超高效液相色谱-串联质谱法测定蜂王浆中糠氨酸

Determination of furosine in royal jelly by ultra-performance liquid chromatography-tandem mass spectrometry

应用超高效液相色谱-串联质谱法建立一种适用于蜂王浆中糠氨酸含量的分析方法。样品用10.6 mol/L盐酸在110℃条件下水解24 h,水解后的样品经C18固相萃取柱净化,3 mol/L盐酸洗脱,纯净水1:1稀释,以2 mM甲酸铵水溶液(含0.01%甲酸)和2 mM甲酸铵甲醇溶液(含0.01%甲酸)为流动相进行梯度洗脱,ACQUITY BEH C18柱上分离,电喷雾离子源(ESI)正离子模式,多反应监测模式(MRM)检测基质匹配,外标法定量。结果显示,糠氨酸在0.00125~0.07500 mg/L范围内线性关系良好,相关系数(r2)为0.997,方法检出限(LOD)和定量限(LOQ)分别为17μg/kg和50μg/kg。在1倍、2倍和10倍LOQ这3个添加水平下,糠氨酸的平均回收率范围为105.8%~119.4%,相对标准偏差(RSD)为0.4%~4.9%(n=6)。采用本方法对20批蜂王浆中的糠氨酸进行了测定,样品中糠氨酸含量在68.0~311.5μg/kg之间。本方法检出限低、灵敏度高、准确性好,适用于蜂王浆中糠氨酸的测定。

A method for the determination of furosine in royal jelly was developed using ultra performance liquid chromatography-tandem mass spectrometry.The sample was hydrolyzed with 10.6 mol/L hydrochloric acid at 110℃for 24 h.Subsequently,the hydrolyzed sample was purified using a C18 solid phase extraction column and eluted with 3 mol/L hydrochloric acid,followed by dilution in a 1∶1 ratio with pure water.Gradient elution was performed using mobile phases consisting of a 2 mM ammonium formate aqueous solution(containing 0.01%formic acid)and a 2 mM ammonium formate methanol solution(containing 0.01%formic acid)on a ACQUITY BEH C18 column.Detected in electrospray positive ionization mode with multiple reaction monitoring(MRM)mode and quantified by matrix matching curve external standard method.The results demonstrated that furosine exhibited excellent linearity within the concentration range of 0.00125-0.07500 mg/L,with a correlation coefficient(r2)value of 0.997.The limits of detection(LOD)and quantitation(LOQ)were determined to be as low as 17μg/kg and 50μg/kg,respectively.Furthermore,the average recovery rates of furosine at levels corresponding to one,two,and ten times LOQ ranged from 105.8%to 119.4%,accompanied by relative standard deviations(RSD)ranging from 0.4%to 4.9%(n=6).This method was successfully employed to determine the content of furosine in twenty batches of royal jelly samples which varied between 68.0μg/kg and 311.5μg/kg.The method has a low detection limit,high sensitivity,good accuracy,and is suitable for the determination of furosines in royal jelly.