摘要
目的:建立一种测定牛初乳保健食品中免疫球蛋白G的反相高效液相色谱法。方法:样品采用磷酸盐缓冲液涡旋振荡提取,经Protein G固相免疫亲和小柱净化后,使用高效液相色谱仪(搭载紫外检测器)检测,紫外波长为280 nm,采用C反相色谱柱(4.6×250 mm, 5μm)分析。流动相为0.1%三氟乙酸-水溶液和乙腈,梯度洗脱分离,外标法定量。结果:色谱分离过程可在10 min内完成,免疫球蛋白G在10~100μg/mL范围内线性良好,相关系数R=0.999 6。检出限为0.5 g/kg,定量限为1.7 g/kg。3个浓度水平的方法加标回收率为92.2~97.5%,方法精密度为0.20~1.18%。样品经前处理后4℃冰箱内保存,在24 h内稳定性良好。结论:该方法特异性强,灵敏度、精确度高,符合方法学验证要求,可快速、准确地对保健食品中免疫球蛋白G进行分析检测。
【Objective】 To establish a reverse high-performance liquid chromatograph method for analyzing immunoglobulin G(IgG) in bovine colostrum supplement. 【Method】 Phosphate buffer solution was used to extract sample by vortex method, and the solution was purified by protein G immunoaffinity column and detected by high performance liquid chromatograph combined ultraviolet detector, the wavelength is280 nm. The separation was performed on reverse Ccolumn(4. 6×250 mm, 5 μm) with 0. 1%trifluoroacetic acid solution-acetonitrile as mobile phase, with gradient elution. IgG was quantified by external standard method. 【Result】 The chromatographic separation could be completed within 10 min. IgG had linear relationship in the range of 10-100 μg/mL, with a correlation coefficient of 0. 999 6. The limit of detection was 0. 5 g/kg, and the limit of quantitation was 1. 7 g/kg. The range of recoveries of 3 concentration levels were 92. 2%-97. 5%.Relative standard deviation of method was 0. 20%-1. 18%. After preprocess method, sample solution were stable within a 4℃ storage condition in 24 h. 【Conclusion】 The method had high specificity, sensitivity and precision, and conform to requirements of methodology validation, could rapidly and accurately analyze IgG in supplement.
作者
罗娇依
刘彤彤
曹进
赵溪
孙姗姗
张旭光
LUO Jiao-yi;LIU Tong-tong;CAO Jin;ZHAO Xi;SUN Shan-shan;ZHANG Xu-guang(National Institutes for Food and Drug Control,Beijing 100050,China;Key Laboratory of Food Quality and Safety of State Administration for Market Regulation,Beijing 100050,China;By-health Co.,Ltd.,Zhuhai 510663,China)
出处
《中国食物与营养》
2022年第9期20-26,共7页
Food and Nutrition in China
