Institute of Sciences of Food Production, National Research Council (ISPA-CNR), via G. Amendola 122/O, 70126, Bari, Italy.
Anal Bioanal Chem. 2018 Sep;410(22):5653-5662. doi: 10.1007/s00216-018-0927-8. Epub 2018 Feb 13.
In recent years, mass spectrometry (MS) has been establishing its role in the development of analytical methods for multiple allergen detection, but most analyses are being carried out on low-resolution mass spectrometers such as triple quadrupole or ion traps. In this investigation, performance provided by a high resolution (HR) hybrid quadrupole-Orbitrap™ MS platform for the multiple allergens detection in processed food matrix is presented. In particular, three different acquisition modes were compared: full-MS, targeted-selected ion monitoring with data-dependent fragmentation (t-SIM/dd2), and parallel reaction monitoring. In order to challenge the HR-MS platform, the sample preparation was kept as simple as possible, limited to a 30-min ultrasound-aided protein extraction followed by clean-up with disposable size exclusion cartridges. Selected peptide markers tracing for five allergenic ingredients namely skim milk, whole egg, soy flour, ground hazelnut, and ground peanut were monitored in home-made cookies chosen as model processed matrix. Timed t-SIM/dd2 was found the best choice as a good compromise between sensitivity and accuracy, accomplishing the detection of 17 peptides originating from the five allergens in the same run. The optimized method was validated in-house through the evaluation of matrix and processing effects, recoveries, and precision. The selected quantitative markers for each allergenic ingredient provided quantification of 60-100 μg/g allergenic ingredient/matrix in incurred cookies.
近年来，质谱（MS）在开发用于多种过敏原检测的分析方法方面确立了其地位，但大多数分析都是在低分辨率质谱仪（如三重四极杆或离子阱）上进行的。在这项研究中，展示了高分辨率（HR）混合四极杆-Orbitrap™ MS 平台在加工食品基质中多种过敏原检测的性能。特别是，比较了三种不同的采集模式：全质量谱、带数据依赖碎片化的靶向选择离子监测（t-SIM/dd2）和平行反应监测。为了挑战 HR-MS 平台，样品制备尽可能简单，仅限于 30 分钟的超声辅助蛋白质提取，然后用一次性尺寸排阻小柱进行净化。在家制曲奇中监测了五种过敏原成分（脱脂牛奶、全蛋、大豆粉、榛子粉和花生粉）的选定肽标记物，该曲奇选择作为模型加工基质。定时 t-SIM/dd2 被发现是灵敏度和准确性之间的良好折衷选择，可在同一次运行中检测到来自五种过敏原的 17 种肽。通过评估基质和加工效应、回收率和精密度，在内部对优化方法进行了验证。每个过敏原成分的选定定量标记物提供了在实际曲奇中 60-100μg/g 过敏原成分/基质的定量。
