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Validation of a simple HPLC method for assay of haplamine and its metabolites in plasma suitable for pharmacokinetic application in rats
Author(s) -
Ea Sompheary,
Aubert Claude,
Giacometti Sarah,
Ciccolini Joseph,
Siv Chhan,
Bun Hot
Publication year - 2008
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.904
Subject(s) - chromatography , chemistry , pharmacokinetics , calibration curve , detection limit , analyte , high performance liquid chromatography , extraction (chemistry) , methanol , pharmacology , medicine , organic chemistry
Abstract A simple HPLC method with ultraviolet detection has been developed and validated for the simultaneous determination of haplamine and its metabolites ( trans / cis ‐3,4‐dihydroxyhaplamine) in rat. A liquid–liquid extraction was used to extract the compounds from rat plasma. The analysis was performed on a C 18 Nucleosil Nautilus column. The mobile phase consisted of water (A) and a mixture of methanol and acetonitrile (85:15; v/v) (B) used in gradient mode (38–40% B for 10 min, 40–58% B for 49 min, 58–38% B for 1 min, and 38% for 5 min) pumped at 1 mL/min. The calibration curves showed good linearity with correlation coefficients greater than 0.999 for the analytes in the investigated concentration range. The lower limit of detection was 0.007, 0.008 and 0.009 µg/mL and the lower limit of quantification was 0.014, 0.017 and 0.018 µg/mL for haplamine, and trans / cis ‐3,4‐dihydroxyhaplamine, respectively. The method was applied to a preliminary pharmacokinetic study in rats. This method proved to meet fully the standards required of experimental pharmacokinetic studies and should be used in further preclinical investigation. Copyright © 2007 John Wiley & Sons, Ltd.