Studies On The HPLC Fingerpring And The Determination Of Chaihu Shugan Decoction

Backgrounds and purposeAccording to the guidance theory of "treat from liver" for epilepsy treatment,research group created Chaihu Shugan Decoction.It consists of fifteen medicine herbs:asdix bupleuri,radix scutellaria,Radix Paeoniae Alba,et al.It has ability to shu gan,rou gan,yang gan,ping gan.Modern medicine research reveal,it has ability to reduce nerve excitability,anti-inflammatory,and adjust the immune function of organism.It has been used on clinic for about thirty years and has remarkable effect on the prevention and treatment of epilepsy.The quality of medicine herbs is closely related to clinical drug efficacy and safety.Chaihu Shugan Decoction hasn’t been done some relative quality control research.The aim of this research is use the HPLC method to study Chaihu Shugan Decoction systematically,to establishing Chaihu Shugan Decoction fingerprint and its main component determination,So that giving comprehensive control from the whole and part of the quality of its effective components,it will be good for the further research and provide the theory basis for clinical medication safety.Method1.To research the fingerprint of Chaihu Shugan Decoction.and determination its main components paeoniflorin,liquiritin,ferulic acid,baicalin,wogonoside,and wogoninThe fingerprint was established by HPLC with Diode Array Detector(DAD)and ZORBAX SB-C8 column(250 mm×4.6 mm,5 μm),gradient eluted with acetonitrile-0.2%phosphoric acid as mobile phase.The detection was at 210 nm,and column temperature was at 25 ℃,the injection volume was 10 μL,the detection time is 65 min.The chromatograms of Ten batches of Chaihu Shugan Decoction were established to evaluating the similarity of Chaihu Shugan Decoction and analyzing the characteristic peaks belong to herb.Establish a methods to Determination main components of Chaihu Shugan Decoction under the same chromatographic condition at the same time,so improve the specificity for quality control of the herbal formula.2.Determination Chaihu Shugan Decoction main components Saikosaponin a,wogonoside and paeoniflorinThe fingerprint was established by HPLC with Diode Array Detector(DAD)and ZORBAX SB-Cg column(250 mm×4.6 mm,5 μm),gradient eluted with acetonitrile-0.2%phosphoric acid as mobile phase.The detection was at 210 nm,and column temperature was at 25 ℃,the injection volume was 10 μL,the detection time is 80 min.Examines the stability,repeatability,and determination of Saikosaponin a and paeoniflorin,provide the basis for quality control of the herbal formula.Results1.The chromatographic fingerprint of Chaihu Shugan Decoction was established,the similarity of the batches of Chaihu Shugan Decoction were more than 0.95.22 common peaks were identified and attributed to the herbs,7 chromatographic peaks were identified.A method for simultaneous determination of paeoniflorin,liquiritin,ferulic acid,baicalin,wogonoside,and wogonin was established.liner relation was good,the average recoveries of components were between 90%to 105%(RSD<3%).2.A method for simultaneous determination of Saikosaponin a,wogonoside and paeoniflorin was established,liner relation was good,the average recoveries of RSD were great,The sample solution was stable within 24 h,and this method was good reproduction,liner relation was good,the average recoveries of components were between 90%to 105%(RSD<3%).ConclusionEstablished the HPLC fingerprint of Chaihu Shugan Decoction and the method for determination its main components,the method was repeatability,good stability,liner relationand the average recoveries of components ware great,which could reflect the overall characteristics of decoction and provide theoretical basis for the further research.