HPLC Qualitative And Quantitative Analysis Of The Chemical Composition Of Shou Taiwan

Objective:To analyze the main material composition of Shou Taiwan and find the optimum preparation method which is suit for Shou Taiwan’s ingredient characteristics;On the basis of previous research to find Shou Taiwan’s unknown active chemical components and to study the quantitative analysis method of possible miscarriage chemical composition.Methods:(1) Use single factor and parallel experiments, cold soak and ultrasound method are investigated different concentrations of methanol and ethanol as solvent,different cold soak time1h,2h,4h,8h,different ultrasonic time30min,1h,2h.Reflux method are investigated different reflux time30min,1h,2h,3h,different concentrations of methanol as solvent. As observed indicators to overall chromatographic peak shape, height and impurity peak,compare cold soak and ultrasound method and reflux method,find the better one.(2) HPLC conditions:Agilent TC-C18(5umx4.6mmx250mm) Column, diode array detector,0.1aqueous phosphoric acid-acetonitrile as the mobile phase gradient elution,flow rate lmL/min,column temperature30℃, injection volume10ul,UV detection wavelength220nm. Use UV spectra and standard comparison to identify the main peaks ingredients of Shou Taiwan.(3)Determine the content of hyperoside, kaempferol, asperosaponinVI of Shou Taiwan by HPLC.Results:(1) Compare the optimal conditions of cold soak and ultrasound method and reflux method,80%methanol soak2h,ultrasound30min,the overall peak shape, the height is preferably and impurities relatively less.(2)Main peaks ingredients of Shou Taiwan identified by UV spectra and standard are7,namely chlorogenic acid, coumaric acid, hyperoside, quercetin, quercetin, kaempferol, asperosaponinVI.(3)Establish the quantity measure methods of hyperoside, kaempferol, asperosaponinVI’s in Shou Taiwan by HPLC. Between the0.53-4.24ug,0.26-2.08ug,4.54-36.32ug, Hyperoside, kaempferol and asperosaponinⅥ have good linear correlation.The correlation coefficients are0.99994,0.99997,0.99999,the average recoveries are91.79%,90.05%,95.21%,RSD are3.698%,2.445%,1.729%(n=6).Conclusions:Comparing cold soak and ultrasound method and reflux method of Shou Taiwan,80%methanol soaking2h,ultrasound30min is better.Seven kinds of materials detected by the finger Print of Shou Taiwan established by HPLC respectively,Coumaric acid, hyperoside, quercetin, kaempferol attributable to semen cuscutae,quercitrin attri-butable to semen cuscutae and taxillus chinensis,Chlorogenic acid attributable to semen cuscutae and dipsacus, asperosaponiVI attributable to dipsacus.Establish the quantity measure methods of flavonoids hyperoside, kaempferol and triterpenoid saponins asperosaponiVI in Shou Taiwan by HPLC at the same wavelength UV detector,the method is accurate and sensitive.By measuring three batches of Shou Taiwan,find the differences of hyperoside content is small,but kaempferol, asperosa--poninⅥ are large,and contents of genuine medicines are higher levels.