Analytical Strategies for LC–MS-Based Untargeted and Targeted Metabolomics Approaches Reveal the Entomological Origins of Honey

Abstract

A comprehensive liquid chromatography-mass spectrometry (LC-MS)-based metabolomics approach was developed to discriminate honey harvested from <i>Apis mellifera ligustica</i> Spinola (<i>A. mellifera</i>) and <i>Apis cerana cerana</i> Fabricius (<i>A. cerana</i>). Based on an untargeted strategy, ultrahigh-performance liquid chromatography electrospray ionization quadrupole orbitrap high-resolution mass spectrometry (UPLC Q-Orbitrap) was combined with chemometrics techniques to screen and identify tentative markers from <i>A. mellifera</i> and <i>A. cerana</i> honey. In targeted metabolomics analysis, a sensitive method of solid-phase extraction followed by ultrahigh-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UPLC-MS/MS) was established for quantifying three markers, and the results showed that 3-amino-2-naphthoic acid and methyl indole-3-acetate could be considered markers of <i>A. cerana</i> honey, as they were present in higher amounts in <i>A. cerana</i> honey than in <i>A. mellifera</i> honey, whereas kynurenic acid was determined to be a marker of <i>A. mellifera</i> honey. This work highlights critical information for the authentication of <i>A. cerana</i> and <i>A. mellifera</i> honey.