Measurement of the limiting equivalent conductivities and mobilities of the most prevalent ionic species of EGTA (EGTA2- and EGTA3-) for use in electrophysiological experiments

Angelo Keramidas, Levin Kuhlmann, Andrew J. Moorhouse, Peter H. Barry

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Abstract

In many experimental biological situations, chelating agents like EGTA (ethylene glycol-bis-(β-amino-ethyl ether) N,N,N',N'-tetra-acetic acid) are commonly used to control or suppress the concentration of divalent ions like Ca2+. The evaluation of liquid junction potentials in electrophysiological measurements, and particularly in patch-clamp situations, requires information about the ions within the solution. Where there is a significant concentration of EGTA present, it is necessary to know the values of the relative mobility of at least the most predominant ionic species of EGTA in order to complete these calculations. EGTA, with four negative charges with different pK(a)s, can therefore exist as four differently charged ions in solution (EGTA-, EGTA2-, EGTA3- and EGTA4-) or as uncharged, although between pH 5.5 and 8 it is almost exclusively EGTA2-. We have measured limiting equivalent conductivities of the most common ionic forms of EGTA (EGTA2-and EGTA3-) encountered at physiological pHs. These were 35.9±0.7 and 56±2.5 S cm2 equiv-1 respectively. Their mobilities relative to K+ were 0.24±0.01 for EGTA2- and 0.25±0.01 for EGTA3-. Thus for typical electrophysiological solutions, the contribution of EGTA to the liquid junction potential should be small (e.g. ~0.4 mV). Copyright (C) 1999 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)41-47
Number of pages7
JournalJournal of Neuroscience Methods
Volume89
Issue number1
DOIs
Publication statusPublished - 1 Jul 1999

Keywords

  • Conductivities
  • EGTA
  • Ion mobilities
  • Liquid junction potentials
  • Patch-clamp

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