HPLC-MS/MS法同时测定功能性饮料中香兰素、甲基香兰素和乙基香兰素的含量

Simultaneous Determination of Vanillin,Methyl Vanillin and Ethyl Vanillin in Functional Beverages by HPLC-MS/MS

目的建立HPLC-MS/MS同时测定功能性饮料中香兰素、甲基香兰素和乙基香兰素含量的方法。方法^(®)采用WatersCORTECS T3色谱柱(2.1mm×100mm,2.7μm),以0.1%甲酸水溶液-甲醇为流动相,梯度洗脱,柱温30℃;流速0.3 ml/min,进样量为2μl。采用质谱检测器,电喷雾正离子模式(ESI^(+)),进行多反应监测(MRM)。按外标法定量。结果香兰素、甲基香兰素和乙基香兰素分别在5.493~175.787,5.131~164.179,5.019~160.619ng/ml浓度范围内线性良好,相关系数(r)分别为0.9997,0.9999和0.9999;方法检出限分别为8,7,7μg/kg;定量限分别为15,14,14μg/kg;平均回收率分别为96.8%,86.7%,88.9%,RSD值分别为4.0%,2.0%,5.2%(n=6)。仪器的精密度和方法的重复性RSD均小于10%。结论该方法简单高效、重复性好,可用于检测功能性饮料中香兰素、甲基香兰素和乙基香兰素的含量。

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,2.7μm),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 2μl.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 8,7,7μg/kg and 15,14,14μg/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.