Study On The Extraction Process And Quality Standard Of Zangyaoganning Capsules

Chronic liver disease including viral hepatitis,non-alcoholic fatty liver disease,alcoholic liver disease,metabolic liver disease,drug induced liver disease,autoimmune liver diseases,liver cirrhosis,liver cancer and other common disease.With the speeding up of modern life rhythm,incidence of chronic liver disease is higher and higher in our country and as a rising trend.Because of the long course,it has become one of the focus of global public health.At present,researchers at home and abroad made a large number of clinical and experimental research of this disease by advanced science and technology and medicine method.However,the effect is not satisfactory.Therefore,the development of a new drug with effective in treatment and minor side effect is extremely urgent.Zangyaoganning capsules are derived from the clinical experience of Tibetan medicine.Consists of Chrysosplenium nudicaule Bunge,Swertia mussotii Franch,Dracocephalum tanguticum Maxim,Herpetospermum pedunculosum Wall,Holarrhena antidysenteriaca Wall.ex A.DC,Adhatoda vasica Nees and other Tibetan medicine.They were widely used in the treatment of various liver diseases in Tibetan areas.The original prescription is powder with a large dose,its production technology lag behind and standard of quality is low,it is not convenient in use and carry.Under the premise of retaining efficacy and characteristics of the original prescription,the purpose of this research is to study the extraction process and quality standards of ganning capsules using the modern technology,hope to develop a new drug with a reasonable preparation technology and stable and controllable quality in order to relieve the suffering of the patients with liver disease.Chrysosplenium nudicaule Bunge is a major component of ganning capsules,but the chemical composition of it is less reported,efficacy material base is unclear.We studied the chemical composition of it and determine the content of the main chemical composition by quantitative analysis of multi-components by single marker(QAMS).As a result,eleven compounds was isolated from Chrysosplenium nudicaule Bunge.They are p-Hydroxyacetophenone,Chrysosplenoside D,Pendulin,Chrysosplenoside B,Chrysosplenol D,cucurbitacin D,Chrysosptertin B,Penduletin,Arvenin Ⅰ,2β-glucosyloxy-16,20,22-trihydroxy-9-methyl-19-norlanosta-5,24-diene-3,11-dione and other one new compound.The eleven compounds were isolated from this plant for the first time.Among them,the contents of Penduletin,Chrysosplenoside D,Pendulin and Chrysosplenoside B were 5.84,3.51,2.76 and 10.52 mg·g-1.These results laid a foundation for the extraction process study.Swertia mussotii Franch is another major component of ganning capsules.In order to control the quality of ganning capsules better,We set up a method of content determination of sweroside and mangiferin in Swertia mussotii Franch.The mobile phase was composed of methanol(A)-0.8 % acetic acid(B)with gradient elution(0~30 min,20%A→45%A;30~60 min,45%A→100%A)at the flow rate of 1.0 mL·min-1.The detection wavelength was 254 nm,the column temperature was 30 ℃ and the sample size was 10 μL.There were good linearities in the range of 23.24~69.72 μg·mL-1 for swertiamarin and 110.12~330.36 μg·mL-1 for mangiferin.The contents of swertiamarin and mangiferin are 1.32 mg·g-1 and 5.85 mg·g-1.The method is simple and reliable with a good repeatability,not only provides a reliable detection index for the next extraction technology research but also offer a quantitative basis for the research of quality standard.According to the physical and chemical properties of effective components of each medicinal materials in the prescription,we preliminary designed four kinds of extraction process,studying the effect of different extraction process on the pharmacodynamics through the model of acute liver injury induced by carbon tetrachloride.At the same time,we determined the extract yield,transfer rate of mangiferin,the content of oleanolic acid and total flavonoids of different extraction process in order to choose out the best one.Then,we designed orthogonal test to select the best ethanol-extracting process.The preparation technology was determined finally as follow: Chrysosplenium nudicaule Bunge,Swertia mussotii Franch,Dracocephalum tanguticum Maxim,Herpetospermum pedunculosum Wall,Holarrhena antidysenteriaca Wall.ex A.DC,Adhatoda vasica Nees with 14 times the amount of 65% ethanol reflux extraction 2 times,each time 2 h;The extract and other medicinal powder were mixed evenly and then add 45% soluble starch.Afterwards,add 70% ethanol to made of soft material,use 20 mesh granulating and drying in the 60~70 ℃.At last,use 20 mesh sorting particles and add 5% magnesium stearate to adjust liquidity,put it into the capsule shell.In the study of quality standard,we checked the water content,disintegration time limit and load difference of ganning capsules,they were complied with relevant regulations of the Chinese pharmacopoeia.We also studied the identification of Dracocephalum tanguticum Maxim,Herpetospermum pedunculosum Wall and Adhatoda vasica Nees by thin-layer chromatography.The spot has good separation,negative control without interference,proves that the selected methods have strong specificity.Besides,we made a content determination of mangiferin in the ganning capsules by high performance liquid chromatography(HPLC).In the end,we limit the content of swertiamarin and mangiferin in the Zangyaoganning capsules not less than 0.1mg and 0.7 mg per gram.In conclusion,the prescription composition of Zangyaoganning capsules is unique,with the innovative theory basis and reasonable preparation technology.It is feasible in the actual production.The quality standard is simple,reliable and reproducible.In a word,the ganning capsules has a good prospect of clinical application.