**2. Experimental methods used in this study**

A random fresh blood sample collected in EDTA tube was obtained from Hamad Medical Corporation (HMC), Doha, Qatar. The blood samples were obtained from HMC blood donor center in Doha, Qatar. Institutional review board (IRB) approval was obtained from HMC for using the donor's blood for the purpose of research. This research has two model samples (the hemolysate and the purified hemoglobin). The principal materials were: iron (III) solution, iron chelators (deferoxamine and deferiprone), and glutathione. Dry-ice acetone bath, hot water bath, centrifuge, UV-VIS spectrophotometer (absorption spectroscope), and disposable PD-10 desalting columns kit.
