Department of Complex Prescription of Traditional Chinese Medicine, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, PR China; Jiangsu Key Laboratory of TCM Evaluation and Translation Research, China Pharmaceutical University, Nanjing 211198, PR China.
Department of Complex Prescription of Traditional Chinese Medicine, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, PR China; Centre of Pear Engineering Technology Research, State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, PR China.
Anal Chim Acta. 2017 Sep 1;984:124-133. doi: 10.1016/j.aca.2017.07.023. Epub 2017 Jul 14.
Natural products, such as rosmarinic acid and apigenin, can act as xanthine oxidase inhibitors (XOIs) as well as superoxide anion scavengers, and have potential for treatment of diseases associated with high uric acid levels and oxidative stress. However, efficient simultaneous screening of these two bioactivities in natural products has been challenging. We have developed a novel method by assembling a multi-hyphenated high performance liquid chromatography (HPLC) system that combines a photo-diode array, chemiluminescence detector and a HPLC system with a variable wavelength detector, to simultaneously detect components that act as both XOIs and superoxide anion scavengers in natural products. Superoxide anion scavenging activity in the analyte was measured by on-line chemiluminescence chromatography based on pyrogallol-luminol oxidation, while xanthine oxidase inhibitory activity was determined by semi-on-line HPLC analysis. After optimizing multiple elements, including chromatographic conditions (e.g., organic solvent concentration and mobile phase pH), concentrations of xanthine/xanthine oxidase and reaction temperature, our validated analytical method was capable of mixed sample analysis. The final results from our method are presented in an easily understood visual format including comprehensive bioactivity data of natural products.
天然产物，如迷迭香酸和芹菜素，可作为黄嘌呤氧化酶抑制剂（XOIs）和超氧阴离子清除剂，具有治疗与高尿酸水平和氧化应激相关疾病的潜力。然而，在天然产物中高效地同时筛选这两种生物活性一直具有挑战性。我们开发了一种新方法，通过组装多联式高效液相色谱（HPLC）系统来实现，该系统结合了光电二极管阵列、化学发光检测器和具有可变波长检测器的 HPLC 系统，以同时检测天然产物中同时具有 XOIs 和超氧阴离子清除活性的成分。分析物中的超氧阴离子清除活性通过基于邻苯三酚-鲁米诺氧化的在线化学发光色谱法进行测量，而黄嘌呤氧化酶抑制活性则通过半在线 HPLC 分析进行测定。在优化了包括色谱条件（例如有机溶剂浓度和流动相 pH）、黄嘌呤/黄嘌呤氧化酶浓度和反应温度在内的多个因素后，我们验证的分析方法能够进行混合样品分析。最终结果以易于理解的可视化格式呈现，包括天然产物的综合生物活性数据。
