Study Of Traditional Chinese Herb By Capillary Electrophoresis And High Performance Liquid Chromatography

Improvement of separation ability and selectivity has been a challenging research work in separation science. In the field, the high performance capillary electrophoresis (HPCE) and high performance liquid chromatography(HPLC) are the two most important tools. Because of the higher efficiency, the more rapid analytical speed, the less sample quantity and the lower cost, HPCE has been considered as an indispensable technique in separation science , and its enormous potentiality has been displayed in various fields such as nutrition science, clinical diagnosis and so on. And recently, more and more Chinese herbs have been analyzed by HPCE .In this dissertation, high performance capillary electrophoresis (HPCE) was employed to analyze traditional Chinese herb, and various amino acids in leaves of glycyrrhiza uralensis and in wall painting of Dunhuang was successfully separated by high performance liquid chromatography (HPLC).The dissertation consists of five chapters:Chapter 1 A comprehensive review on the recent technique and application of CE in traditional Chinese herbal medicine was taken with 142 references.Chapter 2 CZE was used to analyze active constituents in sophora tonkinensis gapnep under the condition of optimum buffer system at pH4.5, which is composed of 0.04mol/L citric acid and 50% methanol. The relative standard deviations of migration time and peak area was 1.39%-2.14% and 3.45% -6.13% respectively. The migration behaviors of the constituents were studied and extraction effect of methanol was compared with that of the chloroform.Chapter 3 The content of active components of sophora tonkinensis gapnep was determined by Micellar Electrokinetic Chromatography(MEKC) under the optimized buffer solution composed of 30 mmol/L sodium dodecylsulfate, 20 mmol/L sodium borate and 40% methanol.The relative standard deviations of migration time and peak area was 1. 42%-2. 55% and 4. 05%-5. 64% respectively. Meanwhile, the effect of concentration of buffer, SDS andmethanol on migration behaviors was also studied. The effect of extraction between in methanol and in chloroform was compared.Chapter 4 The protein in leaves of Glycyrrhiza Uralensis was hydrolyzed with 6mol/L HCI under the protection of vacuum and nitrogen. Amino acids of hydrolysis of protein was analyzed by reversed-phase high performance liquid chromatography with pre-column derivation using o-phthalaldehyde(OPA) as a derivation reagent. The limits of detection for all ammo acids were over the range 0. 02-0. 08 u g/L.Recoveries of all amino acids were also studied.Chapter 5 The protein and peptides in cement material of wall painting of Dunhuang was hydrolyzed with 6mol/L HCl under the protection of vacuum and nitrogen. Amino acids of hydrolysis of protein and peptides was analyzed by reversed-phase high performance liquid chromatography with pre-column derivation using o-phthalaldehyde(OPA) as a derivation reagent. The procedure of gradient elution was investigated and the limits of detection for all amino acids were over the range 0.02-0.08 u g/L.Recoveries of all amino acids were also investigated meanwhile.