Liu Bingrui, Kongstad Kenneth T, Qinglei Sun, Nyberg Nils T, Jäger Anna K, Staerk Dan
Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen , Universitetsparken 2, DK-2100 Copenhagen, Denmark.
J Nat Prod. 2015 Feb 27;78(2):294-300. doi: 10.1021/np5009416. Epub 2015 Feb 13.
The crude methanol extract of Pueraria lobata was investigated by dual high-resolution α-glucosidase inhibition and radical scavenging profiling combined with hyphenated HPLC-HRMS-SPE-NMR. Direct analysis of the crude extract without preceding purification was facilitated by combining chromatograms from two analytical-scale HPLC separations of 120 and 600 μg on-column, respectively. High-resolution α-glucosidase and radical scavenging profiles were obtained after microfractionation of the eluate in 96-well microplates. This allowed full bioactivity profiling of individual peaks in the HPLC chromatogram of the crude methanol extract. Subsequent HPLC-HRMS-SPE-NMR analysis allowed identification of 21 known compounds in addition to two new compounds, i.e., 3'-methoxydaidzein 8-C-[α-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside and 6″-O-malonyl-3'-methoxydaidzin, as well as an unstable compound tentatively identified as 3'-de-O-methylpuerariafuran.
采用双高分辨率α-葡萄糖苷酶抑制和自由基清除分析，并结合联用的HPLC-HRMS-SPE-NMR技术，对葛根的粗甲醇提取物进行了研究。通过分别对120μg和600μg柱上进样量进行两次分析规模的HPLC分离，将色谱图合并，实现了对未经预先纯化的粗提取物的直接分析。将洗脱液在96孔微孔板中进行微分离后，获得了高分辨率的α-葡萄糖苷酶和自由基清除图谱。这使得能够对粗甲醇提取物的HPLC色谱图中的各个峰进行完整的生物活性分析。随后的HPLC-HRMS-SPE-NMR分析除了鉴定出两种新化合物，即3'-甲氧基大豆苷元8-C-[α-D-芹菜呋喃糖基-(1→6)]-β-D-吡喃葡萄糖苷和6″-O-丙二酰基-3'-甲氧基大豆苷之外，还鉴定出21种已知化合物，以及一种暂定为3'-去-O-甲基葛根呋喃的不稳定化合物。
