本研究目的有二，一為開發含氯黴素的魚肉基質前處理及定量分析方法，以符合歐盟規範之標準，第二是以台灣養殖魚類出口大宗吳郭魚來瞭解吳郭魚體內氯黴素排除的情形。將組織均質後以乙酸乙酯萃取，並用正己烷去脂，液相層析系統是用0.5mM醋酸胺和甲醇作為移動相，層析管柱為C18的逆相管柱，質譜儀偵測部分選用的是負離子。回收率方面，鰓、腸、肝臟及魚肉分別是68.8、73.3、82.7、85.5％，偵測極限分別是0.05、0.14、0.07、0.03 ng/g，魚肉CCα、CCβ為0.05、0.07 ng/g。藥物殘留實驗方面，分為藥浴和餵食添加兩部分，藥浴濃度為45.8 mg/L氯黴素，藥浴時間為1小時，鰓、腸及魚肉分別於第193、673、385小時後測不到殘留，肝臟於第673小時仍測得0.08 ng/g。餵食添加部分以5000 mg/kg添加於飼料中，而實際測得濃度為354.5 mg/kg，魚肉於第480小時後測不到殘留，肝臟及腸於第648小時仍分別測到0.09 ng/g及0.16 ng/g。無論是藥浴或者是餵食添加，肝臟累積的氯黴素濃度均為最高。高濃度時各部位排除速率比低濃度時來的快。

There are two purposes in this research, one is to develop the new method which can be used for detection and quantification of chloramphenicol in fish, and the method is according to Commission Decision 2002/657/EC. The other is to study about the depletion of chloramphenicol in Tilapia which is the main aquacultural product in Taiwan. Homogenized fish tissues were extracted with EtOAc and defatted with Hexane. HPLC separation was conducted on RP18 column in 0.5mM ammonium acetate – MeOH. Chloramphenicol was determinated by LC-ESI-MS-MS in negative mode. The recoveries of chloramphenicol from fish gill, gut, liver and muscle were 68.8, 73.3, 82.7 and 85.5%. The limit of detection was 0.05, 0.14, 0.07 and 0.03 ng/g. Decision limit(CCα) of fish muscle was 0.05 ng/g and detection capability(CCβ) was 0.07 ng/g. Chloramphenicol was administered by oral and by bathing respectively in Tilapia. Chloramphenicol was given by bathing at 45.8 mg/kg for one hour. No chloramphenicol was demonstrated in gill, gut and fish muscle on 193th hour, 673th hour, and 385th hour respectively after the beginning of this study. There was 0.08 ng/g chloramphenicol in liver after 385th hour. Chloramphenicol was given orally to Tilapia at 354.5 mg/kg daily for 3 days. No chloramphenicol demonstrated in fish muscle on 480th hour. There were 0.09 ng/g and 0.16 ng/g in liver and gut respectively. No matter oral experiment or bathing experiment, th concertration of chloramphenicol in liver is higher than in the other, the depletion rate in high concentration was faster than in low concentration.

中文摘要…………………………………………………………………… I
英文摘要…………………………………………………………………… II
目錄…………………………………………………………………………III
表目錄………………………………………………………………………VII
圖目錄 ………………………………………………………………… VIII

一、前言…………………………………………………………………… 1

二、材料方法……………………………………………………………… 11
(一) 藥品及溶液配製 ………………………………………………… 11
1.藥品部分 ………………………………………………………… 11
2.試劑水 …………………………………………………………… 11
3.試劑配製 ………………………………………………………… 12
（二）儀器與設備 ……………………………………………………… 12
（三）分析方法 ………………………………………………………… 12
1.標準品配製 ……………………………………………………… 12
2.化合物滯留時間及定量、定性離子與離子比 ………………… 13
(1)滯留時間………………………………………………………… 13
（2）定量離子與定性離子 ………………………………………… 13
（3）離子比 ………………………………………………………… 14
3.萃取方法 ………………………………………………………… 14
4.方法偵測極限與CCα、CCβ…………………………………… 16
（四）畜養及取樣流程………………………………………………… 16
1. 藥浴部分 ……………………………………………………… 16
2. 餵食部分 ……………………………………………………… 17

三、結果與討論…………………………………………………………… 18
（一）儀器條件 ………………………………………………………… 18
1.液相層析部分 …………………………………………………… 18
2.質譜儀部分 ……………………………………………………… 18
3.儀器討論 ………………………………………………………… 25
（二）萃取方法 ………………………………………………………… 27
1.萃取溶劑比較 …………………………………………………… 27
2.溶劑量比較 ……………………………………………………… 29
3.乾燥溫度比較 …………………………………………………… 29
4.去脂溶劑量比較………………………………………………… 32
（三）方法確效 ………………………………………………………… 34
1.偵測極限、CCα、CCβ ………………………………………… 34
2.定性 ……………………………………………………………… 36
3.檢量線及回收率 ………………………………………………… 36
4.各萃取方法比較 ………………………………………………… 40
(四) 藥浴殘留實驗 ………………………………………………… 41
（五）餵食殘留實驗 ………………………………………………… 54

四、結論 ………………………………………………………………… 61

五、建議 ………………………………………………………………… 62

六、參考文獻……………………………………………………………… 63

七、附錄 ………………………………………………………………… 66
（一）CCα(decision limit)、CCβ(detection capability)計算公式
………………………………………………………………… 66
（二）CCα(decision limit)、CCβ(detection capability)原始數據
………………………………………………………………… 67
（三）藥浴實驗各時間鰓中氯黴素殘留量…………………………… 68
（四）藥浴實驗各時間腸中氯黴素殘留量…………………………… 69
（五）藥浴實驗各時間肝臟中氯黴素殘留量………………………… 70
（六）藥浴實驗各時間魚肉中氯黴素殘留量………………………… 71
（七）餵食實驗各時間腸中氯黴素殘留量…………………………… 72
（八）餵食實驗各時間肝臟中氯黴素殘留量………………………… 73
（九）餵食實驗各時間魚肉中氯黴素殘留量………………………… 74

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