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采用 UPLC-MS/MS 同时检测子宫肌瘤治疗用消瘤汤(FM1523)中的四种生物活性化合物。

Application of UPLC-MS/MS to simultaneously detect four bioactive compounds in the tumour-shrinking decoction (FM1523) for uterine fibroids treatment.

机构信息

School of Chinese Medicine, LKS Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong.

Department of Pharmacology and Pharmacy, LKS Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong.

出版信息

Phytochem Anal. 2019 Jul;30(4):447-455. doi: 10.1002/pca.2827. Epub 2019 Mar 27.

Abstract

INTRODUCTION

The Chinese medicine formulation, tumour-shrinking decoction (TSD, FM1523), which consists of 15 natural medicines, is used for uterine fibroids (UFs) therapy and possesses excellent clinical therapeutic effect.

OBJECTIVE

To develop a sensitive and validated analytical method for the simultaneous quantification of four crucial bioactive compounds including isorhamnetin-3-O-neohesperidoside, curcumin, peimine and tetrahydropalmatine in the principal formulation of this decoction.

METHODS

An ultra-performance liquid chromatography coupled tandem mass spectrometry (UPLC-MS/MS) with an electrospray ionisation (ESI) source in multiple reaction monitoring (MRM) mode was conducted to investigate these bioactive compounds in the TSD. The chromatographic separation was performed on a C18 column when the flow rate was adjusted at 0.2 mL/min with gradient elution of acetonitrile-water with 0.1% formic acid. Accelerated solvent extraction (ASE) method with higher extraction efficiency was employed for TSD sample pre-treatment.

RESULTS

The linearity, limit of detection (LOD) and limit of quantification (LOQ) were determined for this analytical method. The mean recoveries of the compounds were determined between 100.23% and 104.02% with satisfactory relative standard deviation (RSD) in the ranges of 2.65% to 3.81%. The precision was evaluated by intra-day and inter-day tests, which revealed RSD within the ranges of 1.21% to 2.14% and 1.24% to 2.32%, respectively.

CONCLUSION

The bioactive compounds of TSD samples were successfully quantified via UPLC-MS/MS with MRM mode. This study could help to evaluate the pharmacokinetic study of TSD during clinical applications and present a facile strategy for quantifying bioactive compounds in traditional Chinese Medicine decoction.

摘要

简介

中药方剂消瘤汤(TSD,FM1523)由 15 种天然药物组成,用于治疗子宫肌瘤(UFs),具有极佳的临床疗效。

目的

建立一种同时定量分析该方剂中 4 种重要生物活性化合物(异鼠李素-3-O-新橙皮苷、姜黄素、巴马汀和延胡索乙素)的超高效液相色谱串联质谱(UPLC-MS/MS)分析方法。

方法

采用电喷雾离子源(ESI)在多反应监测(MRM)模式下,对 TSD 中的生物活性化合物进行 UPLC-MS/MS 分析。采用 C18 柱,以乙腈-水(含 0.1%甲酸)为流动相,梯度洗脱,流速为 0.2 mL/min,进行色谱分离。采用加速溶剂萃取(ASE)法提取 TSD 样品,以提高提取效率。

结果

建立了该分析方法的线性、检测限(LOD)和定量限(LOQ)。化合物的平均回收率在 100.23%至 104.02%之间,相对标准偏差(RSD)在 2.65%至 3.81%之间,均令人满意。日内和日间精密度试验的 RSD 分别在 1.21%至 2.14%和 1.24%至 2.32%之间。

结论

采用 UPLC-MS/MS 结合 MRM 模式成功定量测定了 TSD 样品中的生物活性化合物。该研究有助于评估 TSD 在临床应用中的药代动力学研究,并为中药方剂中生物活性化合物的定量提供了一种简便的策略。

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