婴幼儿配方乳粉中VB_(12)的微量测定  

The micro-detection for vitamin B_(12) in infant formula milk powder

作　　者：刘小青[1] 黄建飞[1] 黄静敏[1] 陈晶[1] 兰全学[1] LIU Xiao-qing HUANG Jian-fei HUANG Jin-min LAI Xiu-juan MA Fang-cao WU Jia-xuan LAN Quan-xue(Shenzhen Academy ofMetrology ＆ Quality inspection, Shenzhen 518131)

机构地区：[1]深圳市计量质量检测研究院,深圳518131

出　　处：《中国食品添加剂》2016年第12期193-196,共4页China Food Additives

摘　　要：目的：建立婴幼儿配方乳粉中VB12的快速微量检测方法。方法：样品加水复原后,95℃水浴提取30 min;0.2 ng/m L标准品贮备液混合0.5%的海藻糖无菌过滤分装后真空冷冻冻干,并随后制备成系列标准曲线浓度;VB12测定培养基无菌过滤后以1/200的比例加入2.0 MCF的菌种储备液;随后向一次性八连小管中加入150μL VB12测定培养基及样品和标准品,密封条件下37℃培养22~24 h,然后采用酶标仪读值。在建立的方法基础上分别做重复性实验和加标回收实验。结果：采用该方法建立的VB12的微量测定法线性R达到0.9993,重复性测试RSD在1.51%~5.39%,测量效果好,回收率在93.6%~106%内,试验灵敏度高。结论：建立了婴幼儿配方乳粉中VB12的快速微量检测方法,且该方法简化了检测步骤,缩短了检测周期,摒弃了GB5413.14-2010和商品化维生素检测试剂盒的缺点,结合两者优点,具有明显优势,适合在基层实验室推广。Objective ：To establish a rapid micro-detection method of vitamin B12 in infant formula milk powder. Method ：The sample were in 95℃ water bath extraction after rehydration;the mixture of 0.2 ng/m L Standard reserve liquid and 0.5 % trehalose was treated with vacuum freeze lyophilization and further prepared series of standard curve of concentration.;the 2.0 MCF strains were in 1/200 proportion added into sterile vitamin B12 assay medium;Whereafter,the same volume of 150 u L sample and standard were pipetted into sterile vitamin B12 assay medium,the mixture were cultured in 37℃ and then measured by microplate reader. Based on the method,repetitive experiments and standard addition test were performed. Results ：By using this method,correlation coefficient of the standard curve was 0.9993,RSD of parallel tests was between 1.51% and5.39%,and recovery rate of standard addition test was between 93.6% and 106%. Conclusions ：This paper has established a rapid micro-detection method of vitamin B12 in infant formula milk. The method was greatly shortened the time and easy to operate. Compare with GB5413.14-2010 and kit method,the method was combining both advantages and was suitable for application in the laboratory.

关 键 词：婴幼儿配方乳粉 VB12 微量测定 GB5413.14-2010 酶标仪 

分 类 号：TS202.3[轻工技术与工程—食品科学] TS252.51[轻工技术与工程—食品科学与工程]