A solid phase extraction/liquid chromatography-tandem mass spectrometric(SPE/LC-MS/MS) method was established for the simultaneous determination of 2′-methoxyacetophenone(2′-MAP),2-methoxybenzoic acid(2-MBA),3-phenyllactic acid(3-PA) and 4-hydroxyphenyllactate(4-HPA).The honey sample was dissolved in 1% formic acid solution,then purified with HLB SPE.The separation of the analytes was performed on a Thermo Hypersil Gold C18(50 mm×2.1 mm,1.9 μm) column with 0.1% formic acid solution and acetonitrile containing 0.1% formic acid as mobile phases by gradient elution.The four compounds were detected simultaneously in electrospray negative and positive ionization modes under multiple reaction monitoring(MRM) mode.The results showed that the calibration curves were linear in the range of 0.01-1.0 mg /L for 2′-MAP,2-MBA and 4-HPA,and 0.1-10.0 mg/L for 3-PA with correlation coefficients(r2) higher than 0.99.The limits of detection(LODs) and quantitation(LOQs) were in the ranges of 0.1-1.0 mg/kg and 0.5-5.0 mg/kg,respectively.The average spiked recoveries for honey samples ranged from 79.4% to 98.3% with relative standard deviations(n=6) of 3.2%-8.7%.This method was used to validate 114 hive honey samples from 12 varieties collected in different locations in New Zealand and 50 commercial honey samples from four different countries.With the charateristics of sensitivity,accuracy and rapidness,the method could provid a reference for the identification of monoflower,multiflower and non-manuka honey. |