Coulometric determination of NAD(+) and NADH in normal and cancer cells using LDH, RVC and a polymer mediator

Research output: Contribution to journalArticle

Abstract

An electrochemical method for the measurement of NAD(+) and NADH in normal and cancer tissues using flow injection analysis (FIA) is reported. Reticulated vitreous carbon (RVC) electrodes with entrapped L-lactate dehydrogenase (LDH) and a new redox polymer containing covalently bound toluidine blue O (TBO) were employed for this purpose. Both NAD(+) and NADH were estimated coulometrically based on their reaction with LDH. The latter was immobilized on controlled pore glass (CPG) by cross-linking with glutaraldehyde and packed within the RVC. The concentrations of NAD(+) and NADH in the tissues, estimated using different electron mediators such as ferricyanide (FCN), meldola blue (MB) and TBO have also been compared. The effects of flow rate, pH, applied potential (versus Ag/AgCl reference) and adsorption of the mediators have also been investigated. Based bn the measurements of NAD(+) and NADH in normal and cancer tissues it has been concluded that the NADH concentration is lower, while the NAD(+) concentration is higher in cancer tissues. Amongst the electron mediators TBO was found to be a more stable mediator for such measurements. (C) 1999 Elsevier Science B.V. All rights reserved.

Details

Authors
  • F Torabi
  • K Ramanathan
  • P. O Larsson
  • L Gorton
  • Katarina Svanberg
  • Y Okamoto
  • B Danielsson
  • M Khayyami
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Atom and Molecular Physics and Optics
Original languageEnglish
Pages (from-to)787-797
JournalTalanta
Volume50
Issue number4
Publication statusPublished - 1999
Publication categoryResearch
Peer-reviewedYes