高效液相色谱法测定牛乳中α-乳白蛋白

Determination of α-lactalbumin in milk products by high performance liquid chromatography

利用常规的C18色谱柱的高效液相色谱建立测定牛乳中主要过敏蛋白α-乳白蛋白含量的方法。色谱条件:色谱柱Alltima-C18(4.6 mm×200 mm,5μm),柱温为45℃,UV检测波长215 nm,流动相A为含0.1%三氟乙酸的超纯水,流动相B为乙腈:超纯水:三氟乙酸=400:100:0.5,采用梯度洗脱方法,流速0.8 mL/min。结果表明:α-乳白蛋白的线性范围分别为50～1 000μg/mL(r=0.990 9);α-乳白蛋白平均回收率为97.27%,RSD=0.76%(n=5);该方法简便、准确,适合于乳制品中α-乳白蛋白含量的测定。

A method for the determination ofα-lactalbumin in milk products by high performance liquid chromatography has been established.The chromatography conditions as follow： Alltima-C18 conlum（4.6 mm×200 mm,5 μm）,UV detection wavelengs 215 nm.This method consisted of a linear gradient of the two mobile phases of 0.1% trifluoroacetic acid in water and 0.5% trifluoroacetic acid and 80% acetonitrile in water at a flow rate of 0.5 mL/min,The conlum temperature was 25 ℃.The result showed that for α-lactalbumin the calibration curve was linear in the range of 50 μg/mL～1 000 μg/mL（r=0.990 9）.The average recovery percent was 97.27,the RSD was 0.76%（n=5）.The methods is suitable for contents analysis of α-lactalbumin in milk products.