超高效液相色谱-串联质谱测定干红葡萄酒中18种花色素单体方法的建立

Establishment of Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry Method for Determination of 18 Individual Anthocyanins in Wine

目的:利用超高效液相色谱-串联质谱技术同时测定干红葡萄酒中18 种花色素单体。方法:将酒样用5%酸化甲醇溶液(含1%甲酸,V/V)稀释500 倍后,滤液经Phenyl-Hexyl色谱柱梯度洗脱,电喷雾离子源正离子模式检测,多反应监测,气帘气压力30 psi,离子喷雾电压5 500 V,离子源温度500 ℃,分析时间20 min。结果:18 种花色素单体在10.0、20.0、50.0 μg/L添加水平的加标回收率在95%~100%之间,R2大于0.999,相对标准偏差在1.71%~2.87%之间,检出限在0.2~3.5 μg/L之间,定量限在0.4~11.5 μg/L之间。结论:该方法前处理简便、灵敏度高、分析时间短,精密度和准确性良好,可用于干红葡萄酒中18 种花色素单体的同时检测。

Objective: To simultaneously determine 18 anthocyanin monomers in red wine using ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). Methods: Wine samples were diluted 500 times with acidified methanol solution containing 1% formic acid(V/V) before being filtrated. The filtrate was separated by gradient elution chromatography on a phenyl-hexyl column. Detection was carried out using an electrospray ionization source in the positive ion mode with multiple reaction monitoring(MRM). The mass spectrometric conditions were as follows: curtain gas 30 psi, ion spray(IS) voltage 5 500 V, and ion source temperature 500 ℃. The analysis took 20 min. Results: The recoveries of the analytes at spiked levels of 10.0, 20.0 and 50.0 μg/L were between 95% and 100%, and the calibration curve for each analyte exhibited a correlation coefficient(R^2) of greater than 0.999. The precision, expressed as relative standard deviation(RSD), ranged between 1.71% and 2.87%. The limits of detection(LOD) were between 0.2 and 3.5μg/L, and the limits of quantitation(LOQ) were between 0.4 and 11.5μg/L. Conclusion: This method was characterized by simple pretreatment, high sensitivity, short analysis time, shorter time and good precision and accuracy and could be applied to detect individual anthocyanins in red wine.