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Quantitative Determination of Biuret using Ion Exchange Column Chromatography and Ninhydrin¹

University of Illinois, Urbana³

Abstract

Techniques using ion exchange chromatography and the ninhydrin reaction for quantitative determination of biuret in urine were evaluated. Samples of standard biuret solutions were passed through the physiological column for neutral and acidic amino acids on a Beckman Model 120B amino acid analyzer.

Conditions of 33° C. and pH 4.25 buffer resulted in acceptable separation of the biuret peak from peaks of other compounds in sheep urine. Quantitative analysis was made at concentrations of 0.25 mg./ml. to 2.0 mg. biuret/ml. The presence of biuret was detected at 0.1 mg./ml.

A modified procedure of "washing" the sample through with 0.2 N NaOH solution permitted the use of 55° C. and pH 5.18 buffer, which resulted in an increase in the area of the biuret peak of approximately 50 percent.

Results indicated that both of the procedures tested can be used to determine biuret content of urine; however, the procedure using 33° C. and pH 4.25 buffer yielded more consistent values than the 55° C. and pH 5.18 buffer.

¹ Supported in Part by Federal Funds (Hatch 20-336) and by grants-In-Aid from Grace Chemical Company, Memphis, Tennessee, and Dow Chemical Company, Midland, Michigan.

² Present Address: Animal Science Dept., U. of Idaho, Moscow.

³ Department of Animal Science.