超高效液相色谱-串联质谱法测定蜂蜜中五种水溶性维生素的含量

Determination of five water-soluble vitamins in honey by ultra performance liquid chromatography-tandem mass spectrometry

建立了蜂蜜中维生素B 1、维生素B 2、维生素B 5(泛酸)、维生素B 6(吡哆醇)和维生素B 7(生物素)同时测定的超高效液相色谱-串联质谱的分析方法。样品经20 mmol/L甲酸铵溶液提取,HLB固相萃取柱净化,采用HSS T3色谱柱分离,以甲醇和20 mmol/L甲酸铵为流动相进行梯度洗脱,采用电喷雾离子源正离子模式,多反应监测进行检测,外标法定量。该方法在5~200 ng/mL范围内线性关系良好(R ^(2)>0.99),5种维生素检出限均为20μg/kg,定量限均为50μg/kg。在空白蜂蜜样品中进行50、250、500μg/kg这3个水平的加标回收实验,方法的平均回收率为90.2%~108.8%,精密度小于15%。该方法简便、高效、准确性好、抗干扰能力强,可用于蜂蜜中5种水溶性维生素的同时检测。

An ultra performance liquid chromatography coupled with tandem-mass spectrometric(UPLC-MS/MS)method was developed for simultaneous determination of thiamine(B 1),riboflavin(B 2),pantothenic acid(B 5),pyridoxine(B 6),and biotin(B 7)in honey.The samples were extracted with 20 mmol/L ammonium formate solution,and cleaned-up with a HLB solid phase extraction(SPE)column.The extracts were separated on a HSS T3 column with a gradient elution using methanol and 20 mmol/L ammonium formate as mobile phases.Mass spectral acquisition was performed in the positive ion mode by applying multiple reaction monitoring(MRM)with external standard method.The results showed that five water-soluble vitamins showed good linearities in the range of 5-200 ng/mL with correlation coefficients 0.99.The limits of detection(LODs)and limits of quantification(LOQs)for the five water-soluble vitamins were 20μg/kg and 50μg/kg respectively.The recoveries of analytes from blank honey at three levels of 50,250 and 500μg/kg ranged from 90.2%to 108.8%with relative standard deviations less than 15%.This rapid and sensitive method is suitable for the determination of five water-soluble vitamins in honey.