EXPERIMENT 7

SOIL ANALYSIS
AVAILABLE PHOSPHORUS

A Laboratory Report Submitted in Partial Fulfilment of the Requirements in the Environmental

Chemistry Laboratory

Jazer John T. Lirazan

November 14, 2018

Chem 189 Laboratory

Environmental Chemistry Laboratory
Department of Chemistry
University of the Philippines- Visayas
 I. Introduction

Phosphorus plays an important role in plant health and growth; it encourages root
development, increases the ratio of grain to straw, and increases resistance to disease, among
other things. Some soil phosphorus is in a form available to plants, but much of it is tied up in the
organic matter pool or bound tightly to mineral particles. The soil pH has a large influence on
phosphorus availability and solubility. Some prairie soils are quite low in available phosphorus,
and more may need to be added for adequate crop growth. However, it can be over-applied in
the form of fertilizers, manures, and other forms. Excess phosphorus that is washed into
waterways can lead to overgrowth of algae and other microorganisms, depleting oxygen, which
results in eutrophication, and fish death. Knowing the level of phosphorus in farmland soil can
be very important. As part of soil analysis, a soil sample from the College of Arts and Sciences
garden, Miagao, Iloilo was subjected to phosphate analysis using the molybdate method.

The term available phosphorus refers to the inorganic form occurring in soil solution
which is almost exclusively ‘Orthophosphate’. This Orthophosphate occurs in several forms and
combinations. The phosphate contributions in soil are governed by hydrogenous equilibria in
which it takes part. This can be represented as,

Phosphorus absorbed in soil phase ⇌ P in soil solution ⇌ Precipitated P

The phosphorus absorbed by plants from soil comes from the soil solution in which it
exists as an inorganic Orthophosphate ion H2PO4-, HPO42- and PO43-. The most accessible ion is
H2PO4-. The quality of phosphorus accessible to the plants is influenced by a series of soil
properties. We can se the phosphorus cycle in Figure 1 below.

Figure 1: The Phosphorus Cycle

 Molybdate method is used in the experiment since Phosphate(V) ions react with
ammonium molybdate to produce a colored complex. The reaction is carried out in an acidic
solution containing excess ascorbic acid (vitamin C) to prevent the complex from slowly oxidizing.
You can use this reaction for the quantitative analysis of low concentrations of PO 4. This colored
solution will be subjected to UV Vis analysis for quantitative interpretation and to solve for the
available phosphorus present in the sample.

This experiment aims to determine the available phosphorus in the soil sample by
molybdate method and relate the calculated result with the fertility and its impact to the
environment.

II. Methods

Preparation of Reagents

Different reagents must be prepared for the experiment, the class was divided into
different groups and each group was assigned with a reagent. The experimenters were assigned
to prepare a 2.5 M sulfuric acid solution. A 14 ml of 18 M sulphuric acid was diluted to 100 ml
using a distilled water, the solution was then allowed to cool in a glass bottle.

Preparation of the standard Curve

The standard curved has been prepared by the class monitor, where a phosphate
calibration standard in the range of 0.02-0.03 ppm P and was pipetted into different volumes of
the standard solution of 100 ml volumetric flask, each solution was then added by a 10 ml of
mixed reagent and was diluted to mark. The solutions were let to stand for 20 mins and was
measure having an absorbance of 880 nm. The absorbance against concentration generated by
each solution was then recorded and plotted solving its slope, Y int using least square methods.

Determination of Available Phosphorus

A 2.5 g or of air-dried soil was measure using 125 mL Erlenmeyer Flask. The container was
added with 0.5 g of activated carbon and a 50 ml or extraction reagent was added. The solution
was shake for 30 mins using reciprocating shaker and by hands. The solution was then filtered,
and the filtrate was collected. A 10 ml of the extract was pipetted out from the flask and was
transferred into a volumetric flask and a 30 ml of distilled water was added. 10 ml of mixed
reagents was then added, and the solution was let stand for 20 minutes and was measure with
the same absorbance to the standard curve 880 nm and the and the available phosphorus was
calculated after the absorbance of each samples were obtained.
 III. Results and Discussion

Phosphorus is one of the most essential elements for the growth of plants and animals. It
plays a critical role in cell development and a component of biochemical molecules such as
adenosine triphosphate, deoxyribonucleic acid, and lipids (The phosphorus cycle, 2013).
Phosphorus usually exists in phosphates, PO4-3. Phosphates are present in three forms:
orthophosphate, polyphosphate, and organically bound phosphates. Orthophosphates are
produced by natural processes and found in sewages. These phosphates are the only form plants
and microorganisms can use. Polyphosphates are used for treating boiler waters and found in
detergents. Organic phosphates are important in nature and are frequently involved in energy
transfer reactions (Oram, 2014). These phosphates are converted to orthophosphates through
bacterial action.

Phosphorus is a limiting nutrient for terrestrial productivity and it plays a key role in net
carbon uptake in terrestrial ecosystems (Lal & Stewart, 2016). Phosphorus has limited availability
due to the restriction by the rate of soil weathering. The largest reservoir of phosphorus is in
sedimentary rocks. So then, it is generally recycled to various extends in ecosystems depending
on climate, soil type, and ecosystems depending on climate. The phosphorus cycle usually does
not involve a gas phase because phosphorus and its various compounds are not gases in Earth’s
normal temperatures and pressures (The phosphorus cycle, 2013).

The release of phosphates through weathering is the very first step of the phosphorus
cycle. When it rain, the phosphates are distributed throughout to soils and waters. Plants take
up the available phosphorus and the food chain is responsible in the distribution of this nutrient
to the other levels of consumers. If the plant was not taken up by consumers, the nutrients go
back to the soil when the plants are being decomposed. For animals, including us, the phosphates
may return to the soil through excreta or through final decomposition after death.

Plant roots absorb phosphorus from soil solution. The phosphorus moves to the root
surface through diffusion. For instance, mycorrhizal fungi enhance the uptake of plants especially
in acidic soils where phosphorus level is low. For plants, energy transfer reactions, development
of reproductive structures, crop maturity, root growth, and protein synthesis involve phosphorus
(Soil Nutrient Management for Maui County, 2017).

This experiment focuses to determine the available phosphorus level of the soil from
College of Arts and Sciences garden, Miagao, Iloilo. The sample was collected on November 11,
2018. The sample was sieved from the previously dried soil. The ascorbic acid – molybdate
method was used in this analysis. Ammonium molybdate and antimony potassium tartrate react
in an acid medium with dilute solutions of phosphorus to form an antimony-phospho-molybdate
complex. This complex is reduced to an intensely blue-colored complex by ascorbic acid. Only
orthophosphate forms a blue color in this test. Polyphosphates and some organic phosphorus
compounds may be converted to the orthophosphate form by sulfuric acid hydrolysis. Utilizing
spectrophotometer, absorbance is being measured at 880 nm as the maximum absorbance for
phosphate. According to Beer Lambert’s Law, the absorbance of electromagnetic radiation by a
given species is directly proportional to the concentration of the analyte:
𝐴 = Ԑ𝑏𝐶

where A is the absorbance, ε is the molar absorptivity, b is the path length and C is the
concentration of analyte. Because ε and b are fixed under experimental conditions the result is a
linear relationship between absorbance and concentration (UV-Visible Spectroscopy, 2013).

Firsthand, a calibration curve was generated using standard solutions of phosphates. A

reagent blank was also prepared to correct the absorbance readings of the instrument. This also
corrects some interferences including the matrix, reflection loss, and scattering losses in solution.
The reported equation of the calibration curve is 𝑦 = 0.7953𝑥 − 0.0005 with R2 = 0.9999. The
%RSD is 2.411%, indicating that the data are highly precise. The accuracy of the measurements
can be affected by factors like judgment in the calibrations by the experimenters and efficiency
of the apparatuses and instruments used. The sample was collected in a single location, not
pooled. This data cannot represent the available phosphorus level of the entire garden. The
nutrient levels may different from one point to another since the garden was inclined and water
running from it can be a factor in phosphorus distribution and the garden.
Table 1. Summary of Results

Weight of the Concentration

Trials Absorbance %RSD
Sample [g]
[ppm PO4 -3]

1 2.5098 0.07 9.25

2 2.5232 0.068 8.94 2.41%

Average 9.09

The optimal phosphorus level of soils is 10 – 20 ppm (Three Fruit Research and Extension
Center, 2004). Less than 10 ppm is classified as low; 10 – 20 ppm is medium; 20 – 40 ppm is high;
and greater than 40 ppm is excessive. So then, the level of the phosphorus, which is 9.09 ppm, in
the soil is low. This is because the soil sample is being stored inside the locker for several days
which volatile substance may have evaporated and contributes to its low phosphorus level.

Sorption and desorption dictate the availability of phosphorus (Soil Nutrient Management
for Maui County, 2017). P-sorption occurs when orthophosphates bind tightly to soil particles.
Factors which increases P-sorption includes soil mineral type, amount of clay, pH and
temperature. Mineralogy of the soil has a great effect on P-sorption. Amorphous materials in
volcanic soils increase the P-sorption capacities. Increasing the amount of clay in the soil
increases the surface area for phosphate sorption to take place. Presence of aluminum and iron
oxides in low pH lowers the availability of phosphate in soil solutions. P-sorption increases as
temperature increase. However, presence of other anions, including silicates, carbonates,
sulfates, arsenate, and molybdate, which compete with phosphate in anion exchange sites,
decreases the P-sorption. Organic matter increases P availability by forming complexes,
displacing phosphate by organic anions, coating aluminum and iron oxides, and releasing
phosphorus through mineralization reactions of its components.
 Humans can alter the phosphorus cycle by cutting tropical rain forests and using
agricultural fertilizers (The phosphorus cycle, 2013). Cutting forest increases the levels of
phosphorus at certain time but heavy rains carry this nutrient to other locations, usually in bodies
of water. The phosphorus is originally from the plants and it is being made available burning or
composition. Eutrophication and algal bloom may be a problem to aquatic ecosystems which
have domino effect to other environmental problems including the death of other organisms.
The effect of faulty application of fertilizers also follows this mechanism.

People are concerned in the effects of phosphorus deficiency to crops. Purpling of leaf
indicates the P deficiency (Unites States Department of Agriculture). Furthermore, their harvest
is greatly affected because of this. So then, they manage the availability of phosphorus in
agricultural lands. The major P-management strategies include the adjustment of pH and correct
application of P-fertilizers (Unites States Department of Agriculture). The highest solubility for
calcium phosphates is around 7.2 so then in this pH, the amount of available phosphorus is in
peak (Thomason, 2002). Using lime to increase the pH of acidic soils is recommended. Application
by small amounts of P fertilizers which is scheduled properly reducing the possibility of nutrient
loss caused by runoffs. P tie-up decreases the availability of phosphates for plants and this can
be reduced by banding or injecting P fertilizer or liquid manure. Placing P fertilizers where the
roots are most active is highly recommended, but application in roots is highly discouraged
because high concentration of phosphorus is toxic to roots.

IV. Conclusion

Soil sample was collected from College of Arts and Sciences garden, Miagao, Iloilo and
subjected to phosphate determination. The average phosphate level of the soil was 9.09 ppm
which is low according to the standards. Phosphorus exists in nature, yet humans can contribute
in its high level. Large amount of phosphorus may be caused by runoffs from farms and livestock,
and from balding forests. The end points of these runoffs are usually bodies of water, so then
eutrophication may occur. Deficiency of phosphorus is also a concern in agriculture. Farmers
must have P-management strategies which include correct application of fertilizers and pH
adjusters.
 V. References

Lal, R., & Stewart, B. A. (2016). Soil Phosphorus. CRC Press.

Oram, B. (2014). Phosphates in the Environment. Retrieved August 30, 2017, from Water
Research Center: http://www.water-research.net/index.php/phosphates

Soil Nutrient Management for Maui County. (2017). Phosphorus. Retrieved 2007, from
University of Hawai'i at Manoa:
https://www.ctahr.hawaii.edu/mauisoil/c_nutrients02.aspx

The phosphorus cycle. (2013, July 30). Retrieved August 30, 2017, from Science Learning Hub:
https://www.sciencelearn.org.nz/resources/961-the-phosphorus-cycle

Thomason, W. (2002). Understanding Phosphorus Behavior in Soils. Retrieved 2017, from Noble
Research Institute: https://www.noble.org/news/publications/ag-news-and-
views/2002/january/understanding-phosphorus-behavior-in-soils/

Three Fruit Research and Extension Center. (2004, July 9). Tree Fruit Soil and Nutrition.
Retrieved 2017, from Washington State University:
http://soils.tfrec.wsu.edu/webnutritiongood/soilprops/soilnutrientvalues.htm

Unites States Department of Agriculture. (n.d.). Phosphorus. Retrieved 2017, from Soil Quality
Kit - Guides for Educators:
https://www.nrcs.usda.gov/Internet/FSE_DOCUMENTS/nrcs142p2_053254.pdf

UV-Visible Spectroscopy. (2013). Retrieved August 30, 2017, from

https://www.soest.hawaii.edu/oceanography/courses/OCN633/Fall%202013/OCN633_
UV-Vis.pdf
VI. Appendix

Table 2. Absorbances of the Standard Solution

Standardization of Phosphate

Concentration Corrected
PPM
(mL/100mL) Absorbance
0 0 0
0.4 0.02 0.016
0.8 0.04 0.029
1.2 0.06 0.051
1.6 0.08 0.059
2 0.1 0.079
4 0.2 0.161
6 0.3 0.237

Figure 1. Calibration Curve in Phosphate Analysis

Absorbance VS Contration
0.25

0.2

0.15 y = 0.7953x - 0.0005

Absorbance

R² = 0.999
0.1

0.05

0
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35
-0.05
Concentration
Calculations

a. Concentration

Trial 1

𝑦 = 𝑚𝑥 + 𝑏

𝑦 = 0.7593𝑥 − 0.0005

0.070 = 0.7593𝑥 − 0.0005

0.7593𝑥 = 0.0705

𝑥1 = 0.092848676𝑝𝑝𝑚

𝐷𝑣 𝐸𝑣
𝑝𝑝𝑚 𝑃 𝑖𝑛 𝑠𝑜𝑖𝑙 = 𝑝𝑝𝑚 𝑃 𝑖𝑛 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 × ×
𝐴𝑣 𝑊𝑡. 𝑆𝑎𝑚𝑝𝑙𝑒

50 𝑚𝑙 50 𝑚𝑙
𝑝𝑝𝑚 𝑃 𝑖𝑛 𝑠𝑜𝑖𝑙 = 0. 0928 × × = 9.25 𝑝𝑝𝑚
10 𝑚𝑙 2.5098 𝑔

Trial 2

𝑦 = 𝑚𝑥 + 𝑏

𝑦 = 0.7593𝑥 − 0.0005

0.068 = 0.7593𝑥 − 0.0005

0.7593𝑥 = 0.0685

𝑥1 = 0.0902146
 𝐷𝑣 𝐸𝑣
𝑝𝑝𝑚 𝑃 𝑖𝑛 𝑠𝑜𝑖𝑙 = 𝑝𝑝𝑚 𝑃 𝑖𝑛 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 × ×
𝐴𝑣 𝑊𝑡. 𝑆𝑎𝑚𝑝𝑙𝑒

50 𝑚𝑙 50 𝑚𝑙
𝑝𝑝𝑚 𝑃 𝑖𝑛 𝑠𝑜𝑖𝑙 = 0.0902146 × × = 8.9352 𝑝𝑝𝑚
10 𝑚𝑙 2.5232 𝑔

b. Average Concentration

𝑥1 + 𝑥2
𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛𝑎𝑣𝑒𝑟𝑎𝑔𝑒 =
2

9.25 𝑝𝑝𝑚 + 8.9352𝑝𝑝𝑚

= 2

= 9.0935 𝑝𝑝𝑚

c. %RSD
𝑠 0.21927 𝑝𝑝𝑚
%𝑅𝑆𝐷 = × 100 = × 100 = 2.411%
𝑥 9.0935 𝑝𝑝𝑚