Comprehensive workflow encompassing discovery, verification, and quantification of indicator peptide in snail mucin using LC-quadrupole Orbitrap high-resolution tandem mass spectrometry

• Published in: Food research international Vol. 180; p. 114054
• Main Authors: Moon, Sung-Kwon, Jeong, Eun-Jin, Tonog, Genevieve, Jin, Cheng-Min, Lee, Jeong-seok, Kim, Hoon
• Format: Journal Article
• Language: English
• Published: Canada Elsevier Ltd 01.03.2024
• Subjects: Achatina fulica; Data-dependent acquisition; Method development; Mucins; Parallel reaction monitoring; Peptide quantification; Peptides - chemistry; Peptidomics; Tandem Mass Spectrometry - methods; Workflow; DDA; LOD; Parallel reaction monitoring; MRM; PRM; Peptidomics; Peptide quantification; LOQ; SRM; Data-dependent acquisition; CV; Method development; BPC; FWHM; ESI; SM; UHPLC; AGC; EIC; FS-ddMS/MS; Achatina fulica
• ISSN: 0963-9969; 1873-7145
• DOI: 10.1016/j.foodres.2024.114054

[Display omitted] •Creation of peptide profile in snail-derived mucin via DDA untargeted method.•Identification and quantification of indicator peptide (TEAPLNPK) in snail mucin.•Development of PRM targeted method for verifying and quantifying peptide.•Proposal of a comprehensive workflow for indicator peptide derived from snail mucin. Peptidomics analysis was conducted using high-resolution tandem mass spectrometry (MS2) to determine the peptide profile of snail-derived mucin extract (SM). The study was also aimed to identify an indicator peptide and validate a quantification method for this peptide. The peptide profiling and identification were conducted using discovery-based peptidomics analysis employing data-dependent acquisition, whereas the selected peptides were verified and quantified using parallel reaction monitoring acquisition. Among the 16 identified peptides, the selected octapeptide (TEAPLNPK) was quantified via precursor ion ionization (m/z 435.2400), followed by quantification of the corresponding quantifier ion fragment (m/z 639.3824) using MS2. The quantification method was optimized and validated in terms of specificity, linearity, accuracy, precision, and limit of detection/quantification. The validated method accurately quantified the TEAPLNPK content in the SM as 7.5 ± 0.2 μg/g. Our study not only identifies an indicator peptide from SM but also introduces a novel validation method, involving precursor ion ionization and quantification of specific fragments. Our findings may serve as a comprehensive workflow for the monitoring, selection, and quantification of indicator peptides from diverse food resources.