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

Torabi, F. ; Ramanathan, K. LU ; Larsson, P.-O. LU ; Gorton, L. LU ; Svanberg, K. LU ; Okamoto, Y. ; Danielsson, B. LU and Khayyami, M. (1999) In Talanta 50(4). p.787-797
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... (More)
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. (Less)
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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Reticulated vitreous carbon, flow injection analysis, Lactate dehydrogenase, Toluidine Blue
in
Talanta
volume
50
issue
4
pages
787 - 797
publisher
Elsevier
external identifiers
  • scopus:0033571162
ISSN
1873-3573
DOI
10.1016/S0039-9140(99)00134-4
language
English
LU publication?
yes
id
64ce2a8b-9834-4a40-99d8-27291d13eae7 (old id 2259624)
date added to LUP
2016-04-04 09:10:09
date last changed
2024-05-30 11:23:28
@article{64ce2a8b-9834-4a40-99d8-27291d13eae7,
  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.}},
  author       = {{Torabi, F. and Ramanathan, K. and Larsson, P.-O. and Gorton, L. and Svanberg, K. and Okamoto, Y. and Danielsson, B. and Khayyami, M.}},
  issn         = {{1873-3573}},
  keywords     = {{Reticulated vitreous carbon; flow injection analysis; Lactate dehydrogenase; Toluidine Blue}},
  language     = {{eng}},
  number       = {{4}},
  pages        = {{787--797}},
  publisher    = {{Elsevier}},
  series       = {{Talanta}},
  title        = {{Coulometric determination of NAD<sup>+</sup> and NADH in normal and cancer cells using LDH, RVC and a polymer mediator}},
  url          = {{https://lup.lub.lu.se/search/files/5250485/2297666.pdf}},
  doi          = {{10.1016/S0039-9140(99)00134-4}},
  volume       = {{50}},
  year         = {{1999}},
}