Detection of proteinuria using SSA

(Sulphur salicylic acid)

By
Abdulrauf yahaya
Department of Biochemistry, Sokoto State University Sokoto, Nigeria
 INTRODUCTION
• Most plasma proteins are too large to pass through the glomeruli of the kidney.
• The small amount of protein which does filter through is normally reabsorbed back into the blood by
the kidney tubules.
• Only trace amounts of protein (less than 150 mg per 24 h) can therefore be found in normal urine.
These proteins include Tamm-horsfall protein (Maximum-40%), Albumin (20%), Immunoglobulins,
Hormones, Enzymes and Mucopolysaccharides.
• When more than trace amounts of protein are found in urine, this is termed proteinuria.
• Detection of proteinuria is an important indicator of renal disease because protein has a very low
maximal tubular rate of reabsorption.
• Sulphosalicylic Acid Test is a good method for detection of protenuria
 Principle
• Proteins are precipitated by 5-sulphosalicylic acid. Any resulting turbidity
will give an estimation of the amount of protein present in the urine which
can be subjectively quantitated visually or more precisely quantitated
using photometry. Cells and casts in the urine must be removed by
centrifuging before carrying out the test. The test can detect albumin,
hemoglobin, myoglobin, and Bence Jones proteins.
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 Requirements and reagents
Requirements
• Specimen:
• Random urine
Apparatus:
• Centrifuge, general purpose
• Graduated pipettes/micropipettes
• Measuring Cylinder
• Test Tubes
• pH paper/ pH meter
Reagents:
• 5-sulphosalicylic acid solution-3%
• Glacial acetic acid-10%
 Procedure
• The pH of a portion of urine was checked, if it is alkaline or neutral. And added
10% acetic acid solution, drop by drop, until it is just acidic (about pH 6).
• Filtration or centrifuging the urine (5 minutes, 2000-3000 rpm) was conducted
in cloudy urine.
• 2ml clear urine in a test tube were taken.
• And added 2 ml 5-sulphosalicylic acid solution and mixed. Avoided shaking
and examination for turbidity against a dark background was done.
 Results and interpretation
• Grade the turbidity as follows:
• Sulphosalicylic-Acid-Test
• Negative : No cloudiness
• Trace: Faint turbidity.
• 1+ : definite turbidity
• 2+ : Heavy turbidity but no flocculation
• 3+ : Heavy turbidity with light flocculation.
• 4+ : Heavy turbidity with heavy flocculation.
Interpretation
 Results and discussion
Patient Result Interpretation
1 1+ definite turbidity

2 1+ definite turbidity
3 2+ Heavy turbidity with light
flocculation.

4 1+ definite turbidity
5 1+ definite turbidity
5 3+
6 1+ definite turbidity
 Conclusion
• The method is a reliable and easy one for determination of protenuria
 References

• World Health Organization (1986). Methods recommended for essential clinical chemical and haematological
tests for intermediate hospital laboratories/Working Group on Assessment of Clinical Technologies. In
Methods recommended for essential clinical chemical and haematological tests for intermediate hospital
laboratories/Working Group on Assessment of Clinical Technologies.
• Ridley J.W. (2018) Procedures for Complete Urinalysis/Confirmation Testing. In: Fundamentals of the Study
of Urine and Body Fluids. Springer, Cham. https://doi.org/10.1007/978-3-319-78417-5_10
• Cheesbrough, M. (1981). Medical laboratory manual for tropical countries (Vol. 1). M. Cheesbrough, 14
Bevills Close, Doddington, Cambridgeshire, PE15 OTT..
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