Development and validation of a HILIC-UV method for the determination of nucleotides in fish samples

Food Chem. 2018 May 15:248:70-77. doi: 10.1016/j.foodchem.2017.12.040. Epub 2017 Dec 13.

Abstract

The aim of this work was the development of a simple, novel and accurate method for the determination of adenosine triphosphate (ATP) and its first five catabolites: adenosine diphosphate (ADP), adenosine monophosphate (AMP), inosine monophosphate (IMP), inosine (Ino) and hypoxanthine (Hx), in fish tissue, based on hydrophilic interaction liquid chromatography (HILIC). For this purpose, a stationary phase for polar and hydrophilic compounds (ZIC-pHILIC) was used. The effect of different chromatographic parameters and the molecular mechanism based on the van't Hoff plot were examined. The t-test and Dixon's Q-test were applied in order to examine statistical differences and outlier values. The recovery of the method ranged between 82.7% and 127% and the %RSD values were lower than 10% for all analytes determined. The method was applied in frozen sea bream samples stored at 0-4 °C. The Ki-, G-, H- and F values were calculated for the estimation of the level of fish freshness.

Keywords: Freshness indicators; HILIC; Nucleotides; Sea bream.

Publication types

  • Validation Study

MeSH terms

  • Adenine Nucleotides / analysis*
  • Adenosine Diphosphate / analysis
  • Adenosine Monophosphate / analysis
  • Adenosine Triphosphate / analysis
  • Animals
  • Chromatography, Liquid / methods*
  • Fish Products / analysis
  • Fishes*
  • Hydrophobic and Hydrophilic Interactions
  • Hypoxanthine / analysis
  • Inosine Monophosphate / analysis
  • Sea Bream

Substances

  • Adenine Nucleotides
  • Inosine Monophosphate
  • Hypoxanthine
  • Adenosine Monophosphate
  • Adenosine Diphosphate
  • Adenosine Triphosphate