Study On The Law Of Cracking Of Saponins In Panax Notoginseng And The Study Of Its Chemical Characteristics

ObjectivePanax notoginseng (Burk.) F.H. Chen or Sanqi in Chinese is a well-known Chinese medicinal herb, which has been used for thousands of years in China due to its good hemostatic effect. P. notoginseng is widely cultivated in Yunnan and Guangxi provinces. Genuine medicinal meterials with special characteristics of TCM, is recognized as high quality medicine. For P. notoginseng, the genuine place is Wenshan county of Yunnan province. The genuine P. notoginseng possesses unique quality characteristics, which are the key to distinguish from other origins. Yet, P. notoginseng research of origin factors related to its chemical composition system is not very deep. The common method to determine the content of saponins in the P. notoginseng is HPLC, which limits the determination of low content composition. Because of the continuous cropping obstacles of cultivation, the cultivating area has been enlarged, from the traditional areas to outer new areas. So, whether the expansion of the planting area is reasonable, and what is the difference between the genuine place and new areas need to be researched.This study included four parts as below:(1) To summarize the fragmentation pathways characteristics of various types of saponin based on the UPLC-MS determination of many saponins reference, and establish the mass spectrometry database for identify of unknown structural compounds; (2) Collection of numerous samples of P.notoginseng from different origins, to establish a determination method for the main components of Panax saponins, explore the correlation of compositions and origins.(3) To determine the main heavy metals in P. notoginseng, then analyse the region difference of heavy metal contents; (4) To determine the trace elements in P. notoginseng and establish elements fingerprint. Thus, all the saponins, heavy mentals, trace elements can reflect the characteristics of different regions in P.notoginseng.Methods1. UPLC-Q-TOF/MS was used for the determination of 17 kinds of saponins reference, The fragmentation behavior of these ginsenosides were researched in negative ion mode by MS/MS, all the ginsenosides could be distinguished and identified, and the rules of the fragmentation pathway was produced to determine sugar numbers, positions, linkages, and isomers.2. P. Notoginseng samples were extracted by reflux, and HPLC-MS was applied to establish a method for quantitative determination of five main saponin components, and all samples were determined. The trace ingredients of saponins were deduced.3. The method of wet digestion and inductively coupled plasma emission spectroscopy (ICP-AES) were used to quantified six kinds of heavy metals As, Cu, Hg, Cd, Pb and Cr for all the notoginseng samples.4. By microwave digestion, using ICP-AES technique, seven kinds of trace elements Ca, Cu, Fe, Mg, Mn, Se and Zn were determined for all samples of notoginseng.Results and conclusion1. The relationships between structural characteristics and fragmentation ways have been summarized for 17 standards. All the saponins were perfectly separated with high resolution and short time by the use of UPLC. Lots of diagnostic product ions useful for structure characterization have been summarized, which will be of great significance for the research of compounds identification, especially for P. notoginseng.2. The contents of ginsenoside Rg1, Re, Rd, Rb1, notoginseng R1 in P. notogingseng were determined using UPLC-MS, the content was o.93%,2.86%,0.64%,2.78%,1.61% respectively. Five of total saponins content in everage was 7.48%. Fourther more, all the 28 chromatographic peaks were identified by the fragment pathways. Studing on correlation of saponins, the ginsenoside Rb1-Rg1; ginsenoside Rbi-Rd1; ginsenoside Rd-Rg11; ginsenoside Rd-Re showed a significant positive correlation.3. Six heavy metals, including As, Cu, Hg, Cd, Pb and Cr in P. notoginseng were determined by ICP-AES, the results showed that the content of As and Cd exceeded the limit of the standard and the percentage-was 32.4%and Cd 29.7%, respectively. The occurrence of contained heavy metals has been discussed.4. Seven trace elements, including Ca, Cu, Fe, Mg, Mn, Se, Zn in P. notoginseng were determined by ICP-AES, feature maps was established, and the analysis results show that there exists difference between genuine areas and new areas in the contents of Zn and Mn. Besides, content data show that Fe and Cu, Mn was significantly positively related in the contents.In this study, modern technology such as UPLC-Q-TOF-MS, ICP-AES are used to determine and analysis the saponins contents, trace elements contents and heavy metal elements contents of lots of P. notoginseng samples. In this study, we have thoroughly analyzed genuine P. notoginseng, overcome the shortcomings of previous studies which are not deep. The results showed that there is no significant difference between genuine areas and infamous area of the type and content of saponin, trace elements and heavy metal elements. Combined with the research of this experiment with the P. notoginseng leaf samples corresponding parts of the DNA sequencing and analysis extract also showed there is no significant genuineness genetically. The reason may be all the seed sources of P.notoginseng are come from wenshan,Yunnan province. Therefor, the P.notoginseng planted in the scope expanded to neighboring Wenshan, Yunnan province is reasonable on the chemical characteristics.The study on chemical composition of authentic ingredients provides a feasible research ideas.