基于TLC-生物自显影-DART-MS-自动分离纯化系统集成的天然脂肪酶抑制剂的分离分析新策略

Obesity has become one of the top ten diseases that seriously threaten human health, and anti-obese or regulating body weight is a hot topic area of scientific research. Anti-obese drugs from TCM are considered as safer and more effective than Western medicine. The project is designed for a new strategy for interpretation of the anti-obese active components（lipase inhibitor） in Chinese medicines complex samples, by integrating Thin layer chromatography - Bioautography - Real-time direct analysis- Mass spectrometry- AutoPurification system（(TLC-Bioautography-DART-MS- AutoPurification）. As the first step，TLC-Bioautography will be conducted to indicating specific bioactivities of the spots in an in situ manner; Then the positive spots will be induced to a DART–MS for rapid identification of target constituents. A parallel sample of the same extractive of the crude drug will be detected by use of LC-MSn to display a characteristic fingerprinting profile．The intended fractions is subjected to the HPLC-MS-AutoPurification system for high-throughput preparation of the target compounds, which will be structurally identified and further assayed for bioactivities. This research will be benefit for rapid discovery of natural lipase inhibitors, providing lead compounds for new anti-obese drug research. Meanwhile, it will provide a new idea and new model for the rapid screening and separation of other natural active ingredients.

肥胖已成为严重威胁人类健康的十大疾病之一，如何有效减少和控制肥胖已成为当前面临的重要科学研究课题。从中药等天然产物中探寻减肥药物，具有西药无法比拟的安全性和有效性。本项目拟采用薄层色谱-生物自显影－实时直接分析－质谱联用技术，建立中药复杂体系中减肥活性成分的快速分离分析新策略。首先运用薄层色谱-生物自显影方法，实现色谱分离与活性检测的同步表征；进而运用原位、实时、直接（DART）-高分辨质谱分析，对目标成分进行快速鉴定；进一步运用LC-MSn进行特征离子捕获，建立反映特定中药内在品质的特征指纹图谱；再运用质谱导向的高效液相自动纯化系统，对目标成分进行高通量制备纯化并进行活性验证。该方法的建立与应用,一方面可快速发现减肥活性成分,为新型减肥药物研究提供先导化合物。另一方面,也为其他天然活性成分的快速筛选和制备提供思路和方法。

肥胖严重威胁人类的健康，从中药等天然药物中探寻作用于外周抑制脂肪酶吸收这个靶点的减肥药物，具有西药无法比拟的安全性和有效性。因此，本项目建立了薄层色谱-生物自显影－实时直接分析－质谱联用技术与生物活性直接相关的筛选和评价方法，能够更快速准确的从中药中定位、分离和鉴定减肥降脂活性成分。. 随后探索该TLC-DART-MS联用方法在中药多种成分类型中的应用；将其成功运用到传统中药三七中具有脂肪酶抑制活性成分的筛选、分离和结构解析。接着运用LC-MSn进行特征离子捕获，建立反映特定中药内在品质的特征指纹图谱；运用质谱导向的高效液相自动纯化系统，对目标成分进行高通量制备纯化、结构鉴定。同时，采用细胞3T3-L1-PPAR进行体外活性验证，一共有10个化合物具有较好的活性。. 该项目的完成为薄层色谱生物自显影技术与原位电离质谱联用，充分将薄层色谱生物自显影从三七复杂体系中直观分离、锁定和评价活性成分的特点与原位电离质谱直接解吸活性斑点结构的特点有机结合，快速筛选获得三七中具有减肥降脂的活性成分。该方法的建立及应用将为中药等天然药物减肥降脂活性成分的快速筛选和鉴定提供新的思路和新的方法。

内容获取失败，请点击重试