摘要
[目的]建立二维液相色谱法同时测定乳制品中乳铁蛋白及免疫球蛋白G的方法。[方法]样品经提取后直接进样,以Pharmacia HI-Trap Protein G柱为第一维色谱柱,磷酸盐缓冲溶液和甘氨酸缓冲液为流动相,分离目标蛋白与其他杂质。以BioCore SEC-300(4.6 mm×250 mm,5μm)为第二维色谱柱,水和磷酸盐缓冲溶液为流动相,分离乳铁蛋白和免疫球蛋白G。[结果]乳铁蛋白和免疫球蛋白G在0~1000 mg/L范围内线性良好,相关系数均>0.99。乳铁蛋白在固态样品和液态样品中的定量限分别为20,13 mg/kg。加标回收率为91.0%~107.0%,相对标准偏差为2.4%~5.3%。免疫球蛋白G在固态样品和液态样品中的定量限分别为34,7 mg/kg。加标回收率为91.2%~104.0%,相对标准偏差为2.1%~4.7%。[结论]试验建立的在线二维液相色谱系统,前处理简单、自动化程度高、分析效率高、准确度高、重复性好,适用于乳制品中乳铁蛋白及免疫球蛋白G含量测定。
[Objective]A method of two-dimensional liquid chromatograohy was established to simultaneously determine the content of lactoferrin and immunoglobulin G in dairy products.[Methods]The pretreated samples were directly injected into the constructed system.Target protein and other effects were separated on a Pharmacia HI-Trap Protein G as the first-dimensional column using a mobile phase consisting of sodium phosphate buffer and glycocoll buffer by gradient elution lactoferrin and immunoglobulin G were separated on a BioCore SEC-300(4.6 mm×250 mm,5μm)as the second-dimensional column using a mobile phase consisting of water and sodium phosphate buffer by gradient elution.[Results]The results showed that the standard curves of Lactoferrin and Immunoglobulin G were linear with R>0.99.The quantitation limits of Lactoferrin in solid samples was 20 mg/kg while in liquid samples was 13 mg/kg,the recovery rate were 91.0%~107.0%and the RSD were 2.4%~5.3%.The quantitation limits of Immunoglobulin G in solid samples was 34 mg/kg while in liquid samples was 7 mg/kg,the recovery rate were 91.2%~104.0%and the RSD were 2.1%~4.7%.[Conclusion]The online two-dimensional liquid chromatography system established in the experiment has the advantages of simple pretreatment,high automation,high analysis efficiency,high accuracy and good repeatability,and is suitable for the determination of lactoferrin and immunoglobulin G in dairy products.
作者
陈钦
周朗君
陈梓敬
刘梦妮
林森煜
CHEN Qin;ZHOU Langjun;CHEN Zijing;LIU Mengni;LIN Senyu(Guangzhou Inspection Testing and Certification Group Company Limited,Guangzhou,Guangdong 511447,China;National Quality Testing Center for Processed Food(Guangdong),Guangzhou,Guangdong 511447,China)
出处
《食品与机械》
CSCD
北大核心
2024年第9期61-65,98,共6页
Food and Machinery
基金
广东省市场监督管理局科技项目(编号:2020kj44)。
关键词
乳铁蛋白
免疫球蛋白G
在线二维液相色谱法
乳制品
lactoferrin
immunoglobulin G
online two-dimensional liquid chromatography
dairy products
