Application and High Throughput Screening of DPPH Free Radical Scavenging Activity by Using 96-Well Plate

96-well plate를 이용한 DPPH free radical 소거활성 측정과 그 응용

  • Choi, Jung-Sup (Korea Research Institute of Chemical Technology) ;
  • Oh, Jung-Im (Korea Research Institute of Chemical Technology) ;
  • Hwang, In-Taek (Korea Research Institute of Chemical Technology) ;
  • Kim, Sung-Eun (Division of Applied Biotechnology, College of Agriculture, Chonbuk National University) ;
  • Chun, Jae-Chul (Division of Applied Biotechnology, College of Agriculture, Chonbuk National University) ;
  • Lee, Byung-Hoi (Korea Research Institute of Chemical Technology) ;
  • Kim, Jin-Seok (Korea Research Institute of Chemical Technology) ;
  • Kim, Tae-Joon (Korea Research Institute of Chemical Technology) ;
  • Cho, Kwang-Yun (Korea Research Institute of Chemical Technology)
  • 최정섭 (한국화학연구원 생물기능연구팀) ;
  • 오정임 (한국화학연구원 생물기능연구팀) ;
  • 황인택 (한국화학연구원 생물기능연구팀) ;
  • 김성은 (전북대학교 응용생물공학부) ;
  • 전재철 (전북대학교 응용생물공학부) ;
  • 이병회 (한국화학연구원 생물기능연구팀) ;
  • 김진석 (한국화학연구원 생물기능연구팀) ;
  • 김태준 (한국화학연구원 생물기능연구팀) ;
  • 조광연 (한국화학연구원 생물기능연구팀)
  • Published : 2003.06.30

Abstract

A 96-well plate was applied to determine the DPPH free radical scavenging activity using 107 plant-specific enzyme inhibitors and 100 unknown plant-originated extracts. The final optimum volume was $250{\mu}L$ containing $100{\mu}M$ DPPH ethanolic solution at pH 7.8. In this condition, the radical scavenging activities were significantly increased by two known antioxidants consisting of ascorbate and a-tocopherol in a concentration-dependent manner. Among the 107 inhibitors, ampicillin and gallic acid showed 90.2% and 92.6% antioxidant activity at $100{\mu}M$, respectively, and these results were consisted with previous findings. In the tested 100 natural materials at $50{\mu}g/mL$, antioxidant activity of AT-407 resulted in the highest of 90.1%, and 10 extracts including AT-388 and AT-443 showed over 70%. Our results suggest that the use of 96-well plate for determining DPPH free radical scavenging activity would be a suitable method to select antioxidant-like substances of both synthetic compounds and natural products.

Keywords

DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical;96-well plate;antioxidants;plant-specific enzyme inhibitor;plant crude extracts

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