【摘要】 通过研究免疫亲和柱性能、免疫亲和柱净化时上样液温度、洗脱液种类等因素对乳粉中免疫球蛋白G（immunoglobulin G,IgG）含量检测的影响，建立一种乳粉中IgG检测的高效液相色谱法。乳粉样品用磷酸缓冲液进行提取，提取液经免疫亲和柱富集纯化，洗脱液采用C₄反相色谱柱（250 mm×4.6 mm,3.5μm）分离，高效液相色谱-紫外检测器检测，外标法定量。结果表明，方法适用于不同IgG含量的乳粉，在10～500μg/mL线性范围内标准曲线相关性良好，方法检出限为80 mg/kg，定量限为300 mg/kg，在300、600、3 000 mg/kg 3个加标水平下，加标回收率和相对标准偏差均满足GB 5009.295—2023《食品安全国家标准化学分析方法验证通则》要求。该方法简单高效、结果准确性高，能为新功能乳制品营养价值评价及产品宣称提供精准的技术支持。更多还原

【Abstract】 A high performance liquid chromatography（HPLC） method for the detection of immunoglobulin G（IgG） in milk powder was established by studying the effects of immunoaffinity column performance, temperature of the sample solution during immunoaffinity column cleanup, and the type of eluent on the detection of the IgG content in milk powder. Samples were extracted with phosphate buffer solution, and the extract was purified by immunoaffinity column chromatography. The eluate was separated on a C₄ reversed-phase column（250 mm × 4.6 mm, 3.5 μm）, detected using HPLC-ultraviolet detector, and quantified by an external standard method. The results showed that the method was applicable to milk powder with different IgG contents. The standard curve exhibited good linearity in the concentration range of 10–500 μg/mL. The detection limit of the method was 80 mg/kg, and the quantification limit was 300 mg/kg. The recoveries and precision expressed as relative standard deviations（RSDs） at three spiked levels（300, 600 and 3 000 mg/kg） were both aligned with the requirements of the Chinese national food safety standard GB 5009.295-2023. This method is simple, efficient and accurate, providing precise technical support for the evaluation of the nutritional value of new functional dairy products and product claims.更多还原