TAILIEUCHUNG - Handbook of practical biochemistry: Part 2

(BQ) Continued part 1, part 2 of the document Handbook of practical biochemistry has contents: Determination of total protein and albumin, determination of urine chloride, determination of titrable acidity and ammonia in urine, determination of phosphorus,. and other contents. Invite you to refer. | Determination of Total Protein and Albumin 15A. DETERMINATION OF SERUM TOTAL PROTEIN AND ALBUMIN Proteins are polymers of L-amino acids. There are numerous proteins in our body concerned with different functions. Here we shall discuss plasma serum proteins. Plasma and serum are both fluid part of the blood. Plasma is the supernatant obtained upon addition of an anticoagulant to the blood where as serum is the supernatant obtained when the plain blood specimen is allowed to clot. Hence serum is devoid of fibrinogen where as plasma contains fibrinogen. Plasma serum proteins comprise a complex mixture of different proteins. The important proteins present in the plasma serum are 1. Albumin 2. Globulin 3. Conjugated proteins such as lipoproteins 4. Fibrinogen absent from serum Important biological functions of different kinds of proteins are given below 1. Oncotic pressure - albumin 2. Transport of molecules - albumin bilirubin transferrin iron 3. Hormone function - TSH 4. Coagulation - fibrinogen 5. Defense - antibodies 6. Nutritional - albumin 7. Catalytic - enzyme proteins Liver is involved in the synthesis of albumin fibrinogen prothrombin other clotting factors and other several proteins coming under alpha and beta globulins. Determination of total protein method used Biuret method. Specimen Serum or plasma. Principle Proteins form purple coloured complex with cupric ions in alkaline solution. The biuret test is given by those substances containing two carbamyl groups CONH joined either directly or by a single nitrogen or carbon atom. The purplish violet colour is due to the formation of a copper coordination complex see Fig. 2A-12 . The molecule should have a minimum of two peptide bonds to give copper coordination complex that impart violet color to test mixture. Reagents Required 1. Sodium chloride Normal saline 2. Biuret reagent 3. Protein standard 5 g Procedure Set tubes labeled as T S and B. For further steps see Fig. 15A-1. Determination of Total Protein

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• Practical biochemistry
• Urine chloride
• Titrable acidity
• Determination of phosphorus
• Determination of calcium
• A given solution
• Determination of blood sugar
• Principles of colorimetry
• Biologically important substance
• Vasudevan practical biochemistry
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