以丹磺醯衍生化法搭配線上固相萃取-液相層析串聯質譜儀進行多重濫用藥物之同時分析

本研究發展以同步連線固相萃取搭配高效能液相層析串聯質譜儀，進行尿液中濫用藥物及其代謝物分析。尿液檢體經酵素水解後直接稀釋，並經由自動連線固相萃取管柱與層析管柱後，進入三段四極柱質譜儀內偵測。研究首先針對多重濫用藥物，包含安非他命類、愷他命類、鴉片類、古柯鹼、大麻代謝物、類大麻合成物質、苯二氮平類與新興濫用藥物等61種濫用藥物及其代謝物之分析建立LC-MS/MS 尿液廣測檢驗之分析方法，並與真實樣品之免疫初篩與氣相層析質譜儀結果進行比對。最後進行真實尿液檢體之分析，透過本研究開發之方法，了解戒治者真實用藥情況，以作為治療之參考。其次，以戒治者檢體常見之濫用藥物及其代謝物為範疇，包含：安非他命類、愷他命類、嗎啡類、大麻、替代療法藥物以及PMMA等13種，利用丹磺醯衍生法可以與胺類以及酚類官能基快速反應之特性，再結合同步連線固相萃取與高效能液相層析串聯質譜儀，建立更靈敏之分析平台，並進行方法確效與乾紙尿片之評估。結果顯示：衍生化反應只需要5分鐘，而衍生後酚類化合物(THC、THCA)的感度有30倍以上的提升，胺類化合物有5倍之提升。在真實尿液檢體與乾紙尿片檢體的比對，也發現衍生方法之可行性。總之，由於同步連線固相萃取可以大幅進樣，縮短前處理所消耗的時間，降低基質干擾，且相較於一般的固相萃取與液液萃取有較好的回收率，而LC-MS/MS 更提供了足夠的靈敏度與分析之藥物廣度，因此大幅減少前處理所需花費的時間與人力，使本方法具備了分析大量尿液樣本的優勢。

This research analyzed drug abuse and its metabolite in urine by online-Solid Phase Extraction with High-performance liquid chromatography Tandem Mass Spectrometer . The urine sample directly dilute after hydrolysis, and then detect by triple quadrupole mass spectrometers. The research establish the analyze method of urine test against Taiwan''s common drug abuse, and compare the consequence to the immunoassay and gas chromatography-mass spectrometry(GC-MS) .Then develop for the simultaneous detection of 61 multiple drugs and its metabolites that contains amphetamines, opiates, opiates derivatives, ketamines, new designed drugs, benzodiazepines, cocaine, cannabis and designed cannabis. In order to understand the situation of the real drug abuse, finally, analysis of real urine specimen collected from patients who received methadone maintenance therapy. Because online-Solid Phase Extraction can significantly shorten the time of pre-treatment and reduce matrix effect, and compared to the solid-phase extraction and liquid-liquid extraction has a good recovery, and LC-MS / MS also provides a sufficient sensitivity. Therefore, to reduce the time and human resource required to spend on pre-treatment make this method has the advantage of analyzing a large number of urine samples. At present , urine is the most widespread sample for drug testing. Generally, immunoassay testing use in the first step, and then the positive sample should be examined by confirmatory testing. But urine sample must store at low temperature for two year and liquid is not convenient on the stored or transported. Therefore, if can use dried filter paper, just like dried blood spots will be better. In this research, the common abuse drug was chosen, including amphetamine, methamphetamine, MDA, MDMA, PMMA, morphine, codeine, ketamine, norketamine, Buprenorphine, Methadone, THC, THCA, totally 13 compound. Dansyl chloride derivative can react with phenol and amine to enhance ionization at source of mass spectrometry. The results are demonstrated that dansylation was improved not only the sensitive but also the separation efficiency. Dansylation can significant increase sensitive of phenol compound like THC and THCA, and can be expected usefully for dried filter paper with real sample and hope this method can be extended to more drug on drug abuse test.