| Abstract: | 有鑑於毒品的濫用方式與品項的氾濫與複雜化，傳統的尿液檢驗易造成防治上的漏洞，因此發展毛髮、唾液等「另類生物檢體」的檢驗方法將能輔助尿液檢體檢驗，以加強檢驗的嚇阻能力。然而另類生物檢體之藥物濃度低於尿液，因此限制了其在微量檢體以及低濃度藥物的檢測方法的發展。
本論文主要目的為發展毛髮與唾液的濫用藥物檢驗方法，項目包含國際間常見之多重濫用藥物：包括安非他命、甲基安非他命、MDMA、MDA、嗎啡、可待因、6-乙醯嗎啡、愷它命及原愷他命、COC、BZE等13種濫用藥物及其代謝物。為了提升在液相層析質譜法 (LC-MS) 偵測的靈敏度，本論文以dansyl chloride管柱前衍生化策略改變分析物的離子化特性。結果顯示： AP、MA、MDA、MDMA、THC、THC-COOH、MOR、6-AM可被dansyl chloride成功衍生，並能提升酚類化合物在正離子APCI上的偵測靈敏度。
之後研究進一步以ESI進行微量毛髮多重濫用藥物同時分析之方法開發，進行方法確效評估，與真實微量毛髮檢體之分析 (1 mg)，並與氣相層析-質譜 (GC-MS) 方法進行比較：經dansyl chloride 衍生的LC-MS方法與GC-MS的定量結果具有高度的一致性。
在唾液檢體方面，由於THC-COOH在唾液中濃度較低，導致目前尚未有足夠靈敏的LC-MS方法被開發出來，因此本研究也利用dansyl chloride衍生化方法應用於唾液的大麻檢測中。在方法開發及確效後，此方法能在6件THC為陽性 (大於2 ng/mL) 的真實檢體中檢測到THC-COOH，這是我們目前所知在LC-MS上最靈敏的方法。
另一方面，由於毛髮檢驗的前處理過程複雜，且耗時甚久。因此，本論文以脫附電噴灑游離化方法 (DESI) 方法，嘗試更直接的偵測方式，於毛髮中進行濫用藥物快速偵測。我們改裝了實驗室的LCQ classic 離子阱質譜儀，並加裝可直線移動之載物平台，稱為掃描式-DESI介面。掃描式-DESI先以羅丹明B進行噴灑溶劑推送速率、噴灑電壓、以及載物平台移動速率的參數最佳化。之後以濫用藥物標準品進行靈敏度評估，其最低偵測極限在安非他命、MDA、原愷他命為 5 pg/mm2，甲基安非他命、MDMA為與愷他命為10 pg/mm2，六乙醯嗎啡為20 pg/mm2，嗎啡為 50 pg/mm2。此介面應用於真實高濃度毛髮檢體的快速直接分析上，並可同時進行毛髮中藥物足跡掃描，藥物含量縱深分布分析。
Due to the drug abuse with items increasing and complicate, traditional urine test could easily lead to the loopholes of drug testing, and therefore need to develop alternative biological specimen testing methods to enhance the deterrent capability of drug test. However, drug concentration in alternative biological specimens was lower than in the urine, thus the method developments of reduced-size sample analysis and trace concentration drug detection were limited.
The purpose of this thesis is to develop sensitive hair and oral fluid testing method for the determination drugs of abuse. The 13 international common drug and its metabolites were selected, including amphetamine (AP), methylamphetamine (MA), methylenedioxy amphetamine (MDA), methylenedioxy methmphetamine (MDMA), Tetrahydrocannabinol (THC), tetrahydro cannabinolcarboxylic acid (THCA), morphine (MOR ), 6-acetyl morphine (6-AM), codeine (COD), cocaine (COC), benzoylecgonine (BZE), that ketamine (K), norketamine (NK).
In order to elevate the sensitivities on liquid chromatography - mass spectrometry (LC-MS), the strategy of pre-column chemical derivatization with dansyl chloride was employed in this thesis to modify the properties of analytes. The results showed: after dansyl chloride derivatization, the AP, MA, MDA, MDMA, THC, THCA, MOR, 6-AM could be derivated successfully. In addition, the sensitivity on phenolic compounds could be enhanced significantly on positive APCI.
Subsequent study further applied to simultaneous analysis of multi-drug abuse in reduced-size hair sample on ESI. The method was validated and applied to the real reduced-size sample hair (1 mg). Compared to the developed method on gas chromatography - mass spectrometry (GC-MS), the quantification results showed high consistence on LC-MS after dansyl chloride derivatization.
On oral fluid testing, due to the low concentration of THC-COOH in oral fluid, caused there is no LC-MS based method was developed with sufficient sensitivity. Thus, this LC-MS after derivatization method was applied to the real cannabis abusers oral fluid sample detection. Six suspect real samples could be further detected the THC-COOH when THC was positive (> 2 ng/mL). This is our current knowledge on the most sensitive LC-MS method.
On the other hand, due to the complication of preparation process on hair test, caused the time waste. Thus, desorption electrospray ionization (DESI) was applied to direct and fast detection abused drug in hair. We modified the laboratory LCQ classic ion trap mass spectrometer, and to install a straight line moving loading platform. The interface was called scanning-DESI. The parameter on spray solvent flow rate, spray voltage, and sample plate moving rate of scanning-DESI was optimized with rhodamine B. Then the sensitivity to drug abuse standards was evaluated. The result showed the lowest detection limit for AP, MDA and NK were 5 pg/mm2, MA, MDMA and K were 10 pg/mm2; for 6-AM was 20 pg/mm2; for morphine was 50 pg/mm2. The interface was applied to direct and fast detection on real high concentration hair sample, and further used on hair drug footprint scanning, and in-depth analysis of the drug distribution. |
