HPLC with Column Switching Coupled to APCI–MS for Pharmacokinetic Study of Amygdalin in Rabbit Plasma
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A fast and validated assay was established for the pharmacokinetic study of amygdalin in Armeniacae Semen in rabbit. The method involved column switching (CS) enrichment, separation, post-column derivatization, and atmospheric pressure chemical ionization (APCI) mass spectrometric detection. Plasma sample was enriched by CS using a MAYI-ODS as the first column. Analytes of interest were isolated and analyzed on a second column of Zorbax SB-C18. To detect amygdalin in plasma samples, a T-piece was connected between the HPLC outlet and the APCI source to add a mixture of dichloromethane and methanol to the eluent by an isocratic pump. Calibration graphs showed good linearity over a range of 1.0–1,280 ng mL−1. The detection limit was 0.2 ng mL−1. The intra- and inter-day accuracies were within 3.9%. The method was successfully applied to a study of the pharmacokinetics of amygdalin after an intravenous injection of amygdalin extracts to rabbits with a dose of 400 mg kg−1. The results indicate that amygdalin is a one-compartment open model with a first order absorption phase.
KeywordsColumn liquid chromatography Column switching APCI–MS detection Pharmacokinetics Amygdalin
This work was supported by grants from the National Natural Science Foundation of China (No. 30470633, 30270554), External Intercourse and Cooperation Item (No. 30511120267), the Research Fund for the Doctoral Program (No.20050698012) and the Cultivation Fund of the Key Scientific and Technical Innovation Project, Ministry of Education of China (No.705045).
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