基于超高效液相色谱串联四极杆飞行时间高分辨质谱分析牛肉丸中的大豆蛋白,以5条大豆源特征肽作为定量标志物,对样品中提取的蛋白经胰蛋白酶酶解、固相萃取柱净化,在电喷雾离子源的正离子模式下进行电离,准多重反应监测模式采集目标碎片离子,结合外标法进行定量。结果表明,5条大豆源特征肽在20～800 μg/L范围内的线性关系较好,相关系数均大于0.99,定量限0.17～0.62 μg/L,基质效应95.8%～112.4%。低、中、高3个水平的加标回收率为84%～112%,处于可接受的范围,相对标准偏差均小于10%,方法重复性好。市售的14种不同价格的牛肉丸样品中可检测出5条大豆源特征肽,其检出比例为50%,含量为1.07～122.04 mg/kg,随着牛肉丸制品价格的升高,其混入5条大豆源特征肽的含量总体呈下降趋势。该方法灵敏度高、回收率好、结果稳定,可利用碎片离子有效排除假阳性,可用于牛肉丸中大豆蛋白的准确鉴别与定量测定。
王军
,
黄俊杰
,
陈济钦
,
温丽萍
,
吴海锋
,
郭鑫
,
何颖霖
,
林奕欣
,
王忠合
. 基于超高效液相色谱-四级杆飞行时间质谱定量测定牛肉丸中的大豆蛋白[J]. 食品与发酵工业, 2026
, 52(3)
: 382
-390
.
DOI: 10.13995/j.cnki.11-1802/ts.044552
The contents of marker peptides derivated from soy proteins were quantitatively analyzed by ultra-high performance liquid chromatography coupled with quadrupole time-of-flight high-resolution mass spectrometry.The extracted proteins from samples were enzymatically digested by trypsin, purified by solid-phase extraction columns, and then ionized in positive ion mode of electrospray ion source.The targeted fragment ions were collected in pseudo-multiple reaction monitoring mode.The external standard method was used for quantification.Results showed that the linear relationship of 5 marker peptides of soy protein was good within the range of 20-800 μg/L, with a correlation coefficient greater than 0.99.The limit of quantification was 0.17-0.62 μg/L.The matrix effect of the protein extract in beef balls was 95.8%-112.4%.The recovery rates were 84%-112% at low, medium and high levels, which were within the acceptable range.The relative standard deviations of multiple determinations were less than 10%, and the repeatability was good.5 of marker peptides from soy protein were detected in 14 commercially available beefball samples on the market.The detection proportion was 50%, and the content of marker peptides was 1.07-122.04 mg/kg.With the increase of the price of beef balls, the content of marker peptides decreased.This method showed high sensitivity, good recovery rate, stable results, and effectively eliminated false positives by using fragment ions,which was applicable for identification and quantitative determination of soy protein in beef balls.
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