高效液相色谱法(HPLC)测定牛乳中β-乳球蛋白.pdf

高效液相色谱法 (HPLC) 测定牛乳中α -乳白蛋白 Determination of α-lactalbumin in milk products by high performance liquid chromatography ( HPLC) 关荣发 1，黄光荣 1 ，贾振宝 1，戴贤君 1，叶兴乾 2 1 1 1 1 2 GUAN Rong-fa , HUANG Guang-rongn , JIA Zhen-bao , DAI Xian-jun , YE Xing-qian （1.中国计量学院生命科学学院， 杭州 310018; 2.浙江大学食品科学与营养系， 杭州 310029 ） (1. College of Life Sciences, China Jiliang University ，Hang Zhou 310018; 2. Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310029) 摘要 : 建立了利用常规的 C18 色谱柱的高效液相色谱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, 5 μm) , 柱温为 45 ℃, UV 检测 波长 2l5 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 0.8mL/min 。结果表明 : α-乳白蛋白的线性范围分别为 50 μg/mL-1000 μg/mL( r=0.9909); 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。 关键词 : 高效液相色谱法 ; α-乳白蛋白 ; 乳制品 ; 测定 Abstract: A method for the determination of α-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 wavelengts 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 μ g/mL ~1000 μg/mL( r=0.9909). 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 mi