Studies On Concomitant Components Of The Extract From Xinjiang Pregnant Mare Urine

Objective: Use fine separation fill MCI GEL CHP20P and Sephadex LH-20 to separate other compositions from pregnant mare urine, except the conjugated estrogens. Identify the structure of those compositions and determine the content of the isolated compounds that was the extract from pregnant mare urine which was through MCI GEL CHP20P.Methods: 1)Use MCI GEL CHP20P as separation materials and methanol - water as the solvent elution with pregnant mare urine and got the different fractions. 2)Use Sephadex LH-20 as the separation materials and methanol - water as well as methanol - the chloroform as solvent elution fractions to make a re-separation of each fraction of distillate. 3)The isolated compounds in the NMR spectra of hydrogen, carbon and two-dimensional spectroscopy spectra were analysised on the structural aspect. 4)The two compounds isolated from the pregnant mare urine were assayed. 5) Monitored extract from mare urine of the progesterone production process of the various components of intermediate.Results: 1) Use MCI GEL CHP20P as separation materials and methanol - water as eluting solvent, formononetin was separated in part of 80% . 2)By Sephadex LH-20 for the separation materials and methanol - water as eluting solvent, in 20% part of was isolated benzoic acid, in 100% part was isolated three compounds, namely: biochanin A, daidzein, genistein. 3)The linear range of formononetin was 0.08 to 0.56μg/ml , linear range of biochanin A was 0.09 to 0.62μg/ml, the average recoveries were 99.5% and 99.6%, respectively RSD is 0.56% and 0.78%. 4)The various intermediates in the production process of the extract from pregnant mare urine, exception of B samples, other samples contained daidzein and genistein, C and G samples contained samples of formononetin, only G samples containing biochanin A;these four components was removed through the MCI GEL CHP20P column.Conclusion: 1)MCI GEL CHP20P, Sephadex LH-20 as a fineness separation materials, in other components except pregnant mare urine combination of conjugated estrogens were separated by the different solvent elution screening to determine methanol -A water and Alcohol - chloroform is the best eluting solvent gradient elution, isolated the benzoic acid, formononetin,biochanin A, daidzein, genistein of the extract from pregnant mare urine . The results showed that Sephadex LH-20 on the other components of pregnant mare urine have a good separation, which can achieve a better effect. 2)We have assayed formononetin and biochanin A, reproducible, high stability, high precision, provide the basis for concomitant components of the extract from pregnant mare urine. 3)Refined the various intermediates of different stages of pregnant mare urine extract in the production process as well as detected the MCI GEL CHP20P column extract of the pregnant mare urine, have been more satisfied with the results confirmed and practical separation method for the industrial production in the future of the intermediates and products lay the foundation for the ingredients.