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dc.contributor.authorLin, Chia-Yingen_US
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dc.description.abstractA simple and sensitive method using micro solid-phase extraction (µSPE) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for determination trace amount of tetramethylammonium hydroxide (TMAH) in mouse serum was evaluated in this thesis. The heated electrospray ionization (HESI) with positive mode of triple quadrupole mass spectrometry was used to analyze tetramethylammonium hydroxide. Tetramethylammonium was extracted from serum by µSPE with a mixed-mode cation exchange sorbent and 0.1% trifluoroacetic acid in acetonitrile used as the eluting solvent. The volume of sample used was 15 µL. The linear range of the method was 10-2500 ng/mL with coefficient of determination at 0.9944. The limit of detection and limit of quantitation was 1.9 and 6.3 ng/mL, respectively. The intra-day and inter-day precisions with relative standard deviations were less than 2.5 and 9.8 %, respectively. The recovery of the method was greater than 95.6 %. As the matrix effect was unconsidered from the results of post-column infusion and post-extraction spiked. The feasibility of applying the proposed method to determine tetramethylammonium in serum was examined by analyzing real mouse serum sample of TMAH-exposured. The concentration of tetramethylammonium detected was range from 1.1 µg/mL to 9.1 µg/mL. The results of this study reveal the proposed method offers low ng/mL level sensitivity and high selectivity for determination of tetramethylammonium in mouse serum.en_US
dc.description.abstract本研究主要是利用微量固相萃取法結合液相層析串聯質譜儀,分析老鼠血清中氫氧化四甲基銨(Tetramethylammonium hydroxide, TMAH)含量。使用混合相陽離子交換(Oasis® MCX)吸附劑之微量固相萃取管萃取血清中四甲基銨,並探討於萃取過程中沖提溶劑種類與沖提溶劑體積對萃取效果之影響,以三段四極矩液相層析串聯質譜儀之加熱式電灑游離法於正離子模式偵測四甲基銨。實驗結果顯示,利用200 µL含0.1%三氟乙酸之乙腈溶液,可得到最佳萃取效率,且僅需15 µL血清樣品即可進行分析。此技術所得相對回收率於不同濃度均高於95.6%,同日(intraday)及異日(interday)之精密度以RSD%表示均低於2.5%及9.8%,於血清中四甲基銨分析線性範圍為10~2500 ng/mL,偵測極限值(LOD)為1.9 ng/mL,定量極限值(LOQ)為6.3 ng/mL。評估基質效應對四甲基銨偵測之影響,以管柱後注入法(post−column infusion)可得基質成分在層析時間中對四甲基銨離子化效率沒有影響;以萃取後添加法(post−extraction spiked)評估基質成分於µSPE-LC-HESI-MS/MS可得基質效應(ME)為95.2%。並將此技術應用於經氫氧化四甲基銨塗覆於皮膚表面之小白鼠血清中,均可有效測得老鼠中四甲基銨濃度為1.1 µg/mL~9.1 µg/mL。zh_TW
dc.description.tableofcontents中文摘要 Ⅰ 英文摘要 Ⅱ 目錄 Ⅳ 圖目錄 Ⅶ 表目錄 Ⅹ 一、緒論 1 1.1 前言 1 1.2 氫氧化四甲基銨 2 1.3 氫氧化四甲基銨中毒事件 4 1.4 質譜儀儀器原理 5 1.4.1 串聯質譜儀 (tandem mass spectrometer, MS/MS) 6 1.4.2 四極矩質量分析器之原理 (quadrupole mass analyzer) 9 1.4.3 電灑游離法 (electrospray ionization, ESI) 13 1.5 前處理技術 17 1.6 基質效應 24 1.7 研究目標 27 二、實驗 29 2.1 標準品與試劑 29 2.1.1 標準品 29 2.1.2 試劑 29 2.2 藥品配製 30 2.2.1 儲存標準溶液配製 30 2.2.2 分析物標準溶液及血清工作溶液配製 30 2.2.3 檢量線血清工作溶液配製 30 2.2.4 品質管制 (quality control, QC) 血清樣品配製 31 2.3 實驗器材與儀器設備 31 2.3.1 微量固相萃取裝置 31 2.3.2 液相層析質譜儀 31 2.3.3 液相層析條件 34 2.4 微量固相萃取之實驗步驟 35 2.5 液相層析質譜儀參數之探討 35 2.5.1 Varian 1200L質譜儀電壓參數之探討 35 2.5.2 Varian 1200L質譜儀離子源之探討 36 2.5.3 TSQ Quantum Ultra EMR質譜儀離子源參數之探討 36 2.6 微量固相萃取條件之探討 37 2.7 血清中基質效應之影響 38 2.8 方法確效之探討 38 2.9 真實樣品的測定 39 三、結果與討論 40 3.1 液相層析質譜分析 40 3.2 液相層析質譜儀離子化參數之探討 43 3.2.1 Varian 1200L之電灑游離法參數探討 43 霧化氣體壓力 (Nebulizing gas pressure)之探討 45 乾燥氣體壓力 (Drying gas pressure)之探討 45 乾燥氣體溫度 (Drying gas temperature)之探討 47 界面腔溫度 (Housing temperature)之探討 47 探針電壓 (Needle voltage)之探討 49 鞘電壓 (Shield voltage)之探討 49 3.2.2 TSQ Quantum Ultra EMR質譜儀游離化法參數探討 52 3.3 微量固相萃取法之條件探討 55 3.3.1 沖洗溶劑種類之探討 55 3.3.2 沖提溶劑種類之探討 57 3.3.3 沖提溶劑添加酸種類之探討 58 3.3.4 沖提溶劑添加酸濃度之探討 58 3.3.5 沖提溶劑體積之探討 61 3.4 血清中基質效應之影響 65 3.5 µSPE−LC−MS/MS分析血清中四甲基銨之萃取回收率 67 3.6 µSPE−LC−MS/MS分析血清中四甲基銨之線性範圍及偵測極限 67 3.7 µSPE−LC−MS/MS分析血清中四甲基銨之精密度 69 3.8 真實樣品中四甲基銨之偵測 73 四、結論 76 五、參考文獻 77zh_TW
dc.subjecttetramethylammonium hydroxideen_US
dc.subjectmatrix effecten_US
dc.titleDetermination of tetramethylammonium hydroxide in mouse serum by micro solid-phase extraction combined with liquid chromatography tandem mass spectrometryen_US
dc.typeThesis and Dissertationzh_TW
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