Protein Binding of [S]-Perillyl Alcohol in HSA using High-Performance Frontal Analysis

HPFA를 이용한 HSA와 [S]-Perillyl Acohol의 단백질 결합력

  • 송명석 (인하대학교 공과대학 화학공학과) ;
  • 왕덕선 (인하대학교 공과대학 화학공학과) ;
  • 구윤모 (초정밀분리연구센터) ;
  • 노경호 (인하대학교 공과대학 화학공학과)
  • Published : 2003.04.01


An on-line frontal analysis HPLC system was developed to determine the unbound concentration of (S)-perillyl alcohol, an potential anti-cancer agent, in human serum albumin (HSA) solution, The analysis was performed on a Develosil 100 Diol 5 (10 cm x 4.6 mm I.D.) high-performance frontal analysis (HPFA) column. Sodium phosphate solution was used as the mobile phase (pH 7.4, ionic strength 0.17) at a flow rate of 1 $m\ell$/min. UV wavelength was set at 205 nm. A injection volume of 600${mu}ell$ was chosen to ensure the compound eluted formed a zonal peak with a plateau. By Scatchard analysis, it was found that the binding constant(K) and binding number(n) of (S)-perillyl alcohol to molecular HSA were 2.05 x $10^6$ [$M{-1}$], 0.00428, respectively.

의학, 약학적으로 작용이 뛰어난 약리성분이 인체 내에서 상처 부위에 얼마나 빨리 도달하여 치료를 하는지를 알기 위하여 혈장 단백질과 약리성분의 결합력을 연구하는 새로운 연구 분야가 HPFA이다. 본 연구는 인체 내에 존재하는 혈장단백질과 항암제로서 알려져 있는 (S)-perillyl alcohol의 결합력과 결합 매개변수를 구하기 위하여 on-line frontal analysis HPLC system을 적용하였다. Develosil 100 Diol 5 (10 cm $\times$ 4.6 mm I.D.)의 HPFA 컬럼을 이용하였고, 이동상은 인산완충용액(pH = 7.4, I = 0.17)을 이용하였다. UV wavelength 205 nm에서 실험을 수행하였고, 주입 부피는 본 실험 조건인 혈장 단백질이 350 rM일 경우에 최대의 혈장 단백질과 결합되지 않은 약리성분의 농도를 갖게 되는 600 ${\mu}\ell$으로 정하였다. Scatchard analysis를 통한 연구 결과로 혈장 단백질인 HSA와 (S)-perillyl alcohol의 결합 매개변수(K)와 단위 HSA에 대한 S-POH의 결합 위치의 수(n)는 각각 K : 2.05 $\times$ $10^{6}$ [M$^{-1}$], n : 0.00428로 실험적으로 구하였다



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