A.M.M. ATTIA^#, W.M. ABOULTHANA, S.W. AZIZ

Biochemistry Department, National Research Centre, 33 Bohouth Street, Dokki, Giza, Egypt,

Our recent multi-component spectrophotometric method for determination of hemoglobin derivatives has been developed theoretically and experimentally to get more accurate and precise concentrations of sulfhemoglobin, methemoglobin, carboxyhemoglobin and oxyhemoglobin in the blood of humans and rats. Theoretically, the equations used for determining the hemoglobin derivatives were derived by using more recent millimolar absorptivity matrices and are based on the theory of multi-component spectrophotometric analysis and the mathematical Gaussian elimination method for matrix calculation. New millimolar absorptivities at λ = 576 and 630 nm have been usedfor the first time to derive these equations. Experimentally, the method has been standardized with respect to the pH and temperature of hemoglobin solution. The results on methemoglobin percentage showed high correlations (r = 0.997 and r = 0.998) with our old multi-component method and the chemical method, respectively. Also, the results of oxyhemoglobin determined by our new method showed a high correlation (r = 0.999) with those determined by our old multi-component method. Moreover, the results of total hemoglobin concentration determined by our new method showed a high correlation (r = 0.998) with the chemical method. These results indicate the high accuracy of our new method. Furthermore, the percent values of sulfhemoglobin, carboxyhemoglobin and oxyhemoglobin showed a coefficient of variation of 1.52%, 5.85% and 0.137%, indicating the high reproducibility and precision of our new method. The method is non-expensive, highly sensitive, accurate, precise and reproducible and have the advantages of small sample volume, simplicity and speed and can be computerized.

Key words: carboxyhemoglobin, hemoglobin, methemoglobin, sulfhemoglobin, oxyhemoglobin, human, rats.

Corresponding author’s e-mail: yousatef@yahoo.com