New multicomponent spectrophotometric method for estimation of five hemoglobin derivatives in normal and traffic policemen

ABSTRACT

Several methods for the determination of five hemoglobin derivatives [oxy - carboxy - sulf - met - deoxy] hemoglobin based on multicomponent spectrophotometric analysis had been described. Most of these methods are applicable for the determination of the physiological concentration of both deoxy-Hb and oxy0Hb, and the oxygen saturation of blood samples. However, these methods suffered from some sources that were not easily overcome e.g. turbidity, interference due to pigments and negligible of other hemoglobin derivatives. The new technique has been applied in the simultaneous determination of SHb. Met-Hb, HbCO and the remaining functional Hb in the oxy-Hb form, with a conventional spectrophotometer. In this method, the measurements for the latter prepared extremely diluted solutions, with a light-path length 1cm. The 16 millimolar absorptivities of HbO[2], HbCO, Met-Hb and SHb, determined previously, at four wavelengths [500, 569, 577 and 620nm] that represent Met-Hb, HbCO, HbO[2], SHb respectively, were substituted into four linear equations of the type described by the theory of multicomponent spectrophotometric analysis with the four unknown concentrations of Hh pigments. This linear system of equations was solved by mathematical manipulation using the Gaussian elimination method for matrix calculation giving the unknown Hb derivatives concentrations