臺灣博碩士論文加值系統

本研究用(S)-甲基安非他命老鼠單株抗體((S)-(+)-methamphetamine mouse monoclonal antibody )修飾矽膠，以之填充毛細管而成毛細免疫親和性管柱（capillary immunoaffinity columns; CIACs），兩支CIACs(規格為：內徑250 μm、外徑363 μm和長度13 cm)輪流交替使用，同時行萃取及偵測的工作。尿液樣品只需加MA-d14作內標（internal standard; IS），並經0.45 μm濾膜過濾，即注入檢測系統，移動相為醋酸鹽水溶液，檢測過程中不用有機溶劑，對操作人員健康和環境污染不造成任何傷害，此法在20分鐘內，直接分離且定量檢測尿液中甲基安非他命(methamphetamine；MA)、亞甲雙氧甲基安非他命 (methylenedioxymethamphetamine；MDMA)和副甲氧基甲基安非他命（paramethoxymethamphetamine；PMMA）等光學異構物。用50、250和500 ng/mL三種不同濃度尿液樣品，測其二次質譜，以各物種特徵碎片離子畫其萃取離子層析圖(extracted ion chromatogram; EIC)，從各物種EIC圖中峰面積與IS的EIC峰面積之比值，得定量檢測上述五種成份之檢量線，此法在不同天檢測添加五種不同濃度之上述五種成份之尿液樣品五次，結果得知：最低偵測濃度 (limit of detection; LOD)和最低定量濃度（limit of quantitation; LOQ）分別介於7.7~11.9 ng/mL和25.6~39.5 ng/mL之間，相對標準偏差（relative standard deviation; RSD）及相對誤差(relative error; RE) 分別在25﹪及15﹪以下，結果顯示此法具高精密度(precision)和準確度(accuracy)，可實際用於尿液中上述五種成分之檢測。

This study developed a method capable of direct quantitation of the enantiomers of methamphetamine (MA) , 3, 4-methylenedioxy methamphetamine (MDMA; ecstasy) and paramethoxymethamphetamine (PMMA) in urine samples by preparing capillary immunoaffinity columns (CIACs) and applying the CIACs to a liquid chromatography electrospray ionization-mass spectrometry system (LC/MS). Two CIACs were prepared by packing (S)-methamphetamine monoclonal mouse antibody immobilized silica into two capillary columns (I. D. 250 μm, O.D. 365 μm and length 13 cm), and these two CIACs were used for extracting analytes from urine samples and detecting the analytes by the LC/MS system alternatively. Aqueous ammonium acetate buffers were used as the mobile phase. Urine samples spiked with different concentrations of S(+)-MA , racemic MDMA and racemic PMMA were added with fixed concentration of racemic deuterated methamphetamine (MA-d14) as internal standard (IS), filtered through a membrane and injected into the LC/MS system without any further pre-treatment. Protonated molecular ion of MA (m/z 150), MDMA (m/z 194), PMMA (m/z 180) and MA-d14 (m/z 164) were isolated and further fragmented, the respective product ions, m/z 119 for MA, m/z 163 for MDMA, m/z 149 for PMMA and m/z 130 for MA-d14 were collected for quantitative determination. The results exhibited that, between-day, accuracy and precision (n = 5) of the method are fairly good.

第一章 前言 1
1-1研究背景 1
1-2 研究動機 1
1-3 研究範圍 1
1-4 研究所面對之困難與解決之道 2
第二章 文獻探討 3
2-1 MA、MDMA和PMMA光學異構物之藥理 3
2-2 安非他命類藥物之光學異構物分離 4
第三章 實驗部份 5
3-1 實驗材料 5
3-2製備填充材料 5
3-3製備毛細免疫親合性管柱 6
3-4製備樣品 6
3-5-1層析方法一 6
3-5-2層析方法二 7
3-6 MS之設定參數 7
第四章 結果與討論 8
4-1安非他命類藥物之選擇 8
4-2 CIAC鑑定AP、EPHEDRINE和MDA之光學異構物 8
4-3 CIAC鑑定HMMA 9
4-4 CIAC鑑定MDEA 9
4-5管柱內PH值變化 9
4-6 CIACS在PH7.0時對(S)-(+)-MA之最大吸附量 10
4-7 MA定性分析 10
4-8 利用IAC從甲醇中萃取 MDMA和PMMA之光學異構物並分離和收集 10
4-9 MDMA和PMMA之光學異構物的定性分析 11
4-10 (S)-(+)-MA、MDMA及PMMA之光學異構物和(R)-(－)-FENFLURAMINE之定量 11
4-11 (S)-(+)-MA、MDMA與PMMA之光學異構物和 R(－)-FENFLURAMINE之檢量線（CALIBRATION CURVES） 12
4-12 最低偵測濃度（LOD）及最低定量濃度（LOQ）的評估 12
4-13 準確度（ACCURACY）與精密度（PRECISION）的評估 12
4-14 CIACS壽命探討 13
4-15 線上萃取與離線萃取 13
第五章 結論 14
參考文獻 15
附錄 18

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