液相層析串聯質譜應用於人體尿液中濫用藥物及
血清中除草劑的分析研究

中文摘要生物性檢體管制藥物之確認檢驗，是為了判定濫用與否，此確認實驗在濫用藥物尿液檢驗實驗室是非常基本且重要的工作，相關檢驗方法，國際皆以二種以上不同原理之方法鑑驗，我國採用免疫分析法做初步檢驗，另以氣相層析質譜儀(GC/MS)作為確認檢驗，而於確認檢驗中，因受限毒品成份之揮發性及儀器之感度，需要做萃取濃縮及衍生工作，增加實驗時間及所需耗材; 同樣的，生物檢體的農藥檢驗，可判定生物中毒與否，做為急診醫療參考及法醫毒物之法庭證據，而常見使用之高水溶性農藥除草劑，因揮發性低，無法直接使用氣相層析質譜儀進行分析。綜合上述藥、毒物檢驗，其零敏度、多重檢出能力及時效性要求皆高，傳統鑑定已漸不敷需求。本研究藉高敏感度液相層析串聯質譜(LC/MS-MS)進行分析生物檢體中的毒品及農藥成分，分三部分進行研究，第一部分挑選五十種管制藥物進行基本資料的建立，並挑選其中二十種做人體尿液多重藥物液相層析串聯質譜儀分析，建立一次可同時鑑定生物檢體內多重管制藥物之方法，定量極限LOQ可低至1.0ng/mL，並成功實測濫用者尿液檢體。第二部分則為建立苯氮平類藥物液相層析串聯質譜術分析方法，以及探討其於人體內代謝狀況，針對尿液檢體氟硝西泮(Flunitrazepam)及硝甲西泮(Nimetazepam)及其代謝物進行分析，並以硝甲西泮之代謝指標物7-aminonimetazepam進行分析，可區別服用硝西泮(Nitrazepam)或硝甲西泮，且藥物及其代謝物之LOQ可低至0.1ng/mL，即使代謝快速之原態藥物，亦可被準確測得。第三部分針對水溶性農藥除草劑進行分析，建立以液相層析串聯質譜術同時分析嘉磷塞(Glyphosate)、固殺草(Glufosinate)及巴拉刈(Paraquat)、二刈特(Diquat)等不同類型除草劑，並成功應用於人類血清檢體之檢測，LOQ可低至數個ng/mL。本研究利用液相層析串聯質譜儀分析高感度，檢品不需濃縮及衍生，直接以藥物原形方式做確認，透過簡易之自動固相萃取，取代繁複且回收率不佳之液相萃取，不但一次可進行多重藥物鑑定，並能透過生物性檢體代謝物鑑定，尋找指標代謝物進行檢測，解決原態藥物殘留量低，不易由傳統檢驗方法被檢出之困境，可得節省溶劑且縮短萃取、儀器分析鑑驗時間、減少可能之系統污染且同時獲得可靠鑑驗結果等效益。本研究成功建立濫用藥物及被廣泛使用水溶性除草、殺蟲劑等多重成分人體內血清濃度鑑定方法，並經由鑑定藥物於體內代謝物形式，建立藥物鑑定方法，達成濫用藥物尿液鑑定，農藥除草劑毒物血清等毒、藥物鑑定，滿足此等藥、毒物於急診醫學、毒品鑑定及法醫毒物學快速、準確之鑑定需求。

關鍵字

液相層析串聯質譜；濫用藥物；除草劑；尿液；血清

並列摘要

Abstract The concentration of abused drug in human urine is measured for the determination of drug of abuse. The confirmation test is a basic and very important work in a Drug Urine Testing Lab. There are two more methods have been choosen for the testing. The EIA(Enzyme Immunoassay Analysis) method is regulated as a screening test and then the GC/MS (gas chromatography/ mass spectrometry)for the confirmation test in many countries. In the confirmation test, the low volality of abuse drugs and their metabolites in urine which have insufficient sensitivities in the GC/MS analysis. There have extra extraction and derivatization works to be overcome which increase the difficulities of the analysis. The same condition is in the determination of the herbicides. Especially in the determination of water soluable herbicides in human serum for the purpose of emergency medicine and forensic evidence. The highly water soluble herbicides which have low valality and increase the difficulties of analysis by GC/MS. However, The traditional method likes GC/MS can’t meet the sensitivity, the simultaneously detection ability and the fast detection requirements for these kinds of drugs and highly water soluble herbicides. In this study, the LC/MS-MS( Liquid Chromatography/ Mass Spectrometry) was introduced to develop methods to determine these kinds of abuse drugs or highly water soluable hebicides in the biofluid of human body. Here LC/MS was used as the main instrument to determine drugs in biofluids for three parts of the fileds. The first, fifty oftenly kinds of abuse drugs in schedule I to IV were choosen for analysis and established the basic mass fragmentation in mass spectroscopy. Twenty of these drugs were selected as the simultaneously detection in human urine of LC/MS. Second, we focus on the benzodiazepines drugs and determined the drug of abuser by the presence of the metabolites in human urine of the fast metabolization cases. The third parts the highly soluble herbicides we determined in human serum by LC/MS. The drugs, however, whether the chemical substance is used in human or not, can be directly analyzed by the high sensitive LC/MS without specimen treatment likes deravative procedure. The developed methods which the simultaneously determinaton of drugs of abuse and along with indicator drugs in fast metabolized case which were analyzed to be achieved by saving the analysis time and the solvents with accuracy. In this study, the developed methods were evaluated to meet the analysis requirements of those abuse drugs and herbicides in human body for determination of those chemicals in emergency medcine, drug of abuse and forensic toxicology.