Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Determination of Brassinolide by HPLC equipped with Fluoresence Detector in Rice(Oriza sativa L.)

HPLC 형광분석법을 통한 벼에서 Brassinolide의 검정

  • Kim, In-Seon (Department of Agricultural Chemistry, Chonnam National University) ;
  • Lee, Kang-Bong (Department of Agricultural Chemistry, Chonnam National University) ;
  • Suh, Yong-Tack (Department of Agricultural Chemistry, Chonnam National University) ;
  • Morgan, E.D. (Department of Chemistry, Keele University) ;
  • Shim, Jae-Han (Department of Agricultural Chemistry, Chonnam National University)
  • Published : 1996.02.29
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Abstract

To determine brassinolide in rice(Oriza sativa L.) using HPLC equipped with fluoresence detector, a highly sensitive fluorescence reagent. 1-cyanoisoindole-2-m-phenylboronic acid, was synthesized from the reaction of o-phthaldehyde, m-phenylboronic acid and KCN, then was reacted with brassinolide. The formation ratio of brassinolide boronate exhibited 90% up at the ratio of $20\;:\;1({\mu}g/{\mu}g)$ of 1-cyanoisoindole-2-m-phenylboronic acid and brassinolide respectively. The detection limit of brassinolide boronate with fluoresence detector was 0.16 ng. Brassinolide was detected in heading stage(biomass : 10 g) and panicle formation stage(biomass : 100 g) of the rice(Oryza sativa L.) with quantity of $0.8\;{\mu}g\;and\;0.2\;{\mu}g$respectively. However, brassinolide was not detected in blooming and elongation stage.

벼의 생육 단계별 brassinolide의 검정을 위하여 HPLC 형광분석법을 개발하였다. 형광분석에 민감한 1-cyanoisoindole-2-m-phenylboronic acid를 o-phthalaldehyde와 m-phenylbornic acid 및 KCN과 반응시켜 합성하여 brassinolide의 1, 2-diol과 유도체화 시켰다. Brassinolide $1\;{\mu}g$에 대한 1-cyanoisoindole-2-m-phenylboronic acid의 brassinolide boronate의 형성정도는 $20\;{\mu}g$에서 90% 이상의 유도체화를 보였고 형광검출법에 의한 brassinolide의 검출한계는 0.16 ng이었다. 벼의 생육단계별 brassinolide는 분얼기 및 신장기에서는 검출이 되지 않았으며 유수형성기에는 $0.8\;{\mu}g$(생체 10g)이 그리고 출수개화기에는 $0.2\;{\mu}g$(생체 10g)이 검출되었다.

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