Primary Study On The Quality Standard Of Ganneng Dripping Pills And Its Effective Compositions In Resisting Hepatic Fibrosis

GDP is made from the effective part of the TLHC.It can protect liver and reduce the replication of hepatitis B virus and treat chronic hepatitis B.Research shows that TLHC can significantly lower the expression of TGF-?₁,reduce the content of HA and HYP and inhibit the synthesis and deposition of liver fiber.It has a good effect of resisting HF.The quality control of GDP is mainly based on TLC identification and quantitative determination of herpetrione.The quality standard detection index is single,which cannot fully control the quality of GDP and ensure efficacy.In the early stage,our group established the HPLC method for the simultaneous determination of7 compolances of Herpetospermum caudigerum,but it has not been applied because of lacking of control materials which is difficult in preparation.In recent years,the combination of fingerprint and QAMS not only comprehensively reflects the quality of drugs,but also enables simultaneous determination of multiple components through a single control products.It effectively solves the problem of control products'expensiveness and lack.This method has the advantages of convenience,rapidity and cheapness,and has been widely used in a variety of traditional Chinese medicines.In this paper,the following study were conducted with the aim of studying the quality standards of GDP:Firstiy,the high pressure preparation method was used to separate and purify the TLHC.The purity of samples were tested by area normalization method,and the compounds were identified by UV,IR,NMR and MS.Eight compounds were prepared from the TLHC.They were identified as dehydrodiconiferyl alcohol?BLG-3?,herpetotriol I?BLG-4?,herpetolide A?BLG-5?,herpetrione?BLG-6?,herpetin?BLG-7?,Herpetetrone?BLG-10?,herpetone?BLG-11?and herpetenone?BLG-12?,of which herpetotriol I is a new compound.Secondly,the quality standard of GDP was studied.Using herpetrione as an indicator component,TLC was used for qualitative identification.It was shown that sample and the reference substance had the same color spot in the same position,while the negative sample was without interference.According to the relevant requirements of the 2015 edition of Chinese Pharmacopoeia,the four General Rule of the 0108 pill,GDP were tested for weight differences,dissolution time and microbial limits.The test results met the requirements of the Pharmacopoeia.Using herpetrione as an internal reference,a QAMS for the simultaneous determination of 8 components?dehydrodiconiferyl alcohol,herpetotriol I,herpetolide A,herpetrione,herpetin,herpetetrone,herpetone and herpetenone?was established,and 5 batches of GDP were tested.This method was stable and reliable.The fingerprint of GDP was established.The similarity analysis of fingerprint of 5 batches of GDP was carried out by the similarity evaluation software of Chinese Medicine chromatogram fingerprint.The fingerprint common pattern of 5 batches of GDP was established by using median,and 14 common peaks were identified.After comparison with the control products,8components were identified.The similarity of the 5 batches of GDP was more than0.99.Finally,according to the results of the determination of GDP,the higher concentrations of herpetrione and herpetone were selected as the research subjects.CCK-8 method was used to detect the different treatment time of different concentrations of herpetrione and herpetone on the HSC-T6.The effect of HSC-T6 on the apoptosis rates were detected by flow cytometry.The results showed that herpetrione and herpetone had obvious inhibitory effects on HSC-T6 proliferation,which was proportional to the concentration and time.The apoptosis rates increased with the increase of herpetrione's and herpetone's concentrations respectively.This shows that herpetrione and herpetone have the function of resisting HF.