Towards the establishment of a comprehensive food inspection system: an approach to utilise de novo peptide sequencing from MS/MS spectra

preprint · SSRN Electronic Journal · 2026

preprint · SSRN Electronic Journal · 2026. Hiromasa Mitsui et al. Ensuring food safety and preventing food fraud require analytical methods capable of identifying unknown and…
Date 2026-02-13
Type preprint
Venue SSRN Electronic Journal
Publisher Elsevier BV (SSRN)
Contribution downstream-application
DOI 10.2139/ssrn.6234726

Abstract

Ensuring food safety and preventing food fraud require analytical methods capable of identifying unknown and unexpected contaminants in foods. This study developed a proteomics-based strategy centered on de novo sequencing, in which peptide sequences are directly determined from MS/MS spectra without reliance on protein databases. To obtain high-confidence peptide sequences, mass spectrometry conditions were optimized using tryptic digests of K562 cells. Among the tested platforms, ZenoTOF 7600 operated with a Zeno trap accumulation time of 25 ms and 80 MS/MS events yielded the highest number of peptide sequences, averaging 1,057 peptides with an Average Local Confidence score of ≥99. The resulting peptide sequences were used for species identification and quantitative analysis, with database searching applied as a complementary step. Application of this strategy to oyster extract-supplemented soy sauce enabled identification of species associated with added ingredients and the manufacturing process, demonstrating its potential for detecting unknown adulteration and unexpected contamination.

Authors

  1. Hiromasa Mitsui · Kazusa DNA Research Institute
  2. Ryo Konno · Kazusa DNA Research Institute
  3. Takeshi Shibata · SCIEX
  4. Ushio Takeda · SCIEX
  5. Yusei Okuda · Kazusa DNA Research Institute, Kitasato University
  6. Daisuke Nakajima · Kazusa DNA Research Institute
  7. Osamu Ohara · Kazusa DNA Research Institute
  8. Yusuke Kawashima · Kazusa DNA Research Institute

Methods and tools

  • Food inspection de novo peptide screening: Database-free food inspection workflow: de novo sequencing of MS/MS spectra on a ZenoTOF 7600 for species identification and quantitation in food extracts, with database search as a complementary step, aimed at unknown contaminants and food fraud.
  • PEAKS: Commercial DP-based de novo

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