中国中药杂志

2021, v.46(24) 6435-6446

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基于HPLC-Q-TOF-MS和UPLC-QQQ-MS/MS的东革阿里生物碱成分分析
Qualitative and quantitative analysis of alkaloids in Eurycoma longifolia by HPLC-Q-TOF-MS combined with UPLC-QQQ-MS/MS

陈秀明;林斌;方东升;
CHEN Xiu-ming;LIN Bin;FANG Dong-sheng;Fujian Provincial Key Laboratory of Screening for Novel Microbial Products, Fujian Institute of Microbiology;

摘要(Abstract):

采用高效液相色谱-串联四极杆飞行时间质谱(HPLC-Q-TOF-MS)结合超高效液相色谱-三重四极杆串联质谱(UPLC-QQQ-MS/MS)对东革阿里提取物中的总生物碱进行定性和定量分析。HPLC-Q-TOF-MS采用Welch Ultimate XB-C_(18)色谱柱(4.6 mm×250 mm, 5μm),以乙腈(内含0.1%甲酸)-0.1%甲酸水为流动相进行梯度洗脱。UPLC-QQQ-MS/MS采用Agilent Eclipse Plus C_(18)色谱柱(2.1 mm×50 mm, 1.8μm),以乙腈(内含0.1%甲酸)-0.1%甲酸水为流动相进行梯度洗脱。质谱分析均采用电喷雾正离子模式采集。HPLC-Q-TOF-MS通过对照品比对及高分辨质谱数据解析,鉴定了东革阿里中17种生物碱成分并解析了其裂解规律。UPLC-QQQ-MS/MS定量分析东革阿里中3种生物碱成分,在考察的浓度范围内线性关系良好(r≥0.999 7),回收率为108.8%~110.2%,RSD为2.9%~5.3%。该方法快速、可靠、高效,能较全面地反映东革阿里中的生物碱成分及其含量,可为深入阐明东革阿里的药效物质基础及其质量标准研究提供参考。
A sensitive and efficient method was established and validated for qualitative and quantitative analysis of total alkaloids from the extract of Eurycoma longifolia by high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS) combined with ultra-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry(UPLC-QQQ-MS/MS). The HPLC-Q-TOF-MS conditions are as follows: Welch Ultimate XB-C_(18) column(4.6 mm×250 mm, 5 μm) with acetonitrile(containing 0.1% formic acid)-0.1% formic acid in water as mobile phase for gradient elution. The UPLC-QQQ-MS/MS conditions are as below: Agilent Eclipse Plus C_(18) column(2.1 mm×50 mm, 1.8 μm) with acetonitrile(containing 0.1% formic acid) and 0.1% formic acid in water as mobile phase for gradient elution. MS data were collected by electrospray ionization in positive ion mode. According to the comparison with reference standards and the accurate masses of molecules, a total of 17 alkaloids in E. longifolia extract were identified by HPLC-Q-TOF-MS. The UPLC-QQQ-MS/MS quantitative analysis result of 3 alkaloids showed that the linear ranges of them were good(r≥0.999 7) and the overall recoveries ranged from 108.8%-110.2%, with RSDs of 2.9%-5.3%. The method is accurate, reliable, and efficient, which can comprehensively reflect the constituents and content of alkaloids in E. longifolia. The result can serve as a reference for further elucidating its therapeutic material basis and quality control.

关键词(KeyWords): HPLC-Q-TOF-MS;UPLC-QQQ-MS/MS;定性分析;定量分析;生物碱;东革阿里
HPLC-Q-TOF-MS;UPLC-QQQ-MS/MS;qualitative analysis;quantitative analysis;alkaloid;Eurycoma longifolia

Abstract:

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基金项目(Foundation): 福建省属公益类科研院所基本科研专项(2017R1009-8)

作者(Author): 陈秀明;林斌;方东升;
CHEN Xiu-ming;LIN Bin;FANG Dong-sheng;Fujian Provincial Key Laboratory of Screening for Novel Microbial Products, Fujian Institute of Microbiology;

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