成分分析

UPLC-MS/MS法同时测定经典方剂槐花散中7个成分含量*

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  • 江西中医药大学中药资源与民族药研究中心,南昌 330004
第一作者 Tel:18738956158;E-mail:979889604@qq.com
**钟国跃 Tel:(0791)87118889;E-mail:zgy1037@163.com;梁 健 Tel:(0791)87118873;E-mail:ocean719@163.com

收稿日期: 2022-03-31

  网络出版日期: 2024-06-24

基金资助

*国家自然科学基金项目(81560641);江西自然科学基金项目(20192ACB20016)

Simultaneous determination of 7 components in classic formula Huaihua powder by UPLC-MS/MS*

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  • Research Center for Traditional Chinese Medicine Resources and Ethnic Minority Medicine, Jiangxi University of Chinese Medicine, Nanchang 330004, China

Received date: 2022-03-31

  Online published: 2024-06-24

摘要

目的: 建立超高效液相色谱串联质谱(UPLC-MS/MS)法同时测定经典名方槐花散中芦丁、胡薄荷酮、川陈皮素、柚皮苷、槲皮苷、新橙皮苷、槲皮素7个有效成分的含量。方法: 采用ACQUITY UPLC HSS T3C18色谱柱(100 mm×2.1 mm,1.8 μm),以0.1%甲酸水(A)-甲醇(B)为流动相进行梯度洗脱,流速为0.4 mL·min-1,柱温为35 ℃。质谱采用电喷雾离子化(ESI)源,正负离子模式,雾化气流速为3 L·min-1,加热气流速为10 L·min-1,滞留时间为45 ms,离子源温度为300 ℃,加热块温度为400 ℃,脱溶剂管温度为250 ℃,干燥气流速为10 L·min-1;多反应监测模式(MRM)进行定量分析,其中芦丁、胡薄荷酮、川陈皮素采用正离子模式,监测离子对为m/z 611.15/303.00、153.15/81.10、403.15/373.10;柚皮苷、槲皮苷、新橙皮苷和槲皮素采用负离子模式,监测离子对为m/z 579.15/271.15、447.10/300.15、609.15/301.15、301.05/151.05。结果: 槐花散中芦丁、胡薄荷酮、川陈皮素、柚皮苷、槲皮苷、新橙皮苷、槲皮素7个有效成分在各自线性范围内线性关系良好,相关系数均>0.999 5;精密度、稳定性、重复性试验的RSD均<5.0%;平均回收率分别为97.7%、103.3%、92.3%、100.7%、101.5%、101.0%、100.2%,RSD均<5.0%。6批槐花散中上述7个成分的含量范围分别为44.30~47.00、2.01~2.10、0.31~0.33、18.53~19.80、0.79~0.85、17.64~18.57、1.29~1.35 mg·g-1结论: 该方法快速、灵敏,专属性高,可为槐花散的质量控制提供方法,并为该方的药效学及药代动力学研究提供参考。

本文引用格式

师艺玮, 王洪玲, 黄慧莲, 高燕萍, 李娜芝, 钟国跃, 梁健 . UPLC-MS/MS法同时测定经典方剂槐花散中7个成分含量*[J]. 药物分析杂志, 2023 , 43(2) : 219 -226 . DOI: 10.16155/j.0254-1793.2023.02.05

Abstract

Objective: To develop an UPLC-MS/MS method for the simultaneous determination of seven active components in Huaihua powder, including rutin, (+)-R-pulegone, nobiletin, naringoside, quercitrin, neohesperidin and quercetin. Methods: An ACQUITY UPLC HSS T3C18 (100 mm×2.1 mm,1.8 μm) column was used with 0.1% formic acid in water (A) and methanol (B) as mobile phases. A gradient elution was employed with flow rate of 0.4 mL·min-1 and column temperature of 35 ℃. Electrospray ionization (ESI) source was used. Samples were determined in both positive and negative ion modes. MS parameters were 3 L·min-1 for nebuliser gas flow, 10 L·min-1 for heater gas flow, 45 ms for dwell time, 300 ℃ for source temperature, 400 ℃ for heater block temperature, 250 ℃ for desolvation tube temperature, and 10 L·min-1 for dry gas flow. Multiple-reaction monitoring (MRM) was used to perform mass spectrometric quantification. Rutin, (+)-R-pulegone and nobiletin were measured by precursor/product pairs of m/z 611.15/303.00, 153.15/81.10 and 403.15/373.10 in positive ion mode. Naringoside, quercitrin, neohesperidin and quercetin were measured by precursor/product pairs of m/z 579.15/271.15, 447.10/300.15, 609.15/301.15, 301.05/151.05 in negative ion mode. Results: Rutin, (+)-R-pulegone, nobiletin, naringoside, quercitrin, neohesperidin and quercetin had good linearity in their respective linear ranges with all linear correlation coefficients greater than 0.999 5 and RSDs of the precision, repeatability and stability less than 5.0%. The average recoveries were 97.7%, 103.3%, 92.3%, 100.7%, 101.5%, 101.0% and 100.2%, respectively, with RSDs lower than 5.0%. The contents ranges of rutin, (+)-R-pulegone, nobiletin, naringoside, quercitrin, neohesperidin and quercetin in 6 batches of Huaihua powder were 44.30-47.00, 2.01-2.10, 0.31-0.33, 18.53-19.80, 0.79-0.85, 17.64-18.57 and 1.29-1.35 mg·g-1. Conclusion: The proposed method is rapid, sensitive and specific, which could offer an analytical method for the quality control of Huaihua powder, reference and insights for its pharmacodynamic and pharmacokinetic studies.

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