基于衍生化的葛根治疗2型糖尿病的羰基化合物的分析技术研究

Composition of traditional Chinese medicine (TCM) is complicated, and the content of most of them is very low. While it’s more difficult to detect when they enter the body. So the detection of ingredients with low content in TCM and their metabolites in vivo become the bottleneck of material basic research for TCM. So it’s necessary to develop a highly sensitive analysis method. Liquid chromatography - mass spectrometry(LC-MS) is widely used in material basic research of TCM because of its high sensitivity. However, The ionization efficiency of carbonyl compounds is low and difficult to detect in the mass spectrum. In our previous studies, we found that Girard's Reagent P (GP) can introduce a permanent charged moiety of quaternary ammonium of GP into the carbonyl compounds, which can dramatically enhanced the detection sensitivities of carbonyl compounds in MS analysis. Based on this, in this work, we used GP as the derivatization reagent and Pueraria Lobata as the research object, who has the curative effect in the treatment of type 2 diabetes (T2DM). And established a analysis method which has the advantage of high sensitivity, high selectivity and high flux. Through this method and combined with statistical analysis, we screened out the biomarkers of carbonyl compounds in T2DM and the carbonyl compounds of Pueraria Lobata in the blood in the treatment of T2DM. And preliminary explan the material basis and functional mechanism of Pueraria Lobata in the treatment of T2DM. This method expanded the visual field of active material analysis of Pueraria Lobata and laid the foundation for the study of the medicinal substance of Pueraria Lobata. And it enrich the functional mechanism of Pueraria Lobata in the treatment of T2DM. At the same time, it provides a methodology for the analysis of carbonyl compounds in TCM.

中药所含成分复杂，大多含量低，进入体内后更难以检测。中药低含量成分及其体内代谢物的检测成为制约中药物质基础研究的瓶颈，亟需高灵敏度的检测方法。LC-MS以其高灵敏度被广泛用于中药物质基础研究，但羰基类化合物离子化效率低，在质谱中难以检测。前期研究发现吉拉德P试剂(GP)可通过向羰基化合物中引入季铵基团来提高其质谱检测灵敏度。基于此，本项目拟以GP为衍生化试剂，以疗效确切的治疗2型糖尿病(T2DM)的中药葛根为研究对象，建立一种高灵敏度、高选择性、高通量的检测羰基化合物的分析方法，并结合统计学分析筛选出T2DM的内源性羰基化合物生物标志物及葛根治疗T2DM的羰基化合物的入血成分，初步阐释葛根治疗T2DM的物质基础和作用机制。该方法拓宽了中药葛根活性物质分析的视野，为葛根药效物质基础研究奠定了基础，并丰富了其治疗T2DM的作用机制。为中药含羰基化合物的分析提供方法学支撑。

中药所含成分复杂，大多含量低，进入体内后更难以检测。中药低含量成分及其体内代谢物的检测成为制约中药物质基础研究的瓶颈，亟需高灵敏度的检测方法。LC-MS以其高灵敏度被广泛用于中药物质基础研究，但羰基类化合物离子化效率低，在质谱中难以检测。前期研究发现吉拉德P试剂(GP)可通过向羰基化合物中引入季铵基团来提高其质谱检测灵敏度。基于此，本项目以GP为衍生化试剂，以疗效确切的治疗2型糖尿病(T2DM)的中药葛根为研究对象，建立了一种高灵敏度、高选择性、高通量的检测羰基化合物的分析方法。该方法对羰基化合物分析有两个优势：（1）通过 GP 衍生，向羰基化合物中引入了季铵.结构，提高了羰基化合物的离子化效率，从而提高了正离子扫描模式下质谱检测灵敏度。（2）GP 衍生后，衍生产物的二级谱图中都含有一个 m/z 80.1 的碎片，基于此特点，可以从众多分析物中筛选出羰基化合物，提高了羰基化合物的选择性。利用建立的方法及结合统计学分析共筛选出19个潜在的T2DM的内源性羰基化合物生物标志物及73个葛根治疗T2DM的羰基化合物的入血成分，其中包括36个是异黄酮类化合物。初步阐释了葛根治疗T2DM的物质基础和作用机制。该方法拓宽了中药葛根活性物质分析的视野，为葛根药效物质基础研究奠定了基础，并丰富了其治疗T2DM的作用机制。为中药含羰基化合物的分析提供方法学支撑。