本研究分成兩部分，一為開發三苯甲烷類化合物前處理及定量分析的方法，其中包含萃取溶劑種類、萃取溶劑的酸鹼度、萃取次數、乾燥溫度、去脂溶劑多寡、基質差異等因子之影響來提升萃取的效率，以達到歐盟法規之標準。第二是進行此開發的分析方法之方法確效，以確認此分析方法的可行性。
分析方法中，萃取是用氰甲烷4.5 ml加上0.5 ml 的McIlvaines buffer萃取兩次，液相層析系統是用0.5 mM醋酸銨然後用醋酸調整到pH 3.8的水相，以及添加0.1%甲酸的氰甲烷，層析管柱為C18的逆相管柱，質譜儀偵測部分用的是陽離子選擇模式。
方法確效方面，每化合物的(標準偏差/平均滯留時間)都小於0.05，而離子比也都維持在穩定的狀態且都符合歐盟規範，檢量線斜率相關係數介於0.998 ~ 0.999。CCα(decision limit, 判定極限值)，孔雀綠為0.16 ± 0.07 μg/kg，還原型孔雀綠為 0.15 ± 0.04 μg/kg，結晶紫為0.20 ± 0.13 μg/kg，還原型結晶紫為0.23 ± 0.12 μg/kg；CCβ(detection capability, 就是偵測能力)，孔雀綠為0.20 ± 0.09 μg/kg，還原型孔雀綠為0.18 ± 0.05 μgkg-1，結晶紫為0.24 ± 0.16 μg/kg，還原型結晶紫為0.29 ± 0.15 μg/kg。

There are two purposes in this research, one is the development of the new method which can be used for detection and quantification of triarylmethanes in fish tissues. The other is that we confirmed validation and utility of triarylmethanes by the method that is according to Commission Decision 2002/657/EC. Homogenized fish tissues were extracted twice with acetonitrile and defatted with n-hexane. HPLC separation was conducted with the RP-18 column. The mobile phases consisted of 0.5 mM ammonium acetate buffer (pH 3.8, adjusted with acetic acid)– ACN (contained 0.1% formic acid) solution. Triarylmethane was determined by LC-ESI-MS/MS in positive mode.
The correlation coefficients of calibration curves with triarylmethane in fish tissues were 0.998 ~ 0.999. The decision limits (CCα) were 0.16 ± 0.07 μg/kg(MG), 0.15 ± 0.04 μg/kg(LMG), 0.20 ± 0.13 μg/kg(CV) and 0.23 ± 0.12 μg/kg(LCV), and detection capabilities (CCβ) were 0.20 ± 0.09 μg/kg(MG), 0.18 ± 0.05 μg/kg(LMG), 0.24 ± 0.16 μgkg-1(CV) and 0.29 ± 0.15 μg/kg(LCV).

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

壹.前言 ……………………………………………… 1
一.孔雀綠 ……………………………………………4
二.結晶紫………………………………………………8
三.水產品檢驗技術……………………………………9
四.研究目的……………………………………………10

貳.材料及方法 …………………………………………11
一.藥品及溶液配製…………………………………… 11
1.藥品 …………………………………………………11
2.溶劑 …………………………………………………12
3.氣體 …………………………………………………13
4.試劑水 ………………………………………………13
5.標準品配製 …………………………………………14
6.試劑配製 ……………………………………………15
二.器材與設備 …………………………………………16
1.器材 …………………………………………………16
2.設備 …………………………………………………16
三.分析方法 ……………………………………………17
1.樣品處理 ……………………………………………17
2.LC-MS/MS操作條件…………………………………19
四.方法最佳化 …………………………………………31
1.萃取溶劑的酸鹼度 …………………………………31
2.萃取溶劑與萃取次數…………………………………32
3.乾燥溫度………………………………………………32
4.去脂次數的影響………………………………………32
5.基質的差異……………………………………………32
五.方法確效 ……………………………………………33
1.化合物定量、定性離子及滯留時間與離子比………33
2.檢量線 ………………………………………………36
3.真實（Trueness）與精密度（Precision） ………36
4.CCα與CCβ …………………………………………37
5.試藥樣品的安定性 …………………………………38

參.結果與討論 …………………………………………40
一.方法最佳化 …………………………………………40
1.萃取溶劑………………………………………………40
2.乾燥溫度………………………………………………47
3.去脂次數的影響………………………………………48
4.基質的差異……………………………………………49
二.方法確效 ……………………………………………50
1.化合物定量、定性離子及滯留時間與離子比………50
2.檢量線…………………………………………………51
3.真實（Trueness）與精密度（Precision) ………52
4.CCα與CCβ …………………………………………57
5.試藥樣品的安定性……………………………………58

肆.結論 …………………………………………………70

伍.參考文獻 ……………………………………………72

陸.附綠 …………………………………………………76
附錄一. CCα與CCβ計算 ………………………………76
附錄二. 萃取溶劑的酸鹼度數據 ………………………79
附錄三. 萃取溶劑與萃取次數的差異 …………………82
附錄四. 乾燥溫度 ………………………………………84
附錄五. 去脂次數的影響 ………………………………86
附錄六. 基質的差異 ……………………………………88
附錄七. 方法確效的十五組數據 ………………………90
附錄八. 試藥樣品的安定性數據 ………………………94

Commission Decision 96/23/EC 2002. Off. J.Eur. Commun., L221: 8.

Commission Decision 2002/657/EC 2003. off. J.Eur. Commun., L71: 17.

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食品中動物用藥殘留量檢驗方法 – 孔雀綠及其代謝物之檢驗草案(Draft method of test for veterinary drug residues in foods – test of malachite green and its metabolite) 行政院衛生署97年1月11日署授食字第0971800008 號公告。

行政院農業委員會漁業署 – 漁業年報(1959 ~ 2007)。