Preparation Of A Bacteriostatic Preparation And Anti-oxidation Quality Control Of Two Freeze-dried Powders

Objective:1.To prepare a liquid bacteriostatic preparation containing rhubarb active ingredient emodin,aloe emodin and gallic acid active ingredient and investigate its antibacterial effect in vitro.2.Establish anti-oxidation methods of Yiqi Fumai for injection（lyophilized）and salvia polyphenolic acid for injection by using DPPH method,and initially establish quality control standards.3.Establish a protective method of Yiqi Fumai for injection（lyophilization）on H9C2 cardiomyocyte injury,and measure the protective effect of cardiomyocyte injury on 30 batches of samples,and establish quality control standards.Methods:1.The active ingredients emodin and aloe-emodin were extracted from rhubarb,and the results were characterized by high performance liquid chromatography.Investigate the compatibility of emodin,aloe-emodin and gallic acid against seven common bacteria（Staphylococcus aureus,Streptococcus mutans,Streptococcus sanguis,Actinomyces viscosus,Porphyromonas gingivalis,Streptococcus cousin,Actinomyces nervii）,the optimal ratio of the three components was determined,and the three were combined into a liquid bacteriostatic preparation.2.Use the UV absorption method,investigate the color change of1,1-diphenyl-2-trinitrophenylhydrazine（DPPH）after adding Yiqi Fumai for injection（lyophilization）,and pass the UV spectrophotometer quantitatively measure the changes before and after the reaction;study the modeling time and dosage of H₂O₂on H9C2 cardiomyocyte injury,determine the best modeling method,and conduct linear,reproducible,precision,and stability methodological investigations;benefit from injection the accompanying samples of qi fumai（lyophilized）were used as a control,and the percentage of CCK-8 cell viability was used as an index to measure its relative protective effect;in order to establish the quality standard of Yiqi fumai for injection（lyophilized）.3.The UV absorption method was used to investigate the color change of DPPH solution after adding salvia polyphenolic acid for injection,and the change value before and after the reaction was quantitatively measured by a UV spectrophotometer.Results:1.（1）The extracts were determined to be emodin and aloe-emodin by HPLC and standard control.（2）All three had a certain inhibitory effect on the seven common bacteria investigated.The optimal concentration combination of the three were 8 mg/100 m L of emodin,8 mg/100 m L of aloe emodin,and 400 mg/100 m L of gallic acid.（3）A liquid bacteriostatic preparation was prepared in accordance with the optimal ratio of the three,and the bacteriostatic effect was improved after the compatibility was tested by the bacteriostatic test.2.The linear range of Yiqi Fumai for injection（lyophilization）was 0-12.5 mg/m L（r²=0.9924）,the relative standard deviations（RSD）of precision were 1.1%and 0.9%,and the repeatable RSD values??were respectively 1.0%and 1.2%.The reference was stable within 35 minutes at room temperature.This method had good durability.Based on half the antioxidant capacity of the accompanying reference,the quality standard was set to 50%-150%.Salvia polyphenolic acid for injection had a linear range of 8μg/m L-32.02μg/m L（r²=0.9976）,the relative standard deviation（RSD）of precision was 0.9%,the repeatable RSD value was 1.2%,and the room temperature of the reference was stable in the next 35 minutes,and the method had good durability.With reference to the relative antioxidant capacity of the accompanying reference substance,the quality standard was set to 30%-80%.3.When the modeling time was 0.5 h and the dosage of H₂O₂was 0.2 m M,the degree of cell damage was about 60%,and the modeling effect is relatively stable;the concentration of 30 batches of Yiqi Fumai（lyophilized）for injection is 5 mg/m L.At the same time,its protective effect on H9C2cardiomyocyte injury caused by H₂O₂was between 84.4%-96.8%,which had a good cytoprotective effect.Conclusions:1.The bacteriostatic preparations prepared after the combination of emodin,aloe-emodin and gallic acid have a good inhibitory effect on seven common bacteria.2.DPPH method can be used to determine the antioxidative capacity of Yiqi Fumai（lyophilized）for injection and salvia polyphenolic acid for injection,and the method has strong operability,simplicity and convenience.3.The established method is suitable for the research on the protective effect of Yiqi Fumai for injection（lyophilization）on H9C2 cell damage,and can be used as a preliminary quality standard for evaluating Yiqi Fumai for injection（lyophilization）.