A Strategy for Pinpointing Natural Bioactive Components Using Two-Dimensional Bioassay Profilings Combined with Comprehensive Two-Dimensional Countercurrent Chromatography x High-Performance Liquid Chromatography
A Strategy for Pinpointing Natural Bioactive Components Using Two-Dimensional Bioassay Profilings Combined with Comprehensive Two-Dimensional Countercurrent Chromatography x High-Performance Liquid Chromatography
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DOI：
10.1021/acs.analchem.2c02196
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发表时间：
2022-09-08
影响因子：
7.4
通讯作者：
Tong, Shengqiang
中科院分区：
文献类型：
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作者：
Xu, Ping;Wang, Xiang;Tong, Shengqiang
受二维(2D)核磁共振波谱解释的启发，提出了一种从复杂天然产物中精确定位生物活性成分的有效策略。采用离线综合逆流色谱(CCC)×高效液相色谱(HPLC)建立二维化学图谱，通过一维(D-1)CCC淋洗液和二维(D-2)HPLC淋洗液的生物测定获得二维生物测定图谱。然后，将2D化学色谱图和2D生物测定图谱进行匹配，以从复杂基质中精确定位生物活性天然成分。因此，可以有效地分析、分离和确定复杂基质中的生物活性成分。这一实验方案以传统中草药虎杖为例得到了成功的验证。Et Zucc.通过酪氨酸酶抑制实验、α-葡萄糖苷酶抑制实验、DPPH自由基清除实验和ABTS(中心点+)脱色实验验证了该2D策略的可行性。从虎杖中成功地定位和鉴定了8个天然抑制物。2‘’-O-没食子酸酯(10)和香草苷B(20)均为首次筛选鉴定为天然酪氨酸酶抑制剂。同时，香草苷B(20)也是所有分离的成分中最强的α-葡萄糖苷酶抑制剂。大多数化合物都表现出更高的自由基清除活性。与传统的基于一维色谱分离的方法相比，本文提出的二维方法将更加精确、高效、方便地从复杂基质中筛选和分离生物活性物质。
Inspired by the interpretation of two-dimensional (2D) nuclear magnetic resonance spectra, an efficient strategy was proposed for pinpointing bioactive components from complex natural products. An off-line comprehensive countercurrent chromatography (CCC) x high-performance liquid chromatography (HPLC) was employed to achieve a 2D chemical chromatogram, and 2D bioassay profilings were obtained from bioassays of the eluent of the first dimension (D-1) CCC and the eluent of the second dimension (D-2) HPLC. Then 2D chemical chromatograms and 2D bioassay profilings were matched for pinpointing bioactive natural components from complex matrices. Thus, bioactive components in a complex matrix could be efficiently analyzed, separated, and bioactivity-determined. This experimental scheme was successfully demonstrated with a traditional medicinal herb Polygonum cuspidatum Sieb. et Zucc. The feasibility of this 2D strategy was verified with tyrosinase inhibition assay, alpha-glucosidase inhibition assay, DPPH radical scavenging assay, and ABTS(center dot+) decolorization assay. Eight natural inhibitors were successfully pinpointed and identified from P. cuspidatum. Both pieceid-2 ''-O-gallate (10) and vanicoside B (20) were screened and identified as natural tyrosinase inhibitors for the first time. Meanwhile, vanicoside B (20) was also found as the strongest alpha-glucosidase inhibitor among all the isolated components. Most of the compounds exhibited much higher radical scavenging activities. Compared with traditional methodology based on one-dimensional chromatographic separation, the present 2D strategy would be more precise, efficient, and convenient to screen and separate bioactive compounds from complex matrices.