Quantitative Determination of Ticlopidine Hydrochloride in Human Plasma by High-performance Liquid Chromatography-Electronspray Ionization Mass Spectrometry

Abstract

A sensitive, selective and simple high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) was developed and validated for the quantification of ticlopidine hydrochloride (CAS 53885-35-1) in human plasma using loratadine (CAS 79794-75-5) as internal standard (IS). Following liquid-liquid extraction, the analyte and the IS were extracted from plasma samples by n-hexane : iso-propanol (95:5, v/v), separated by HPLC on a commercially available column (150 mm > 2.0 mm ID, 5 μm) with a mobile phase of acetonitrile : 10 mmol/L ammonium acetate buffer solution (85:15, v/v) and analyzed on a quadrupole mass spectrometer with ESI interface operating in the positiveion mode. The correlation coefficient of the calibration curve was linear (r² > 0.99) over the concentration range of 1 – 1000 ng/mL for ticlopidine hydrochloride. The intra- and inter-batch precisions were less than 15 % of the relative standard deviation and the accuracy ranged from 85 to 115 % in terms of percent accuracy. The limit of detection (LOD) of ticlopidine hydrochloride was 0.5 ng/mL. The extraction recovery of ticlopidine hydrochloride was more than 80 %. The proposed method enables the unambiguous identification and quantification of ticlopidine hydrochloride for pharmacokinetic, bioavailability or bio-equivalence studies.

Key words

Anti-platelet agents - CAS 53885-35-1 - Ticlopidine hydrochloride, bioequivalence, HPLC-ESI-MS, pharmacokinetics

• References

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