- 學位論文
透過同位素標誌技術建立超靈敏之合成卡西酮分析方法
Development of ultra-sensitive method using chemical isotope labeling to determine synthetic cathinones with liquid chromatography-triple quadrupole mass spectrometry
摘要
近年來新興濫用藥物成長快速，其中合成卡西酮已經成為主要的濫用藥物之一且種類繁多，因此使分析合成卡西酮變得更加重要也具有挑戰性。目前偵測合成卡西酮的方法主要仰賴液相層析搭配串聯式質譜儀，其偵測極限一般可達0.1~0.5 奈克/毫升(ng/ml)，也可以同時偵測多種合成卡西酮。為求準確定量，同位素內標準品常被用來校正基質效應產生之偏差。然而，並非所有新興合成卡西酮都能在市面上找到相對應之內標準品，並且大部份可以獲得的內標準品也非常昂貴，這也使得合成卡西酮的準確定量受到阻礙。化學同位素標記法可以提升逆相層析滯留效果與偵測靈敏度。丹磺醯肼(dansyl hydrazine)已經用來標記許多化合物如酮類、固醇類、有羰基或醛基的物質，並可提升偵測度。合成卡西酮皆具有貝他位置酮基，適合丹磺醯肼衍生提升其偵測靈敏度。同時，藉由使用不同穩定同位素之丹磺醯肼(C12 與C13)，可以合成同位素內標準品做為絕對定量之用。 本研究針對其衍生效率，在反應時間、溫度與試劑種類進行最佳化。衍生後產物透過串聯式質譜儀就各項質譜參數進行離子對之最佳化，取得最佳之多重反應監測離子對後則進行層析條件最佳化。最終分析條件經過一系列之方法確效，包括線性、準確度、精確度及穩定度。我們使用31個合成卡西酮來檢視衍生化策略，其中有10個合成卡西酮的偵測極限達到飛克/毫升 (fg/ml) 等級，確校結果顯示判定係數皆大於0.996，準確度介於78~123%，精確度之相對標準偏差值皆小於18%。藉由化學同位素標記法，我們開發出更高靈敏度且可以同時準確定量31個合成卡西酮的方法。同時，透過大體積稀釋尿液模擬極低濃度環境，如社區汙水系統，用以評估此一方法之應用性。這一方法預期可應用於汙水系統並提供監測特定地區的非法濫用藥物使用狀況。
並列摘要
New psychoactive substances (NPS) have grown rapidly in the past decade. Among them, synthetic cathinones, a group of beta-ketone phenylalkylamine derivatives, had become one of the major abused species. The fast-growing synthetic cathinone market thus poses an analytical challenge. Current methods for detecting synthetic cathinones mostly relied on liquid chromatography coupled to tandem mass spectrometry techniques, which are able to reach to a limit of detection (LOD) of 0.1–0.5 ng/mL and analyze numerous analogues simultaneously. For accurate quantification, isotope-labeled standards (ISTD) are necessary to counter matrix effect among samples. However, commercial isotope-labeled internal standards for each compound are expensive and not always available, thus makes absolute quantification more challenged. Chemical isotope labeling (CIL) methods have been developed to improve retention and sensitivity. Dansyl hydrazine (DnsHz) has been applied to compounds with various moieties including ketones, carbonyls, aldehydes and can greatly improve detectability. Employing heavy version of DnsHz (C13), one-on-one internal standards can be successfully synthesized and absolute quantification can be achieved. In this study, we employed CIL strategy to develop analytical method for determining ketone-containing synthetic cathionones with synthesis of their stable isotopes for absolute quantification. We optimized several parameters that could influence the derivatization yield including reaction time, reaction solvent and reaction temperature. Multiple reaction monitoring (MRM) transitions for derivatized products at optimal conditions were then optimized in terms of product ions and various MS parameters. Separation was performed by C18 column after optimizing LC conditions. Method validation was performed by evaluating linearity, accuracy, precision and stability on 31 synthetic cathinones. Our results showed that LODs of 10 species can reach fg/ml level. The values of coefficients of determination (R2) were all above 0.996. The accuracy and precision of all analytes were in the range of 78~123% and <18%, respectively. Besides, we employed diluted urine to simulate trace conditions such as sewage system for sewage-based epidemiology (SBE) to test the applicability of this approach. With chemical isotope labeling, we developed a method with better sensitivity and can accurately quantify 31 synthetic cathinones simultaneously. This method could be applied to sewage system and help monitor appearance of illicit drugs in a certain area.
參考文獻
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