液质联用法测定深度水解乳清蛋白婴儿配方中的酪蛋白过敏原
Determination of Casein Allergens in Extensively Hydrolyzed Casein Infant Formula by Liquid Chromatography-Tandem Mass Spectrometry.
机构信息
Perrigo Nutritionals, Research and Development, 147 Industrial Park Road, Georgia, VT 05468, USA.
Perrigo Nutritionals, Scientific Affairs, 652 Peter Jefferson Parkway, Charlottesville, VA 22911, USA.
出版信息
J AOAC Int. 2021 Mar 5;104(1):172-179. doi: 10.1093/jaoacint/qsaa142.
BACKGROUND
The use of hypoallergenic infant formulas and the need for reliable tests to determine the presence of residual antigens have increased in parallel.
OBJECTIVE
An LC-MS method for quantitation of casein was validated using incurred samples and a matrix-matched external standard curve.
METHOD
Powdered infant formula samples were extracted in a buffer of sodium deoxycholate and ammonium bicarbonate at 60°C and filtered through 7 kDa desalting columns. Samples were digested overnight with trypsin and precipitated with acid prior to analysis of marker peptides by tandem mass spectrometry.
RESULTS
Based on three marker peptides, the linear range for casein was 1.8-42 μg/g of powdered infant formula with an LOQ of 1.8 μg/g. The determination coefficients (R2) for each curve were ≥0.99 for casein peptides. Method repeatability was ≤22% RSD and intermediate precision was ≤23% RSD; recovery of casein from incurred material (2-20 µg/g) ranged from 78% to 118%.
CONCLUSIONS
An LC-MS/MS method was developed and validated for confirmation of casein allergens in hypoallergenic infant formula.
HIGHLIGHTS
A method was developed to accurately and reliably quantify casein allergens in extensively hydrolyzed casein infant formula by LC-MS without the need for custom peptide standards.
背景
低变应原婴儿配方食品的使用和对确定残留抗原的可靠检测需求同步增加。
目的
使用加标样品和基质匹配的外部标准曲线，验证用于定量检测酪蛋白的 LC-MS 方法。
方法
将婴儿配方粉样品在 60°C 的脱氧胆酸钠和碳酸氢铵缓冲液中提取，并用 7kDa 脱盐柱过滤。样品用胰蛋白酶消化过夜，用酸沉淀后，通过串联质谱分析标记肽。
结果
基于 3 个标记肽，酪蛋白的线性范围为 1.8-42μg/g 婴儿配方粉，定量限为 1.8μg/g。每条曲线的决定系数（R2）均≥0.99。方法重复性的 RSD 不超过 22%，中间精密度的 RSD 不超过 23%；从加标材料（2-20μg/g）中回收的酪蛋白回收率为 78%-118%。
结论
开发并验证了 LC-MS/MS 方法，用于确认低变应原婴儿配方食品中的酪蛋白过敏原。
要点
开发了一种无需定制肽标准品，通过 LC-MS 准确可靠地定量检测高度水解酪蛋白婴儿配方食品中酪蛋白过敏原的方法。
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