RPHPLC method for the estimation of glibenclamide in human serum
J. S. S. College of Pharmacy, Ootacamund - 643 001, India
S D Rajendran J. S. S. College of Pharmacy, Ootacamund - 643 001 India E-mail: [email protected]
A high performance liquid chromatographic method is described for estimation of glibenclamide in human serum. After precipitation with methanol, the separation of glibenclamide and internal standard was accomplished using reversed phase chromatography. The mobile phase, a combination of acetonitrile and 25 mM phosphate buffer (pH 3.5) at 3:2 ratio was run isocraticaly through a C18 analytical column. The UV detection was done at 253 nm for glibenclamide. Analytical run time was less than 12 min. Mean recovery was 92% for 0.5 µg/ml concentrations. The assay exhibited good linear relationship between peak area ratios and serum concentration. Quantification limit was at least 25 ng/ml of glibenclamide and accuracy and precision were over the concentration range of 50-500 ng/ml. Assay was successfully applied to the measurement of glibenclamide in serum for therapeutic drug monitoring.