HPLC-QAMS法同时定量分析归芪补血口服液中8种成分含量

Simultaneous quantitative analysis of eight components in Guiqi Buxue Oral Liquid by HPLC-QAMS

关键词:  高效液相一测多评法  归芪补血口服液  相对校正因子  党参炔苷  紫丁香苷  毛蕊异黄酮葡萄糖苷  芒柄花苷  9,10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷  去氢土莫酸  去氢茯苓酸  茯苓酸

Key words:  HPLC-QAMS  Guiqi Buxue Oral Liquid  relative correction factor  lobetyolin  syringin  calycosin 7-O-β-D-glucopyranoside  ononin  9,10-dimethoxy-pterocarpane-3-O-β-D-glucoside  dehydrotumulosic acid  dehydropachymic acid  pachymic acid

摘要:

[目的] 建立高效液相色谱一测多评法（HPLC-QAMS）同时定量分析归芪补血口服液中党参炔苷、紫丁香苷、毛蕊异黄酮葡萄糖苷、芒柄花苷、9，10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷、去氢土莫酸、去氢茯苓酸和茯苓酸含量。[方法] 色谱柱为Agilent Zorbax SB-C18柱（250 mm×4.6 mm，5 μm），柱温为25℃。流动相为乙腈-0.05%磷酸水溶液，梯度洗脱，流速为1.0 mL/min。检测波长分别为254 nm（检测党参炔苷、紫丁香苷、毛蕊异黄酮葡萄糖苷、芒柄花苷、9，10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷）和210 nm（检测去氢土莫酸、去氢茯苓酸和茯苓酸）。进样量为10 μL。以毛蕊异黄酮葡萄糖苷为内参物，建立党参炔苷、紫丁香苷、芒柄花苷、9，10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷、去氢土莫酸、去氢茯苓酸和茯苓酸的相对校正因子，计算各成分含量，并与外标法（ESM）检测结果进行对比，验证所建立的HPLC-QAMS法的准确性和可行性。[结果] 党参炔苷、紫丁香苷、毛蕊异黄酮葡萄糖苷、芒柄花苷、9，10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷、去氢土莫酸、去氢茯苓酸和茯苓酸分别在1.91～47.75、0.64～16.00、1.57～39.25、0.98～24.50、1.40～35.00、0.76～19.00、0.58～14.50、1.16～29.00 μg/mL（r≥0.999 1）范围内线性关系良好。平均加样回收率及相对标准偏差（RSD）分别为98.2%（0.97%）、97.0%（1.05%）、100.1%（0.76%）、98.7%（1.32%）、99.1%（1.43%）、97.2%（0.89%）、97.3%（1.38%）和98.4%（1.69%）。HPLC-QAMS法计算结果与ESM实测结果无明显差异。[结论] 所建立的HPLC-QAMS法为归芪补血口服液现有质量标准提供了补充和完善。

Abstract:

[Objective] To establish an high performance liquid chromatography-quantitative analysis of multi-components by single marker(HPLC-QAMS) method for simultaneous determine of lobetyolin, syringin, calycosin 7-O-β-D-glucopyranoside, ononin, 9, 10-dimethoxy-pterocarpane-3-O-β-D-glucoside, dehydrotumulosic acid, dehydropachymic acid and pachymic acid in Guiqi Buxue Oral Liquid.[Methods] The determination was performed on an Agilent Zorbax SB-C18 column(250 mm×4.6 mm, 5 μm) with a gradient elution of acetonitrile-0.05% phosphoric acid as mobile phase at the flow rate of 1.0 mL/min. The detection wavelength were set at 254 nm for lobetyolin, syringin, calycosin 7-O-β-D-glucopyranoside, 9, 10-dimethoxy-pterocarpane-3-O-β-D-glucosideand ononin, 210 nm for dehydrotumulosic acid, dehydropachymic acid and pachymic acid. The column temperature was 25℃. Using calycosin 7-O-β-D-glucopyranoside as an internal standard, the relative correction factors of lobetyolin, syringin, calycosin 7-O-β-D-glucopyranoside, ononin, 9, 10-dimethoxy-pterocarpane-3-O-β-D-glucoside, dehydrotumulosic acid, dehydropachymic acid and pachymic acid were calculated, after which the content determination was made, and compared with the results of the external standard method(ESM) to verify the accuracy and feasibility of the method.[Results] Lobetyolin, syringin, calycosin 7-O-β-D-glucopyranoside, ononin, 9, 10-dimethoxy-pterocarpane-3-O-β-D-glucoside, dehydrotumulosic acid, dehydropachymic acid and pachymic acid showed good linear relationships within the ranges of 1.91~47.75, 0.64~16.00, 1.57~39.25, 0.98~24.50, 1.40~35.00, 0.76~19.00, 0.58~14.50, 1.16~29.00 μg/mL(r ≥ 0.999 1), whose average recoverie and relative standard deviation(RSD) were 98.2%(0.97%), 97.0%(1.05%), 100.1%(0.76%), 98.7%(1.32%), 99.1%(1.43%), 97.2%(0.89%), 97.3%(1.38%), 98.4(1.69%), respectively. No significant difference of quantitative results was observed between ESM and HPLC-QAMS method.[Conclusion] The HPLC-QAMS method can provide a supplement and improvement method for the existing quality standards of Guiqi Buxue Oral Liquid.