유도체화 반응에 따른 빠르고 용이한 마황 중활성성분의 TLC상 분리
Fast and Easy Separation of Active Ingredients of Ephedra herba Using In Situ Derivatization Reaction in TLC
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초록
마황(Ma huang; Ephedra herba)은 불법적인 체중 조절제, 경기력 향상제 그리고 methamphetamine(Meth)의 합성에 이용되는 물질로 이에 대한 분석은 중요한 사안이다. 신속하고 용이한 마황의 확인을위한 TLC상 동시점적(in situ, Co-spot)방법을 실험하였다. 마황의 액상-액상 추출물과 세 종류의 유도체화 시약들(9-fluorenylmethyl chloroformate; FMOC-Cl, dansyl chloride; DAN-Cl 및 dabsyl chloride; DABCl)을 검출특성이 다른 유도체들의 생성을 위하여 각각 상온 및 70 oC에서 반응시켰다. 마황의 주요 활성성분인 ephedrine (EP)과 pseudoephedrine (PSE)의 유도체는 본 TLC법으로 분리가 명확히 진행되었다. 이러한 분리는 상호 부분입체이성질체인 EP 및 PSE의 유도체화 이전에 비하여 용이하게 확인을 가능하게 하였다. 그리고 마황 분말의 검출한계량을 사용 시약별 및 반응 온도별로 결정하고 비교하였다. 시약 DAN-Cl, FMOC-Cl 사용 시, 검출한계량은 1 mg 이하이었다. 본 TLC방법은 마황에 대한 현장에서의 확인 혹은 법 과학 실험실에서의 예비적인 분석에 도움을 줄 수 있는 있는 방법이라고 사료된다.
Abstract
The analysis of Ephedra herba (Ma huang) is an important issue since Ephedra herba has been involved in the production of illegal substances such as nutritional supplements for weight loss and performance enhancement, and methamphetamine (Meth). A quick and easy thin-layer chromatographic (TLC) co-spot (insitu) method for the identification of Ephedra herba was performed. The liquid-liquid extraction products of Ephedra herba were reacted with each of these three derivatizing reagents (FMOC-Cl, dansyl-Cl (DAN-Cl) or dabsyl-Cl (DAB-Cl)) in situ to generate derivatives having different identification properties at room temperature and 70 oC. Derivatives of ephedrine (EP) and pseudoephedrine (PSE) that are major active ingredients of Ephedra herba were well separated from each other using the TLC method. This separation allowed easy identification of such derivatives compared to the non-derivatized EP and PSE, which comprises a diastereomeric pair. Then, the detection range of the amount of Ephedra herba powder was determined and compared, based on the type of reagents used and the reaction temperature for the current derivatization method. When the reagents DAN-Cl, FMOC-Cl were used to identify Ephedra herba, less than 1 mg of Ephedra herba powder was needed. This TLC method can be a valuable tool to identify Ephedra herba at the scene and helpful as a preliminary analysis at the forensic laboratory.
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