| Gynoslemma pentaphyllum, is a member of Gynostenuna genus in Cucurbitaceae family. It has a variety of effects such as calming the nerves, anti-aging, anti-tumor, and so on. In our previous study, gypenosides were proved to possess significant anti-hepatic fibrosis activity. In present paper, we will focus on the studies of the fingerprint of Gynostenuna pentaphyllum from different sources, the extraction and purification process of gypenosides, and the content determination of main components in gypenosides. In addition, the in vivo metabolites of gypenosides after oral administration were preliminary analyzed and identified by LC-Q-TOF-MS/MS approach. The researches above would lay a foundation for the studies of modern TCM. and the main achievements were shown as follows:1. The herb origin which will be used in the further research was confirmed by the fingerprint study. The fingerprint of ten lots of Gynostemma pentaphyllum from different sources was established first. Gypenoside A (Gyp A) was chosen as the index component, and the overall similarity of herbs was calculated by similarity software. The similiarity of the samples S3and S6were over0.99. which of S1, S4, S5, S7, S8, S9,S10were above0.94. however, only S2was below0.9. Combined with the results of the similarity analyses, principal component contents and consideration of cost, Shaanxi Ankang was chosen for the herb origin for further research and production of’Gynostemma pentaphyllum.2. The methods of extraction and purification of gypenosides from crude drugs were established. The extraction factors such as solvent and time, were investigated by orthogonal test, and then the optimized extraction was confirmed as:,40%ethanol as solvent, the liquid ratio of6:1, extracting for3times and1.5h for each time. The crude extract was subjected to the macroporous absorptive resin column, and then the gypenosides fraction were obtained.3. A method of determination of the contents of two main components in gypenosides by HPLC-ELSD was constructed. The linear equation of gypenoside XLIX was y=1.5489x+5.4436(R=0.9997). The values of RSD for precision, stability, repeatability were0.49%.0.89%,1.13%. respectively. The average sample recovery rate was97.48%. and RSD was1.17%. The linear equation of gypenoside A is y=1.7578x+4.8426(R=0.9996). The values of RSD for precision, stability, repeatability were0.44%,0.93%,1.23%,respectively. The average sample recovery rate was97.85%. and RSD was1.03%. It indicated that this method was reliable and stable, which was further proved to be rational and feasible according to the results of content determination of three batches of gypenosides prepared by our lab.4. The drug-containing plasma and urine of rats after oral administration of gypenosides (including prototype and metabolites) were preliminary analyzed by LC-HSI-Q-TOF-MS/MS method. Gyp XLIX and Gyp A could be detected in both plasma and urine, particularly, six metabolites were identified in urine. The results demonstrated that all of the prototypes of gypenosides can be absorbed into the blood, which could be a reliable basis for the quality control of Gynostemma pentaphyllum. |
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