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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
High-performance liquid chromatography/tandem mass spectrometry for simultaneous determination of four
Yongbing Sun1, Lan Xu, Qi Wang
1National Pharmaceutical Engineering Center for Solid Preparation in Chinese Herbal Medicine, Jiangxi University of Traditional Chinese Medicine, 56 Yangming Road, Nanchang 330006, China.
Abstract:
A specific and reliable HPLC-MS/MS method was developed and validated for the simultaneous determination of four dicaffeoylquinic acids (DCQA): 3,4-DCQA, 1,5-DCQA, 3,5-DCQA and 4,5-DCQA. The analytes were separated on a C₁₈ column (150 mm × 2.1 mm, 1.8 μm) and a triple-quadrupole mass spectrometry equipped with an electrospray ionization (ESI) source was applied for detection. The plasma sample was prepared by a liquid-liquid extraction method and the recovery for the four analytes was around 80%. The calibration curves were linear over a concentration range of 10.6-1060.0 ng/mL for 3,4-DCQA, 19.2-1920.0 ng/mL for 1,5-DCQA, 14.0-2900.0 ng/mL for 3,5-DCQA, 9.7-970.0 ng/mL for 4,5-DCQA. The intra-day and inter-day precision was less than 15% and the relative error (RE) were all within ±15%. The validated method was successfully applied to a pharmacokinetics study in rats after oral administration of the extracts of Ainsliaea fragrans cham (a traditional Chinese herb).
