免疫亲和柱净化-高效液相色谱法测定乳粉中的生物素含量.pdfVIP

现代食品科技 Modern Food Science and Technology 2013, Vol.29, No.3 免疫亲和柱净化-高效液相色谱法测定 乳粉中的生物素含量 1 2 3 3 佟晓芳 ，张永胜 ，陶大利 ，姜金斗 （1.黑龙江省产品质量监督检测研究院，黑龙江哈尔滨 150050）（2.北京博欧实德生物技术有限公司，北京 100107）（3.农业部乳品质量监督检验测试中心，黑龙江哈尔滨 150090） 摘要：建立了免疫亲和柱净化-高效液相色谱法测定乳粉中生物素的方法。该方法样品处理为样品用磷酸盐缓冲液溶解后，样液 通过生物素免疫亲和柱净化、甲醇洗脱液洗脱、浓缩富集后直接测定。采用紫外检测器，波长为215 nm ；C18色谱柱，柱温为35 ℃， 流动相为50 mmol/L磷酸缓冲盐，PH=3.5: 甲醇(85:15,v/v)，流速为1.0 mL ／min。标准曲线回归方程y=343.2+20.797x，相关系数r=0.9958 ； 回收率为90.7% ；RSD%为2.62 。本方法检出限为1.0 μg/kg。该法较国标方法具有样品处理简单方便，灵敏度高，重现性好，分析时间 短等优点，可以满足调制乳粉及婴幼儿配方乳粉中生物素含量的测定。 关键词：免疫亲和柱；乳粉；生物素；高效液相色谱 文章篇号：1673-9078(2013)3-660-663 Determination of Biotin Content in Milk Powder by Immunoaffinity Column Purification-HPLC 1 2 3 3 TONG Xiao-fang , ZHANG Yong-sheng , TAO Da-li , JIANG Jin-Dou (1.Heilongjiang Province Product Quality Supervision and Testing Institute, Harbin 150050, China) (2.Beijing the Biostest Biotechnology Co. , Ltd, Beijing 100107, China) (3.Ministry of Agriculture Dairy Quality Supervision and Testing Center, Harbin 150090, China) Abstract: The Immunoaffinity column purification - high performance liquid chromatographic method was developed in this paper for the determination of biotin in milk powder. Sample was dissolved with phosphate buffer and purified through biotin immunoaffinity column with ethanol, and concentrated to a direct determination after enrichment. UV detect or wavelength was of 215 nm and C18 column was used at 35 ° C. Mobile phase is 50 mmol/L phosphate buffer salt (pH 3.5)-methanol (85:15, v/v) at a flow rate of 1.0mL/mi