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建立了高效凝胶渗透色谱法测定配方奶粉中免疫球蛋白G(IgG)含量的方法。奶粉样品经pH 7的0.05 mol/LNa_2HPO_4溶液溶解,高速离心、过滤,滤液经protein G免疫亲和柱净化。采用Zorbax GF-450凝胶色谱柱(250 mm×9.4 mm,6μm)分离,流动相为pH 7的0.05 mol/L Na_2HPO_4溶液,流速1.0 m L/min,检测波长280 nm。免疫球蛋白G在0.2~5.0 mg/m L的含量范围内线性关系良好,相关系数为0.9991,方法检出限为0.1 mg/g,加标回收率为86.0%~117.7%,相对标准偏差(n=6)为3.8%~7.1%。方法适用于配方奶粉中免疫球蛋白G的定性、定量分析。
A method for the determination of immunoglobulin G (IgG) in milk powder by high performance gel permeation chromatography was established. The milk powder sample was dissolved in 0.05 mol / L Na2HPO4 solution at pH 7, centrifuged at high speed and filtered. The filtrate was purified by protein G immunoaffinity column. The separation was carried out on a Zorbax GF-450 gel column (250 mm × 9.4 mm, 6 μm). The mobile phase consisted of 0.05 mol / L Na_2HPO_4 solution with a flow rate of 1.0 mL / min and a detection wavelength of 280 nm. The linearity of immunoglobulin G in the range of 0.2-5.0 mg / m L was good with a correlation coefficient of 0.9991. The detection limit was 0.1 mg / g and the recoveries were between 86.0% and 117.7%. The relative standard deviations n = 6) was 3.8% to 7.1%. The method is suitable for qualitative and quantitative analysis of immunoglobulin G in formula milk powder.