Improved high-pressure liquid chromatographic-fluorometric assay for measurement of adenosine in plasma

Y. Zhang, J. D. Geiger, W. W. Lautt

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

A high-pressure liquid chromatographic system using fluorescence detection was characterized for the determination of adenosine levels in plasma from anesthetized cat, rat, dog, mouse, rabbit, and guinea pig. The optimal concentration of chloroacetaldehyde necessary to convert physiological levels of adenosine to its fluorescent derivative 1,N6-ethenoadenosine (ethenoadenosine) was in excess of 220 mM. As little as 0.2 pmol of ethenoadenosine could be measured, and detection was linear up to 200 pmol. Derivatization of adenosine into ethenoadenosine was highly dependent on temperature and time. However, ethenoadenosine showed thermal instability in that levels dropped sharply after 30 min at 100°C, 2 h at 80°C, and 24 h at 55°C. Adenosine nucleotides were extracted from plasma samples with an efficiency of >91% to prevent adenosine formation from ATP and AMP that would have otherwise occurred during the derivatization procedure. Plasma levels (μM) of adenosine in venous blood were 0.31 in dog, 0.54 in cat, 0.71 in guinea pig, 1.03 in mouse, 1.04 in rat, and 1.68 in rabbit. Plasma levels of adenosine in arterial blood were not significantly different from levels in venous blood. This method can be used to measure even very low levels of adenosine without interference from nucleotide breakdown.

Original languageEnglish (US)
Pages (from-to)G658-G664
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume260
Issue number4 23-4
DOIs
StatePublished - 1991
Externally publishedYes

Keywords

  • 1,N-ethenoadenosine
  • Cat
  • Dog
  • Fluorometry
  • Mouse
  • Purines
  • Rabbit Guinea pig
  • Rat

ASJC Scopus subject areas

  • Physiology
  • Hepatology
  • Gastroenterology
  • Physiology (medical)

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