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In the course of the study of acute hemolytic anemia and hemoglobinuria during administration of sulfonamide drugs (1), the patients' sera were found to contain a pigment which, like methemo-globin and sulfhemoglobin, showed an absorption in the red region of the spectrum. The characteristic band of this pigment, around X620 mu and adjacent to that of methemoglobin, was not altered by cyanide, although that of methemoglobin was obliterated. Furthermore, comparisons with known mixtures and calculations based upon absorption coefficients for sulfhemoglobin, methemo-globin, and hemoglobin did not correspond with the observed spectrophotometric measurements.2 At this time (1940) Fairley's paper appeared describing a new pigment (3) with practically identical properties named methemalbumin (1937, 1938) (4). The pigment, which did not occur in washed, laked erythrocytes nor in urine, was readily formed by incubating laked human ery-throcytes with human plasma at 370 C. for several days. Our observations were in accord with these. Inasmuch as the sera in cases of acute hemolytic anemia contained hemoglobin, methemoglobin, and apparently methemalbumin (1), it was necessary to obtain absorption coefficients for this new pigment in order to determine its proportion in these sera. Fairley's procedure was, therefore, repeated and the resulting solutions (after centrifuging off the heavy precipitate to be described after further study) were analyzed with the Hardy recording spectrophotometer (5). To establish the identity of the new pigment, two reagents were used: 5 per cent neutralized NaCN 8 and 3 per cent hydrogen peroxide. The cyanide was used first to obliterate the absorption 1 Aided in part by the Dazian Foundation. 2Absorption coefficients for hemoglobin and methemo-globin taken from Fox and Cline (7) and for sulfhemo-globin taken from Drabkin (2). $Ammonium sulfide may be used but this frequently causes confusion by subsequently producing sulfhemo-globin. caused by methemoglobin at x630 mu without changing absorption in this region attributed to methemalbumin. Hydrogen peroxide was next added to obliterate the absorption band of sulf-hemoglobin, if present. In Figure 1 are shown the transmission curves of the new pigment, methemalbumin, together with curves of methemoglobin, sulfhemoglobin, and oxyhemoglobin and the changes that negate the absorption in the region of interest.4 In the case of methemalbumin after the addition of cyanide, the absorption in the red region is decreased slightly owing to the removal of small contaminating amounts of methemoglobin. The subsequent addition of hydrogen peroxide caused no further change. Table I summarizes these findings which correspond to Fairley's …

SPECTROPHOTOMETRY OF FAIRLEY'S NEW BLOOD PIGMENT, METHEMALBUMIN