Study On Establishment Of Quality Standard And Fingerprint Of Panax Ginseng Fruit

Panax ginseng fruit is dry ripening fruit of Panax ginseng C.A. Meyer, which belongs to Araliaceae, Panax. Panax ginseng is a perennial herb, mainly produced in Northeast China. At present, there’s a lot of research about underground parts of Panax ginseng in both here and abroad, however there’s few research about aerial parts, especially fruit. The Chinese Pharmacopoeia (2010Edition) develop and embodies the quality standard of Panax ginseng and Panax ginseng leaf, fruit was not included.This paper establishes quality standard for Panax ginseng fruits, using high-performance liquid chromatography (HPLC) and infrared spectroscopy (IR) established a fingerprint of Panax ginseng fruit and application technology of expressed sequence tags simple sequence repeats (EST-SSR) to mark the sample. Specific methods and results are as follows:Panax ginseng fruit characteristics:Growing conditions, processing and medicinal properties of Panax ginseng fruit have been clarified and described mainly through reviewing the documentation and field work.Inspection item of quality standard:Detection method for moisture, total ash, heavy metals and toxic elements, organochlorine pesticide residues have been established. Formulating Panax ginseng fruit herbs moisture may not be too9%; total ash not more than7%; Pb may not have two out of10,000; Cd shall not two out of10,000; As not two out of10,000; Hg must not have two out of10,000; Cu shall not twenty out of10,000; Organochlorine pesticide residues in plants of BHC not one out of10,000; DDT not one out of10,000; PCNB not one out of10,000.10batches of simples as described above was tested, the results showed they were all in line with the provisions.Identification of ginsenoside:Ginsenoside Re/Rgi/Rbi of Panax ginseng fruit have been identified by using thin-layer chromatography (TLC). Just as standard products, the simples showed the same stripe at the same location on the silica gel plate by using above method.Determination of ginsenoside content:The content of total ginsenoside in Panax ginseng fruit have been determinated by UV spectrophotometry. It showed that the average content of total ginsenoside was14.77%in10batch of simples. Formulating containing total ginsenoside as ginsenoside Re shall be not less than12%in the dried Panax ginseng fruit. The content of ginsenoside Re and Rd in Panax ginseng fruit have been determinated by HPLC after optimized conditions. It showed that the average content of ginsenoside Re/Rd was4.5%and2.5%respectively in10batch of simples. Formulating containing ginsenoside Re shall be not less than3.5%and ginsenoside Rd shall be not less than3.5%in the dried Panax ginseng fruit.HPLC fingerprint:After optimizing, the conditions of HPLC fingerprint has been determined as followed:Agilent C18column (250mmx4.6mm,5μm), the mobile phase gradient Elution of acetonitrile-0.05%phosphorylase, flow rate1.0ml/min, detection wavelength203nm, temperature of35℃. It showed that7characteristic peaks were identified as common peak. All common peaks ingredients were respectively identified as ginsenoside Rgi/Re/Rbi/Rg2/Rc/Rb2/Rd.10batches of samples have been analysed for HPLC fingerprint by using chromatographic fingerprints of traditional Chinese medicine software, showed that the similarity of8samples above0.97.IR fingerprint:IR fingerprint of10simples has been established, the result showed Panax ginseng fruit extracts from different simples have the similar composition. On the IR chromatogram, there was a noticeable characteristic peaks at2000cm-1, which could be a basis for qualitative identification of Panax ginseng fruit.DNA fingerprint:DNA fingerprint of10simples have been established by using EST-SSR marker. Screened18of polymorphic primers from the26EST primers. Using that polymorphic primers,165stripes were amplified from10simples DNA. It showed each primers had an average of7.94amplified alleles, while there were127polymorphic alleles, polymorphic percentage of amplified was76.97%. Results showed that Panax ginseng from different areas have genetic differences, sample relatively close genetic relationships of the same origin. In summary, this study established quality standard and fingerprint of typical authentic herbs Panax ginseng fruit in Jilin province for the first time. The inspection items of characteristics and quality standard have been systematically determined and formulated. This study provided comprehensive, scientific and effective method and basement to quality evaluation and control of Panax ginseng fruit, and then laid the foundation for development of Panax ginseng fruit related products, adding value of Panax ginseng.