Estimation of phosphate and chloride content in a urine sample

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BACKGROUND

Phosphate is a charged particle that contains the mineral phosphorus and one of the most vital electrolyte in the blood chloride that along with other electrolyte like sodium, potassium helps in the maintenance of the fluid amount in the body. Both are the two-essential component of blood whose increase or decrease level can cause problems. So, the basic objective of this test is to estimate phosphate and chloride content in a urine sample.

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REQUIREMENTS

Reagents:      Standard uranium acetate solution: Take a 1 litre volumetric flask and put 35 g of uranium acetate in it. Then in it                         add 3-4 ml of glacial acetic acid and then add water up to level mark.¹

Acetate buffer: In 80 ml distilled water add 10 gm of sodium acetate and to it 10 ml of 30% acetic acid is added and then diluted with distilled water to make it 100 ml.

Indicator solution: Make 10% potassium ferricyanide solution in distilled water.

Standard silver nitrate solution: In a 100 ml volumetric flask add 1.7 g of silver nitrate and by taking a very less amount of water dissolve it and then add more water to bring it to up to level mark.

Standard ammonium thiocyanate solution: In 100 ml volumetric flask, add 0.76 g of ammonium thiocyanate and dilute it with water to bring it up to mark. Then standardize this solution against 0.1 M silver nitrate solution using ferric alum as indicator.²

     Ferric ammonium sulphate

    Concentrated nitric acid

Apparatus:     Glassware

     Beaker

     Conical flasks

     Burette

     Pipette

     Measuring cylinder

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PROCEDURE

Phosphate estimation

In a conical flask add 5 ml urine sample. Then add 25 ml of distilled water and 5 ml of acetate buffer and heat this till boiling. At boiling state take standard solution of uranium acetate dropwise till precipitation stops. To test whether titration is completed or not bring one drop of the mixture with a glass rod and mix with drop of indicator solution above a porcelain tile. When this reaction produces reddish brown colour titration is completed.¹

1 ml uranium acetate = 0.002 g phosphate.

Chloride estimation

Take a 100 ml conical flask and add 5 ml urine sample in it with the help of pipette. Then to it add 5 ml concentrated nitric acid, 10 ml of 0.1M silver nitrate solution 1 or 2 drops of ferric alum and leave it for about 5 min. Perform titration of this mixed solution with 0.1 M ammonium thiocyanate solution till the persistence of red brown colour for about 15 seconds.² The burette reading should be noted and titration is repeated till two readings are obtained.

1 ml of 0.1M AgNO₃=0.00585 gm of Nacl

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CONCLUSION

This is the two rapid and reproducible method of chloride and phosphate estimation in urine sample.

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REFERENCES

1. Determination of the True Chloride Content of Biological Fluids and Tissues. I. Analysis by Chlorine-36 Isotope Dilution. Ernest. Cot love Analytical Chemistry 1963 35 (1), 95-100
   

1. Robinson-Cohen, C., Ix, J. H., Smits, G., Persky, M., Chertow, G. M., Block, G. A., & Kestenbaum, B. R. (2014). Estimation of 24-hour urine phosphate excretion from spot urine collection: Development of a predictive equation. Journal of Renal Nutrition, 24(3), 194-199.