Estimation of inorganic phosphorus by fiske subbarow method

Estimation of inorganic phosphorus by fiske subbarow method

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  ESTIMATION OF INORGANIC PHOSPHORUS BYFISKE SUBBAROW METHOD Dr. Akshata S Mirajkar Assistant Professor Department Of Biochemistry D.Y Patil Medical college Kolhapur
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  INTRODUCTION Abundant mineral Total bodyphosphorus is about 1kg 80% present in bones and teeth Hydroxyapatite in bone – structural support of the body Involved in the release of energy from Fat, Protein, Carbohydrate during metabolism and in the formation of genetic material, cell membranes and many enymes. Buffer Nucleic acids, Nucleotides like ATP, GTP, ADP etc
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  Fiske and Subbarow
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  Reagent 1. 10% TCA 2.Ammonium molybdate 3. Perchloric acid 4. 1,2,4 – Amino Naphthol Sulphonic Acid ( ANSA) 5. Working Phosphate Standard (0.004 mg/ml)
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  Preparation of ProteinFree Filtrate Mix by Inversion and keep aside for 5 minutes.Filter and use filtrate for estimation.
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  Preparation of ProteinFree Filtrate  Trichloroacetic acid is an alkaloidal agent which helps in the precipitation of proteins.  RBC’s are rich in phosphates so they need to be removed before estimation of phosphorus in blood and hemolysed blood sample should be avoided.
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  Procedure
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  1)Test 2)Standard 3)Blank
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  Principle Perchloric acid +Ammonium Molybdate Ammonium chloride + Molybdic acid Molybdic acid + Inorganic Phosphorus Phosphomolybdic acid 1,2,4 Amino Naphthol Sulphonic Acid + Phosphomolybdic acid Blue Coloured Compound ( Read using a Red Filter/620nm)
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  Normal Values Adult =2.5 to 4.5 mg/dl Infant = 4.5 to 8 mg/dl Child = 4 to 6 mg/dl
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  Calculations 𝑇−𝐵 𝑆−𝐵 × Concentration ofStd/ml × Volume of Standard× 100 𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑆𝑒𝑟𝑢𝑚 = 𝑇−𝐵 𝑆−𝐵 ×0.004×5× 100 0.5 = 𝑇−𝐵 𝑆−𝐵 ×4 =--------------------mg/dl Volume of serum  10ml PFF contains 1ml serum So 5ml PFF contains 0.5 ml serum
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  Result The given bloodsample contains ------------------mg/dl of Inorganic Phosphorus. The given sample shows higher/lower/normal values as compared with the Adult/ Infant/ Child range.
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  Interpretation Hyperphosphataemia: Hypoparathyroidism Hypervitaminosis D Chronic Nephritis
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  Interpretation Hypophosphataemia: Hyperparathyroidism Rickets/ Osteomalacia Physiological fallin increased carbohydrate utilization. e.g: effect of insulin in causing the shift of glucose into the cells also enhances the transport of phosphate into cells.
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  Thank you