超高效液相-质谱联用测定不同产地半夏中10种成分含量及质控指标筛选

Determination of 10 components in Pinelliaternata from different habitats by UPLC-MS/MS and selection of quality control indices

目的:建立同时测定半夏中6个核苷类(尿嘧啶、胞苷、尿苷、腺嘌呤、鸟苷、腺苷)和4个有机酸类(酒石酸、柠檬酸、苹果酸、琥珀酸)的超高效液相-质谱联用(UPLC-MS/MS)分析方法,并对不同产地的半夏进行品质评价。方法:色谱条件为Shim-pack XR-ODSⅢ(200 mm×2 mm,2.2μm),流动相A为0.1%甲酸水,B为甲醇,梯度洗脱程序为0→5 min:1%B,5→10 min:1%~3%B,10→25 min:3%~20%B;柱温25℃,流速0.3 mL·min^(-1);质谱采用ESI离子源,多反应监测(MRM)的扫描方式进行检测;最后对结果采用SPSS和SIMCA进行聚类分析和主成分分析。结果:10种待测成分在1~32 ng·mL^(-1)线性关系良好,它们的加样回收率在96.8%~104.1%之间;该方法精密度、重复性和稳定性良好,符合方法学考察要求;24批半夏药材中酒石酸含量为0.22~1.08 mg·mL^(-1)、柠檬酸0.33~1.89 mg·mL^(-1)、苹果酸0.31~1.17 mg·mL^(-1)、琥珀酸0.28~1.17 mg·mL^(-1)、尿嘧啶0.01~0.83 mg·mL^(-1)、胞苷0.60~4.85 mg·mL^(-1)、尿苷0.10~0.26 mg·mL^(-1)、腺嘌呤0.02~0.27 mg·mL^(-1)、鸟苷0.06~0.34 mg·mL^(-1)、腺苷0.01~0.79 mg·mL^(-1)。结论:本研究所建立方法稳定可行,可用于半夏中多成分含量的同时测定,为半夏的质量控制提供新参考。

OBJECTIVE To establish the simultaneous determinations of 6 nucleosides(uracil, cytidine, uridine, adenine, guanosine & adenosine) and 4 organic acids(tartaric acid, citric acid & malic acid & succinic acid) through UPLC-MS/MS analysis and perform quality evaluations of Pinelliaternata from different producing areas.METHODS The chromatographic conditions were Shim-pack XR-ODS Ⅲ(200 mm × 2 mm, 2.2 μm), mobile phase A 0.1% formic acid water, B methanol, gradient elution 0→5 min: 1% B, 5→10 min : 1%-3% B, 10→25 min: 3%-20% B;column temperature 25 ℃, flow rate 0.3 mL·min^(-1);mass spectrometry was detected by ESI ion source MRM scanning. Finally SPSS and SIMCA were employed for cluster and principal component analyses.RESULTS These 10 tested components had an excellent linear relationship between 1-32 ng·mL^(-1) and their sample recovery rates were between 96.8% and 104.1%. Such a method had good precision, reproducibility and stability and it fulfilled the requirements of methodological examinations. The content of tartaric acid in 24 batches of Pinelliaternate was 0.22-1.08 mg·mL^(-1), citric acid 0.33-1.89 mg·mL^(-1), malic acid 0.31-1.17 mg·mL^(-1), succinic acid 0.28-1.17 mg·mL^(-1), uracil 0.01-0.83 mg·mL^(-1), cytidine 0.60-4.85 mg·mL^(-1), uridine 0.10-0.26 mg·mL^(-1), adenine 0.02-0.27 mg·mL^(-1), guanosine 0.06-0.34 mg·mL^(-1) and adenosine 0.01-0.79 mg·mL^(-1).CONCLUSION Both stable and feasible, the above method may be employed for simultaneous determinations of multiple components in Pinelliaternate and providing a new reference for its quality control.