실리카 젤과 알루미나 TLC 상 유도체화 반응을 이용한 암페타민류 유사 화합물의 동정 및 검출한계 비교
Identification and Comparison of Detection Limit of Amphetamine-like Compounds by Derivatization Reaction on Silica gel and Alumina TLC plates
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초록
불법약물인 amphetamine류 화합물에 대한 신속하고 용이한 동정 및 반 정량적인 분석은 법과학상 중요한 쟁점이 되어왔다. 이를 위한 예비실험으로 amphetamine과 구조적으로 유사한 3종의 유기 화합물(1-3)을 실리카 젤 (SiO2)과 알루미나 (Al2O3) 박막 크로마토그래피 (TLC) 상에서 각각 N-(9-Fluorenylmethoxycarbonyloxy)succinimide (FMOC-NHS)와 직접 유도체화 반응을 시켜 얻은 반응 생성물(R1-R3)을 유기합성을 통하여 얻은 대조 물질과 UV 조사 하에서 확인하고 검출한계(LOD)를 비교하였다. 유도체화 시약으로서 FMOC-NHS는 상기 2종의 TLC상에서 1차 및 2차 아민과 반응하여 UV를 강하게 흡수하는 FMOC-group을 함유한 화합물이 생성됨을 확인하였다. 검출한계는 실리카 젤 TLC 실험에서 0.002~0.007 mg/mL (2 μL per spot), 그리고 알루미나 TLC 실험에서는 0.004 ~ 0.1 mg/mL (2 μL per spot) 를 나타냄으로서 실리카 젤 TLC 사용시의 검출한계 농도가 낮게 나타났으며 유도체화 시약의 농도에 다소 의존적이었다. 그러나 알루미나 TLC사용 시, 유도체화 된 화합물들의 분리가 실리카 TLC에 비하여 용이하였다. 그러므로 차후, 검출한계 및 유도체화 조건에 대한 구체적인 조정실험으로 유도체화 시약의 종류, 반응온도, 농도 및 용매에 따른 변수를 찾아야 할 것이다.
Abstract
Rapid and easy identification, characterization and semi- quantitative determination of the presence and amount of amphetamines as illegal drugs in a sample are very important issues in the area of forensic science. A preliminary test was performed on three structurally similar amphetamine-like organic compounds (1-3) which were tested in silica gel and alumina thin-layer chromatography (TLC) using N-(9-fluorenylmethoxycarbonyloxy)succinimide (FMOC-NHS) as a derivatizing reagent. Three reference compounds (R1-R3) were synthesized and used for the identification, comparison of the limit of detection (LOD) of the products from a direct derivatization method on a TLC plate (as visualized under UV). FMOC-NHS as a derivatization reagent generated compounds containing strongly UV-absorbing FMOC- group on the TLC plate after reacting with primary- and secondary amines. In the experiment on silica gel plates the LOD of amines was in the range of 0.002 ~ 0.007 mg/mL (2 uL/spot), and alumina plates the LOD was in the range of 0.004 ~ 0.1 mg/mL (2 uL/spot). In general, lower LODs were achieved on silica gel plates compared to the alumina plates. The LODs of the amines tested were dependent on the concentration of the derivatizing reagent. Three derivatized products were shown to be separated well on alumina plate but not on silica gel plate. Further studies on aspects of these experiments, such as the type and concentration of the derivatizing reagent, reaction temperature and its associated solvents should be pursued.
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