Application Research For Separation And Analysis Of Furancoumarins By HPLC And HPCE

Objective:The separation and analysis of the six furanocoumarins active ingredients(psoralen,isopsoralen,imperatorin,phellopterin,cnidilin and isoimperatorin)in Angelica Dahurica Radix and Glehniae Radix were achieved applying high performance liquid chromatography(HPLC)in the first part.A simple and effective method for the determination of the six active ingredients in herbs and their formulations was provided a scientific basis for their quality control.In the second part of this paper,the effect of capillary electrophoresis based on a binary cyclodextrin system on the separation of six furanocoumarin components was studied,and a new efficient and fast capillary electrophoresis method was established and applied to the separation and determination of furanocoumarins in herbs and its formulations.Methods:(1)A Prominence-i LC-2030 high performance liquid chromatograph was used,connected to an Inert Sus Tain C18 column with UV detection.The ultrasonic extraction and gradient elution conditions of the six furanocoumarins were investigated,and the flow rate,solvent and ratio of the gradient elution were studied.(2)A BECKMAN capillary electrophoresis apparatus connected to a diode array detector with an uncoated capillary column(40.2 cm × 75 μm id,effective length 30cm)was used.The factors affecting the separation of the six furanocoumarins were studied: the concentration and p H of buffer solution,the concentration of sulfobutylether-β-cyclodextrin(SBE-β-CD)and 2-hydroxypropyl-β-cyclodextrin(HP-β-CD),and separation voltage and temperature,etc.Results:The results showed that the extraction of the six target analytes was achieved by using 80% methanol as the extraction solvent and ultrasonication at 30 °C for 40 min at 280 W.The target analytes and the extracted matrices were well separated in the actual sample analysis.The HPLC gradient elution conditions: 0～10 min,53%methanol;10～12 min,53%～52% methanol;12～13 min,52%～55% methanol;13～20min,55% methanol;20～35 min,55%～65% methanol;35～65 min,65% methanol.Under the gradient elution conditions,complete separation of the six furanocoumarins active ingredients in Angelica Dahurica Radix and Glehniae Radix could be achieved within 60 min;the RSDs of intra-day precision were 0.1%～0.4%,and the RSDs of interday precision were 0.1%～0.6%,the linear correlation coefficients of each analyte were0.9992～0.9999,and the recoveries of the actual samples were in the range of98.3%～102.0%.The optimal conditions for the simultaneous separation of the six furanocoumarins were obtained as follows: containing 12 m M sulfobutylether-β-cyclodextrin(SBE-β-CD),1.5 m M 2-hydroxypropyl-β-cyclodextrin(HP-β-CD)in 30 m M boric acid solution(p H 7.80)as running buffer solution,separation voltage of 20 KV.Samples were injected into the capillary at 0.5 psi for 5 s and detection wavelength was 246 nm.Under the optimal electrophoretic conditions,complete separation of the six furanocoumarins could be achieved within 7 min;the RSDs of intra-day precision were0.7%～2.0% and inter-day precision were 1.2%～1.8%,and the linear correlation coefficients of each analyte were 0.9992～0.9999,and the recoveries of the actual samples were in the range of 98.8%～101.8%.Conclusion:In this paper,high performance liquid chromatographic method was developed to realize the separation and determination of six furanocoumarins in actual samples,which provides a new method for the quality control of Angelica Dahurica Radix,Glehniae Radix and their formulations.In this paper,a capillary electrophoresis method based on a binary cyclodextrin system was developed to achieve rapid and effective separation of six furanocoumarin compounds and can be used for the determination of the six furanocoumarin active ingredients in Angelica Dahurica Radix and its formulations,which provides a reference method for the rapid analysis of furanocoumarin-like components in real samples.