| Objective:Vulvar Lichen Sclerosus（VLS）is a chronic inflammatory non-neoplastic skin lesion mediated by lymphocyte.The pathogenesis of VLS is unclear.Hydrocortisone,skin relaxation and other glucocorticoids are commonly used in clinic,and some traditional Chinese medicine clinical experience prescriptions are used to treat VLS.The compound pearl ointment is the clinical experience prescription of Beijing Obstetrics and Gynecology Hospital affiliated to Capital Medical University for the treatment of vulvar lichen sclerosis.It has been used for more than 20 years and has a definite effect.However,in the original prescription,the pig bile containing different concentrations can not be accurately quantified,so in the new prescription,the compound pearl cream is prepared by replacing the pig bile with pig bile powder.In this paper,the moulding process of compound pearl paste was systematically studied,and the stable and feasible technological route suitable for industrial production was screened.The quality standard and preliminary stability were also discussed.In addition,the quality control of Compound Pearl ointment and its main medicine porcine gall powder is studied in depth,so as to contribute to the comprehensive evaluation of Compound Pearl ointment.Method:The research content of this subject mainly includes the optimization of forming process,pilot scale-up study,quality standard study,stability study and other related research.The specific methods of each part are as follows:1.Forming technology research:According to the prescription,compound pearl powder was prepared by equal incremental method for reserve;the suitability of water-soluble matrix,emulsion matrix and oil-based matrix was investigated with the appearance,uniformity of dispersion and stability of paste as evaluation indexes;the optimal formulation of oil-based matrix was screened by D-optimal mixed standard design,and the physical stability was taken as evaluation index;The processing parameters of the compound pearl paste were studied in detail,including the melting temperature of the matrix and the dosing temperature,and the preparation process of the compound pearl paste was optimized.2.Pilot study:According to the prescription and preparation technology,8 batches of pilot test samples were prepared according to the size of 5 kg per batch.3.Quality standard research:①Qualitative identification:microscopic identification of compound pearl paste,calcined red stone,calcite and cassia;identification of calcium salts in paste;TLC identification of pig bile powder and borneol in paste;chemical identification of calcium salts;②Quantitative identification:quantitative determination of taurocholodeoxycholic acid in paste by HPLC,and determination of Borneol in paste by gas chromatography.The quality standard of compound pearl paste was determined and drafted.4.Preliminary stability study:According to the accelerated stability and long-term stability test conditions in the 2015 edition of Chinese Pharmacopoeia,the stability of the pilot preparation of compound pearl paste was investigated.The survey items include:character,homogeneity,content（taurine,porcine gallbladder powder,borneol,borneol,porcine bile powder thin layer:porcine deoxycholic acid,cholic acid,geo deoxycholic acid,borneol thin layer:borneol,iso-borneol）.5.Determination of cholic acid in 4 kinds of combined gallbladder powder and compound pearl ointment:A method for simultaneous determination of 4 conjugated cholic acids in pig bile powder and a pig bile powder characteristic map based on glycine deoxycholic acid were established by high performance liquid chromatography.The contents of 4 conjugated cholic acid in the prepared and compound pearl ointment were determined.Result:1.The determination of moulding process:①To determine the equivalent and increasing mixing method of compound pearl powder:first mix pearl powder with artificial musk,then mix it with calcined gypsum,calcined red stone grease,borneol,alum,calcined Cassia mixture,and finally mix with pig bile powder,then screen over 120 mesh.②Forming process:The ointment prepared by water-soluble matrix and emulsion matrix could not be formed well,and the compound pearl ointment was finally determined to be oil-based matrix.The specific process is as follows:octadecanol 10g,lanolin 10g,white vaseline 68.8 g,ethyl p-hydroxybenzoate 0.1 g were heated and melted.Compound pearl powder 11.065 g was added at about 55℃.After stirring,the temperature was immediately lowered to 350C.After stirring for 15 minutes,the mixture was filled.2.Pilot scale-up study:The preparation of 8 batches of Compound Pearl ointment was finished.The results showed that the appearance quality was good,the ointment rate was above 95%,the RSD value of the taurine porcine deoxycholic acid and borneol was less than 3%,the content of taurocholodeoxycholic acid was 1.66 mg·g-1,the average borneol content was 5.31mg·g-1,and the thin layer chromatography could identify the pig bile powder and borneol in the paste.The production data show that the process conditions are reasonable and feasible,and basically meet the requirements of large-scale production.3.Study on Quality Standards:The results of microscopic identification of Compound Pearl plaster,calcined gypsum,calcined red stone and calcite Cassia are good.The results of chemical identification of calcium salts are good,and the thin layer chromatography of pig bile powder and borneol is good.The quality standard of porcine gall powder is determined to be less than 1 mg·g-1 without taurine,or less than 0.10%.The quality standard of borneol is less than 4.25 mg·g-1,that is,not less than 0.43%.4.Preliminary stability study:Long-term and accelerated stability experiments of compound pearl paste were carried out.The results showed that the properties of compound pearl paste were good.The content of taurine,porcine deoxycholic acid and borneol,which are the index components,all meet the requirements of the draft standard.It shows that the quality of the compound paste is better in June,and the content of the index components is in line with the requirements.The ointment is more stable.5.Determination of cholic acid in 4 kinds of combined gallbladder powder and compound pearl ointment:A method for simultaneous determination of 4 conjugated cholic acids in pig bile powder by high performance liquid chromatography was established:the stability,repeatability,precision and recovery of samples were all met by acetonitrile and 0.03 mol ·L-1 sodium dihydrogen phosphate.The 7 peaks in the 12 batches of pig bile powder were identified as common characteristic peaks,and the similarity of characteristic maps was over 0.9.The value of RSD maintained at 3%or less was less than 3%.This method was used to determine the 29 batches of marketed porcine gall powder preparations.The content of 4 kinds of conjugated cholic acid in Compound Pearl ointment is as follows:the average content of taurocholodeoxycholic acid is 1.66±0.02 g（n=8）,and the content of glycine deoxycholic acid is 18.30±0.24 g（n=8）,the average content of taurocholodeoxycholic acid is 1.10±0.03 g（n=8）,and the average content of sodium glycine deoxycholic acid sodium is 6.46±0.07g（n=8）Conclusion:The forming process of Compound Pearl ointment was determined and verified by 8 batches of pilot-scale tests.The qualified product ointment was prepared.The quality standard of compound pearl paste was established and the preliminary stability study was carried out.The results showed that the sample was stable during the investigation period.A high performance liquid chromatographic method for simultaneous determination of 4 conjugated bile acids in pig bile powder and a pig bile powder characteristic map based on glycine deoxycholic acid were established.The contents of 4 conjugated cholic acid in Compound Pearl ointment were calculated. |
PDF Full Text Request