형광검출 역상 액체크로마토그래피에 의한 Histamine의 정량

Determination of Histamine by Reverse-Phase High Performance Liquid Chromatography with Fluorescence Detection

Histamine을 정확하고 신속하게 정량하기 위해 9-Fluorenylmethyl chloroformate를 형광유도체화제로 하여 역상 HPLC법으로 정량하였다. 히스타민을 형광유도체화할 때 반응액의 pH, 반응시간, 형광유도체화제의 농도 등 최적 반응 조건을 검토하였다. 이 방법으로 히스타민을 분석한 결과 $0.1{\sim}0.5{\mu}g/ml$의 농도범위에서 상관계수가 0.992인 양호한 직선성을 나타내였으며 검출한계는 $0.01{\mu}g/ml$였다.

A rapid and simple method for the determination of histamine by reverse-phase high performance liquid chromatography with fluorescence detection was established. 9-Fluorenylmethyl chloroformate(FMOC) was used as fluorescent derivative reagent. The optimum conditions for the derivatization such as pH, reaction time and the concentration of FMOC were investigated. Linearity of calibration curve was obtained between $0.1{\mu}g/ml$ and $0.5{\mu}g/ml$(r=0.992) and the limit of detection was $0.01{\mu}g/ml$.