摘要
目的:建立一种肝素亲和柱-高效液相色谱法(High Performance Liquid Chromatography,HPLC)快速检测乳粉中乳铁蛋白的分析方法。方法:样品经水溶液超声提取,调节pH,采用50 mmol/L磷酸氢二钠溶液(A)和1 mol/L氯化钠溶液(B)作为流动相进行梯度洗脱,采用HPLC进行测量,检测器采用280 nm波长。结果:乳铁蛋白在上述色谱条件下能完全分离。乳铁蛋白线性范围为5.54~110.9μg/mL,相关系数r=0.999966,方法检出限为12 mg/100g,定量限为28.5 mg/100g;平均回收率97.5%~101.9%,相对标准偏差(relative standard deviation,RSD)<3.0%。该方法检测结果与酶联免疫吸附法(ELISA)基本一致。结论:该方法操作简便,精密度和回收率高,重复性好,可以用于测定乳粉中乳铁蛋白。
Objective :To establish a method for determination of lactoferrin in milk powder samples by heparin affinity column-high performance liquid chromatography(HPLC). Methods :The samples were ultrasonically extracted with aqueous solution,adjusted the pH,using a gradient elution program of 50 mmol/L disodium phosphate aqueous solution(A)and 1 mol/L sodium chloride aqueous solution(B). HPLC was used for the measurement with 280 nm wavelength. Results :Lactoferrin was completely separated in the chromatographic conditions. The linear range was 5.54 ~ 110.9μg/mL,with the correlation coefficient r=0.999966. The limit of detection was 12mg/100g,and the limit of quantification was 29mg/100g. The average recoveries were 97.5%-101.9%,with relative standard deviation(RSD)< 3.0%. The detection results of this method were basically consistent with the ELISA method.Conclusion :This method is simple,accurate and reproducible,which can be used for the determination of lactoferrin in milk powder.
作者
苏昭仑
张颖
贺瑞坤
SU Zhaolun;ZHANG Ying;HE Ruikun(BY-HEALTH CO.,LTD.,Zhuhai 519040)
出处
《中国食品添加剂》
CAS
北大核心
2023年第2期289-294,共6页
China Food Additives
