Dansyl chloride를 이용한 아민 계 화합물의 TLC판 상 동정 및 검출한계
Identification and Detection Limit of Amine Compounds with Dansyl Chloride on TLC Plates
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초록
Dansyl chloride (DNS-Cl, D)가 아민 계 화합물과 반응하여 생성된 DNS 유도체의 높은 형광성을 이용하여 검출한계의 향상과 용이한 동정을 위한 예비실험을 실시하였다. 그 결과, 구조적으로 유사한 3종의 아민 계 화합물(1-3) 을 실리카 젤(SiO2) 박막 크로마토그래피(TLC) 상에서 DNS-Cl과 직접 유도체화 반응시켜 얻은 반응생성물을 UV (λmax.= 365 nm) 조사 하에서 확인하고 검출한계(LOD)를 각각비교하였다. 유도체화 시약으로서 DNS-Cl은 TLC상에서 1차 및 2차 아민과 반응하여 형광을 방출하는DNS-group을 함유한 화합물이 생성됨을 확인하였다. 검출한계 농도는 0.001 mg/mL (2 μL per spot)를나타내어 FMOC-group의 부착(labeling) 사용 시의 검출한계 농도와 유사하게 나타났으며 유도체화 시약의 농도에 의존적이었다. 3종의 생성된 유도체들은 실리카 젤 TLC상에서 분리가 잘 되었다. 차후, 검출한계 및 유도체화 조건에 대한 구체적인 조정실험을 통하여 유도체화 시약의 농도, 반응온도 및 용매에따른 변수들을 찾아야 할 것이다.
Abstract
As a preliminary step to improve detection limit and easy identification using the highly fluorescentcharacteristics of derivatized organic compounds with dansyl chloride (DNS-Cl), three structurally similar aminecontainingorganic compounds (1-3) were reacted on silica gel thin-layer chromatography (TLC) with DNSClas a derivatizing reagent. The new products from a direct derivatization method on a TLC plate (as visualizedunder long wavelength UV365 light) were used for the identification and their limit of detection (LOD) wascompared. DNS-Cl as a derivatization reagent generated compounds containing highly fluorescent dansyl-groupon the TLC plate after reacting with primary- and secondary amines. In the experiment on TLC plates theLOD of amines was in the range of 0.01 ~ 0.001 mg/mL (2 uL/spot) which was similar to that of FMOCderivatizedamines. The LODs of the amines tested were dependent on the concentration of the derivatizingreagent. Three derivatized products were shown to be separated well on silica gel plate. Further studies onaspects of these experiments, such as concentration of the derivatizing reagent, reaction temperature and itsassociated solvents should be pursued.
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