Study On The Preparation And Quality Control Of Anemoside B4 Suppository For Children

Objective:At present,there are few reports about Anemoside B4 preparation,in view of its significant anti-inflammatory effect,it can be considered to develop drug preparation for children’s inflammation.The purpose of this study was to study the preparation technology,quality standard of AB4suppository for children.By adding absorption enhancers to promote the absorption of AB4 in rabbits,so as to improve the bioavailability of AB4suppository in vivo,and provide laboratory data for the research and development of AB4 preparation.Methods:1.Determination of oil-water partition coefficient of AB4 and investigation of its stability in intestinal floraObjective to establish an HPLC method for the determination of AB4,and to investigate the methodology of the method.The oil-water partition coefficients of AB4 in water and buffer solutions with different p H values were measured,and the absorption of AB4 in intestinal tract and mucosa was primarily explored.The stability of AB4 in buffer solution with p H value of 7.3 and the effect of intestinal flora on the stability of AB4 were investigated.2.Research on formulation screening and preparation technologyA method for the determination of AB4 in rabbit plasma was established by UPLC-MS/MS.AB4 suppository was prepared by hot melting method.After AB4suppositories of different prescriptions were given through the rectum of rabbits,and AB4 normal saline solution was injected into the ear vein.The pharmacokinetic characteristics of the suppositories in rabbits were investigated,calculate the absolute bioavailability,the best absorption enhancer and its dosage were selected,and the optimal prescription was determined.After the prescription was determined,the preparation process of suppository was optimized through single factor investigation to determine the best preparation process.3.Study on the quality standard of suppositoryThe study on the quality standard of AB4 suppository.The TLC was used to qualitatively identify the negative,suppository and reference substance.According to the Chinese Pharmacopoeia,the formulation methodology was established by HPLC method,and the quality of AB4suppository was evaluated,the main contents were appearance,weight difference,melting time and dissolution.Results:1.Determination of oil-water partition coefficient of AB4 and investigation of its stability in intestinal floraIn vitro analysis method showed that the linear relationship of AB4concentration was good,the precision and repeatability were less than 1%,and the recovery rate was between 98%and 102%,which met the requirements of Pharmacopoeia.The results of oil-water partition coefficient show that AB4 has good water solubility and poor fat solubility.The Lg P value of AB4 is-0.03～-0.20 in the range of p H4～5,AB4 has a certain effect on intestinal flora within 2 hours.2.Research on formulation screening and preparation technologyThe plasma concentration of AB4 in rabbits was determined by UPLC-MS/MS,the detection mode was ESI^-1,the scanning mode was multi reaction monitoring（MRM）,the detection ion pair:AB4:m/z:1219.6-749.4,and the internal standard was Ginsenoside Rg1:m/z:823.4-643.3.The results showed that the linear relationship of the method was good,and the precision and stability met the requirements.It can be used for the determination of AB4 in rabbit plasma and the quantitative analysis of AB4 in rabbit plasma.The optimal formulation was determined by dissolution and pharmacokinrtics:AB4 30mg,34^#semi-synthetic fatty acid glycerol ester as matrix,and 0.5%chitosan as absorption enhancer.Preparation process:firstly,added AB4 with100 mesh sieve to the mortar for grinding,and then the 34^#semi-synthetic fatty acid glycerol ester was added in equal amount,stirring while adding,and then add 0.5%Chitosan,grinded until the mixture was uniform,heated and melted in a water bath at 45°C,until it was 2/3,took out,continued to stir,adjusted p H≈4～5 with glacial acetic acid,cooled naturally to 30°C,and then injected at medium speed,scrape off the overflow part after solidification,after 10min at room temperature,the mold was opened.3.Study on the quality standard of suppositoryThis product is milk white torpedo suppository with smooth appearance,uniform color distribution and moderate hardness.The weight difference limits and melting time of suppositories prepared in the laboratory met the requirements of 2015 edition of Chinese Pharmacopoeia.The content of AB4 suppository was determined by HPLC with the mobile phase of acetonitrile water（32-68）;the detection wave length was201 nm;the flow rate was 1.0 m L·min^-1;the column temperature was 30°C;the injection volume was 10μL.The method met the requirements and could be used to analyze the content of AB4.Conclusion:The method verfication results of the AB4 in vivo and vitro analysis methods meet the determination requirements.The pharmacokinetic data showed that the bioavailability of 0.5%chitosan AB4 suppository is high in rabbits.According to the best prescription process and preparation technology,the suppository was milky white,smooth surface,moderate hardness and fast dissolution rate in vitro.