Development and Application of Capillary Electrophoresis Method in the Analysis of Multiple Amino Acids
| Autor: | Yu-Hsiang Ho, 何宇祥 |
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| Rok vydání: | 2006 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 94 The problem in real sample analysis is usually to determine trace target analytes coexisted in complicate matrices. To address the difficulties, high resolution method with good sensitivity is basically required. In this study, multiple amino acids were derivatized with a highly chromophoric reagent (2-naphthoxy)acetyl chloride. The resulting naphthoxy derivatives were analyzed by high resolution capillary electrophoresis with ultraviolet detection. The analytical derivatization makes the amino acid derivatives sensitive to a UV detector as well as easily resolvable by micellar electrokinetic capillary chromatography (MEKC). The main results of the study are summarized as follows: 1. (2-Naphthoxy)acetyl chloride was easily synthesized by reacting (2-naphthoxy)acetic acid with oxalyl chloride and it was first used to derivatize multiple amino acids for their quantitative analysis. 2. Multiple amino acids were derivatized with (2-naphthoxy)acetyl chloride in alkaline media, and the resulting derivatives were analyzed by MEKC (borate 35 mM, pH 9.50 and SDS 150 mM) with UV detection (230 nm). Seventeen amino acid derivatives (from alanine, arginine, aspartic acid, cystine, glutaminc acid, glycine, histidine, isoleucine, leucine, lysine methionine, phenylalanine, serine tryptophan, tyrosine and valine) can be completely separated in 40 min. The detection limits of the analytes were in the range of 3-8 μM (S/N = 3, injection 5 s, 1.0 psi). The precision (relative standard deviation, RSD) and accuracy (relative error, RE) of the method for intra- and inter-day analyses of the analytes were respectively below 4.8% and 4.9%. 3. Application of the method to the analysis of amino acids in real samples proved feasible including pharmaceutical preparation, sports drink, amino acids fortified liquor, and fruit juice. Further development of the method for analyzing related amino acids in biosamples for clinical study could be interesting. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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