反相高效液相色谱法同时检测多种乳清蛋白成分

Determination of components of whey protein by reversed-phase HPLC method

建立一种快速、简便鉴定和定量检测乳清蛋白中牛乳铁蛋白(bLf)、转基因人乳铁蛋白(rLf)、α-乳白蛋白(α-La)和β-乳球蛋白(β-Lg)的方法。对样品进行离心脱脂和酸法去除酪蛋白,水洗乳脂和酪蛋白以提高α-乳白蛋白、β-乳球蛋白和乳铁蛋白(Lf)的回收率。乳清通过高效液相色谱检测,采用Proteonavi反相C4色谱柱,以体积分数为0.1%的三氟乙酸水溶液和乙腈水溶液为流动相,梯度洗脱,流速0.5mL/min,柱温35℃,检测波长280nm。结果显示:此方法可以有效分离牛α-乳白蛋白、牛β-乳球蛋白、牛乳铁蛋白和转基因人乳铁蛋白。标准品线性关系良好,平均回收率在正常范围内。

The aim of this study was the rapid determination of α-lactalbumin（α-La）,β-lactoglobulin（β-Lg）,Lactoferrin（Lf）.After removing fat from whey protein by centrifugation and precipitating caseins by adjusting pH,the whey contents were detected by Dikma Proteonavi Chromatographic column with 0.1%（volume fraction） TFA solution and acetonitrile solution by 0.5 mL/min flow rate,gradient elutio and column temperature was stetted at 35 ℃.Detection was carried out at the wavelength 280 nm.The results illustrate that the separation of α-La,β-Lg and Lf is available.The linear realationship of standard samples is good,and the recovery of the standard samples is in the normal range.The method which is simple,fast and accurate can determine the content of α-La,β-Lg and Lf of breast milk,milk and transgenic milk.