以液相層析串聯質譜儀搭配直接樣品注射開發高通量之多重藥物分析方法

High throughput multi-drugs analysis using LC-MS/MS with direct sample injection

江孟翰(Meng-Han Chiang)

指導教授 ： 張耀仁

中山醫學大學/醫學院/醫學分子毒理學研究所/碩士(2011年)

摘要

本篇研究開發了以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。尿液檢體酸水解後直接稀釋，並經由管柱層析後，進入三段四極柱質譜儀內偵測。研究首先針對台灣常見之濫用藥物，建立尿液檢驗之分析方法，並與真實樣品之免疫初篩 (immunoassay) 與氣相層析質譜儀 (gas chromatography-mass spectrometry, GC-MS) 結果進行比對。隨後擴展至包含了安非他命類、愷他命類、鴉片類、苯二氮平類與新興濫用藥物等33種濫用藥物及其代謝物分析。最後以真實尿液檢體檢驗方法之可行性。
由於高效能液相層析串聯質譜儀 (high performance liquid chromatography-tandem mass spectrometry, LC-MS/MS) 提供了足夠的靈敏度，因此使用直接稀釋法，大幅減少前處理所需花費的時間與人力。實驗結果發現：與免疫初篩相較，大幅降低假陽性之機率，而與 GC-MS 確認檢驗結果相近。由於簡便前處理的直接進樣方法和同時檢測多種濫用藥物的能力，使本方法具備了分析大量尿液樣本的優勢，可推廣至需要分析大量尿液樣本之常規藥物檢驗實驗室。

關鍵字

尿液檢驗 ； 液相層析串聯質譜儀 ； 濫用藥物

並列摘要

A high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method is presented for the determination of 33 multiple drugs and their metabolites including amphetamines, ketamines, opiates, benzodiazepines, and new popular drugs in urine sample. After urine samples were acid hydrolysis, then direct diluted and introduced into LC-MS/MS. Determination was based on the retention time, one precursor ion and two daughter ion obtained by multiple reaction monitoring (MRM) at an individually selected collision energy. The feasibility of applying the proposed method to determine 33 drugs was examined by analyzing real urine samples.
As the high sensitivity of LC-MS/MS, this approach is greatly simplified the processes of sample preparation, and increases the efficiency of analysis significantly. The results also demonstrate the suitability of this approach for high throughput determination of the various drugs of abuse in urine. It will be welcome for routine drug-testing laboratory.

並列關鍵字

urine testing ； LC-MS/MS ； drugs of abuse

參考文獻

2. World Drug Repot 2010. 2010, United Nations Office on Drugs and Crime (UNODC).

4. Gilman AG, R.T., Nies AS, Taylor P, editors., Goodman and Gillman's the pharmacological basis of therapeutics, 8th edn. 1990.

5. Kelly, T.H., et al., Behavioral response to diazepam in a residential laboratory. Biol Psychiatry, 1992. 31(8): p. 808-22.

6. Perez-Reyes, M., et al., Clinical effects of daily methamphetamine administration. Clin Neuropharmacol, 1991. 14(4): p. 352-8.

7. Cho, A.K. and J. Wright, Pathways of metabolism of amphetamine and related compounds. Life Sci, 1978. 22(5): p. 363-72.