Simultaneous Determination of Vanillin,Methyl Vanillin and Ethyl Vanillin in Functional Beverages by HPLC-MS/MS
Objective To establish a method for the simultaneous determination of vanillin,methyl vanillin and ethyl vanillin in functional beverages by HPLC-MS/MS.Methods The determination was performed on a Waters CORTECS® T3 column(2.1 mm×100 mm,2.7 μm),using 0.1%formic acid-water solution and methanol at the flow rate of 0.3 ml/min as mobile phase of gradient elution.The column temperature was 30℃,and the injection volume was 2 μl.For quantitative analysis,the MS detector was used,and electro-spray positive ion mode(ESI+)and multi reaction monitoring(MRM)mode were adopted.Results Vanillin,methyl vanillin and ethyl vanillin showed good linear relationships within the ranges of 5.493-175.787,5.131-164.179,5.019-160.619 ng/ml,and the correlation coefficients were 0.9997,0.9999 and 0.9999,respectively.The LODs and LOQs of vanillin,methyl vanillin and ethyl vanillin were 8,7,7 μg/kg and 15,14,14 μg/kg,respectively.The average recoveries were 96.8%,86.7%and 88.9%,and the RSDs were 4.0%,2.0%and 5.2%,respectively(n=6).The RSDs of precision and repeatability tests were both less than 10%.Conclusion This method is simple,efficient and repeatable.It can be used for the determination of vanillin,methyl vanillin and ethyl vanillin in functional beverages.
HPLC-MS/MSvanillinmethyl vanillinethyl vanillinMRMexternal standard method