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Clinical Chemistry 21: 398-401, 1975;
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Clinical Chemistry, Vol 21, 398-401, Copyright © 1975 by the American Association for Clinical Chemistry

Improved Biuret Procedure for Routine Determination of Urinary Total Proteins in Clinical Proteinuria

Eugene W. Rice 1

1 Pathology Department, Allentown Hospital Association, Allentown, Pa. 18102.

This communication describes and evaluates an improved routine methodology for quantitating clinical proteinuria. Based on investigations of Piscator and of Savory et al., a modified Tsuchiya's reagent (ethanolic HCl-phosphotungstic acid) is used to precipitate proteins at 56 °C, followed by biuret spectrophotometry at 540 nm. The accuracy of the proposed procedure was assessed by comparisons with results obtained by using an ultrafiltration membrane that retains solutes with an average molecular weight in excess of 10 000 for separating of urinary proteins before they are measured with the biuret reaction. Precision of the method (coefficient of variation) is typically 2-3%.


Key Words: Tsuchiya's reagent (ethanolic HCl-phosphotungstic acid) • spectrophotometry • ultrafiltration

Submitted on June 10, 1974
Accepted on December 30, 1974




The following articles in journals at HighWire Press have cited this article:


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G. L. Hortin and B. Meilinger
Cross-Reactivity of Amino Acids and Other Compounds in the Biuret Reaction: Interference with Urinary Peptide Measurements
Clin. Chem., August 1, 2005; 51(8): 1411 - 1419.
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T. Marshall and K. M. Williams
Total Protein Determination in Urine: Elimination of a Differential Response between the Coomassie Blue and Pyrogallol Red Protein Dye-binding Assays
Clin. Chem., March 1, 2000; 46(3): 392 - 398.
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P. L. M. Lynch, J. Savory, and D. M. Haverstick
Urine Total Protein Measurement with the Vitros Dry Reagent Technology: Modification of Diluent to Resolve Positive Bias of Diluted Samples
Clin. Chem., March 1, 1998; 44(3): 674 - 675.
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Copyright © 1975 by the American Association for Clinical Chemistry.