High Sensitivity High Performance Liquid Chromatography Electrospray Tandem Mass Spectrometry Determination of Terfenadine in Human Plasma

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Abstract
A highly sensitive and selective HPLC-MS-MS method was developed for the determination of terfenadine in human plasma. The analytes, terfenadine and terfenadine related compound A (internal standard), were extracted from human plasma by methyl t-butyl ether/methylene chloride/n-butyl chloride (3:2:1). The organic solvent is evaporated to dryness and reconstituted in 100 μL of acetonitrile/0.02 M ammonium acetate buffer pH 3.5 (50:50). Chromatographic separation is achieved on a TSK. gel ODS-80TS column with a mobile phase composed of acetonitrile/1% formic acid/0.01 M ammonium acetate pH 4.0 (85:13:2). The analytes were detected by HPLC in conjunction with electrospray tandem mass spectrometry. The assay was linear in the concentration ranges of 0.2 to 50 ng/mL. The analysis of pooled quality controls demonstrated excellent precision with relative standard deviations being less than 5.2%. The method is accurate with all intraday and over-all mean values being less than 5.3% from theoretical.