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Correction for Hemolysis in a Photometric Method for Assaying Sulfobromophthalein in the Blood of Cattle in Tests for Hepatic Function¹

Department of Dairy Science, New Jersey Agricultural Experiment Station, Sussex

ABSTRACT

Varying degrees of hemolysis of blood have interfered with photometric measurements of the dye sulfobromophthalein (Bromsulphalein or BSP) in studies of hepatic function. In the usual procedures plasma blanks are measured in acidic media, since dye colors are developed in basic media. However, the optical density (O.D.) of hemoglobin (as hematin), when measured in an acidic medium, differs from that obtained in a basic medium. Similarly, varying degrees of hemolysis as between plasma blank (no dye) and plasma dye tubes do not allow proper blank correction when either acidic or basic plasma blanks are used.

In developing the improved method, plasma blank O.D. of a series of blood samples containing no dye, but with varying degrees of induced hemolysis, were measured both in acidic and in basic media. Regression equations were derived to predict the basic plasma blank O.D. from acidic plasma blank O.D. When such predicted basic plasma blank O.D. readings were applied to plasma samples containing known amounts of dye but varying amounts of hematin, accurate estimates of dye contents were obtained. Also, when the improved photometric method was used in performing actual hepatic function tests in calves and cows, marked improvement resulted in the precision or repeatability of the measures as compared to the usual procedures.

It is suggested that this new procedure for photometric analysis be used only in those instances where hemolyzed plasma samples are present in the series of samples to be analyzed.

¹ Paper of the Journal Series, New Jersey Agricultural Experiment Station, Rutgers—The State University, New Brunswick. This work was supported in part by a grant from Merck Sharp & Dohme Research Laboratories, Division of Merck & Company, Rahway, New Jersey.

² Present address: Department of Dairy Husbandry, University of Missouri, Columbia.