药学学报, 2019, 54(12): 2303-2307
引用本文:
叶晓芸, 郭青, 郭斌, 谭力, 黄青, 张艳海, 杨丹丹, 施海蔚. HPLC-CAD法对黄杨宁片原料中环维黄杨星D和有关物质的定性定量研究[J]. 药学学报, 2019, 54(12): 2303-2307.
YE Xiao-yun, GUO Qing, GUO Bin, TAN Li, HUANG Qing, ZHANG Yan-hai, YANG Dan-dan, SHI Hai-wei. Qualitative and quantitative analysis of cyclovirobuxine D and related substances by HPLC-CAD in the active pharmaceutical ingredient of Huangyangning tablets[J]. Acta Pharmaceutica Sinica, 2019, 54(12): 2303-2307.

HPLC-CAD法对黄杨宁片原料中环维黄杨星D和有关物质的定性定量研究
叶晓芸1, 郭青2, 郭斌1, 谭力2, 黄青2, 张艳海3, 杨丹丹4, 施海蔚2
1. 南京中医药大学, 江苏 南京 210023;
2. 江苏省食品药品监督检验研究院, 江苏 南京 210019;
3. 赛默飞世尔科技(中国)有限公司, 上海 201206;
4. 中国药科大学, 江苏 南京 211198
摘要:
首次建立高效液相色谱-电喷雾检测(HPLC-CAD)法对黄杨宁片原料中环维黄杨星D及有关物质进行定量分析,并用高效液相色谱串联四极杆-静电场轨道阱高分辨质谱仪联用技术(HPLC-Q-Exactive)对CAD检测中出现的有关物质进行了定性研究。采用XBridge Amide色谱柱(4.6 mm×250 mm,5 μm)分离;乙腈-100 mmol·L-1甲酸铵溶液(85:15)(混合溶液用甲酸调pH 2.8)为流动相等度洗脱,流速1.1 mL·min-1,柱温30℃;CAD雾化温度35℃,气压62.2 psi;新建立的方法能检测到除环维黄杨星D外的5个有关物质。方法学结果显示,环维黄杨星D检测限为12.588 ng,定量限为28.323 ng;平均回收率为95.74%,RSD为1.79%(n =6)。3个厂家12批原料样品中环维黄杨星D含量为79.94%~88.49%,平均值82.20%。CAD对非挥发性物质具有响应均一性,以环维黄杨星D作为对照品外标法计算5个有关物质含量,结果在15.99%~22.15%,平均值20.10%。该方法对于无发色团的黄杨生物碱类物质具有检测信号齐全和微量组分检测灵敏度高的优势,成分间分离度良好,能与MS兼容,可应用于黄杨宁片原料中成分的定性定量分析。
关键词:    HPLC-CAD      黄杨宁片原料      环维黄杨星D      有关物质      定性定量      HPLC-Q-Exactive     
Qualitative and quantitative analysis of cyclovirobuxine D and related substances by HPLC-CAD in the active pharmaceutical ingredient of Huangyangning tablets
YE Xiao-yun1, GUO Qing2, GUO Bin1, TAN Li2, HUANG Qing2, ZHANG Yan-hai3, YANG Dan-dan4, SHI Hai-wei2
1. Nanjing University of Traditional Chinese Medicine, Nanjing 210023, China;
2. Jiangsu Institute of Food and Drug Control, Nanjing 210019, China;
3. Thermo Fisher Scientific Corporation, Shanghai 201206, China;
4. China Pharmaceutical University, Nanjing 211198, China
Abstract:
We have developed a new method using HPLC-CAD (charged aerosol detector) for the quantitative analysis of cyclovirobuxine D and related substances in the API of Huangyangning tablets. The related substances were further studied by HPLC-Q-Exactive coupled with hybrid quadrupole-orbitrap mass spectrometry. A HILIC column of XBridge Amide (4.6 mm×250 mm, 5 μm) was used, and the mobile phase was composed of acetonitrile and 100 mmol·L-1 ammonium formate (85:15), which was adjusted to pH 2.8 with formic acid. Isocratic mode elution was adopted at a flow rate of 1.1 mL·min-1. The column temperature was set at 30℃. For CAD, the temperature of atomization and gas pressure were respectively set at 35℃ and 62.2 psi. This method detected and quantified five related substances to cyclovirobuxine D. The results showed that the LOD and LOQ of cyclovirobuxine D was 12.588 ng and 28.323 ng, respectively with an average recovery of 95.74% (RSD=1.79%, n=6). The content of cyclovirobuxine D in 12 batches of API samples provided by three manufacturers was from 79.94% to 88.49%, with an average value of 82.20%. The total content of the five related substances was from 15.99% to 22.15% with an average value of 20.10%, using an external standard method with cyclovirobuxine D as the reference and according to the CAD uniform response to non-volatile substances. The newly developed HPLC-CAD method has advantages in terms of the comprehensiveness of signals from Buxus alkaloids without UV absorption and with high sensitivity to its trace-related substances; the method yields good separation between the components and is compatible with mass spectrometry. It is applicable for the accurate quantitative analysis of main components and related substances in the API of Huangyangning tablets.
Key words:    HPLC-CAD    active pharmaceutical ingredient of Huangyangning tablet    cyclovirobuxine D    related substance    qualitative and quantitative analysis    HPLC-Q-Exactive   
收稿日期: 2019-06-12
DOI: 10.16438/j.0513-4870.2019-0469
通讯作者: 郭青,Tel:86-25-86251121,E-mail:guoqing850@sohu.com
Email: guoqing850@sohu.com
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