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一种用于同时测定银杏叶中选定黄酮类化合物的超高效液相色谱-电喷雾电离串联质谱法的开发与验证

Development and validation of an ultra high performance liquid chromatography electrospray ionization tandem mass spectrometry method for the simultaneous determination of selected flavonoids in Ginkgo biloba.

作者信息

Pandey Renu, Chandra Preeti, Arya Kamal Ram, Kumar Brijesh

机构信息

Sophisticated Analytical Instrument Facility, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India; Academy of Scientific and Innovative Research (AcSIR), New Delhi, India.

出版信息

J Sep Sci. 2014 Dec;37(24):3610-8. doi: 10.1002/jssc.201400853. Epub 2014 Nov 4.

DOI:10.1002/jssc.201400853
PMID:25280343
Abstract

A rapid and sensitive ultra high performance liquid chromatography electrospray ionization tandem mass spectrometry method was developed and validated for the simultaneous determination of 13 flavonoids in leaf, stem, and fruit extracts of male and female trees of Ginkgo biloba to investigate gender- and age-related variations of flavonoids content. Chromatographic separation was accomplished on an Acquity UPLC BEH C18 column (50 mm × 2.1 mm id, 1.7 μm) in 5 min. Quantitation was performed using negative electrospray ionization mass spectrometry in multiple reaction monitoring mode. The calibration curves of all analytes showed a good linear relationship (r(2) ≥ 0.9977) over the concentration range of 1-1000 ng/mL. The precision evaluated by an intra- and interday study showed RSD ≤ 1.98% and good accuracy with overall recovery in the range from 97.90 to 101.09% (RSD ≤ 1.67%) for all analytes. The method sensitivity expressed as the limit of quantitation was typically 0.25-3.57 ng/mL. The results showed that the total content of 13 flavonoids was higher in the leaf extract of an old male Ginkgo tree compared to young female Ginkgo trees.

摘要

建立了一种快速灵敏的超高效液相色谱-电喷雾电离串联质谱法,并进行了验证,用于同时测定银杏雌雄树叶片、茎和果实提取物中的13种黄酮类化合物,以研究黄酮类化合物含量与性别和年龄相关的变化。在Acquity UPLC BEH C18柱(50 mm×2.1 mm内径,1.7μm)上5分钟内完成色谱分离。采用负电喷雾电离质谱在多反应监测模式下进行定量分析。所有分析物的校准曲线在1-1000 ng/mL浓度范围内显示出良好的线性关系(r(²)≥0.9977)。通过日内和日间研究评估的精密度显示,所有分析物的RSD≤1.98%,准确度良好,总体回收率在97.90%至101.09%之间(RSD≤1.67%)。以定量限表示的方法灵敏度通常为0.25-3.57 ng/mL。结果表明,与年轻雌银杏树相比,老龄雄银杏树叶片提取物中13种黄酮类化合物的总含量更高。

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