Enrichment Of Total Flavonoids From Rosa Xanthina Lindl And Establishment Of HPLC Fingerprint

Objective:The Rosa xanthina Lindl of Shanxi province is rich in natural resources, it is a kind of sustainable development and utilization,green, with the advantages of Shanxi characteristic resources.To provide comprehensive control the quality of the Rosa xanthina Lindl abundant quality evaluation information, based on the extraction process of flavonoids from Rosa xanthina Lindl,to study the separation and purification of flavonoids with macroporous adsorption resin,and established HPLC fingerprint.Make sure the selected samples of the antithrombotic compounds and bioactive fraction from Rosa xanthina Lindl have homogeneous qualities,It has provided a reference for industrial.Methods:On the basis of optimum extraction process,enrichment and purification of flavonoids from Rosa xanthina Lindl were studied using eight kinds of macroporous resins( NKA-9、HPD100、FL-1、FL-2、FL-3、DM130、AB-8 and D101) in order to select optimal resin.The concentration of sample solution、the largest sample weight、 the concentration of eluent and the dosage of eluent were investigated to acquire the optimum separation and purification process.Aim to establish HPLC method for quality control of Rosa xanthina Lindl.The experiment investigated 6 kinds of chromatographic column of the Thermo Syncronis C18column(4.6×150mm,5μm),Thermo Hypersil GOLD column(4.6×250mm,5μm),Agilent Eclipse XDB-C18 column(4.6×150mm, 5μm),Inertsil ODS-SPcolumn(4.6×250mm,5μm),Agilent ZORBAX SB-C18 column(4.6×250mm,5μm),Thermo BDS HYPERSILC18 column(4.6×250mm, 5μm),and 6 kinds of mobile phase elution system as methanol-H2 O solution, acetonitrile-H2 O solution, methanol-0.2% phosphoric acid solution,methanol-0.2% HCOOH solution,acetonitrile-0.2% phosphoric acid solution,methanol-acetonitrile-0.2% phosphoric acid solution,as well as the different column temperature and wavelength.Optimized the gradient elution procedure.optimized the preparation methods of test sample solution in terms of extraction manner,extraction time and extraction solvent.Results:The static tests indicated that D101 resin was appropriate and its optimized technology conditions were as followings: The concentration of sample solution 1:12, the largest sample weight was 2.6g Rosa xanthina lindl /g resin, the volume of 70% ethanol(3BV)as eluant.The desorption rate reached 95%, the recovery rate of total flavonoids from Rosa xanthina lindl was 72%.With these paramaters,the purity of Rosa xanthina lindl extract was 24.87%. The total flavonids from Rosa xanthina lindl can be effectively purificated by D101 macroporous adsorption resin.The HPLC Fingerprint conditions of Rosa xanthina lindl: the chromatographic column:Inertsil ODS-SP column(4.6 × 250 mm, 5μm); mobile phase:methanol-acetonitrile-0.1% phosphoric acid solution; column temperature: 25℃; the flow rate: 1.0 mL/min; the sample injection volume: 10μL; detection wavelength: 254nm;gradient elution.The preparation methods of test sample solution:10.0 g Rosa xanthina lindl powder is weighed, 50% methanol 200 mL, reflux extraction about 120 min, suction filtration when it was hot, stress concentration to dry which keep a temperature about60 ℃, ultrasonic dissolving with 50 ml methanol, wait until it is cooling, we filter it by a0.22 μm filtration membrane, then the test sample is made. The methodological evaluation showed that the relative retention time of precision、repeatability and reproducibility were under 1%, and the relative peak area were under 3%.Conclusion:In the issue, we optimized the process conditions of separation and purification of total flavonoids from Rosa xanthina lindl with macroreticular adsorption resin,and established the HPLC fingerprint of Rosa xanthina lindl. this method can be used to purple the reflecting Rosa xanthina lindl species. We can understand the internal difference by system evaluation. A fingerprint quality evaluation method for quality control of Rosa xanthina lindl is established.This study has practical value and theoretical significance.