대한약리학회지 (The Korean Journal of Pharmacology)
- 제24권2호
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- Pages.233-240
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- 1988
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- 0377-9459(pISSN)
생체의 Polyamine-분석을 위하여 FNBT-유도체를 이용하는 간편하고 특이적이며 예민한 Isocratic RP-HPLC 분석법과 재생성 흰쥐-간의 Polyamine-대사의 변동에 관한연구
A Simple, Sensitive, and Specific HPLC Analysis of Tissue Polyamines using FNBT Derivatization: Its Application on the Study of Polyamine Metabolism in Regenerating Rat Liver
- Choi, Sang-Hyun (Department of Pharmacology, Korea University College of Medicine) ;
- Kim, Hyung-Gun (Department of Pharmacology, Korea University College of Medicine) ;
- Park, Hong-Ik (Department of Pharmacology, Korea University College of Medicine) ;
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Chun, Boe-Gwun
(Department of Pharmacology, Korea University College of Medicine)
- 발행 : 1988.12.30
초록
복잡한 시료-전처리과정을 배제하여 간편하고 선택적이며 예민한 polyamine 측정방법으로써 4-fluoro-3-nitrobenzotrifluoride (FNBT)유도체 및 단일이동상을 이용하는 역상의 고속액체 크로마토 그래피 (HPLC) 방법과 아울러 이 방법을 이용한 흰쥐의 재생성간내 polyamine대사의 변동에 관한 관찰-결과를 요약하면 다음과 같다. 1. 간내-polyamine은 0.4M perchloric acid에 추출한 후 Spragg와 Hutchings의 방법에 따라서 FNBT로 N-2'-nitro-4'-trifluoromethylphenyl polyamine (NTP-polyamine) 유도체를 만들고 이를HPLC로 분석 하였다. 2. HPLC-분석은 ERC ODS-1161 (3 um,
A simple and selective isocratic HPLC method for the analysis of tissue polyamine contents is described and applied on the study of the changes of the hepatic polyamine contents after partial hepatectomy in male rats. The hepatic polyamines are extracted with 0.4 M perchloric acid containing 2 mM disodium EDTA, and then the extract is redissolved in 100 ul of 1 M sodium carbonate and incubated with 300 ul of FNBT-dimethylsulfoxide (1: 100) mixture. The N-2'-nitro-4'-trifluoromethylphenyl drivatives of polyamines are separated through a ERC-ODS column in an isocratic mode with an acetonitrile-water (80:20) mobile phase within 20 min. per a sample, while monitoring the effluent at 242 nm. This improved method which could detect subnanogram of each polyamines is highly specific and reproducible as evidenced by the application of it on the study of the changes of polyamine contents in the regenerating rat liver after partial hepatectomy.