HPLC-MS/MS法测定芪葵颗粒中25个成分的含量<sup>*</sup>

陆明霞, 欧阳冰琛, 廖健城, 桂婉玉, 向杰, 邹建东, 李长印

doi:10.16155/j.0254-1793.2022.10.04

药物分析杂志 >

2022 , Vol. 42 >Issue 10: 1708 - 1719

DOI: https://doi.org/10.16155/j.0254-1793.2022.10.04

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HPLC-MS/MS法测定芪葵颗粒中25个成分的含量^*

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南京中医药大学附属医院/江苏省中医院,临床药理科,南京 210029

第一作者 Tel:(025)86587807;E-mail:2802108710@qq.com

**Tel:(025)86587807;E-mail:fsyy00612@njucm.edu.cn

收稿日期: 2022-01-18

网络出版日期: 2024-06-24

基金资助

*江苏省自然科学基金面上项目(BK20201502);“333高层次人才培养工程”科研资助项目(BRA2020384);江苏省卫健委医学科研项目面上项目(H2019092);江苏省中医院高峰人才项目(y2018rc22,y2021rc31);江苏高校优势学科建设工程资助项目(苏政办发[2018]87号);南京中医药大学中医学优势学科三期项目开放课题(ZYX03KF030)

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Simultaneous determination of 25 components in Qikui granules by HPLC-MS/MS^*

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Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Hospital of Traditional Chinese Medicine, Clinical Pharmacology, Nanjing 210029, China

Received date: 2022-01-18

Online published: 2024-06-24

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摘要

目的: 建立高效液相色谱-串联质谱(HPLC-MS/MS)法测定芪葵颗粒中25个成分(7,2’-二羟基-3’,4’-二甲氧基异黄烷、L-苯丙氨酸、毛蕊异黄酮-7-O-葡萄糖苷、芦丁、2,3,5,4’-四羟基二苯乙烯-2-O-葡萄糖苷、美迪紫檀苷、异黄芪皂苷Ⅱ、杨梅素、无水柠檬酸、紫檀烷、黄芪皂苷Ⅲ、黄芪甲苷、黄芪皂苷Ⅰ、环黄芪苷Ⅱ、异黄芪皂苷Ⅰ、L-色氨酸、黄芪皂苷Ⅱ、异槲皮苷、刺芒柄花苷、金丝桃苷、没食子酸、槲皮素、毛蕊异黄酮、芒柄花素、大黄素)的含量。方法: 采用Agilent Poroshell 120 SB-C₁₈色谱柱(100 mm×3.0 mm,2.7 μm),以含0.1%甲酸和1 mmol·L^-1甲酸铵的水溶液(A)-甲醇(B)为流动相,梯度洗脱,流速0.4 mL·min^-1,柱温45 ℃,进样体积2 μL;采用电喷雾离子源(ESI),多反应监测(MRM)模式,负离子模式扫描。结果: 25个成分在测定的质量浓度范围内线性关系良好,相关系数(r)均＞0.995 0,精密度、重复性、稳定性均良好,平均加样回收率在89.5%~105.7%,RSD≤4.0%;3批芪葵颗粒中上述25个成分的含量范围分别为0.005~0.006、0.102~0.104、0.107~0.155、0.073~0.074、0.016~0.019、1.058~1.120、0.032~0.047、0.040~0.052、5.600~6.617、0.026~0.030、0.009~0.011、0.009~0.013、0.077~0.085、0.018~0.026、0.131~0.154、0.061~0.068、0.032~0.038、0.108~0.127、0.043~0.064、0.167~0.185、0.084~0.110、0.021~0.026、0.186~0.203、0.091~0.094、0.020~0.033 mg·g^-1。结论: 所建立的方法可准确测定芪葵颗粒中多个主要成分,适用于芪葵颗粒制剂的质量评估与控制。

关键词： 高效液相色谱-串联质谱法; 芪葵颗粒; 多反应监测模式; 7,2’-二羟基-3’,4’-二甲氧基异黄烷; L-苯丙氨酸; 毛蕊异黄酮-7-O-葡萄糖苷; 芦丁; 异黄芪皂苷; 2,3,5,4’-四羟基二苯乙烯-2-O-葡萄糖苷; 美迪紫檀苷; 杨梅素; 无水柠檬酸; 黄芪紫檀烷; 黄芪皂苷; 黄芪甲苷; 环黄芪苷; L-色氨酸; 异槲皮苷; 刺芒柄花苷; 金丝桃苷; 没食子酸; 槲皮素; 毛蕊异黄酮; 芒柄花素; 大黄素

本文引用格式

陆明霞, 欧阳冰琛, 廖健城, 桂婉玉, 向杰, 邹建东, 李长印 . HPLC-MS/MS法测定芪葵颗粒中25个成分的含量^*[J]. 药物分析杂志, 2022 , 42(10) : 1708 -1719 . DOI: 10.16155/j.0254-1793.2022.10.04

Abstract

Objective: To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC- MS/MS) method for the simultaneous determination of 25 components (7,2’-dihydroxy-3’, 4’-dimethoxyisoflavan, L-phenylalanine, calycosin-7-O-glucoside, rutin, isoastragaloside Ⅱ, 2, 3, 5, 4’-tetrahydroxyl diphenylethylene-2-O-glucoside, methylnissolin-3-O-glucoside, myricetin, citric acid, 3-hydroxy-9,10-dimethoxypterocarpan, astragaloside Ⅲ, astragaloside Ⅳ, astragaloside Ⅰ, cyclocephaloside Ⅱ, isoastragaloside Ⅰ, L-tryptophan, astragaloside Ⅱ, isoquercitrin, formononetin-7-O-glucoside, hyperoside, gallic acid, quercetin, calycosin, formononetin and emodin)in Qikui granules. Methods: Chromatographic conditions: the chromatographic column was Agilent Poroshell 120 SB-C₁₈(100 mm×3.0 mm, 2.7 μm). The mobile phase was composed of 1 mmol·L^-1 ammonium formate containing 0.1% formic acid(A) and methanol(B) in gradient elution at a flow rate of 0.4 mL·min^-1 and the column temperature was set at 45 ℃. The injection volume was 2 μL. Mass spectrum conditions: the analytes were determined by electrospray ionization source (ESI) combined with multiple reaction monitoring(MRM) in negative ion mode. Results: The linear relationships of 25 components were good in corresponding ranges with correlation coefficients(r) all above 0.995 0. The precision, repeatability and stability were good. The average recoveries varied between 89.5% and 105.7% with RSDs≤4.0%. And the average contents of the above-mentioned 25 components in 3 batches of Qikui granules were 0.005-0.006, 0.104-0.104,0.107-0.155, 0.073-0.074, 0.016-0.019, 1.058-1.120, 0.032-0.047, 0.040-0.052, 5.600-6.617, 0.026-0.030, 0.009-0.011, 0.009-0.013, 0.077-0.085, 0.018-0.026, 0.131-0.154, 0.061-0.068, 0.032-0.038, 0.108-0.127, 0.043-0.064, 0.167-0.185, 0.084-0.110, 0.021-0.026, 0.203-0.186, 0.091-0.094, and 0.020-0.033 mg·g^-1, respectively. Conclusion: The established method can accurately determine several main components in Qikui granules and is suitable for quality evaluation and control of Qikui granules.

Key words： HPLC-MS/MS; Qikui granules; multiple reaction monitoring; 7,2’-dihydroxy-3’,4-dimethoxyisoflavan; L-phenylalanine; calycosin-7-O-glucoside; rutin; isoastragaloside; 2,3,5,4’-tetrahydroxyl diphenylethylene-2-O-glucoside; methylnissolin-3-O-glucoside; myricetin; citric acid; 3-hydroxy-9, 10-dimethoxypterocarpan; astragaloside; cyclocephaloside; L-tryptophan; isoquercitrin; formononetin-7-O-glucoside; hyperoside; gallic acid; quercetin; calycosin; formononetin; emodin

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