Separation And Contents Determination Of Several Traditional Chinese Herbal Medicines

Quantitative analysis is essential for the determination of drug components and drug quality testing.With the development of science and technology,the modern analytical technology and equipment have been applied,such as gas chromatography,high performance liquid chromatography,ion exchange chromatography,et al.With high separation efficiency,short analysis time and low cost,capillary electrophoresis has become one of the most widely used analytical techniques.In this paper,the response surface methodology(RSM)was used to optimize the separation of several compounds from different traditional Chinese medicine(TCM),including Inula britannica L.,Aleuritopteris argentea,Apocynum venetum L.,and Acanthopanax seuticosus.And the content of some components in the sample was determined with capillary electrophoresis,which provided new analytical methods for the quality control of these TCM.Objective:1.To optimize the separation and determination of rutin,quercetin and chlorogenic acid in Inula britannica L.single-factor test and RSM were used.2.To optimize the separation of rutin and quercetin-3-?-D-glucoside and to determine the content of rutin and quercetin-3-?-D-glucoside in Aleuritopteris argentea samples from different areas.RSM with capillary electrophoresis method was used in this study.3.To optimize the separation conditions of acacetin and genistein and to analysis of acacetin and genistein in Apocynum venetum L.,RSM with capillary electrophoresis was used,which provided a reference for further development of Apocynum venetum L.resources.4.To analyze daidzein and quercetin in TCM of Acanthopanax seuticosus,dispersive liquid-liquid microextraction(DLLM)with capillary electrophoresis was utilized.Method:1.The conditions for the capillary electropheretic separation conditions were: 30mmol/L borax-0.5 mmol/L boric acid-4.37 mmol/L BMIM-PF6(pH 9.0)as background electrolyte,applied voltage of 16 kV,hydrostatic injection of 20 s(height difference of 11.5cm),detection wavelength of 354 nm.2.The conditions for the capillary electropheretic separation were: 23 mmol/L borax-1.0mmol/L boricacid-5.34 mmol/L BMIM-PF6(pH 8.7)as background electrolyte and applied voltage of 18 kV,hydrostatic injection of 20 s(the hight of 11.5 cm),detection wavelength of354 nm.3.The conditions for the capillary electropheretic separation were: 35 mmol/L borax-1.0mol/L boric acid-10 mmol/L ?-cyclodextrin(pH 9.0)as background electrolyte and applied voltage of 11 kV,hydrostatic injection of 15 s(the height difference of 11.5 cm),detection wavelength of 214 nm.4.The conditions for the capillary electrophoretic separations were: 25 mmol/L borax-1.0 mol/L boric acid(pH 9.0)as background electrolyte,applied voltage of 16 kV,hydrostatic injection of 5 s(the height difference of 11.5 cm),detection wavelength of 214 nm.Separation and extraction of daidzein and quercetin were carried out by using 77 ?L of ethyl acetate as extractant and 980 ?L of acetonitrile as dispersant.Result:1.Under the optimum conditions,rutin,quercitrin and chlorogenic were well separated within 10 min,and good linearity was found in the range of 2-100,2-250 and 2-200 mg/L for rutin,quercitrin and chlorogenic acid,respectively.The detection limits of rutin,quercitrin and chlorogenic acid were 0.6,0.6 and 0.5 mg/L,respectively.The method was applied to analyze rutin,quercitrin and chlorogenic acid in Inula britannica L.samples and recoveries were 102.1±4.0%,100.9±3.3% and 101.1±4.8%,respectively.2.Under the optimum conditions,rutin and quercetin-3-?-D-glucoside were separated successfully within 10 min,and good linearity was found in the range of 2-200 and 2-250mg/L for rutin and quercetin-3-?-D-glucoside,respectively.The detection limits of rutin and quercetin-3-?-D-glucoside were 0.6 and 0.4 mg/L,respectively.The method was applied to analyze rutin and quercetin-3-?-D-glucoside in Aleuritopteris argentea samples and recoveries were 107.9±4.6% and 102.4±4.8%,respectively.3.Under the optimum conditions,acacetin and genistein were well separated within 10 min,and good linearity was found in the range of 2-200 and 2-400 mg/L for acacetin and genistein,respectively.The detection limits of acacetin and genistein were 0.7 and 0.5 mg/L,respectively.The method was applied to analyze acacetin and genistein in Apocynum venetum L.samples and recoveries were 102.9±3.9% and 100.6±4.8%,respectively.4.A method was established for the determination of daidzein and quercetin by liquid-liquid microextraction and capillary electrophoresis.Good linearity was found in the range of 2-400 mg/L for both daidzein and quercetin.The detection limits of daidzein and quercetin were 0.6 mg/L.The method was applied to analyze daidzein and quercetin in Acanthopanax seuticosus real samples and recoveries were 94.7 ± 4.9% and 94.2 ± 2.7%,respectively.Conclusion:In this study,the single-factor test and RSM were used to optimize the extraction and separation conditions of several TCM with Design-Expert 8.0 software.The developed methods have the advantages of fastness,convenience and low cost and can be used for the quality control of these TCM.