Abstract
An environment-friendly and efficient method for simultaneous determination of five alkaloids (nunciferine, O-nornuciferin, liriodenine, armepavine, and pronuciferine) in rat plasma was established by HPLC-MS/MS associated with micro-solid phase extraction (micro-SPE). The plasma sample was pretreated by using micro-SPE columns filled with polymer materials PEP-2 and eluted by little organic solvent (400 µl acetonitrile). The five alkaloids were separated with acetonitrile and 0.1% formic acid aqueous solution on Eclipse plus C18 column. The mode of positive electrospray ionization was used to measure the analytes in multiple-reaction monitoring (MRM). The determination coefficients (R2) of the five alkaloids were greater than 0.99. The lower limit of quantification (LLOQ) of O-nornuciferin, liriodenine, and armepavine was 0.5 ng·ml−1, and that of nunciferine and pronuciferine was 1 ng·ml−1. The validated method was effectively used for the pharmacokinetics of the five orally administrated alkaloids of lotus leaf extract in rat plasma.
Highlights
Lotus leaf, dry leaf of the lotus, is one of the traditional Chinese herbal medicines compiled in Chinese Pharmacopoeia (Chinese Pharmacopoeia Commission, 2015)
Three alkaloids including O-nornuciferin, liriodenine, and pronuciferine in rat plasma were firstly determined in the pharmacokinetic study of lotus leaf
Specificity The specificity was evaluated by analyzing blank plasma sample, blank plasma added into five alkaloids at level of lower limit of quantification (LLOQ), and real plasma orally administrated lotus leaf extract
Summary
Dry leaf of the lotus (species Nelumbo nucifera Gaertn of genus Nelumbo Adans. of family Nelumbonaceae A.Rich.), is one of the traditional Chinese herbal medicines compiled in Chinese Pharmacopoeia (Chinese Pharmacopoeia Commission, 2015). An efficient method was established by combining HPLC-MS/MS with micro-SPE for simultaneous determination of five alkaloids (nunciferine, O-nornuciferin, liriodenine, armepavine, and pronuciferine) in rat plasma in the pharmacokinetic study of lotus leaf (Figure 1). Three alkaloids including O-nornuciferin, liriodenine, and pronuciferine in rat plasma were firstly determined in the pharmacokinetic study of lotus leaf. Quality control (QC) samples of all analytes were prepared at the lower limit of quantification (LLOQ), low, medium, and high concentration levels (1, 2, 100, and 500 ng·ml−1 for nunciferine; 0.5, 1, 40, and 200 ng·ml−1 for O-nornuciferin; 0.5, 1, 40, and 200 ng·ml−1 for liriodenine; 0.5, 1, 20, and 100 ng·ml−1 for armepavine; 1, 2, 100, and 500 ng·ml−1 for pronuciferine) by dissolving appropriate mixed standard solutions in methanol.
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