Analytical Science and Technology (분석과학)

The Korean Society of Analytical Sciences (한국분석과학회)
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Determination of Biotin by HPLC

고성능 액체크로마토크래피를 이용한 Biotin의 정량

  • Received : 2003.08.18
  • Accepted : 2003.10.21
  • Published : 2003.12.25
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Abstract

A high performance liquid chromatography gradient elution method with fluorescence detection for the determination of biotin in pure form and pharmaceutical preparations has been developed. BrMDMC gives intense fluorescence and the fluorescence was monitored with excitation at 360 nm and emission at 410 nm. The calibration curve for biotin shows good linearity over the range of 5 ~ 400 ng with correlation coefficient of 0.999. The detection limit of biotin was 2 ng and the result of recovery was 98.75% with relative standard deviation of 1.1%.

Biotin을 4-bromomethyl-6,7-dimethoxycoumarin (BrMDMC), 18-crown-6와 $K_2CO_3$를 사용하여 형광성 유도체로 만들어 (들뜸 파장 360 nm, 방출파장 410 nm에서) 기울기 용리 HPLC 분석을 하였다. 검정선의 농도범위는 5 ~ 400 ng, 검출한계는 2 ng, 회수율 98.75%, 그리고 상대표준편차는 1.1% 이었다. 시료의 전처리는 Sep-Pak $C_{18}$ catridge를 이용하였고, 유도체 제조의 중탕온도는 $50^{\circ}C$에서 30분간 시행하였다.

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