Plasma hemoglobin determination : two manual methods compared : an honors thesis (HONRS 499)

Abstract

Plasma hemoglobin levels are clinically significant in the determination of the amount of in vivo hemolysis. In this study, two manual methods for plasma hemoglobin were compared. The current method uses sodium carbonate to measure oxyhemoglobin at three wavelengths: 380 nm, 415 nm, and 450 nm. The second method, from SIGMA Diagnostics, uses 3,3',5,5'-tetramethylbenzidine in a peroxidase reaction; the absorbance is then read at 600 nm. In the peroxidase method from SIGMA Diagnostics, timing plays a very crucial role in obtaining accurate results. The peroxidase procedure from SIGMA Diagnostics is less time consuming than the current method and is therefore more advantageous. The peroxidase method proves to be the better method due to better linearity and accuracy.