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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
REQUIREMENTS:
·
Three test-tube,
·
Colorimeter or
Semi-Automatic Analyzer
·
Total Protein
working reagent,
·
Distilled water,
·
Incubator,
·
Cuvette,
·
Pipette etc.
·
Serum Specimen
PROCEDURE:
Contents |
Blank |
Standard |
Test |
|
Working reagent |
1ml |
1ml |
1ml |
|
Distilled water |
10µl |
- |
- |
|
Standard |
- |
10µl |
- |
|
Serum |
- |
- |
10µl |
|
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
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
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.
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