Multiplicative scatter correction and principal component analysis of UV-Vis absorption spectra during acid hemolysis of erythrocyte suspension

The process of hemolysis of erythrocyte suspension induced by hydrochloric acid (HCl) is investigated by measuring the UV-Vis absorption spectrum in the range 250 nm - 750 nm at interval 1 nm. The spectrum is measured each 15 s to the end of hemolysis. The methods of Multiplicative Scatter Correction (MSC) and Principal Component Analysis (PCA) were applied for analysis of spectra. Multiplicative Scatter Correction of spectra allowed removing effects of scattering and observing the changes related to the hemoglobin absorption in the course of hemolysis - shift of Soret band from 420 to 412 nm, decreasing of two peaks of oxyhemoglobin at 543 and 577 nm and disappearing of globin-hem interaction band at 344 nm. The Principal components of transformed spectra were more informative compared to Principal components of raw spectra and contained features related to absorption of hemoglobin.