Development and validation of two liquid chromatography-tandem mass spectrometry methods for the determination of silibinin and silibinin hemisuccinate in human plasma

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• 作者：Federica Sala ; Pablo Albares ; Milena Colovic ; Stefano Persiani ; Lucio C. Rovati
• 关键词：ACD ; acid citrate dextrose ; HCV ; hepatitis C virus ; IFN ; interferon ; MF ; matrix factor ; MP ; mobile phase ; NMF ; normalized matrix factor ; SOC ; standard of care ; SRM ; selected reaction monitoring ; TNF ; tumor necrosis factor ; UHQ ; ultra high quality
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：15 January, 2014
• 年：2014
• 卷：945-946
• 期：Complete
• 页码：1-9
• 全文大小：783 K

文摘

To investigate the pharmacokinetics of silibinin and silibinin hemisuccinate in human plasma, two high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) methods were developed and validated. The methods require a small volume of sample (100 渭L), and the recovery of the analytes was complete with a good reproducibility (CV% 1.7-9.5), after a simple protein precipitation. Naringenin was used as internal standard. The chromatographic methods provided a good separation of diastereoisomers A and B of both silibinin and silibinin hemisuccinate onto a Chromolith Performance RP18e 100 mm 脳 3 mm column, with a resolution of peaks from plasma matrix in less than 6 min. The methods precision values expressed as CV% were always 鈮?.2% and the accuracy was always well within the acceptable 15% range. Quantification was performed on a triple-quadrupole tandem mass spectrometer by Selected Reaction Monitoring (SRM) mode, in a negative ion mode, via electrospray ionization (ESI). The lower limit of quantitation was set at 5.0 ng/mL (silibinin) and 25.0 ng/mL (silibinin hemisuccinate), and the linearity was validated up to 1000.0 and 12,500.0 ng/mL, for silibinin and silibinin hemisuccinate, respectively, with correlation coefficients (R²) of 0.991 or better. The methods were suitable for pharmacokinetic studies and were successfully applied to human plasma samples from subjects treated intravenously with Legalon^庐 SIL at the dose of 20 mg/kg, expressed as silibinin.