反相高效液相色谱法快速测定乳铁蛋白含量

Rapid determination of lactoferrin by reversed-phase high performance liquid chromatography

中文关键词:  乳铁蛋白  反相高效液相色谱法  快速测定  乳制品

英文关键词:lactoferrin  reversed-phase high performance liquid chromatography  rapid determination  dairy

中文摘要:

      目的 建立一种反相高效液相色谱法(RP-HPLC)快速测定乳铁蛋白在乳原料及乳制品中含量的方法。方法 根据酪蛋白在等电点(pH 4.6)凝聚沉淀特点, 采用RP-HPLC法, 用pH4.6醋酸盐缓冲液溶解分离乳铁蛋白, 用C18色谱柱快速分离, 用流动相A为0.1%三氟乙酸水溶液, 流动相B为含有0.09%(V:V)三氟乙酸的90%(V:V)乙腈水溶液, 进行梯度洗脱, 检测波长214 nm, 流速0.7 mL/min。结果 乳铁蛋白在10～500 mg/L之间呈现良好的线性, 相关系数为0.9999, 平均回收率为98.0%, RSD=0.8% (n=9)。结论 该方法推广性高、快速, 简便、准确, 可用于乳铁蛋白原料、奶粉、牛奶及保健品等中乳铁蛋白的含量检测。

英文摘要:

      Objective To establish a method for the rapid determination of the content of lactoferrin in raw material milk and dairy products by reversed-phase high performance liquid chromatography (RP-HPLC). Methods The RP-HPLC method was used in this method. According to the characteristics that lactoferrin precipitated at the isoelectric point (pH 4.6), lactoferrin was dissolved and separated by acetate buffer with pH 4.6. A C18 column was used for rapid separation of protein, with the mobile phase A of 0.1% trifluoroacetic acid aqueous solution, and the mobile phase B of 90% (V:V) acetonitrile solution with 0.09% trifluoroacetic acid (:V). The detection wavelength was 214 nm and the flow rate was 0.7 mL/min. Results Lactoferrin showed a good linearity between 10~500 mg/L and the correlation coefficient was 0.9999. The average recovery rate was 98.0%, with RSD of 0.8% (n=9). Conclusion The method was wide used, rapid, simple and accurate, it could be used in the determination of lactoferrin in lactoferrin raw material, milk powder, milk and health products.