book of quantitative chemical analysis", 5th Ed Longmans London, 1991.

[8]. M. McCoy, ‟Goodbye Phosphates", Chemical and Engineering News, ACS, vol. 5, pp. 12, 2011

[9]. V. Smith, G. Tilman and J. Nekota, ‟Eutrophication impacts of Excess nutrient in Freshwater, Marine and Terrestrial

Ecosystems", Environ Pol vol. 100, pp. 179, 1999

[10]. S.J. Kharat and S.D. Pagar, ‟Determination of Phosphate in Water Samples of Nashik District Rivers by UV-Visible Spectroscopy",

E-Journal Chem, vol. 6, issue 1, pp. 515-521, 2009

[11]. C.X. Galhardo and J.C. Masini, ‟Spectrophotometric Determination of Phosphate and Silicate by Sequential Injection using

Molybdenum Blue Chemistry", Analytical Chimica Acta vol. 4, issue 17, pp. 191-200, 1999

[12]. M. Jalali, ‟Phosphorus Status and Sorption Characteristics of some Calcareous Soils of Hamadan", Western Iran Environmental

Geology, vol. 53, pp. 365-374, 2007.

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