A.M.M. ATTIA*#, FATMA ABD EL-HAMID ABD EL-SAMEE IBRAHIM*, NOHA AHMED ABD EL-LATIF*, S.W. AZIZ*, AZHAR MOHAMED ELWAN*, W.M. ABOULTHANA*, S.A.A. MOUSSA*,**, M.S. ELALFY***
*Biochemistry Department, National Research Centre, 33 Bohouth Street, Dokki, Giza, Egypt
** Department of Physics, College of Science, Al-Imam Muhammad Ibn Saud Islamic University, Riyadh, Saudi Arabia
*** Pediatric Department, Faculty of Medicine, Ain Shams University, Cairo, Egypt
New multi-component spectrophotometric methods were developed experimentally and theoretically to determine the accurate concentrations of the total hemoglobin (Hb) and its inactive derivatives including methemoglobin (MetHb), carboxyhemoglobin (HbCO) and sulfhemoglobin (SHb)) as well as the active derivative in the form of oxyhemoglobin (HbO2) in the human and mice blood. With respect to the experimental technique, the method of preparation of Hb solutions has been developed, like the separation of erythrocytes ghosts, plasma leukocytes and plasma lipid aggregates and preparation of diluted Hb solutions, in order to overcome the drawbacks of the previous methods. It was found that results of this experimental method revealed values of normal human SHb% in the range (0.263–0.472) and MetHb% (0.174–1.003), HbCO% (0.683–1.365) and HbO2% (97.469–98.810). Furthermore, the results of this method revealed values of normal mice SHb% in the range (0.033–0.091) and MetHb% (0.033–0.357), HbCO% (1.689–3.369) and HbO2% (96.430–98.086). The methods are non-expensive, highly sensitive, accurate and reproducible and have the advantages of small sample volume, simplicity and speed and can be computerized.
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