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基于填充吸附剂微萃取的细胞膜色谱-气相色谱-质谱联用分析方法的建立及其在天然产物挥发性活性成分筛选中的应用

Development of an analytical method coupling cell membrane chromatography with gas chromatography-mass spectrometry via microextraction by packed sorbent and its application in the screening of volatile active compounds in natural products.

作者信息

Li Miao, Wang Sicen, He Langchong

机构信息

The First Affiliated Hospital of Xi'an Jiaotong University, No. 277, Yanta West Road, Xi'an 710061, Shaanxi, China.

School of Pharmacy, Health Science Center, Xi'an Jiaotong University, No. 76, Yanta West Road, Xi'an 710061, Shaanxi, China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2015 Jan 1;974:9-16. doi: 10.1016/j.jchromb.2014.10.019. Epub 2014 Oct 24.

Abstract

Natural products (NPs) are important sources of lead compounds in modern drug discovery. To facilitate the screening of volatile active compounds in NPs, we have developed a new biochromatography method that uses rat vascular smooth muscle cells (VSMC), which are rich in L-type calcium channels (LCC), to prepare the stationary phase. This integrated method, which couples cell membrane chromatography (CMC) with gas chromatography-mass spectrometry (GC-MS) via microextraction by packed sorbent (MEPS) technology, has been termed VSMC/CMC-MEPS-GC-MS. Methodological validation confirmed its specificity, reliability and convenience. Screening results for Radix Angelicae Dahuricae and Fructus Cnidii obtained using VSMC/CMC-MEPS-GC-MS were consistent with those obtained using VSMC/CMC-offline-GC-MS. MEPS connection plays as simplified solid-phase extraction and replaces the uncontrollable evaporation operation in reported offline connections, so our new method is supposed to be more efficient and reliable than the offline ones, especially for compounds that are volatile, thermally unstable or difficult to purify. In application, senkyunolide A and ligustilide were preliminary identified as the volatile active components in Rhizoma Chuanxiong. We have thus confirmed the suitability of VSMC/CMC-MEPS-GC-MS for volatile active compounds screening in NP.

摘要

天然产物是现代药物发现中先导化合物的重要来源。为便于筛选天然产物中的挥发性活性化合物,我们开发了一种新的生物色谱方法,该方法利用富含L型钙通道(LCC)的大鼠血管平滑肌细胞(VSMC)制备固定相。这种通过填充吸附剂微萃取(MEPS)技术将细胞膜色谱(CMC)与气相色谱-质谱联用(GC-MS)的集成方法,被称为VSMC/CMC-MEPS-GC-MS。方法学验证证实了其特异性、可靠性和便利性。使用VSMC/CMC-MEPS-GC-MS对白芷和蛇床子进行筛选的结果与使用VSMC/CMC离线-GC-MS获得的结果一致。MEPS连接起到简化固相萃取的作用,并取代了报道的离线连接中不可控的蒸发操作,因此我们的新方法应该比离线方法更高效、更可靠,特别是对于挥发性、热不稳定或难以纯化的化合物。在应用中,初步鉴定出川芎内酯A和藁本内酯为川芎中的挥发性活性成分。因此,我们证实了VSMC/CMC-MEPS-GC-MS适用于天然产物中挥发性活性化合物的筛选。

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