Orginal Publication Date
MCV/Q, Medical College of Virginia Quarterly
Blood was obtained by cardiac puncture from the Rhesus monkey (Macaca mulatta). Hemolysates were prepared by filtration and centrifugation at 20,000 X g at 4 C. Oxyhemoglobin affinity curves were determined by a simplified spectrophotometric procedure on hemolysates diluted with phosphate buffers at pH of 7.4 and 6.8, and at an ionic strength of 0.3. The spectrophotometric readings were made at a wavelength of 640mμ and at a temperature of 23 C. The oxygen pressure at one-half saturation (P₅₀) for Rhesus homoglobin at a pH of 7.4 was 15 mm Hg, and at the more acid pH of 6.8, 23 mm Hg. Using this spectrophotometric method, as many as eight points can be obtained and plotted, so that an oxyhemoglobin affinity curve can be constructed, in less than three hours.
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