免疫亲和柱净化/色谱技术测定婴幼儿配方乳粉中V_(B12)  

Determination of vitamin B_(12) in infant milk powder by immunoaffinity column purification/chromatography method

作　　者：曹慧[1,2] 陈小珍 朱岩[2] 李祖光[3] 郑军科 祝颖 

机构地区：[1]浙江省质量检测科学研究院,杭州310030 [2]浙江大学化学系,杭州310012 [3]浙江工业大学化学工程与材料学院,杭州310014

出　　处：《食品科技》2014年第9期302-305,共4页Food Science and Technology

基　　金：浙江省质量技术监督系统科研计划(20120210)

摘　　要：建立了婴幼儿配方乳粉中VB12的超高效液相色谱-串联质谱和超高效液相色谱测定方法。样品采用pH=4.0的醋酸钠溶液提取,经免疫亲和柱净化,采用Waters BEH C18色谱柱分离,液相法采用0.03 mol/L的磷酸缓冲盐（pH=3.5）和乙腈为流动相进行等度洗脱,紫外检测器波长为361 nm。质谱法以0.1%的甲酸水和甲醇为流动相进行梯度洗脱,采用电喷雾-正离子模式多反应监测进行定性和定量分析,外标法定量。VB12在10～500μg/L的质量浓度范围内线性关系良好,VB12的检出限均为0.5μg/kg,定量限均为1.5μg/kg,液相方法回收率为90.2%～98.3%,相对标准偏差为1.4%～2.5%,液相色谱-质谱法回收率为89.3%～95.7%,精密度在1.9%～3.1%。该方法简单、灵敏度高、分析时间短、定量准确,适用于婴儿乳粉中VB12的测定。A method was established for determining VB12 in infant milk powder by ultra performance liquid chromatography-tandem mass spectrometry （UPLC/MS/MS） and ultra performance liquid chromatography. The analytes were extracted with ammonium acetate （pH=4.0） and cleaned up with immunoaffinity column. The target analytes were separated by Waters BEH C18 column. The mobile phases of UPLC were 0.03 mol/L potassium dihydrogen phosphate （pH=3.5） and acetonitrile （85：15）. The detection wavelength was 361 nm. The mobile phases of UPLC-MS/MS were acetonitrile and 0.1% formate in water with gradient elution. The analytes were detected by tandem quadrupole mass spectrometry after positive electrospray ionization by multiple reaction monitoring （MRM）. External standard method was used to determine the results. The linear range of the calibration curve was 10-500 μg/L. The limit of detection （LOD） were 0.5μg/kg, and the limit of quantitation （LOQ） were 1.5 μg/kg. The mean recoveries of LC method were 90.2%-98.3%. The relative standard deviation （RSD） were 1.4%-2.5%. The mean recoveries of LC-MS/ MS method were 89.3%-95.7%. The relative standard deviation （RSD） were 1.9%-3.1%. The simplicity, sensitivity and good precision of the method made it be well suitable for determination of VD in infant milk powder.

关 键 词：免疫亲和柱净化 超高效液相色谱-串联质谱 超高效液相色谱 VB12 

分 类 号：TS252.1[轻工技术与工程—农产品加工及贮藏工程]