臺灣博碩士論文加值系統

本實驗以固相萃取法 (Solid phase extraction; SPE) 配合液相層析串聯光電二極體陣列檢測器與四極軸軌道阱質譜儀 (High performance liquid chromatography coupled to diode array detector and quadrupole orbitrap mass spectrometry; HPLC-DAD-Q-Orbitrap) 建立魚乾、魚卵及蝦米製品中 23 種人工合成著色劑之定性與定量技術。將檢體均質後以水、甲醇與四氫呋喃萃取，並利用甲醇鈉與樣品中三酸甘油脂進行交酯化反應形成脂肪酸甲酯，將分層後的上層有機相以Silica gel SepPak plus silica 管柱將脂溶性著色劑分離出來，水相以5 N 鹽酸調整 pH 值至 4，接著以 Oasis PRiME hydrophilic-lipophilic-balanced管柱純化，並以 Acquity UPLC BEH C18 管柱分離 (1.7 μm，內徑 2.1 mm × 100 mm)，移動相為 pH 7.8 之 20 mM 醋酸銨溶液與乙腈進行梯度層析，分析時間為 23 分鐘，光電二極體陣列檢測器分析於 427 nm、500 nm、550 nm 與 620 nm 波長，質譜條件為碰撞能量 30、60 eV，偵測模式以全掃描 (Full scan)，m/z 70 ~ 1050 偵測下進行檢測。魚乾、魚卵及蝦米製品中著色劑回收率介於 99.6% ~ 52.5%，變異係數小於 12.3%，線性範圍介於0.005 ~ 1 mg/kg，且相關係數皆大於 0.995。進一步採樣自台北市、新北市、基隆市及高雄市之魚乾 15 件、魚卵 12 件與 蝦米 12 件共 39 件樣品供上述方法分析，結果魚乾樣品中測得 3 件含有著色劑，但未檢出非法著色劑，魚卵樣品中 4 件檢測出含有著色劑，其中分別有 1 件及 2 件樣品中含有非法著色劑 Azorubine 及 Orange II，12 件蝦米樣品中 12 件皆有檢測到著色劑，其中有 2 件檢測到非法著色劑 Orange II。結果顯示蝦米為最常使用著色劑的樣品，每件樣品皆添加一種以上著色劑，而魚卵及蝦米樣品中甚至檢測出非法著色劑 Azorubine 及 Orange II。結果顯示本檢測方法已可供魚乾、魚卵及蝦米製品中人工合成著色劑定性定量使用。

The simultaneous determination of 23 artificial dyes in dried fish, fish roe and dried shrimp by using high performance liquid chromatography coupled to diode array detector and quadrupole orbitrap mass spectrometry (HPLC-DAD-Q-Orbitrap) was developed in this study. The samples were homogenized and extracted by water, methanol and tetrahydrofuran. Transesterification with sodium methoxide to change triglycerides converted to fatty acid methyl esters. Organic phase was SPE by Silica gel SepPak plus silica column to extracted oil-soluble dyes. Water phase was SPE by Oasis PRiME hydrophilic-lipophilic-balanced column after adjusted pH value to 4 with 5 N hydrochloric acid. The extracts wrer chromatographed by Acquity UPLC BEH C18 column (2.1 mm × 100 mm,1.7 μm) with mobile phase solution of 40 mM ammonium acetate and 2.5% acetonitrile (pH 7.8). The detecting wavelengths of 427, 500, 550 and 620 nm were used and the retention time was 23 minutes for each sample. Ionizing collision energy were 30 and 60 eV. Detection mode was under full scan m/z 70 to 1050. All peaks of chromatography showed good resolution. The recovery rates of each artificial dye in dried fish product ranged from 99.6% to 52.5% with coefficient variation being less than 12.3%. The relationship of concentration of each analyte and responding peak was linear between the range of 0.005 - 1 mg/kg, and all coefficient values are greater than 0.995. It means the developed method are useful for determining 23 artificial dyes in dried fish, fish roe and dried shrimp products. Furthermore, 15 dried fish, 12 dried shrimps and 12 fish roe products were collected from Taipei, New Taipei, Keelung and Kaohsiung for artificial dye analysis. It was found that 3 dried fish products contained legal artificial dye. One fish roe sample contained legal artificial dye, 1 sample contained illegal artificial dye Azorubine and 2 samples contained illegal artificial dye Orange II. All dried shrimp contained more than 1 artificial dyes and 2 dried shrimp samples contained illegal artificial dye Orange II. Result indicated that developed method can apply to artificial dyes detected in dried fish, fish roe and dried shrimp sample.

目錄
中文摘要 2
英文摘要 4
壹、文獻整理 6
一、著色劑 7
二、著色劑鑑定方法 15
三、食品中著色劑萃取方法 17
貳、魚乾、魚卵及蝦米中著色劑分析檢驗方法開發 18
一、前言 19
二、材料與方法 21
三、結果 31
四、討論 35
圖表 38
參、魚乾、魚卵及蝦米中著色劑含量調查 53
一、前言 54
二、材料與方法 55
三、結果 61
四、討論 63
圖表 64
肆、結論 67
伍、參考文獻 68
附錄 80
謝辭 104

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