臺灣博碩士論文加值系統

本研究發展快速 (Quick)、簡單 (Easy)、便宜 (Cheap)、有效 (Effective)、耐用 (Rugged) 及安全 (Safe) 之 QuEChERS 萃取技術結合液相層析串聯質譜術 (Liquid Chromatography-Tandem Mass Spectrometry) 於蔬果中 Diafenthiuron、Dicamba、Fenpyrazamine、Hymexazol、Thiocyclam、Tridemorph 及 Tecloftalam 等七種殘留農藥進行檢測。
實驗結果之 QuEChERS 方法最佳化條件，於離心管中加入 2 g 均質化之蔬果樣品，添加 2 mL 含 1% 醋酸之乙腈溶劑進行液液萃取，渦流震盪 1 分鐘並靜置 10 分鐘後，加入 800 mg 無水硫酸鎂及 200 mg 無水醋酸鈉，蓋上離心管蓋後隨即震盪數次以防鹽類結塊，接著渦流震盪 1 分鐘並以 3000 rpm 離心 5 分鐘後，取上清液 1.2 mL 置於含 80 mg florisil 及 180 mg 無水硫酸鎂的離心管中，渦流震盪 1 分鐘並以 3000 rpm 離心 5 分鐘後，取上清液 0.6 mL，並以氮氣吹乾後加入 200 μL 初始動相溶液進行回溶，通過 0.22 μm PVDF 濾膜後，取得之溶液以 LC-MS/MS 進行上機分析。於此條件下，偵測蔬果樣品中七種農藥之線性範圍為 0.1-1000 ng/g，線性相關係數 (R2) 均在 0.9977 以上，偵測極限介於 0.03-0.12 ng/g。本研究不僅可以有效地降低有機溶劑之使用量，減少樣品前處理時間，具有高靈敏度並可應用於其他蔬果中殘留農藥之分析。

In this study, the QuEChERS (quick, easy, cheap, effective, rugged, and safe) as sample preparation method and combined with liquid chromatography/tandem mass spectrometry (LC-MS/MS) was proposed to determine seven pesticides including diafenthiuron, dicamba, fenpyrazamine, hymexazol, thiocyclam, tridemorph, and tecloftalam in fruits and vegetables.
The extraction efficiency of QuEChERS were systematically studied and the optimal extraction conditions were as follow:2 g of homogenized fruits or vegetables were extracted with 2 mL of acetonitrile containing 1% acetic acid. Then the sample was vortexed for 1 min and left for 10 min. For phase separation, 800 mg anhydrous magnesium sulfate and 200 mg sodium acetate anhydrous were added. The tube was immediately shaken to avoid the caking behavior of salt, vortexed in a Vortex-Mixer for 1 min, and then centrifuged for 5 min at 3000 rpm. A 1.2 mL aliquot of the supernatant was transferred to a centrifuge tube containing 180 mg anhydrous magnesium sulfate and 80 mg florisil. The tube was vortexed for 1 min and centrifuged at 3000 rpm for 5 min. A 0.6 mL of the resulting supernatant was evaporated to dryness under a stream of nitrogen, and reconstituted using 200 μL of initial mobile phase. Finally, the extract was filtered through 0.22 μm PVDF syringe filter and then placed into a glass autosampler vial for LC-MS/MS analysis. In this study, the linear range for seven pesticides was from 0.1 to 1000 ng/g with coefficients of determination (R2) above 0.9977. The limits of detection (LODs) of proposed method ranged from 0.03 to 0.12 ng/g. The results showed the developed method can efficiently decrease the usage of organic solvent, reduce the operation time, and successfully utilized for analyzing of trace pesticides in fruits or vegetables.

謝誌 i
摘要 ii
Abstract iv
目錄 vi
圖目錄 x
表目錄 xiv
第一章 緒論 1
1.1 農藥之介紹 2
1.2 農藥之毒性 3
1.3 七種農藥分析物之偵測方法 7
1.4 樣品前處理技術 9
1.4.1液相-液相萃取法 (Liquid-Liquid Extraction, LLE) 10
1.4.2固相萃取法 (Solid-Phase Extraction, SPE) 11
1.4.3 QuEChERS 萃取技術 14
1.5 質譜技術原理 19
1.5.1串聯質譜儀 (Tandem mass spectrometer, MS/MS) 20
1.5.2質量分析器 23
1.5.3三段四極矩串聯質譜儀 (Triple quadrupole mass spectrometer, QqQ) 24
1.5.4電噴灑游離法 (Electrospray ionization, ESI) 27
1.5.5加熱式電噴灑游離法 (Heated electrospray ionization, HESI) 33
1.6 研究目的 34
第二章 實驗部分 35
2.1 藥品與試劑 35
2.2 藥品配製 36
2.2.1七種農藥 1000 μg/mL儲存溶液 (Stock solution) 之配製 36
2.2.2七種農藥 100 μg/mL儲存溶液 (Stock solution) 之配製 36
2.2.3七種農藥 10 μg/mL 及 5 μg/mL 混合標準溶液之配製 37
2.2.4七種農藥 100 μg/mL 標準水溶液之配製 37
2.2.5七種農藥 10 μg/mL 及 1 μg/mL 混合標準水溶液之配製 37
2.2.6 1 M 甲酸銨水溶液 38
2.2.7 不同比例醋酸之乙腈溶液 38
2.2.8 真實樣品之來源 38
2.3 實驗器材與儀器設備 39
2.3.1實驗器材 39
2.3.2液相層析串聯質譜儀 40
2.3.3液相層析條件 40
2.4 液相層析質譜儀參數之探討 42
2.4.1游離化方式最佳化之探討 42
2.5 QuEChERS 萃取技術之條件探討 42
2.5.1萃取溶劑種類 42
2.5.2萃取溶劑添加酸之含量 43
2.5.3萃取溶液之靜置時間 44
2.5.4無水醋酸鈉之比例 44
2.5.5添加吸附劑之種類 45
2.5.6吸附劑之添加量 46
2.6 方法確效 46
2.7. 真實樣品之分析 47
第三章 結果與討論 49
3.1 質譜儀最佳化參數條件 49
3.2 七種農藥標準品質譜圖之建立 49
3.3 選擇反應偵測模式之探討 51
3.4 游離化方式最佳化之探討 56
3.5 QuEChERS 萃取技術之條件探討 63
3.5.1萃取溶劑種類 63
3.5.2萃取溶劑添加酸之含量 65
3.5.3萃取溶液之靜置時間 65
3.5.4無水醋酸鈉之比例 68
3.5.5添加吸附劑之種類 68
3.5.6吸附劑之添加量 74
3.6 方法確效 78
3.6.1線性濃度範圍、線性方程式、線性相關係數、定量極限與偵測極限 78
3.6.2 精密度評估 80
3.6.3 回收率評估 82
3.7 真實樣品之分析 82
第四章 結論 89
第五章 參考資料 91

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