Determination of abused drugs in urine by QuEChERS combined with liquid chromatography mass spectrometry

本計畫主要建立新前處理技術QuEChERS搭配高靈敏及高選擇性之液相層析串聯質譜術，應用於尿液中微量多重濫用藥物之檢測。所選定的藥物有安非他命、甲基安非他命、MDA、MDMA、MDEA、嗎啡、可待因、美沙東、古柯鹼及代謝物苯甲醯基愛哥寧、愷他命及代謝物去甲基愷他命、氟硝西泮及代謝物7-胺基氟硝西泮、阿普唑他、二氮平及代謝物nordiazepam、勞拉西泮、硝甲西泮、硝西泮、佐配眠、三唑他及其主要代謝物α-hydroxytriazolam、Δ9-四氫大麻酚及大麻代謝物四氫大麻酚-9-甲酸、及七種新興濫用藥物4-甲基甲基卡西酮(喵喵) 、1-pentyl-3-(1-naphthoyl)indole、naphthalen-1-yl-(1-butylindol-3-yl)methanone、2-(2-methoxyphenyl)- 1-(1-pentylindol-3-yl)ethanone)、1,1-Dimethylheptyl-11-hydroxyl–tetrahydro-cannabinol及 2-[(1R,3S)-3-hydroxycyclohexyl]-5-(2-methyloctan-2-yl)phenol與氯安非他命等三十二種化合物。計畫中將先最佳化液相層析串聯質譜術之分離條件、游離化方式及選擇反應偵測之離子斷裂途徑和其最佳碰撞能量，之後再探討QuEChERS萃取方法之最佳化條件，包括萃取溶劑種類、加酸於萃取溶劑中、加醋酸鹽於尿液中及吸附劑種類於尿液樣品之淨化效果。以兩者最佳化條件進行性線範圍、偵測極限、最低定量極限、回收率與精密度等方法確效之評估，最後將應用此方法於真實樣品之分析。此分析方法將作為尿液中快速篩檢常見管制藥品與新興濫用藥物方法之參考。The feasibility of applying QuEChERS combined with liquid chromatography/tandem mass spectrometry (LC/MS/MS) for simultanous determination of various abused drugs and their metabolites in urine is proposed. The target analytes are in the first to fourth controlled drugs of FDA. The analytes are including amphetamine, methamphetamine, MDA, MDMA, MDEA, morphine, codeine, methadone, cocaine, benzoylecgonine, ketamine, norketamine, flunitrazepam, 7-aminoflunitrazepam, alprazolam, diazepam, nordiazepam, lorazepam, nimetazepam, nitrazepam, zolpidem, triazolam, α-hydroxytriazolam, Δ9-tetrahydrocannabinol, 11-nor-9-carboxy-Δ9-tetra- hydrocannabinol, 4-methylmethcathinone, 1-pentyl-3-(1-naphthoyl)indole, naphthalen-1-yl- (1-butylindol-3-yl)methanone, 2-(2-methoxyphenyl)-1-(1-pentylindol-3-yl)ethanone、1,1-Dimethyl- heptyl-11-hydroxy-tetrahydrocannabinol), 2-[(1R,3S)-3-hydroxycyclohexyl]-5-(2-methyloctan-2-yl)- phenol and p-chloroamphetamine. The optimum parameters of extraction and mass spectrometric conditions will be symtematically studied. The parameters of QuEChERS included the kinds of extraction solvent, added acid in extraction solvent, and the kinds of sorbent used will be optimized. The selected reaction monitoring mode of LC/MS/MS will be utilited to determine analytes in urine after sample preparation. The method validation including linear detection ranges, limit of quantitation, limit of detection and reproducibility will be further evaluated. The application of the proposed method to analyze the trace abused drugs in real urine samples will also be examined

The studying of high throughput screening of the common controlled drugs and novel abused drugs in urine by ultra high-performance liquid chromatography/tandem mass spectrometry

本計畫主要是利用極致高效能液相層析法結合串聯質譜術，於高通量快速篩檢尿液中常見第一~第四級管制藥物與新興濫用藥物分析方法之開發與比較。研究中選定常見的第一~第四級管制藥品包括安非他命、甲基安非他命、MDA、MDMA、MDEA、嗎啡、海洛英、古柯鹼及其代謝物苯甲醯基愛哥寧、可待因、美沙東、愷他命及其代謝物去甲基愷他命、氟硝西泮(FM2)、阿普唑他、二氮平、勞拉西泮、硝甲西泮、硝西泮、佐配眠、三唑他、Δ8-四氫大麻酚、Δ9-四氫大麻酚及大麻代謝物四氫大麻酚-9-甲酸，以及新興濫用藥物4-甲基甲基卡西酮(喵喵)與氯安非他命等二十六種藥物為主要分析物。計畫中分別針對極致高效能液相層析法及串聯質譜法進行系統性最佳化探討，並針對靈敏度和準確度與文獻進行比較。計畫中將針對使用填充次二微米(sub-2 μm)以及核殼技術(core-shell)兩種不同填充材質的極致高效能液相層析管柱，進行液相層析分離條件中動相組成與分離沖提梯度，對於二十六種分析物之分離效果進行最佳化之探討。計畫中採用串聯質譜技術中高選擇性反應偵測模式(highly selected reaction monitoring，h-SRM)，進行二十六種分析物之偵測，且針對不同離子化效率進行最優化之討論。方法確效方面，研究將針對極致高效能液相層析法結合串聯質譜術分析尿液檢體中常見第一~第四級管制藥物與新興濫用藥物之線性範圍、最低定量極限、偵測極限、再現性、準確性與回收率進行探討。最後將此分析方法對於真實尿液檢體樣品進行高通量之分析，並評估此方法做為濫用藥物尿液篩檢技術之參考。This project will investigate the feasibility of applying ultra high-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) for simultanous determination of 24 common controlled drugs and 2 novel abused drug in urine. The target analytes are including amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy- methamphetamine (MDMA), 3,4- methylenedioxyethylamphetamine (MDEA), chloroamphetamine, mephedrone, morphine, heroin, cocaine, benzoylecgonine, codeine, methadone, ketamine, norketamine, flunitrazepam, alprazolam, diazepam, lorazepam, nimetazepam, nitrazepam, triazolam, zolpidem, Δ8-THC, Δ9-THC, 11-nor-Δ9-tetrahydrocannabinol- 9-carboxylic acid. Two UPLC columns, packed with sub-2 μm material and core-shell material,will be tested in this study for rapid separation of 26 analytes. The chromatographic conditions including compositions of mobile phase and separation gradient will be systematic optimized. The highly selected reaction monitoring (h-SRM) will be utilized to determine 26 analytes, and the mass spectrometric conditions will be optimized. The validation of proposed method including linear detection ranges, limit of quantitation, limit of detection and reproducibility will be systematic evaluated. The feasibility of applying the proposed method to analyze the trace controlled drugs in real urine samples was also examined

Liquid chromatography-mass spectrometry for fast screening of sedative-hypnotic drugs in urines

本計畫主要是以加熱式電灑游離法結合三段四極式串聯質譜儀，以直接進樣方式分析經簡單處理後的尿液樣品，同時分析尿液中苯二氮平類安眠鎮靜藥物alprazolam、lorazepam、flunitrazepam、midazolam、nimetazepam以及新興濫用藥物zaleplon、zolpidem及zopiclone，將作為尿液樣品初篩的檢驗方法參考。實驗中將利用加熱游離化方式結合高解析四極矩質譜儀降低尿液中基質干擾效應，之後評估利用高選擇性反應偵測模式(highly selected reaction monitoring，h-SRM)直接分析尿液中安眠鎮靜劑，以提高初篩結果的可信度。並將此篩檢應用於真實尿液樣品，其結果並將與一般篩檢方法比較，期提供尿液樣品中快速初篩安眠鎮靜及濫用藥物方法的參考