Isolation of choline and choline esters from Krebs-Ringer solution for gas chromatographic determination

D. Budai, Péter Szerdahelyi, P. Kása

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

A simple and efficient novel method for isolating picomole amounts of choline and choline esters in milliliter volumes of Krebs-Ringer solution has been developed. The procedure is based on the observation that the solubility of choline esters in acetonitrile is 104-105 times higher than that of the inorganic salt constituents of Krebs-Ringer solution. The glucose content of the medium, which prevented the one-step isolation of choline esters based on acetonitrile extraction from its lyophilizate, was removed using Amberlite CG-50 column chromatography. Bound compounds to the column were eluted in 0.25 n HCl and lyophilized. The lyophilizate was extracted with acetonitrile, which was then decanted and eliminated by evaporation to dryness. The resultant glucose and salt-free residue can be assayed by gas chromatography. Total recoveries of added choline and choline esters over the entire isolation procedure, measured isotopically and/or gas chromatographically, were 93 and 97%, respectively. Due to the high and close-to-equal recoveries of choline esters, and the high purity of the final product, this procedure is suitable for estimating acetylcholine and choline in neural tissue perfusates by gas chromatography, as was demonstrated by this method using hippocampal slices.

Original languageEnglish
Pages (from-to)260-266
Number of pages7
JournalAnalytical Biochemistry
Volume159
Issue number2
DOIs
Publication statusPublished - 1986

Fingerprint

Choline
Esters
Gases
Gas chromatography
Gas Chromatography
Salts
Glucose
Recovery
Column chromatography
Krebs-Ringer solution
Solubility
Acetylcholine
Chromatography
Evaporation
Tissue
acetonitrile

Keywords

  • acetonitrile
  • choline esters
  • extraction
  • ion exchange
  • Krebs-Ringer solution
  • recovery

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Isolation of choline and choline esters from Krebs-Ringer solution for gas chromatographic determination. / Budai, D.; Szerdahelyi, Péter; Kása, P.

In: Analytical Biochemistry, Vol. 159, No. 2, 1986, p. 260-266.

Research output: Contribution to journalArticle

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