Assessment of Pru p 1 and Pru p 3 in peach fruit by liquid chromatography-tandem mass spectrometry

摘要

Pru p 1 and Pru p 3 are proteins found in peach (Pruntis persica (L.) Batsch), and they are members of the pathogenesis-related protein 10 (PR-10) and non-specific lipid transfer protein (nsLTP) families, respectively. They are the main allergens of peach. In this paper, we describe a quantitative method for these proteins based on protein absolute quantification with liquid chromatography-tandem mass spectrometry, and LVASPSGGSIIK [~(13)C_6,~(15)N_2] and ISASTNC[CAM]ATVK[~(13)C_6,~(15)N_2] (C[CAM]:carbamidomethyl-modifled C), stable isotope-labelled internal standard peptides. We show that this method exhibits linearity with r~2 > 0.97 in a dose range of 3.1-200 fmol/pL for LVASPSGGSIIK[~(13)C_6,~(15)N_2] and 13-200 fmol/muL for ISASTNC[CAM]ATVK[~(13)C_6,~(15)N_2]), and overall coefficients of variation for multiple tests of 11.6-30.1%. This approach is therefore helpful for screening and breeding low allergenic peach cultivars.