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Clinical Chemistry 19: 1265-1267, 1973;
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Clinical Chemistry, Vol 19, 1265-1267, Copyright © 1973 by the American Association for Clinical Chemistry

A New Micromethod for Determination of Protein in Cerebrospinal Fluid and Urine

Michael A. Pesce 1 and Carl S. Strande 1

1 Columbia Presbyterian Medical Center, Babies’ Hospital, Clinical Chemistry Laboratory, 3975 Broadway, New York, N. Y. 10032.

We developed a new micro-scale procedure for determination of protein in cerebrospinal fluid and urine. The sample (50 µl) is mixed with a trichloroacetic acid—Ponceau S dye solution. The proteins, together with a proportional amount of dye, are precipitated and the red precipitate is dissolved in dilute sodium hydroxide solution, giving a violet-colored solution. This color, linearly proportional to the amount of protein present (up to 150 mg/dl), is measured spectrophotometrically at 560 nm. By varying the sample size alone, protein concentrations up to 1500 mg/dl can be measured. The detection limit for the method is 2 mg/dl of sample. Precipitation of the proteins is independent of temperature and albuminbinding compounds, and is not appreciably affected by the albumin—gamma globulin ratio.


Key Words: dye-protein complex, spectrophotometry of • inter-method comparison

Submitted on July 5, 1973




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