Research Of Cortex Phellodendron Amurensis Centent Determination Method Of Active Ingredients And Extraction Technology

Objective:With assistance of HPLC technique,we determinedthe content of seven kinds of ingredients of phellodendrine,magnoflorine,j atr orrhizine,palmatine,berberine,obaculactone and obacunone from cor tex phellodendron amurensissimultaneously,and discussed the best extracted technology of cortex phellodendron amurensis on the b asis of this analytic method.In addition,we determinated the cont ent of total alkaloids in the extraction from cortex phellodendron amurensis,and established quality control of it.Methods:HPLC was adopted to construct the content of five kinds of alkaloids component and two kinds of limonoids chemical composition form cortex phellodendron amurensis which was determined simultaneously.At this analytic method as premise,Using heating reflux extraction to investigate the influence of four factors of ethanol concentration?extracting time?the volume of solvents and extraction times for the best extracted technology of cortex phellodendron amurensis by orthogonal design.With berberine hydrochloride as reference substance,employing acid dye colorimetry to measure the content of total alkaloids in the extraction form cortex phellodendron amurensis.Results:(1)Applying the means of HPLC to set up the coinstantaneo us content determination method of seven kinds of ingredients fr omcortex phellodendron amurensis.The sample was analysed by Diamonsil C18 chromatographic column(4.6mm×200mm,5?m)an d Nova-PakC 18 Guard-PakTM protect column,and the mobile ph ase was acetonitrile-0.1%Phosphoric acid water gradient elution.The flow velocity was 1.0mL·min-1.The column temperature wa s 30?.The detection wavelength of phellodendrine,obaculactone,obacunone,magnoflorine,jatrorrhizine,palmatine and berberine resp ectively was 207nm,207nm,213nm,223nm,348nm,348nm,348nm.Th e range of 3.55?35.5,9.78?97.8,0.579 8?5.798,7.25?72.5,25.2?252,14.66?146.6,5.738?57.38?g·mL-1 for phellodendrin e,magnoflorine,jatrorrhizine,palmatine,berberine,obaculactone and obacunone respectively showed a good linear relationship.A t the same time,the correlation coefficients were greater than 0.999.The relative standard deviation of precision,reproducibility and stability respectively for above seven elements were 1.68%?0.72%?1.38%?0.73%?0.51%?1.13%?0.93%,0.32%?1.52%?0.81%?1.09%?1.54%?2.53%?2.03%and 1.88%?0.68%?1.34%?0.64%?0.87%?1.17%?0.96%.The average recovery for above seven ele ments,respectively,were 98.31%?100.61%?99.68%?97.22%?01.06%?98.36%?100.71%.these evidences suggest that precision,stability,repeatability and recovery rate all meet the requireme nts,this method can be used to the seven component content si multaneous determination form Cortex Phellodendron amurensis.(2)The best extraction process of cortex phellodendron amurensis w as an amount of 6 times of 70%alcohol for 60 minutes and 3 t imes of extractions.The transfer rate of each index component o f-extractive reached 90%.(3)The best condition of employing ac id dye colorimetry to measure the content of total alkaloids in t he extraction from cortex phellodendron amurensis was pH4.0 cit raate-7.5ml disodium hydrogen phosphate buffer with bromopheno 1 blue solution 1.5ml and 10ml chloroform extracting one times,and the detection wavelength was 414.5nm.The range of 0.02?0.1 mg/ml for beberine hydrochloride had advantages in good linear relationship,and the correlation coefficients were greater than 0.999.The regression equation was y=6.6x-0.0168.Therelative stan dard deviation of reproducibility and average recovery for the te st solution of extractive of cortex phellodendron amurensis was 1.15%and 99.80%.At the same time,this analysis method was gr eatly stabilization.Conclusion:We Applyed the means of HPLC to set up the coinstantaneous content determination method of seven kinds of ingredients from cortex phel lodendron amurensis in this study.On the basis of this analytic method,the best extracted technology of cortex phellodendron amurensis was confirmed.In the meantime,we established a content determination method of the total alkaloids in the extraction from cortex phellodendron amurensis.These researches provide the basis for further perfect quality control and industrial production of cortex phellodendron amurensis.