The Study On The Determination Of The Saponin’s Content And Fingerprint Of Flos Lonicerae Confusae

Objective: Flos Lonicerae Confusae is a common Chinese medicinal material in our country.Review of the past research, the quality evaluation of Flos Lonicerae Confusae is mostly measured by the content of chloro-genic acid.But this method doesn’t reflect its quality in an all-round way. The ivy saponins have proved to be other important constituents and the content of them has fortune to be a part of the quality control the same as chlorogenic acid. The number of peaks and peak height in traditional Chinese medicine (TCM) fingerprint represent the type and the amount of chemical composition,reflecting and evaluating the immanent quality of Chinese medicinal materials systematically.As a result, the research to the content determination method of ivy saponin and set up the fingerprint of Flos Lonicerae Confusae are of great significance to the quality control and comprehensive evaluation.To establish a method of direct determination of hederagenin after acid hydrolysis and determination after pre-column derivatization, comparing the advantages and disadvantages of the two methods; Establish the HPLC fingerprint, get the reference fingerprint,the using the method setted to evaluate the collected samples.Methods:1. The research of the determination of hederagenin content:(1) The sample was hydrolyzed with hydrochloric acid-ethanol for4hours at80℃and using chloroform to extract the hederagenin;the determination was performed on Waters Symmetry(?) C18(150mm x4.6mm,5μm) column with methanol-0.4%glacial acetic acid (80:20) as mobile phase, the flow rate was1.0mL·min-1,column temperature was set at30℃,the detected wavelength was210nm.(2) Pre-column derivatization method:on the basis of the above, using pyridine-benzoyl chloride as the pre-column derivatized reagent to deri-vate the hederagenin,the using the HPLC method to determinate the content of the hederagenin.The column was Waters Symmetry (?) C18(150x4.6mm,5μm);acetonitrile-0.2%phosphoric acid aqueous solution (90:10) was used to be mobile phase;the flow rate is1.0mL/min;the column temperature was30℃;the detection wavelength was setted at230nm.Results:1.To establish the HPLC method of direct measurement and determination after pre-column derivatization of the hederagenin’s content.The determination results of the two methods were approxi- mate,but the determination result using pre-column derivatization method is a bit higher.2. Our research has established the fingerprint of Flos Lonicerae Confus-ae herbs.Using the method to evaluate the10batches of different origin and11batches from Longhui,Hunan.The evaluation of similarity of10batches of different origin was above0.957and the11batches from Longhui,Hunan was above0.969.Conclusion:Our study has used HPLC method to determine the content of hederagenin after acid hydrolysis directly and determination aftering being derivatization. The two methods that we established to determine the content of hederagenin have good precision and repeatability,the results are accurate and can be used to control the content of saponin; The established fingerprint is stable and has good reproducibility.It has high,, degree of separation and can use to conduct a comprehensive quality control of Flos Lonicerae Confusae herbs.