基于UPLC-Q TOF MS/MS和HPLC的祛瘀散结胶囊的化学成分分析及其多成分含量测定<sup>*</sup>

甘金月, 王宝林, 丘海芯, 刘振杰, 陈道峰, 高红伟, 奉建芳, 唐红珍, 覃喜军

doi:10.16155/j.0254-1793.2024.01.03

药物分析杂志 >

2024 , Vol. 44 >Issue 1: 23 - 34

DOI: https://doi.org/10.16155/j.0254-1793.2024.01.03

成分分析

基于UPLC-Q TOF MS/MS和HPLC的祛瘀散结胶囊的化学成分分析及其多成分含量测定^*

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1.广西中医药大学,南宁 530299;
2.复旦大学,上海 201203;
3.广西优势中成药与民族药开发工程技术研究中心,南宁 530299

第一作者 Tel:15578169093;E-mail:1162429136@qq.com

^**Tel:18877177612;E-mail:114525914@qq.com

修回日期: 2023-12-18

网络出版日期: 2024-06-21

基金资助

^*国家自然科学基金(NO.81860270, NO.81860711);广西中医药大学2017引进博士启动基金项目(2017BS031);广西科技基地与人才专项(桂科AD19245124);中国-东盟传统医药发展研究中心开放课题(0501801726)

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Analysis of chemical components and determination of multi-component contents in Quyusanjie capsules based on UPLC-Q TOF MS/MS and HPLC^*

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1. Guangxi University of Chinese Medicine, Nanning 530299, China;
2. Fudan University, Shanghai 201203, China;
3. Guangxi Engineering Technology Research Center of Advantage Chinese Patent Drug and Ethnic Drug Development, Nanning 530299, China

Revised date: 2023-12-18

Online published: 2024-06-21

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摘要

目的:应用液质联用法分析祛瘀散结胶囊的化学成分,构建祛瘀散结胶囊中有效成分的含量测定方法。方法:采用超高效液相色谱-三重四极杆飞行时间质谱联用(UPLC-Q TOF MS/MS)技术,色谱柱为Hypersil Gold C₁₈(100 mm×2.1 mm,1.9 μm),以乙腈(A)-0.1%甲酸水(B)为流动相,梯度洗脱,体积流量0.4 mL·min^-1,柱温40.0 ℃,质谱数据采集为负离子模式扫描;通过数据库匹配、元素组成和碎片结构分析,鉴定祛瘀散结胶囊中的主要化学成分。另外,采用HPLC,色谱柱为Ultimate^®AQ-C₁₈(250 mm×4.6 mm,5 μm),以乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱,体积流量1 mL·min^-1,柱温25 ℃,检测波长203 nm;以外标法计算,测定了11个不同批次祛瘀散结胶囊中的柚皮苷、新橙皮苷、三七皂苷R₁、人参皂苷Rg₁、人参皂苷Rb₁的含量,并以人参皂苷Rg₁为内参物,建立一测多评法。结果:从祛瘀散结胶囊中鉴定出29个化合物;外标法测得柚皮苷、新橙皮苷、三七皂苷R₁、人参皂苷Rg₁、人参皂苷Rb₁的含量分别为0.484~1.097、0.341~0.618、1.685~2.399、5.748~8.386、3.868~5.898 mg·g^-1,一测多评法测得的柚皮苷、新橙皮苷、三七皂苷R₁、人参皂苷Rb₁的含量分别为0.516~1.153、0.372~0.667、1.794~2.580、4.373~6.690mg·g^-1,一测多评法计算值与外标法实测值之间相对误差≤8.9%。结论:UPLC-Q TOF MS/MS法能快速鉴定祛瘀散结胶囊的化学成分;建立的外标法稳定可靠,可用于祛瘀散结胶囊的质量控制;以人参皂苷Rg₁为内参物建立的一测多评法有较好的可行性,适用于祛瘀散结胶囊日常生产的含量测定。

关键词： 祛瘀散结胶囊; 化学成分分析; 含量测定; 一测多评; 柚皮苷; 新橙皮苷; 三七皂苷R₁; 人参皂苷; 超高效液相色谱-三重四极杆飞行时间质谱; 高效液相色谱

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甘金月, 王宝林, 丘海芯, 刘振杰, 陈道峰, 高红伟, 奉建芳, 唐红珍, 覃喜军 . 基于UPLC-Q TOF MS/MS和HPLC的祛瘀散结胶囊的化学成分分析及其多成分含量测定^*[J]. 药物分析杂志, 2024 , 44(1) : 23 -34 . DOI: 10.16155/j.0254-1793.2024.01.03

Abstract

Objective: To analyze the chemical constituents of Quyusanjie capsules by LC/MS, and establish a method for the determination of active ingredients in Quyusanjie capsules. Methods: Using UPLC-Q TOF MS/MS technology, the Hypersil Gold C₁₈ column(100 mm×2.1 mm,1.9 μm) was used, the mobile phase was acetonitrile(A) and 0.1% formic acid in water(B) with gradient elution, at a flow rate of 0.4 mL·min^-1, the the column temperature was 40.0 ℃, and the mass spectrometry data was collected by negative ions mode scanning. Through database matching, elemental composition and fragment structure analysis, the main chemical substances in Quyusanjie capsules were identified. HPLC was used to qualitatively analyze the chemical components of Quyusanjie capsules. The Ultimate^® AQ-C₁₈column(250 mm×4.6 mm, 5 μm) was used, the mobile phase was acetonitrile(A)-0.1% phosphoric acid(B) with gradient elution at the flow rate of 1.0 mL·min^-1, the column temperature was 25 ℃, and the detection wavelength was 203 nm. The content of naringin, neohesperidin, notoginsenoside R₁, ginsenoside Rg₁, and ginsenoside Rb₁ in 11 different batches of Quyusanjie capsules were determined using external standard method. QAMS method was established using ginsenoside Rg₁ as the internal reference. Results: Twenty-nine compounds were identified from Quyusanjie capsule. The contents of naringin, neohesperidin, notoginsenoside R₁, ginsenoside Rg₁ and ginsenoside Rb₁ measured by external standard method were 0.484-1.097 mg·g^-1, 0.341-0.618 mg·g^-1, 1.685-2.399 mg·g^-1, 5.748-8.386 mg·g^-1, 3.868-5.898 mg·g ^-1, respectively. Measured with the QAMS method, the contents of naringin, neohesperidin, notoginsenoside R₁ and ginsenoside Rb₁ were 0.516-1.153 mg·g^-1, 0.372-0.667 mg·g^-1, 1.794-2.580 mg·g^-1, 4.373-6.690 mg·g^-1, respectively. The relative error between the calculated values of the QAMS method and the measured value of the external standard method was less than 8.9%. Conclusion: UPLC-Q TOF MS/MS method can quickly identify the chemical components of Quyusanjie capsules. The established external standard method is stable and reliable, and can be used for the quality control of Quyusanjie capsules. The method of QAMS has good feasibility and is suitable for the determination of the daily production of Quyusanjie capsules.

Key words： Quyusanjie capsules; chemical composition analysis; content determination; QAMS; naringin; neohesperidin; notoginsenoside R₁; ginsenoside; UPLC-Q TOF MS/MS; HPLC

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