Abstract: In this paper, a reversed-phase high-performance liquid chromatography method was proposed to determine five major proteins including α-lactalbumin, β-lactoglobulin, glycomacropeptide, bovine serum albumin, and immunoglobulin G in whey protein powder. After dissolution and ultrasonication, samples were separated on ChromCore 300 C4-T chromatographic column (4.6 mm × 150 mm, 5 μm) with gradient elution using a mobile phase consisting of 0.1% (V/V) aqueous trifluoroacetic acid (TFA) and 90% acetonitrile (containing 0.09% TFA). These analytes were detected using a diode array detector scanning in the wavelength range from 200 to 700 nm at 214 nm. Under the optimal experimental conditions, the five major bovine whey proteins were well separated within 30 min. The average recoveries from spiked samples were in the range of 83.5%–95.3% with relative standard deviations (RSDs) of 2.03%–7.49%. The limit of detection was 0.25% (m/m) for all analytes. The established method is rapid, accurate and sensitive that enables the simultaneous detection of the major bovine whey proteins in whey protein powder.

Key words: reversed-phase high-performance liquid chromatography; whey protein powder; α-lactalbumin; β-lactoglobulin; glycomacropeptide; bovine serum albumin; immunoglobulin G

摘要： 研究一种运用反相高效液相色谱技术同时快速检测乳清蛋白粉中α-乳白蛋白、β-乳球蛋白、糖巨肽、牛血清白蛋白、免疫球蛋白G 5 种主要蛋白含量的方法。样品经溶解、超声后，采用ChromCore 300 C4-T色谱柱（4.6 mm×150 mm，5 μm）分离，用0.1%三氟乙酸溶液和90%乙腈溶液（含0.09%三氟乙酸）体系作为流动相进行梯度洗脱，采用二极管阵列检测器，在200～700 nm波长处进行扫描，214 nm波长处进行检测。在此实验条件下，5 种主要蛋白可在30 min内有效分离，平均加标回收率为83.5%～95.3%，相对标准偏差为2.03%～7.49%，5 种蛋白的检出限均为0.25%（m/m）。建立的方法可实现对乳清蛋白粉中5 种主要蛋白的同时检测，具有快速、准确、灵敏的特点。

关键词: 反相高效液相色谱；乳清蛋白粉；α-乳白蛋白；β-乳球蛋白；糖巨肽；牛血清白蛋白；免疫球蛋白G