碩士 / 國立屏東科技大學 / 食品科學系 / 94 / The objects of this study were to compare the analytical methods of HPLC and LC-MS for the antibacterial drugs- nitrofuran and its metabolites residues in aquacultural fish and to apply ELISA method to detect AOZ residues in fish tissue. Nitrofuran, which including FZD, FTD, NFZ, and NFT, had been widely used in animal feed additives or aquacultural fish due to their efficiency and cheap price. But, because of the carcinogenic and mutagenic toxicity, its application on animals for food producing in EU was prohibited since 1993. The nitrofurans are characterized by the rapid metabolism with short half-life, but, its metabolites, AOZ, AMOZ, SC, and AH, which are much stable in tissue bounded with protein. Hence, it will be more meaningful to detect the metabolites to monitor the illegal use of nitrofurans.
About HPLC method, samples were extracted by adding 1 to 1 ratio of 0.2 % metaphosphate methanol and deionized water solution and ethyl acetate. Mobile phase was mixed with acetonitrile and 0.05 M NaH2PO4 by 20 to 80 ratio at pH 5.1, and could be completely conducted within 20 minutes. The limit of quantification was 0.02 ppm, the recovery was ranged from 63.8- 75.5 %, and the CV was 2.8-7.4 %.
About LC-MS analysis, the derivatisation should be done previously by adding 0.125 M HCl and 2-NBA overnight for 16 hours. Then, SPE cartridge was used to clean up and eluted by ethyl acetate. The gradient mobile phase was used, and total analysis could be finished within 30 minutes. The limit of quantification for AMOZ and AOZ were 0.25 ppb, SC and AH were 0.50 ppb, respectively. The recovery was 63.2-77.3 %, and CV was 4.8-8.4 %. Trying to understand whether the different treatment had the effect on the residues or not, the fish samples were stored for 10 days under -18℃ and heated by microwave for 6 minutes, respectively, and the results revealed both the treatments had not the significant effects no destroying metabolite residues.
About AOZ-ELISA analysis, the detection limit was 0.2 ppb, recovery was from 82.6- 98.2 %, CV was 0.8-2.9 %, and the total analysis was within 90 minutes. Therefore, AOZ-ELISA kit would be recommended for the screening work on large sample size.
| Identifer | oai:union.ndltd.org:TW/094NPUST253002 |
| Date | January 2006 |
| Creators | Hsing-Hsiang Cheng, 鄭幸祥 |
| Contributors | Chi-Ching Yung, 楊季清 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 73 |
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