Simultaneous Determination of 26 Endogenous Components in Honey by Automated Solid Phase Extraction Coupled with Ultra-high Performance Liquid Chromatography-Tandem Mass Spectrometry

An automated solid phase extraction coupled with ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of 26 endogenous components in chaste, acacia and rape honey. The samples were extracted with phosphate buffer solution, purified by solid-phase extraction using an Oasis HLB cartridge, and separated on an Agilent Eclipse Plus RRHD C₁₈ column (2.1 mm × 100 mm, 1.8 µm) by gradient elution using a mobile phase comprising of 0.03% formic acid in water and acetonitrile. The analytes were detected by using an electrospray ion source in the negative ion mode with dynamic multiple reaction monitoring (dMRM), and quantified by an external standard method. Results showed that good linear relationships were obtained for the 26 analytes in their respective concentration ranges with correlation coefficients (r²) greater than 0.996. The limits of detection (LOD) and quantitation (LOQ) ranged from 1.0 to 10.0 and 2.5 to 25.0 μg/kg, respectively. The average recoveries from spiked samples were between 81.9% and 101%, with relative standard deviations (RSD) of 3.4%–9.8%. The UPLC-MS/MS method has the advantages of rapidity as well as high efficiency and accuracy, which can provide technical support for the quality control and identification of honey.