Determination Of Carboxyhemoglobin With Derivative Spectrophotometry

Objective:To study the derivative spectrogram characters of carboxyhemoglobin and establish a method for the quantitative determination of carboxyhemoglobin in blood with derivative spectrophotometry.Methods:COHb and Hb standard solutions were prepared with the pretreatment of carbonic oxide or oxygen ventilation followed by adding Na2S2O4. The derivative spectrophotometry was operated with Shimadzu UV-265 visible-ultraviolet spectrophotometer.Results:Absorption spectrum showed the maximum absorption wavelengthes (λmax) were 420nm for COHb and 430nm for Hb respectively, and both absorption spectrum of COHb and Hb were seriously overlaped. So that it was impossible to quantitative determine COHb in blood with routine spectrophotometry. However, it was found that the value of 1st derivative spectrophotometry at 427nm could be applied to test the COHb in blood quantitatively with zero-crossing technique. This method could effectively...