UHPLC-MS/MS法同时测定啤酒中4种嘌呤

Determination of Four Purines in Beer by UPLC-MS/MS

建立一种采用超高效液相色谱-串联质谱(UHPLC-MS/MS)法同时测定啤酒中4种嘌呤类生物碱(腺嘌呤、鸟嘌呤、次黄嘌呤、黄嘌呤)的方法。啤酒样品脱气后,经三氟乙酸和甲酸高温水解,采用Agilent Poroshell 120EC-C_(18)(3.0 mm×150 mm,2.7μm)色谱柱进行分离,以甲醇-水作为流动相体系梯度洗脱,在电喷雾电离源(ESI)正离子模式下,以多反应监测模式(MRM)进行定性定量分析。结果表明,4种嘌呤组分在0.5~250μg/L的质量浓度范围内线性关系良好,相关系数(r^(2))均大于0.999,检出限(LOD)为6.0~12μg/L,定量限(LOQ)为20~40μg/L。在腺嘌呤5.04~15.12 mg/L、鸟嘌呤12.76~38.28 mg/L、次黄嘌呤1.32~3.96 mg/L和黄嘌呤2.52~7.56 mg/L加标水平下,平均回收率为80.5%~98.2%,相对标准偏差(δ_(RSD),n=6)为1.3%~4.2%。该方法前处理简单、分析快速、灵敏度高、准确性高,适用于啤酒中4种嘌呤的同时测定。

A method was developed for the determination of four purines(adenine,guanine,hypoxanthine and xanthine)in beer by ultra high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS).The beer sample was degassed and hydrolyzed with trifluoroacetic acid and formic acid,and the chromatographic separation was carried out on an Agilent Poroshell 120 EC-C_(18) column(3.0 mm×150 mm,2.7μm)using methanol-water as mobile phase by gradient elution.The four purines were qualitatively and quantitatively determined by ESI+-MS/MS under multi-reaction monitoring mode(MRM).The results showed that good linear ranges for four purines were obtained from 0.5μg/L to 250μg/L with correlation coefficients(r^(2))larger than 0.999,and the limits of detection(LODs)were within 6.0-12μg/L,while the limits of quantitation(LOQs)were within 20-40μg/L.The mean recoveries of four purines were in the range of 80.5%-98.2%at spiked levels of 5.04-15.12 mg/L for adenine,12.76-38.28 mg/L for guanine,1.32-3.96 mg/L for hypoxanthine and 2.52-7.56 mg/L for xanthine,with relative standard deviations(δ_(RSD),n=6)of 1.3%-4.2%.The optimized method was proved to be simple,rapid,highly sensitive,accurate,and suitable for the determination of four purines in beer.