1.

Blood Urea estimation by Diacetyl Monoxime (DAM) Method

Reagent Preparation: 1. Mixed acid reagent: Mix 200 ml of conc. Sulphuric acid with 800 ml of dist water. Add 0.5 ml of Ferric chloride solution (dissolve 5gm of ferric chloride in 100ml of Orthophosphoric acid and 150 ml of dist water). 2. Mixed color reagent: Reagent A: 2% Diacetyl monoxime ReagentB: 0.5% Thiosemicarbazide Mix 67ml of Reagent A and 67ml of Reagent B and make up 1 litre with dist water 3. Stock urea standard: Dissolve 600 mg of Urea in 100 ml of Benzoic acid(1gm/l) Conc of std is equivalent to 100 mmol/L

2. Total protein estimation by BIURET Method

Reagent preparation: 1. Sodium hydroxide: 6 mmol/L 240 gm of NaOH in 1 Litre of dist water. 2. Biuret reagent: Dissolve 3gm of copper sulphate in 500ml of dist water. Add 9gm of potassium sodium tartrate and 5gm of potassium iodide, when completely dissolved add 100ml of NaOH (6mmol/L) and make upto 1 Litre with dist water.

3. Albumin estimation by BROMOCRESOL GREEN method

Reagent preparation: 1. Succinic Acid: 50gm/L prepare as required. 2. Sodium hydroxide: 10 gm/L prepare as required. 3. BCG Dye: Dissolve 8.85 gm of Succinic acid, 108 mg of BCG dye and 100mg of sodium Azide in about 900 ml of dist water, add 4 ml of Brij-35(30%). Adjust to pH 4.2 using small amount of NaOH (10gm/L) Or Succinic Acid(50gm/L) if necessary. Make up 1 litre with dist water.

4. Estimation of Serum Alkaline phosphatase by BOWERS Mc COMB Method

Reagent preparation: 1. Carbonate buffer: (pH: 10.15) Dissolve 3.18 gm of NaCO3(Sod Carbonate) and 1.68 of NaHCO3(sod Bicarbonate) in 1 litre of dist. Water. 2. Buffer Substrate: Dissolve 450 mg of 4-amino antipyrane and 1.1 gm of phenyl phosphate disodim salt in 1 liter of carbonate buffer. 3. Colour reagent: Dissolve 13 gm Boric acid and 1.9 gm of potassium ferri cyanide in 1 litre of dist water. 4. Stock phenol std: Dissolve 100mg of phenol in 100 ml of 0.1 N HCl. That stock is equivalent to 100 KA.

5. Serum amylase estimation by Starch Iodine Method:

Reagent preparation: 1. Buffer starch substrate: a. Dissolve 22.6 gm of disodium hydrogen phosphate and 12.5 gm of potassium dihydrogen phosphate in about 500 ml of NaCl solution(0.15 mol/L). Heat to boil. b. Mix separately 0.4 gm of soluble starch in 10 ml to form a paste. Add the paste to the boiling mixture rinsing the beaker with NaCl solution. c. Allow to cool, add 5ml of formaldehyde solution and dilute to about 990 ml NaCl solution. pH of this sol should be 7.00.1

2. Stock Iodine: Dissolve 3.57 gm of potassium iodate and 45 gm of potassium iodide in about 100ml of dist water. Slowly mix and add 9 ml of conc HCl. Make up to 1 litre with dist water. Working iodine: 1ml stock iodine+9ml dist water