Research On Extraction And Separation Technology Of Juglanthraquinone C In Juglans Mandshurica

The present study is to development a systematic preparation method Juglanthraquinone C(JC)from Juglans mandshurica Maxim by combining the traditional extraction technology and modern instrument purification process.The process for extraction,separation and refining purification of JC were explored,and the important technological parameters of conditions were optimized.Our study provided experimental guidance and data support for the large scale production and exploration.At the same time,the method for determination of JC was established,which provides a reliable method for tracking and monitoring of the extraction,separation and purification process of JC.The results of experiment are shown as follows.The detection method of JC was established,and the methodology validation was also completed.HPLC was performed on a Extend-C18(4.6×150mm i.d,5μm)analytical column with mobile phase consited of methanol-0.1% formic acid(85:15)at the flow rate of 0.8 mL/min.The detection wavelength was 230 nm.JC had good linearity in the range of 1.22 μg/mL-19.52 μg/mL(R=0.9996).With the yield of JC in extraction as control criteria,the extraction conditions of ultrasonic assisted extraction were investigated through one-single factor experiments and response surface methodology.The optimal extraction conditions were shown as below.Extraction with 61.61% ethanol,the ratio of solvent to material 10.44(v/w),extraction time 47.55 min,extraction temperature 30 ℃,the ultrasonic power 320 w,extraction of 5 times.Through model validation,the productivity of JC was 324.15μg/g under the condition by ultrasonic.Purification process can be divided into following steps: Liquid-liquid extractions(LLE),medium and low pressure liquid chromatography were used for preliminary separation and purification.Then the further purification for JC was made by versamide absorption resin.In the end of refine for JC was silica-gel column chromatography.After these processes,the purity of sample was more than 90%.