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Niazi and State (1) recently reported that when human serum was heated with the diphenylamine reagent of Dische (2, 3) a purple color developed. They also observed that in certain disease states, e.g., carcinoma, pulmonary tuberculosis and rheumatic fever, the purple color reaction was of considerably greater intensity than in the serum of normal persons. We observed this color reaction when testing saline extracts of bovine tonsils for the presence of desoxyribonucleic acids using the Dische diphenylamine reagent. Subsequent fractionation of the tonsil extracts revealed that the substance which gave the purple color reaction with diphenylamine was a mucoprotein with an unusually low isoelectric point (pI. < 2.0) (4). In confirmation of certain findings of Niazi and State (1) we have observed that serum from rheumatic fever and rheumatoid arthritis patients gives a color of greater intensity than does the serum from normal individuals. The resulting purple solution has one absorption band in the visible spectrum, with a maximum at 530 mju. This same absorption band is present when either serum or our purified mucoprotein from beef tonsils (4) is tested. In this report we shall present observations with respect to the reaction, and suggest modifications in technique. We have applied the method to rheumatic fever sera, the results of which are included.

THE PURPLE COLOR REACTION GIVEN BY DIPHENYLAMINE REAGENT. I. WITH NORMAL AND RHEUMATIC FEVER SERA 1