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A new multi-component spectrophotometric method was developed experimentally and theoretically to determine the accurate serum concentrations of the total bilirubin (TB), oxyhemoglobin (HbO2), and methemalbumin (Mha) in healthy human adults and neonates with hemolytic jaundice. With respect to the experimental technique, the method of preparation of serum solution has been developed, like the use of distilled water as a solvent and centrifugation of serum solutions to clear the sample turbidity. The results of TB were compared to the diazo-assay. Theoretically, the formulas used for the calculation of the major components (TB, HbO2, and Mha) in human sera have been derived based on the theory of multi-component spectrophotometric analysis and the mathematical Gaussian elimination method for matrix calculation. The method of multi-component spectrophotometry, suggested in this study for determination of TB, showed % error (3.1%–4.9%), indicating the high accuracy of the method. The small coefficients of variation (CV = 3.65%–5.1%) indicate the high precision of the method. The results showed higher values of serum TB (p < 0.00005), HbO2 (p < 0.001), and Mha concentrations (p < 0.00005) in neonates, when compared to adults. The method is highly sensitive and accurate. It is inexpensive, precise, reproducible, and has the advantages of simplicity, speed, and can be computerized.

A multi-component spectrophotometric method for simultaneous determination of total bilirubin, oxyhemoglobin, and methemalbumin in human sera