Comprehensive Quality Evaluation Research On Lonicera Japonica Thunb

Objective: This research project is mainly aimed to study on Lonicera japonicaThunb by modern analytical instrument, to comprehensively evaluate the quality ofLonicera japonica Thunb,to optimize the extraction process of chlorogenic acid and Luteoloside inLonicera japonica Thunb.to establish a UPLC method for Simultaneous determination ofchlorogenic acid, rutin, Luteoloside in Lonicera japonica Thunb from different areas. Thiswould be an alternative method for content determination and quality evaluation of Lonicerajaponica in Pharmacopoeia of the People,s Republic of China(2010.Vol Ⅰ) Throughdetermination of the index component of Lonicera japonica from different flowering period,different months and different places, to provide a scientific reference for theharvesting period of Lonicera japonica and scientific reference for the quality of Lonicerajaponica from different months and different areas. To establish the HPLC and UPLCfingerprint profile of Lonicera japonica to evaluate the overall quality of the pros and consof it.Method: To optimize the extraction process of chlorogenic acid and Luteoloside inLonicera japonica by single factor and orthogonal experiments. Determinating the content ofchlorogenic acid, rutin and Luteoloside in Lonicera japonica by HPLC and UPLCrespectively. Establishing the HPLC and UPLC fingerprint chromatographic profile ofLonicera japonica. Performing cluster analysis and similarity analysis and comparingdifferences between the two fingerprints.Results: The extraction process was optimized by single factor and orthogonalexperiments：Being soaked30min with120times70%ethanol, extracted45min withultrasound (power350W, frequency40KHz). HPLC was used condition of chlorogenic acid.The chromatographic separation was performed on a Agilent ZORBAX XDB C18(4.6×150mm，5.0μm) column with isocratic elution. The mobile phase was0.2%phosphoricacid(A) and acetonitrile(B). The flow rate was1.0ml/min. The detection wavelength was327nm and the column temperature was30℃, injected2.0μL. The linear range ofchlorogenic acid was from39.6μg/mL to297.0μg/mL (r=1.0000). The average recoverywas97.29%(RSD=0.48%).HPLC was used condition of rutin. The chromatographicseparation was performed on an Diamonsil C18(4.6mm×250mm,5μm)column, the mobilephase was0.5%acetic acid and methanol，gradient elution separation. The flow rate was1.0ml/min. The detection wavelength was257nm and the column temperature was30℃,injected10μL. The linear range of rutin was from4.1μg/mL to41.0μg/mL (r=0.9996). The average recovery was98.08%(RSD=0.30%).HPLC was used condition of Luteoloside.The chromatographic separation was performed on an Agilent ZOTBAX XDB-Phenyl C18(4.6mm×250mm,5μm)column, the mobile phase was0.4%acetic acid and acetonitrile，gradient elution separation. The flow rate was1.0ml/min. The detection wavelength was350nm and the column temperature was30℃, injected10μL. The linear range ofLuteoloside was from2.00μg/mL to30.0μg/mL (r=0.9996). The average recovery was97.91%(RSD=0.49%). To establish a UPLC method for Simultaneous determination ofchlorogenic acid, lutin, luteoloside in Lonicera japonica Thunb. UPLC was used. Thechromatographic separation was performed on a C18(50×2.1mm，1.8μm) column withgradient elute. The flowrate was0.2mL/min. The column temperature was30℃and Thedetection wavelength was350nm. The linear range of chlorogenic acid was from15.84μg/mL to316.8μg/mL. The linear range of rutin was from1.64μg/mL to32.8μg/mL.The linear range of luteoloside was from1.60μg/mL to32μg/mL.The recoveries ofchlorogenic acid,lutin and luteoloside are96.57%~99.50%. To establish the HPLC andUPLC fingerprint chromatographic profile of Lonicera japonica Thunb of differentflower and different months.The data from HPLC and UPLC were analyzed by similarityevaluation and system cluster analysis.Get the22peaks of common peaks HPLCfingerprint, get the23peaks of common peaks UPLC fingerprint,and making sure thechlorogenic acid, rutin,Luteoloside,3,5-Odicaffeoylquinic Acid,3,4-Odicaffeoylquinic Acidand4,5-Odicaffeoylquinic Acid peak. The similarity of0.848～0.992. Establish methodsfingerprints of Lonicera japonica Thunb to evaluate the quality.Conclusion: Over the research of extraction process of Lonicera japonica Thunb.To Establish the extraction process chlorogenic acid and Luteoloside as indicators inLonicera japonica Thunb.To establish a UPLC method for Simultaneous determination ofchlorogenic acid, lutin,Luteoloside in Lonicera japonica Thunb. UPLC and HPLC methodsfor the determination of Chlorogenic acid in Lonicera japonica Thunb. are simple, theresults are accurate and reliable. The major advantages of UPLC than HPLC are the speed ofanalysis, the narrower peaks, HUPLC can not only increase separation speed and efficiency,but also reduce solvent analysis time and consumption. UPLC as a new detection methodcan replace HPLC for the quality control of Lonicera japonica Thunb. The UPLC systemenjoys the merits of a higher speed,sensitivity,and resolution, It can be used to control thequality of Lonicera japonica Thunb. Determined the heavy metal and other microelementscontents and the common quality of Lonicera japonica Thunb. was studied.The quality ofLonicera japonica Thunb. was comprehensive evaluated. And we can overall control thequality of Lonicera japonica Thunb.