Chen Yeqing, Yu Hongli, Wu Hao, Pan Yaozong, Wang Kuilong, Liu Liping, Jin Yangping, Zhang Chenchao
School of pharmacy, Nanjing University of Chinese Medicine, Nanjing, P. R. China.
Jiangsu Key Laboratory of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing, P. R. China.
J Sep Sci. 2015 May;38(10):1691-9. doi: 10.1002/jssc.201500100. Epub 2015 Apr 17.
Flos Sophorae and its processed product have been clinically used to treat hemorrhage. In this study, the total ion chromatographic fingerprints of the heating products of total flavonoids in Flos Sophorae were established by high-performance liquid chromatography with tandem mass spectrometry and the hemostatic activities were studied by hemostatic screening tests in vivo. The spectrum-effect relationships between fingerprints and hemostatic activities were investigated using canonical correlation analysis to trace the peaks responsible for the hemostatic effects. The predicted active peaks in fingerprints were isolated by column chromatography and their structures were identified by NMR spectroscopy and mass spectrometry. The hemostatic activities of them were verified by platelet aggregation and procoagulation assays in vitro. Canonical correlation analysis results showed that peak 8 and peak 11 were correlated most closely, thus probably being the main hemostatic compounds. Through column chromatography separation, peak 8 (compound I) and peak 11 (compound II) were obtained with purities of 95.61 and 93.38%, respectively, and were discovered new hemostatic compounds named as huaicarbon A (I) and huaicarbon B (II), respectively. This study provides a universal model to trace the active compounds of other herbs which have bioactivity enhancement after processing by spectrum-effect relationships and column chromatography.
槐花及其炮制品在临床上一直用于治疗出血症。本研究采用高效液相色谱 - 串联质谱法建立了槐花总黄酮加热产物的总离子色谱指纹图谱，并通过体内止血筛选试验研究了其止血活性。利用典型相关分析研究指纹图谱与止血活性之间的谱效关系，以追踪具有止血作用的峰。通过柱色谱法分离指纹图谱中预测的活性峰，并通过核磁共振光谱和质谱鉴定其结构。通过体外血小板聚集和促凝血试验验证了它们的止血活性。典型相关分析结果表明，峰8和峰11的相关性最密切，因此可能是主要的止血化合物。通过柱色谱分离，分别得到了纯度为95.61%和93.38%的峰8（化合物I）和峰11（化合物II），并发现它们分别是名为槐碳A（I）和槐碳B（II）的新型止血化合物。本研究提供了一个通用模型，通过谱效关系和柱色谱法追踪其他经炮制后具有生物活性增强的草药的活性成分。
