藥材成分層析研究

The methods to determine Chinese herbs are usually performed by capillary electrophoresis (CE) and high-performance liquid chromatograph

溫武哲(Wu-Che Weng)

指導教授 ： 許順吉教授

國立臺灣師範大學/理學院/化學系/博士(1999年)

摘要

無資料

關鍵字

HPLC ； CE ； 中藥

並列摘要

The methods to determine Chinese herbs are usually performed by capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC). Both of these methods follow different separation mechanisms. In this study, CE method was used to assay the contents of constituents of Ligustici Rhizoma and Artemisiae Capillaris Herba. Two techniques including CZE and HPLC have been developed to separate anthraquinones and citrus herbs. The goals of this study are to compare the differences and to establish the relationship of analyses and analytical condition between CE and HPLC. Finally, we used the HPLC method to postulate the origin and quality of the citrus herbs.
A reversed electroosmotic flow capillary zone electrophoresis (reversed EOF CZE) and a micellar electrokinetic chromatography (MEKC) were developed to analyze the organic acids and essential oil constituents of Ligustici Rhizoma. In reversed EOF CZE method, a buffer solution containing 8 mM sodium borate, 3 mM sodium dihydrogenphosphate and 9 mM lauryltrimethylammonium chloride and acetonitrile (7: 3) were found to be the most suitable approach to determine the contents of phthalic acid, caffeic acid, protocatchuic acid, folic acid, nicotinic acid, vanillic acid, p-hydroxybenzoic acid, ferulic acid and folinic acid within 12 minutes. In MEKC technique based on sodium dodecyl sulfate was applied to analyze butylphthalide, senkunolide A, ligustilide and butylidenephthalide within 30 minutes.
Using the techniques of MEKC by adding sodium dodecyl sulfate to sodium borate solution and adjusting to pH 9.82 with 0.05 M NaOH, we can separate phenol, o-cresol, m-cresol, p-cresol, 4-ethyl phenol, 6-ethyl phenol, eugenol, capillarisin, scopletin, chlorogenic acid and caffeic acid within 42 minutes. Especially, the isomers of cresol and ethylphenol that can’t be separated in HPLC have been analyzed in this method.
In order to compare the superiority and shortcoming of both methods and to understand the relationship between chemical structures and analysis methods, this study has developed both CE and HPLC methods for the analysis of twelve anthraquinones (anthraquinone, chrysphanol, aloe-emodin, alizarin, purpurin, emodin, sennoside B, sennoside A, anthraquinone 2-carboxylic acid, quinarizalin, rhein and anthraflavic acid). A buffer solution containing 30 mM sodium borate (adjusted to pH=10.56 with 0.05 M NaOH) and CH3CN (9: 1) in CZE or with a linear gradient elution containing 20 mM KH2PO4 (adjusted to pH=2.91with 0.05% H3PO4) and MeOH in HPLC was found to be the best. Contents of six components (chrysphanol, aloe-emodin, emodin, sennoside B, sennoside A, rhein) in Rhei Rhizoma could easily be determined within 39 minutes by CE or 63 minuts by HPLC. The effect of buffers on this separation and the validation of two methods were studies.
The HPLC and CE methods for separating citrus herbs were established. The major components of citrus herbs contained umbelliferone, citropten, imperatorin, narirutin, naringin, naringenin-7-glucoside, hesperidin, neohesperidin, quercetin, naringenin, hesperitin, chrysine, sinensetin, nobiletin, acacetin, tangeretin, 5-demethyl-nobiletin and synephrine. The HPLC analysis was carried out within 60 minutes by using a gradient solvent system of phosphate salt buffer-methanol-acetonitrile. Using the techniques of MEKC by adding sodium dodecyl sulfate to sodium borate solution and adjusting to pH 10.01 with 0.05 M NaOH, we could separate eighteen of the thirteen components within 40 minutes. The effect of buffers on this separation and the validation of two methods were studies.
In addition, we used HPLC method to analyze the citrus herbs collected from market. In this study, we found that the Aurantii Fructus Immaturus belonged to Poncitrus trifoliata RAF, and the Aurnatii Fructus Maturus was derived from Citrus aurantium L. and C. Wilsonii TANAKA. The two kinds of Aurnatii Fructus Maturus can be distinguished by the ratios of HE/NG, HE/NE and NGC/NE. The ratio HE/NG in C. aurantium was higher than 0.49, but less than 0.40 in C. Wilsonii. In addition, the ratio HE/NE was higher than 2.30 for the farmer, but less than 1.31 for the latter; the ratio NGC/NE was higher than 0.21 for the former, and less than 0.02 for the latter. From the data of chemical analysis of an herb’s constituents, we can postulate the origin and quality of the herb.

並列關鍵字

HPLC ； CE ； CHINESE HERBS

參考文獻

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