Determination of Carvedilol in Human Plasma by High-Performance Liquid Chromatography Applying On-Line Deproteination and Column Switching
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An HPLC-column switching method has been developed and validated for the determination of carvedilol in human plasma. 20 μL plasma was injected onto a Brownlee C8 column and the high molecular weight proteins were eluted isocratically. Carvedilol was separated from the majority of the components by a linear acetonitrile gradient. The fraction containing carvedilol was switched on-line onto a LiChrospher RP 18 column and separated isocratically from accompanying compounds applying 0.1 M triethylamine-methanol-acetonitrile = 17: 18:19 (v/v/v). The influence of pH and temperature on retention was investigated on both columns. No interference from other plasma components was observed. Reproducibility of the complete method was better than 4% and the limit of quantitation was 0.8 ng mL−1.
KeywordsColumn liquid chromatography Column switching On-line deproteination Carvedilol
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