estimation of phosphorus and functions.pptx

phosphate
method
normal values
function
disorder

1. ESTIMATION OF SERUM PHOSPHORUS

2. i . AIM To estimate the quantity of inorganic phosphate in given sample or serum

3. ii . METHOD Fiske – Subbarow method

4. iii. PRINCIPLE Inorganic phosphate + Ammonium molybdate + sulphuric acid  Ammonium phosphomolybdate Intensity of color complex is directly proportional to phosphorus in sample which is measured at 340 nm

5. iv. PROCEDURE Take 3 test tubes Label the tubes as Blank(B), Standard (S) and Test(T) Add the reagents according to table 1

6. Mix well and incubate for 5minute Measure the absorbance of sample, standard against reagent at 340nm Table 1 REAGENT BLANK (B) STANDARD (S) TEST (T) Molybdate reagent 1mL 1mL 1mL Distilled water 10uL - - Standard - 10uL - Sample - - 10uL

7. v. CALCULATION OF PHOSPHORUS Concentration of Phosphorus = ------------------------- x 4mg/dL = ---------- mg/dL Table 2 (OD) Test - (OD) Blank in the test = OD (Blank) OD (Standard) OD (Test) (OD) Std - (OD) Blank Concentration of Standard X

8. vi. RESULT Concentration of Serum I norganic Phosphorus in the test is mg/dL

9. vii. NORMAL RANGE Infant = 4.5 – 9 mg/dL Children = 4.5 – 5 mg/dL Adult = 2.7 – 4.5 mg/dL Renal Threshold : 2 mg/dL

10. viii. INTERPRETATION Hyperphosphatemia Hypophosphatemia

11. Causes for Increased serum inorganic phosphorus: 1. Increased absorption of phosphate Excess vitamin D Phosphate infusion 2. Increased cell lysis Chemotherapy for cancer Bone secondaries 3. Decreased excretion of phosphorus Renal impairment Hypoparathyroidism 4. Hypocalcemia 5. Massive blood transfusions 6. Thyrotoxicosis 7. Drugs Chlorothiazide, Nifedipine, Furosemide

12. Causes for Decreased serum inorganic phosphorus: 1. Decreased absorption of phosphate Malnutrition Malabsorption Chronic diarrhea Vitamin D deficiency 2. Intracellular shift Insulin therapy Respiratory alkalosis 3. Increased urinary excretion Hyperparathyroidism Fanconi’s syndrome Hypophosphatemic rickets 4 . Hereditary hypophosphatemia 5. Drugs: Antacids, diuretics, salicylate

13. ix. OTHER METHODS Vanadate-Molybdate - Vanadate and molybdate form a yellow complex with phosphate at acid pH Method of Gomorii Enzymatic methods

14. Functions of phosphate ions Formation of bone and teeth Production of high energy phosphate compounds such as ATP, CTP, GTP Synthesis of nucleoside coenzymes such as NAD and NADP DNA and RNA synthesis, (phosphodiester linkages) Formation of phosphate esters, such as glucose-6-phosphate Formation of phosphoproteins, e.g. casein Activation of enzymes by phosphorylation Phosphate buffer system in blood. The ratio of Na2HPO4: NaH2PO4 in blood is 4:1 at pH of 7.4

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