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Chapter-18 Estimation of serum proteins and albumin/globulin ratio by Biuret method

BOOK TITLE: Laboratory Manual for Practical Biochemistry

Author
1. MK Ganesh
2. A Hemavathi
3. YM Shivaraja Shankara
ISBN
9788184482591
DOI
10.5005/jp/books/10435_19
Edition
1/e
Publishing Year
2008
Pages
4
Author Affiliations
1. KVG Dental College, Sullia, Karnataka, KVG Dental College, Sullia, Karnataka, India
2. KVG Medical College, Sullia, Karnataka
3. KVG Medical College, Sullia, Karnataka, KVG Medical College, Sullia, Karnataka, India
Chapter keywords

Abstract

The principle of biuret reaction is used in this colorimetric method. The cupric ion in an alkaline medium forms a violet colored complex with peptide bond nitrogen of protein. Total proteins (Tp) of serum are determined by treating serum with biuret reagent. The intensity of violet color obtained is proportional to the number of peptide bonds, which inturn is a measure of total protein. In the differential estimation of albumin and globulin, the property of globulins to get precipitated by 28 percent sodium sulphite is made use of. The same serum is taken in another test tube and globulins are precipitated with sodium sulphite solution. The filtrate obtained contains albumin which is treated with biuret reagent. Violet color formed is the measure of serum albumin. Measurement of OD values is made using green filter (540 nm).

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