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Estimations of Serum Total Protein by Biuret method

SUMMARY:

Measurement of total protein is a useful test in a variety of disorders. Decreased total protein concentration can be deleted in defective protein synthesis in the liver, protein loss due to impaired kidney function, Intestinal malabsorption, or nutritional deficiency. Elevated protein levels occur in chronic inflammatory disease, liver cirrhosis, and dehydration

PRINCIPLE:

Proteins together with copper iron form a violet-blue color Complex in an alkaline solution the absorption of the color is directly proportional to the concentration of total protein in the specimen

                             Protein + Cu^{++                        Alkaline pH}                  Blue-Violet Complex                                  

REQUIREMENTS:

·       Three test-tube,

·       Colorimeter or Semi-Automatic Analyzer

·       Total Protein working reagent,

·       Distilled water,

·       Incubator,

·       Cuvette,

·       Pipette etc.

·       Serum Specimen

PROCEDURE:

1.      Measure all content according to the chart in three test tubes.

2.      Mix well and incubate for 10 minutes at 37 ⁰c temperature in incubator

3.      Read the optical density of the test and standard against blank at 546 nm wavelength

Calculation:-

Blood Sugar (g/dl)                      =            Optical Density of Test X Concentration of Standard

                                                                                      Optical Density of Standard

Concentration of Glucose Standard = 6 g/dl  

After Testing,

Optical Density of Test               =            0.28 &,

Optical Density of Standard           =            0.26

Then,

0.28*6

   0.26

Blood Sugar (g/dl)                      =            6.46     

Result: -                                         =            6.46 g/dl 

Normal Value:-   

Total S. Protein             = 6 - 8 g/dl 

S. Albumin                      = 2.5 – 5.0 g/dl   

S. Globulin                    = 2.5 – 3.5 g/dl

S. AG Ration                   = 0.9-2.0

Clinical Significance:-

Decreased total protein concentration can be deleted in defective protein synthesis in the liver, protein loss due to impaired kidney function, Intestinal malabsorption, or nutritional deficiency. Elevated protein levels occur in chronic inflammatory disease, liver cirrhosis, and dehydration.