Spectrophotometric determination of hemoglobin with acid chrome blue K

In Clark-Lub's buffer solution of pH 3.2,acid chrome blue K reacted with hemoglobin to form a complex,the absorbance could be enhanced,and the enhanced absorbance value at the wavelength of 375 nm was proportional to the concentration of hemoglobin.Based on this,a new spectrophotometric method was developed for the determination of hemoglobin.The optimum conditions and effects of foreign ions were investigated.The results indicated that hemoglobin concentration demonstrated good linear correlation in the range of 0～112 μg/mL,and the apparent molar absorptivity e375 was 4.7×105 L·mol-1·cm-1.This method has been applied to the determination of hemoglobin concentration in animal blood and human urine samples with satisfactory results.