Simultaneous determination of nine bioactive components in Bazhen Yimu Pills by HPLC-ESI-MS/MS / 中草药

ABSTRACT

Objective To develop and validate a high performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) method for simultaneously qualitative and quantitative determination of nine major bioactive components (stachydrine hydrochloride, leonurine hydrochloride, paeoniflorin, ferulic acid, liquiritin, lobetyolin, verbascoside, atracylenolide I, and polyporenic acid C) in Bazhen Yimu Pills (BYP). Methods The chromatographic separation was performed on an Waters Atlantis T3 column (150 mm × 4.6 mm, 3.0 μm) with a gradient elution of methanol and 0.1% formic acid in water at a flow rate of 0.5 mL/min, and the injection volume was 10 μL. The nine major bioactive components were detected using an electrospray ionization source in negative ionization mode (ESI-) and quantified by multiple reaction monitor (MRM) scanning at the same time. Results The linear ranges of stachydrine hydrochloride, leonurine hydrochloride, paeoniflorin, ferulic acid, liquiritin, lobetyolin, verbascoside, atracylenolide I, and polyporenic acid C were 0.04-40.00 μg/mL (r = 0.999 2), 0.04-40.00 μg/mL (r = 0.999 3), 1.0-100.0 μg/mL (r = 0.999 1), 0.2-20.0 μg/mL (r = 0.999 6), 0.2-20.0 μg/mL (r = 0.997 5), 0.05-5.00 μg/mL (r = 0.999 4), 0.1-10.0 μg/mL (r = 0.999 4), 0.1-10.0 μg/mL (r = 0.999 2), 0.1-10.0 μg/mL (r = 0.999 2), and the average recoveries were 99.7% (RSD = 0.52%), 98.1% (RSD = 0.64%), 98.5% (RSD = 1.08%), 101.5% (RSD = 1.12%), 99.5% (RSD = 0.39%), 98.4% (RSD = 0.74%), 99.1% (RSD = 0.91%), 101.2% (RSD = 0.54%), and 100.1% (RSD = 0.47%), respectively. The content of nine batches of the nine major bioactive components were 0.423-0.752, 0.505-0.722, 0.613-1.300, 0.102-0.184, 0.195-0.255, 0.021-0.035, 0.034-0.072, 0.039-0.063, and 0.051-0.095 mg/g, respectively. Conclusion The developed method is simple, specific, and sensitive, and it can be applied for the determination of nine major bioactive components and the quality control of BYP collected from different production batches.