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高效液相色谱-电喷雾电离飞行时间质谱法在黄芪及其制剂分析与质量控制中的应用

Application of high-performance liquid chromatography-electrospray ionization time-of-flight mass spectrometry for analysis and quality control of Radix Astragali and its preparations.

作者信息

Qi Lian-Wen, Li Ping, Ren Mei-Ting, Yu Qing-Tao, Wen Xiao-Dong, Wang Yu-Xia

机构信息

Key Laboratory of Modern Chinese Medicines (China Pharmaceutical University), Ministry of Education, Nanjing 210009, China.

出版信息

J Chromatogr A. 2009 Mar 13;1216(11):2087-97. doi: 10.1016/j.chroma.2008.02.095. Epub 2008 Mar 6.

Abstract

A high-performance liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry (HPLC-ESI-TOF-MS) method has been developed for qualitative and quantitative analysis of isoflavonoids and saponins, as well as for the quality control of Radix Astragali and its preparations. The selectivity, reproducibility and sensitivity are compared with HPLC with diode array detection (DAD) and evaporative light scattering detection (ELSD). Limits of detection and quantification fell in ranges of 9-40 and 23-103 ng/mL for 13 analytes with a injection of 10 microL samples, and all calibration curves showed good linear regression (r(2)>0.9938) within the test range. The assay was successfully utilized to analyze the 13 marker components in 20 samples of Radix Astragali products. The quantitative results demonstrated that samples from different localities and manufacturers showed different quality. Advantages, in comparison with conventional HPLC-DAD-ELSD, are that reliable identification of target compounds could be achieved by accurate mass measurements (<3 ppm) along with characteristic retention time, extracted ions chromatograms using a narrow mass window for quantification ensure that the chromatographic peaks are free from background or co-elutes interference, and the great enhancement in selectivity and sensitivity allows identification and quantification of low levels of constituents in complex Radix Astragali matrixes.

摘要

已开发出一种高效液相色谱-电喷雾电离飞行时间质谱联用(HPLC-ESI-TOF-MS)方法,用于异黄酮和皂苷的定性和定量分析,以及黄芪及其制剂的质量控制。将该方法的选择性、重现性和灵敏度与配备二极管阵列检测(DAD)和蒸发光散射检测(ELSD)的HPLC进行了比较。对于13种分析物,进样10微升样品时,检测限和定量限分别在9-40和23-103纳克/毫升范围内,所有校准曲线在测试范围内均显示出良好的线性回归(r²>0.9938)。该测定法成功用于分析20个黄芪产品样品中的13种标记成分。定量结果表明,来自不同产地和生产商的样品质量不同。与传统的HPLC-DAD-ELSD相比,该方法的优点在于,通过精确质量测量(<3 ppm)以及特征保留时间可实现对目标化合物的可靠鉴定,使用窄质量窗口进行定量的提取离子色谱图可确保色谱峰不受背景或共洗脱干扰,并且选择性和灵敏度的极大提高使得能够在复杂的黄芪基质中鉴定和定量低含量成分。

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