Abstract
Bushen Pills (BSPs), as a traditional Chinese medicine (TCM), is widely used in clinic to enrich Yang, nourish Yin, stem essence, and strengthen kidneys. Two chromatographic methods, liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), were applied to analyze the multiple active components of BSPs in dosage form for quality evaluation and in rat plasma for pharmacokinetics study, respectively. Three active constituents of BSPs, including paeoniflorin (PF), berberine hydrochloride (BBR), and schizandrin (SCH), were simultaneously determined by the established LC-MS method with electrospray ionization (ESI) in positive selected ion monitoring (SIM) mode at m/z 503.1, 336.0, and 455.2. The contents of PF, BBR, and SCH were (6.112 ± 0.166) mg/g, (335.1 ± 14.95) μg/g, and (5.867 ± 0.136) μg/g in BSPs. On this basis, PF and BBR were selected as targeted analytes for the pharmacokinetic study of BSPs in rats. Memantine hydrochloride was used as an internal standard (IS), and the plasma samples were processed by liquid-liquid extraction with ethyl acetate. All the analytes were separated on a C18 reversed phase column, eluted with a mobile phase consisting of acetonitrile-formic acid (0.01%) (25 : 75, v/v), and detected by ESI in the selected ion mode with multiple reaction monitoring (MRM). The target fragment ions were m/z 525.3 ⟶ 449.5 for PF, 336.2 ⟶ 320.2 for BBR, and 180.1 ⟶ 163.1 for IS. The linear ranges of PF and BBR were 5–500 ng/mL and 0.1–20 ng/mL with good linearity (r2 > 0.99). No obvious matrix effect was observed, and acceptable accuracy, precision, recovery, and stability were obtained. The proposed method has been successfully applied to the pharmacokinetic study of BSPs in rats after a single dose.
Highlights
Bushen Pills (BSPs), which was modified from “Dabuyin Pills” initially recorded in Danxi xinfa 500 years ago, is one of the well-known traditional Chinese medicines (TCM) employed to enrich Yang, nourish Yin, stem essence, and strengthen kidneys. It is prescribed in combination with 10 medicinal crude drugs at a certain mass ratio based on the theory of traditional Chinese herbal medical science. e types and ratio of the drugs were as follows: Herba Cynomorii (9.3%), Plastrum Testudinis (9.3%), Radix Rehmanniae
Praeparata (23.3%), Fructus Lycii (14.0%), Radix Asparagi (9.3%), Rhizoma Anemarrhenae (9.3%), Radix Paeoniae Alba (9.3%), Rhizoma Zingiberis (2.3%), Cortex Phellodendri Chinensis (9.3%), and Fructus Schisandrae Chinensis (4.6%) [1, 2]. e formula has the effect of ameliorating clinical symptoms such as kidney water insufficiency, Yin deficiency and Yang hyperactivity, aching and pain waist and knees, dizziness, cough, wet dream, and so on [3]
The MS conditions of the cited analytes were investigated in both positive and negative ion modes. e major m/z ions identified for PF, berberine hydrochloride (BBR), and SCH were 503.1, 336.0, and 455.2 in positive mode
Summary
Bushen Pills (BSPs), which was modified from “Dabuyin Pills” initially recorded in Danxi xinfa 500 years ago, is one of the well-known traditional Chinese medicines (TCM) employed to enrich Yang, nourish Yin, stem essence, and strengthen kidneys. It is prescribed in combination with 10 medicinal crude drugs at a certain mass ratio based on the theory of traditional Chinese herbal medical science. Mass spectrometry, as one of the most widely used analytical methods, with the further development, the identification and the quantification limit of target components has been greatly improved, no matter in vitro or in vivo
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