బయోఅనాలిసిస్ & బయోమెడిసిన్ జర్నల్

Quantification of Ticlopidine in Human Plasma Using Protein Precipitation and Liquid Chromatography Coupled with Tandem Mass Spectrometry

Abstract

SoJeong Yi, Hyun-Suk Shin, Seo-Hyun Yoon, Kyung-Sang Yu, In-Jin Jang, Sang-Goo Shin and Joo-Youn Cho

A simple and rapid method has been developed and validated for determination of ticlopidine in human plasma using protein precipitation and liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS). Ticlopidine was extracted from 20-μL aliquots of human plasma by one-step protein precipitation with 980 μL of acetonitrile containing 10 ng/mL clopidogrel as an internal standard (IS). Chromatographic separation was performed on a reverse-phase Gemini C 18 column (50 mm x 2.0 mm, 5 μm) with an isocratic mobile phase (acetonitrile: 1 mM ammonium acetate in water = 75:25, v/v). Ticlopidine and IS were detected and quantified by tandem mass spectrometry with positive electrospray ionization using multiple reaction monitoring of the transition m/z 264.04 to m/z 154.20 for ticlopidine and m/z 322.40 to m/z 212.20 for IS. This method was linear over the concentrations ranging from 2 to 2000 ng/mL. The accuracy in the inter-batch assay was 92.4-95.6% and the precision was within 6.4% coefficient of variation. The validated method was successfully applied to the human pharmacokinetic study of ticlopidine.