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A high throughput and sensitive ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed for the determination of terbinafine in human plasma. The method employed liquid-liquid extraction of terbinafine and terbinafine-d7 (used as internal standard) from 100 μL human plasma with ethyl acetate: n-hexane (80: 20, v/v) solvent mixture. Chromatography was performed on BEH C18 (50 × 2.1 mm, 1.7 μm) column using acetonitrile-8.0 mM ammonium formate, pH 3.5 (85:15, v/v) under isocratic elution. For quantitative analysis, MS/MS ion transitions were monitored at m/z 292.2/141.1 and m/z 299.1/148.2 for terbinafine and terbinafine-d7, respectively using electrospray ionization in the positive mode. The method was validated according to regulatory guidance for selectivity, sensitivity, linearity, recovery, matrix effect, stability, dilution reliability and ruggedness with acceptable accuracy and precision. The method shows good linearity over the tested concentration range from 1.00-2000 ng/mL (r ≥ 0.9984). The intra-batch and inter-batch precision (% CV) varied from 1.8-3.2 % and 2.1-4.5 %, respectively. The method was successfully applied to a bioequivalence study with 250 mg terbinafine in 32 healthy subjects. The major advantage of this method includes higher sensitivity, small plasma volume for processing and a short analysis time.
This article was published in the following journal.
Name: Biomedical chromatography : BMC
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