Study On The Application Of High-performance Capillary Electrophoresis With Indirect Ultraviolet Absorption Detection

On the basis of direct ultraviolet absorption detection for plant hormones, the substances without photometric activation were researched with indirect ultraviolet absorption detection in high-performance capillary electrophoresis(HPCE), and the targets of the separation were concentrated in the fields of food additive substances and Chinese medicine, such as amino acids and bile acids, Fritillaria L. compounds. In the study, the technique of the indirect detection is a universal detection method for HPCE.Auxins and cytokinins are two sorts of important plant hormones. The normal separation methods (HPLC and GC-MS, etc) are often used in the analysis of single sort of plant hormone. Seven plant hormones with photometric activation, which is four auxins [indole-3-butyric acid (IBA), indole-3-propionic acid (IPA), indole-3-acetic acid (IAA) and 1-naphthylacetic acid (NAA)] and three cytokinins [zeatin (ZT), kinetin (KT) and 6-benzylamin purine (6-BAP)], have been researched by HPCE with direct ultraviolet absorption detection. A new additive, that was the use of glucose and triethanolamine as a mixed additive in SDS, could improve the resolution of two sorts of plant hormones. The separation of seven plant hormones could be completed in 13min under the optimum condition. The mechanisms of the effect of the mixed addition on the resolution in micellar electrokinetic capillary chromatography (MECC) were discussed in detail.Amino acids are the base units of protein as well as the common food additive substances. The normal separation methods (filter paper chromatography, molecular sieve chromatography, HPLC or GC-MS, etc) are often used in the analysis of single amino acid. Three amino acids as L-Ornithine (Orn), L-Proline (Pro) and L-Glutamine (Gin) in the study have none photometric activation. Thus they had to be researched by HPCE with indirect ultraviolet absorption detection using background electrolyte (BGE)with strong photometric activation instead of direct ultraviolet absorption detection. It was by the numbers researched for the effect of the sort and the concentration of BGE on the detected sensitivity of indirect ultraviolet absorption detection and the effect of pH and concentration of potassium dihydrogen phosphate on the HPCE's resolution. The separation of them could be completed in llmin under the optimum condition. The proposed method has successfully been applied to determining the amino acids in foods (Haitian soybean sauce, Tsingtao Beer and Amino acids oral liquid) at first. At the end, the study would provide the new method for the detection standard of food.Bile acids, which belong to drainage sterides, are both of the effective composition in traditional Chinese herbal drugs of animal bile and the surface actitive substants in bodies. The normal separation methods (TLC and HPLC, etc) are also often used in the analysis of single bile acid. The five bile acids without photometric activation, such as Cholic acid (CA), Ursodeoxycholic acid (UDOC), Hyodeoxycholic acid (HDOC), Taurocholic acid(TC), Tauroursodeoxyholic acid (TUDOC), have alike structure, particularly. Among them, UDOC and HDOC are structure-isomer. As a result, it was not only unable to determine them by direct ultraviolet absorption detection but also difficult to separate them by a common separation technology. Instead, they have been researched by HPCE with indirect ultraviolet absorption detection in the study at first. It was proved that the use of high concentrate urea and 10%acolocl in SDS could improve the resolution of them after analyzing by the numbers about the effect of composition, concentration and pH of buffer. The separation of them could be completed in 13min under the optimum condition. At the end, the study would provide the new method for animal bile's detection standard, clinical examine and metabolism mechanism research in bodies.Frit Maria L. compounds, which also belong to drainage sterides, are the effective composition in traditional Chinese herbal drugs of Bulbus Fritillariae thunbergii Miq.. The normal separation methods (TLC or HPLC,etc) are also often used in the analysis of single Fritillaria L. compound. Similarly it was not only difficult to separate them by HPCE but also unable to determine them by direct ultraviolet absorption detection because the three Fritillaria L. compounds without photometric activation, such as Peimine (PM), Peiminine (PMN), Sipeimine (SPM), have alike structure, furthermore. PMN's formula mass (F.M.) has only two hydrogen atom (2H) less than PM's. The five compounds have been researched by HPCE with indirect ultraviolet absorption detection in the study at first. It was simply researched for the effect of the sort of BGE on the detected sensitivity of indirect ultraviolet absorption detection and the effect of the concentration of SDS and borax on the HPCE's resolution. It was separated between two of former composition as PM and PNM, and the third composition as SPM, but both of PM and PNM couldn't still be separated under the initiatory condition. It has been researching for the separation and the optimum in the study so that it would provide the new method for detection standard and clinical examine of Bulbus Fritillariae thunbergii Miq..