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Citation: ZHAN Lina, CHEN Qin, GU Shuqing, DENG Xiaojun. Determination of milk allergen caseins in foods by ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry[J]. Chinese Journal of Chromatography, ;2017, 35(4): 405-412. doi: 10.3724/SP.J.1123.2016.10034 shu

Determination of milk allergen caseins in foods by ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry

1.
Department of Food Engineering, School of Life Science, Shanghai University, Shanghai 200436, China;
2.
Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai 200135, China

Received Date: 11 October 2016

Fund Project: Shanghai Natural Science Foundation (No. 14ZR1450000)

Based on ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry (UPLC-Q/Orbitrap MS) system, a rapid screening and quantitative detection method for milk allergen caseins was developed. After the sample was extracted with protein extraction buffer, a 5 kD ultrafiltration membrane was used to remove small molecule impurities. The extracted protein was then digested by trypsin. The full-scan mass spectra obtained by the data-dependent acquisition (DDA) mode were used for protein identification, and parallel reaction monitoring (PRM) technique was used for quantitative analysis of the target signature peptide. The internal standard peptide and the internal standard substances for every signature peptide were designed and synthesized to reduce the matrix effect and eliminate the loss in sample pretreatment. The proposed method showed a good linear relationship in the range of 5-250 μg/L. The limits of quantitation were in the range of 0.2-5.5 μg/kg. The observed recoveries ranged from 68.8% to 104.4%, and the RSDs were lower than 6%. This method can be used for rapid screening and quantitative analysis of milk allergen casein in juice, jam, bread and breakfast cereals.
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