Study On The Forming Process And Quality Control Method Of Wenyang Tongluo Fengshi Gel Paste

ObjectiveThis topic intends to develop Wenyang Tongluo Rheumatic paste prescription into gel paste,and optimize its matrix prescription;The quality of the paste was evaluated through the examination of appearance,paste content,adhesion,formability and microbial limit.The thin-layer identification method and content determination method of Wenyang Tongluo Fengshui gel paste were established.The quality evaluation method was combined with a number of indexes,and provided the basis for the subsequent release in vitro and transdermal experiments in vitro.Stability test was carried out to investigate the stability of Wenyang Tongluo Rheumatic gel paste and its change condition with time under the influence of temperature,humidity and light,and to improve and optimize the research on its quality control method;The in vitro release and in vitro transdermal properties of the gel paste were preliminarily studied,which provided the experimental basis for the follow-up research of Wenyang Tongluo Rheumatic gel paste.Methods（1）With NP-700:The contents of carbomer,hydroxypropyl methyl cellulose,hydroxyaluminum and drug were the main influencing factors.The coating index,forming index,initial viscosity,holding viscosity and peeling strength of Wenyang Tongluo Fengshui gel paste were determined as evaluation indexes.L₉（3⁴）orthogonal test was used to investigate,and the optimal matrix formula was selected,and validation test and pilot test were conducted.（2）According to the"Chinese Pharmacopoeia"（2020 edition）,the paste content,adhesion,formability,microbiological limit test in Wenyang Tongluo Fengzi gel paste were checked.Thin layer chromatography（TLC）was used to identify angelica Dahuricae,cassia twig,calamus,parsnax,myrrh and menthol in Wenyang Tongluo Rheumatic gel paste.The content of paeoniflorin and paeonol in gel paste was determined by high performance liquid chromatography（HPLC）.The chromatographic method was used inertsil ODS-SPC18 column（150 mm×4.6mm,5μm）with acetonitrile(（A）-water（B）as mobile phase and gradient elution（0-16 min,12%A;16-39min,30%A）,column temperature was 30℃,detection wavelength was 230nm,flow rate was 1.0m L/min,sample size was10μL.（3）Stability studies were conducted with reference to Chinese Pharmacopoeia（2020edition）,the influence factor test（high temperature test,high humidity test,strong light exposure test）,acceleration test（0,6,9）and long-term test（0,3,6,9,12）of Wenyang Tongluo Rheumatic gel paste were conducted.（4）Established HPLC content determination method,so that it is suitable for in vitro release and in vitro transdermal performance research,combined with Franz diffusion cell method,using normal saline as the receiving solution,respectively at the first 1,2,4,8,12,24h sampling,after processing sampling analysis,The cumulative release rate（Q/%）-time（t/h）curve and the cumulative transmittal rate（Q/%）-time（t/h）curve of Wenyang Tongluo Rheumatic gel paste were plotted respectively.The zero-order kinetic model,first-order kinetic model and Higuchi equation model were used to fit the curves,and the results are compared,the cumulative release rate and in vitro transmittal rate of 24h after drug administration were calculated.Results（1）According to the experimental results,the optimal matrix prescription ratio was NP-700:carbomer:hydroxypropyl methyl cellulose:hydroxyaluminum:drug content=250:20:25:25:1600.According to the optimal matrix prescription ratio,Wenyang Tongluo Rheumatic gel paste was prepared,and the verification test was carried out.The results showed that the five indexes of Wenyang Tongluo Rheumatic gel paste prepared with the optimal matrix prescription ratio were stable,and the pilot test was carried out,and the results showed that the indexes met the requirements.（2）Three batches of Wenyang Tongluo Rheumatic gel paste were prepared according to the optimal matrix formula.Based on the test results of the quality control it was tentatively determined that the paste content of this product should not be less than 18g per 100cm².The formability should meet the requirements of the 2020 edition of Chinese Pharmacopoeia.The paste surface of the gel paste should have no flowing phenomenon,and the adhesive force can stick to No.10 steel ball.The total number of aerobic bacteria,mold and yeast was less than 10cfu/cm²,and Staphylococcus aureus and Pseudomonas aeruginosa should not be detected.In the TLC of Angelica Dahuricae,cassia twig,calamus,parsnips,myrrh and menthol,the spots were clear and significant,and had good separation effect.The negative samples had no interference.The regression equations of paeoniflorin and paeonol were Y₁=13303.9X₁-4885.00,R²=0.9998,Y₂=33391.1X₂-7109.15,R²=0.9999,respectively,and the linear relationship between Paeoniflorin and Paeonol was good in the range of 1.752-28.032μg/m L.The average recoveries were 105.68%（RSD=1.93%）and 101.04%（RSD=5.65%）.（3）According to the test results of influencing factors,temperature has a great influence on the paste content,adhesion,formability,paeoniflorin and paeonol contents of the paste,humidity can affect the paeoniflorin and paeonol contents,light can affect the paste content and paeonol content,accelerated test and long-term test have no effect on the indexes of Wenyang Tongluo Fenghi gel paste.（4）The cumulative release rate of paeoniflorin in Wenyang Tongluo Rheumatic gel paste for 24h was 50.81%,and the cumulative transmittance rate for 24h was 19.32%.The fitting degrees of the cumulative release rate（Q/%）-time（t/h）curve and the cumulative transmittal rate（Q/%）-time（t/h）curve to the zero-order kinetic model,first-order kinetic model and Higuchi equation model are 0.8160,0.9397,0.9518 and 0.7984,0.9736,0.9430,respectively.Conclusion（1）The Wenyang Tongluo Rheumatic gel paste prepared according to the optimal matrix prescription ratio optimized in this experiment is convenient for coating,smooth and delicate appearance,uniform color,good adhesion,peeling without residue,and convenient for production.The verification test results of the Wenyang Tongluo Rheumatic gel paste prepared according to the optimal ratio are stable,then the production is further expanded and pilot test is conducted.The three batches of samples all meet the relevant requirements of each index and can be used for production and use;（2）The evaluation requirements of the appearance character,paste content,adhesive force,formability and microbial limit of Wenyang Tongluo Rheumatic gel paste were determined,which could be used as the index of its quality control.The method of thin layer chromatography and high performance liquid chromatography is accurate and stable,and the instrument has good precision,stability and repeatability.It can be used to study the quality control method of Wenyang Tongluo Rheumatic gel paste.（3）Warm Yang Tongluo Rheumatic gel paste has good stability and should be stored at normal temperature and humidity.（4）The in vitro release of paeoniflorin in this gel paste is more consistent with the Higuchi equation,and the in vitro transdermal rule is more consistent with the first-order kinetics,which provides the experimental basis for the study of in vitro release and in vitro transdermal properties of Paeoniflorin.