TITLE:
Rapid and Sensitive Carvedilol Assay in Human Plasma Using a High-Performance Liquid Chromatography with Mass/Mass Spectrometer Detection Employed for a Bioequivalence Study
AUTHORS:
Soo-Hwan Kim, Sang Hun Lee, Hye Jung Lee
KEYWORDS:
Carvedilol, Bioequivalence, HPLC-MS/MS, Validation
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.1 No.3,
November
30,
2010
ABSTRACT: A method for the determination of carvedilol in human plasma was developed using a high-performance liquid chromatography with tandem mass spectrometer (HPLC-MS/MS). Plasma samples were deproteinized using acetonitrile and the supernatant was directly injected onto the HPLC column without any preparative steps. Chromatography was performed on a reversed-phase (C18) column with isocratic mobile phase for 2 min. The calibration curve was linear over the range of 2 to100 ng/ml (R2 > 0.9998) and the lower limit of quantitation (LLOQ) was 2 ng/ml. This method showed acceptable precision and accuracy, good recovery from the plasma matrix, and stability during the analytical procedures. When its application to the bioequivalence test of two carvedilol 25 mg tablet formulations in male healthy 28 volunteers, this validated analysis method was appropriate resulting in the bioequivalence of two formulations: no statistically significant difference was observed between the logarithmic transformed area under curve (AUC) and maximum plasma concentration (Cmax) values of the two formulations. The 90% confidence interval for the ratio of the above mentioned two parameters were within the bioequivalence limit of 0.80-1.25. These results suggested that the HPLC-MS/MS analysis method developed was suitable for the carvedilol analysis in human plasma.