Separation Of Active Components And Analysis Of Curative Effect Of Chinese Herbal Medicine Psyllium Sylvestris In The Treatment Of Postpartum Retention In Cows

Retained fetal membranes(RFM),where the fetal placenta is not expelled within 12 hr after calving,and it is one of the most important reasons for low reproductive efficiency in dairy cows.As a traditional Chinese medicine,semen plantaginis has been found to be effective in the treatment of RFM.Through experimental studies of wang yao et al.,who are our laboratory,determined the effective parts of semen plantaginis for the treatment of RFM,but the effective substances in the effective parts to prevent and control RFM are still not clear.Therefore,this study is to determine the separation system of compounds in the effective parts to cure RFM,and to verify the effect of the compounds on the treatment of RFM.It also provides a theoretical basis for the research and development of drugs with the advantages of high efficiency,little the poisonous side effect and low cost for the treatment of RFM in the cows.In experiment 1,the semen plantaginis by stir-frying with wine was extracted by petroleum ether(PET),dichloromethane(DCM)and ethyl acetate(EAC),and then the separation system was determined by silica gel column chromatography and TLC with different column heights.In experiment 2,the compounds in the effective parts were separated by the separation system determined in experiment 1,and the recovery rate of the compounds was calculated.In experiment 3,thirty cows were randomly divided into six groups,each group five.These cows were fed compounds from experiment 2 for clinical validation.The fetal membranes discharge was observed 48 h after drug administration,and the time of fetal membranes discharge was recorded.the results of dichloromethane parts and ethyl acetate parts showed that the active ingredient s could not be effectively separated at 15 cm column height and the recovery rate at 25 cm colum n height was higher than that at 35 cm.The eluent and developing system of the active ingredient s separated at the dichloromethane parts were determined that:1.PET:EAC,1:2;2.PET:EAC,1:30;3.DCM:methanol(MT),30:1.The column height was 25 cm.The eluent and developing system of the active ingredients separated at the ethyl acetate parts :1.PET:EAC,100:1;2.PET:EAC,30:1;3.DCM:MT,30:1.In experiment 2,the three compounds isolated from dichloromethane were 0.234 g,0.571 g and 3.024 g,respectively,with a total weight of 3.829 g and a recovery ratio of 55.93%.The three compounds isolated from the ethyl acetate were 0.218 g,0.085 g and 0.753 g,respectively,with a total weight of 1.056 g and a recovery rate of 51.39%.In experiment 3,the cure rate of compoundsⅠ,Ⅱ and Ⅲ form dichloromethane parts were 40%,20% and 60%,respectively.The cure rate of compounds Ⅳ,Ⅴ and Ⅵ from the ethyl acetate part were 20%,20% and 40%,respectively.The results of this study showed that the chromatographic column with an internal diameter of 6 cm and a volume of 500 mL were selected,and the silica gel filling height of 25 cm was the best.Dichloromethane parts in the isolated compounds Ⅲ curative effect is distinct,the cure rate is 60%.Compounds Ⅵ from the ethyl acetate part of therapy effect is obvious,the cure rate was 40%.Taken together,our data indicate that compounds Ⅲ and Ⅵ were the main active components for curing RFM with the semen plantaginis by stir-frying with wine.