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超高效液相色谱-串联三重四极杆质谱法测定乳清蛋白粉中α-乳白蛋白和β-乳球蛋白
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杜娟,郑云鹏,董洪涛,张凤霞 1.黑龙江飞鹤乳业有限公司集团实验室2.飞鹤(龙江)乳品有限公司 
中文摘要:建立了超高效液相色谱-串联三重四极杆质谱(UPLC-MS/MS)法测定乳清蛋白粉中α-乳白蛋白(α-La)和β-乳球蛋白(β-Lg)的分析方法。样品经碘代乙酰胺烷基化,再经碱性胰蛋白酶酶解。在Waters ACQUITY UPLC-BEH C18,300 色谱柱上分离,流动相为0.1%甲酸水和01%甲酸乙腈,采用梯度洗脱分离,正离子多反应监测模式(MRM)检测,内标法定量。结果表明,α-乳白蛋白和β 乳球蛋白分别在40~1 000 nmol/L和80~2 000 nmol/L范围内呈良好线性关系,定量下限(S/N=10)均为0.020 g/100 g;加标回收率为84.7%~95.6%,相对标准偏差(RSD,n=6)为1.6%~5.8%。基于上述方法对5个国家的4类乳清蛋白粉(脱盐乳清粉D70和D90、浓缩乳清蛋白粉和α-乳白蛋白粉)进行了含量测定和差异对比。该方法操作简便,定量准确,可用于乳清蛋白粉等蛋白类原料中α-乳白蛋白和β-乳球蛋白的测定。
中文关键词:超高效液相色谱-串联三重四极杆质谱  α-乳白蛋白  β-乳球蛋白  乳清蛋白粉
 
Determination of α Lactalbumin and β Lactoglobulin in Whey Powder by Ultra Perfor mance Liquid Chromatography-Triple Quadrupole Tandem Mass Spectrometry
Abstract:An ultra performance liquid chromatography-triple quadrupole tandem mass spectrometric(UPLC-MS/MS) method was developed for the determination of α-lactalbumin(α-La) and β-lactoglobulin(β-Lg) in whey powder under positive ion multiple reaction monitoring(MRM) mode.The samples were firstly alkylated with iodoacetamide,followed by enzymatic hydrolysis with alkaline trypsin,then separated on a Waters ACQUITY UPLC-BEH C18,300 - column with 0.1% formic acid water solution and 0.1% formic acid acetonitrile solution as mobile phases by gradient elution.Results showed that there were good linear relationships for α-La and β-Lg in the ranges of 40-1 000 nmol/L and 80-2 000 nmol/L,respectively.The limits of quantitation(S/N=10) for α-La and β-Lg were 0.020 g/100 g,respectively.The average recoveries of two analytes from spiked whey powder samples were in the range of 84.7%-95.6%,with relative standard deviations(RSDs,n=6) of 1.6%-5.8%.Based on the above method,the targets in four kinds of whey powder samples,i.e.70% demineralised whey powder,90% demineralised whey powder,whey protein concentrate and α-lactalbumin powder from five countries were determined and contrasted.With the advantages of simple operation,accuracy and reliability,the established method was suitable for the determination of α-La and β-Lg in whey powder.
Key Words:ultra performance liquid chromatography-triple quadrupole tandem mass spectrometry(UPLC-MS/MS)  α-lactalbumin(α-La)  β-lactoglobulin(β-Lg)  whey powder
引用本文:杜娟,郑云鹏,董洪涛,张凤霞.超高效液相色谱-串联三重四极杆质谱法测定乳清蛋白粉中α-乳白蛋白和β-乳球蛋白[J].分析测试学报,2020,39(7):900-905.
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