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Selection of possible marker peptides for the detection of major ruminant milk proteins in food by liquid chromatography-tandem mass spectrometry

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Abstract

The aim of this work was the determination of peptides, which can function as markers for identification of milk allergens in food samples. Emphasis was placed on two casein proteins (α- and β-casein) and two whey proteins (α-lactalbumin and β-lactoglobulin). In silico tryptic digestion provided preliminary information about the expected peptides. After tryptic digestion of four milk allergens, the analytical data obtained by combination of reversed-phase high performance liquid chromatography and quadrupole tandem mass spectrometry (LC-MS/MS) led to the identification of 26 peptides. Seven of these peptides were synthesized and used for calibration of the LC-MS/MS system. Species specificity of the selected peptides was sought by BLAST search. Among the selected peptides, only LIVTQTMK from β-lactoglobulin (m/z 467.6, charge 2+) was found to be cow milk specific and could function as a marker. Two other peptides, FFVAPFPEVFGK from α-casein (m/z 693.3, charge 2+) and GPFPIIV from β-casein (m/z 742.5, charge 1+), occur in water buffalo milk too. The other four peptides appear in the milk of other species also and can be used as markers for ruminant species milk. Using these seven peptides, a multianalyte MS-based method was developed. For the establishment of the method, it was applied at first to different dairy samples, and then to chocolate and blank samples, and the peptides could be determined down to 1 ng/mL in food samples. At the end, spiked samples were measured, where the target peptides could be detected with a high recovery (over 50%).

Possible marker peptides from four milk allergens for the detection of ruminant milk proteins

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Acknowledgements

The authors thank the Christian Doppler Research Association and RomerLabs for funding the Christian Doppler Laboratory ‘Rapid Test Systems for Allergenic Food Contaminants’. We thank also the government of Lower Austria for financial supporting of this work.

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Authors and Affiliations

  1. Department IFA-Tulln, Center for Analytical Chemistry, Christian Doppler Laboratory for Rapid Test Systems for Allergenic Food Contaminants, University of Natural Resources and Life Sciences, Konrad Lorenz Str. 20, 3430, Tulln, Austria

    Parisa Ansari, Norbert Stoppacher, Judith Rudolf, Rainer Schuhmacher & Sabine Baumgartner

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  1. Parisa Ansari
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  2. Norbert Stoppacher
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  3. Judith Rudolf
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  5. Sabine Baumgartner
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Correspondence to Sabine Baumgartner.

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Ansari, P., Stoppacher, N., Rudolf, J. et al. Selection of possible marker peptides for the detection of major ruminant milk proteins in food by liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 399, 1105–1115 (2011). https://doi.org/10.1007/s00216-010-4422-0

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  • DOI: https://doi.org/10.1007/s00216-010-4422-0

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