Extraction,Purification And Analysis Of Total Flavonoids In Lysimachia Christinae Hance

Christina Loosestrife Herb has been long used as a chinese traditional medicine,which is from fresh or dry plant ofLysimachia christinae Hance,family Primulaceae.This medicinal plant is mainly produced in SiChuan and ZheJiang Province;It is used for jaundice,red urine, liver and gall stone.It has effects of Heat-removing,Water-promoting,Dampness-lessening and Stranguria-eliminating,Modern researchs show total flavonoids of Christina Loosestrife Herb can prevents formation and recurrence of calcium oxalate stone,increases coronary flow, and has prominent anti-inflammatory effect.However,,these is no evaluation of the quality of the medicinal plant in Chinese Pharmacopoeia 2005.Our research use HPLC method to identify the content of the main active constituents of Christina Loosestrife Herb,try to establish a qualification standard for Christina Loosestrife Herb.Flavonoids are the main components of Christina Loosestrife Herb.According to some related literatures,Isorhamnetin,Quercetin and Kaempferol are the most familar contents.We are to design an experiment to extract and purify Christina Loosestrife Herb total flavonoids by microwave-assisted extraction(MAE) and macroporous resins,then HPLC is used to make a content mensuration of Quercetin and Kaempferol,therefore,the Christina Loosestrife Herb's quality we can judge. To extract Christina Loosestrife Herb total flavonoids,an orthogonal table has been used to find the best procedure for MAE of flavonoids.The optimun condition for procedure is extracting 30min at 60℃with 8 folds of 50%ethanol.The ethanol concentration was significant among all factors on the test,extraction temperature was in the next place,while extraction time and folds were unconspicuous.The flavonoids contents in Christina Loosestrife Herb is 5.58%by MAE,80%more than traditional distillation extraction method,which only made 3.09%yield.Comparing with traditional extraction,The advantages of MAE in every important factors of yield,time and consumption of solvent et.After extraction,the Christina Loosestrife Herb total flavonoids have been purified by macroporous resins.The D 101,AB-8 and NKA-9 macroporous adsorbents were investigated to separate and purify Christina Loosestrife Herb total flavonoids,evaluating by adsorption capacity,elution ratio and product purity.Among the three types of macroporous adsorbents, the NKA-9 type shows better property with the adsorption capacity of 51.2 mg·mL^-1 and the elution ratio of 96.5%.According to the result of experiment,The best method condition is: extract sample concentration of 20 mg·mL^-1,6 times washing water and elution reagent of 50%. The purity of Christina Loosestrife Herb total flavonoids product can reach 60.8%after separation and purification.The adsorbent can be used 4 times repeatedly.Moreover,to establish a kinetic model for this separation process,the curves of dynamic adsorption-elution process have been pitted,and the kinetic equation has been present as y=u(1—e^-kx).Content mensurate of Quercetin and Kaempferol in Christina Loosestrife Herb total flavonoids by HPLC after separation and purification.The optimum procedure is:separation of these components was achieved on a C₁₈ Nova—pak column with gradient elution using methanol—water—phosphate acid(500:500:1) as mobile phase,constant flow rate of 1.0 mL·min^-1,column temperatue was 25℃and UV detection was set at 360nm Retention time of Quercetin is approximately 5.1 min and 8.8min of Kaempferol.The two calibration curves shows good linearity(r²ï¼ž0.999) with test ranges.Intra-assay precision of both components are 0.31%and 0.27%,the average recovery rates botained are in the range of 101～105%.Both components in the experiment of repeatability with RSD are 2.95%and 1.67%.,and they can keep stable in 24 hours in the experiment.The peak shape and resolution are both satisfied in the content mensuaration,therefore,it can be a method for other flavonols' mensuarations.By this research we find a method to qualify Christina Loosestrife Herb.