关键词: 超高效液相色谱-串联质谱, 过敏原, 卵白蛋白, 胰蛋白酶, 特征肽段?

Abstract: An ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the detection of the egg allergen ovalbumin in food in this investigation. Egg white was trypsinized and the resulting hydrolysate was purified by ultrafiltration, and then separated by Easy-nLC 1000 nano liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry with acetonitrile-0.1% (V/V) formic acid water as the mobile phase. By comparison with the Uniprot database, the characteristic peptide GGLEPINFQTAADQAR(GR-16) with good specificity and sensitivity was selected. Then, the UPLC-MS/MS analysis was performed using positive ion electrospray ionization (ESI+) in the multi-reaction monitoring (MRM) mode. The method showed a good linear relationship in the concentration range of 5–5 000 ng/mL with a limit of quantification (LOQ) of 0.1 μg/g. The average recoveries ranged from 77.22% to 98.55% with relative standard deviations (RSDs) lower than 9.97%. This method has high specificity and good sensitivity and is also time-saving, and thus it can provide technical support for the accurate determination of egg allergens in food product.

Key words: ultra-high performance liquid chromatography-tandem mass spectrometry, allergen, ovalbumin, trypsin, characteristic peptides