[关键词]
[摘要]
目的 建立基于超高效液相色谱-四极杆/静电场轨道阱高分辨质谱(ultra performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry,UPLC-Q-Orbitrap HRMS)的妇可靖胶囊中多成分(没食子酸、丹参素、阿魏酸、迷迭香酸、丹酚酸B、木犀草素、芹菜素、芦荟大黄素、大黄素、丁烯基苯酞、欧当归内酯A)的定量分析方法。方法 液相采用Acquity UPLC®; BEH C18色谱柱(50 mm×2.1 mm,1.7 μm),以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱,体积流量为0.2 mL/min;质谱采用HESI离子源,利用其高分辨的特点采用一级Full mass扫描的方式进行定量;并将定量测定结果导入多元数据处理软件SIMCA14.0中进行质量评价分析。结果 在优化的色谱质谱条件下,没食子酸、丹参素、阿魏酸、迷迭香酸、丹酚酸B、木犀草素、芹菜素、芦荟大黄素、大黄素、丁烯基苯酞、欧当归内酯A分别在0.15~1.50、0.80~8.00、1.50~15.00、0.04~0.40、0.015~0.150、0.10~1.00、0.004~0.040、0.004~0.040、0.02~0.20、0.01~0.10、0.04~0.40 μg/mL线性关系良好(r ≥ 0.999 5);精密度、重复性及稳定性良好(RSD ≤ 4%);加样回收率在98%~102%,RSD均小于3%;所测成分在各批次样品中的质量分数依次为没食子酸26.36~31.03 μg/g、丹参素178.85~210.79 μg/g、阿魏酸320.91~343.16 μg/g、迷迭香酸2.84~3.09 μg/g、丹酚酸B 3.55~4.25 μg/g、木犀草素27.33~32.36 μg/g、芹菜素1.89~2.13 μg/g、芦荟大黄素0.47~0.60 μg/g、大黄素3.17~3.57 μg/g、丁烯基苯酞1.99~2.54 μg/g、欧当归内酯A 7.51~8.53 μg/g;分析结果表明大多数批次药物质量较为稳定,其中丹参素和阿魏酸对药物质量具有较大影响,可对其进行重点监控以保证药物批次质量。结论 建立的定量方法灵敏度高且准确性好,方法学考察结果符合测定要求,可用于妇可靖中多种活性成分的快速测定;并为其质量评价提供新的科学依据和参考。
[Key word]
[Abstract]
Objective To establish a quantitative analysis method of multiple active components (gallic acid, tanshinol, ferulic acid, rosmarinic acid, salvianolic acid B, luteolin, apigenin, aloe-emodin, emodin, butylidenephthalide, and levistilide A) in Fukejing Capsule (FC) based on ultra performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS). Methods The column was BEH C18 (50 mm×2.1 mm, 1.7 μm) and the mobile phase was consisted of acetonitrile-water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min with gradient elution; Mass spectrometer conditions:Heated electrospray ionization source (HESI) and full mass quantify methods were used to perform the determination, and the results of the contents were imported into the muti-data processing soft ware SIMCA14.0 to make a quality evaluation. Results Under the optimized conditions, gallic acid, tanshinol, ferulic acid, rosmarinic acid, salvianolic acid B, luteolin, apigenin, aloe-emodin, emodin, butylidenephthalide, and levistilide A all showed good liner relationship (r ≥ 0.999 5) in the range of 0.15-1.50, 0.80-8.00, 1.50-15.00, 0.04-0.40, 0.015-0.150, 0.10-1.00, 0.004-0.040, 0.004-0.040, 0.02-0.20, 0.01-0.10, and 0.04-0.40 μg/mL, respectively; The results of the accuracy, the repeatability and the stability all reached the standards (RSD ≤ 4%); The recoveries ranged from 98%-102% and RSDs were below 3%; The result of the content ranges in different batches were 26.36-31.03 μg/g (gallic acid), 178.85-210.79 μg/g (tanshinol), 320.91-343.16 μg/g (ferulic acid), 2.84-3.09 μg/g (rosmarinic acid), 3.55-4.25 μg/g (salvianolic acid B), 27.33-32.36 μg/g (luteolin), 1.89-2.13 μg/g (apigenin) A, 0.47-0.60 μg/g (aloe-emodin), 3.17-3.57 μg/g (emodin), 1.99-2.54 μg/g (butylidenephthalide), and 7.51-8.53 μg/g (levistilide A). The analysis results showed that the quality of the most batches was stable, the tanshinol and ferulic acid had a great influence on the quality of the medicine, and could be monitored to ensure the quality of different batches. Conclusion The methods established in this paper have a high sensitivity and accuracy; The results of the methodology conform to the relevant requirements and the methods can rapidly determinate the multiple active components in FC; The research also provides a new scientific basis and reference for the quality assessment at the same time.
[中图分类号]
[基金项目]