Allergen Targets

Milk

Peanut

Soy

Tree nuts

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Food Matrix

Chocolate

A rapid solid-phase extraction combined with liquid chromatography-tandem mass spectrometry for simultaneous screening of multiple allergens in chocolates

Gu, S., Chen, N., Zhou, Y., Zhao, C., Zhan, L., Qu, L., Cao, C., Han, L., Deng, X., Ding, T., Song, C., Ding, Y.

Food Control (2018), 84, 89--96 DOI: 10.1016/j.foodcont.2017.07.033

Abstract

Allergen derived from food is a world-wide problem to specific consumers. The regulation for accurate control of risk demands a fast and sensitive method for comprehensive screening and confirmation for potential allergens in foods. In this work, an optimized and sensitive LC-MS/MS method for multiplex detection of eight allergenic ingredients in chocolate matrix is proposed. Improved throughput and enhanced sensitivity were obtained by introduction of a rapid solid-phase extraction step using MonoSpin PBA spin column. The most intense and reliable peptide markers were first identified by untargeted survey experiment using quadrupole-orbitrap high resolution mass spectrometer, and subsequently employed to design a multi-target MRM method, based on the most intense transitions recorded for each selected precursor peptide. The MRM quantitative analysis was performed on triple quadrupole mass spectrometer with optimized sensitivity and reproducibility. Protein sequence coverage of 70.8–93.3% were obtained for confirmation of each allergenic ingredient and two or three most sensitive peptide markers were selected in order to retrieve quantitative information. Limits of quantification shown as 0.2–0.4 μg/g for milk, 1.0–4 μg/g for soy, 2.5–4 μg/g for peanut, 1–3 μg/g for tree nuts respectively as well as recoveries ranged from 60.1(±2.7)% to 92.4(±5.6)% for chocolate samples proved the excellent capabilities of the exploited sample treatment method combined with the LC-MS/MS analysis. Finally, this method was successfully applied to the investigation of multiple allergens in commercially available chocolates of different brands aiming to ascertain possible discrepancies between allergen content and food allergen labelling.

Reference raw data

Reference ID	Food	Protein	Peptide
80	Almond	Pru du 6	ADFYNPQGGR
80	Hazelnut	Cor a 9	ADIYTEQVGR
80	Cashew	Ana o 2	ADIYTPEVGR
80	Milk	Bos d 10	ALNEINQFYQK
80	Almond	Pru du 6	ALPDEVLANAYQISR
80	Cashew	Ana o 2	AMTSPLAGR
80	Milk	Bos d 11	AVPYPQR
80	Walnut	Jug r 2	DFLAGQNNIINQLER
80	Soy	Gly m 5	DSYNLHPGDAQR
80	Soy	Gly m 5	DSYNLQSGDALR
80	Pistachio	Pis v 2	EGQLVVVPQNFAVVK
80	Milk	Bos d 10	FALPQYLK
80	Walnut	Jug r 2	FFDQQEQR
80	Milk	Bos d 9	FFVAPFPEVFGK
80	Peanut	Ara h 3	FNLAGNHEQEFLR
80	Almond	Pru du 6	GNLDFVQPPR
80	Peanut	Ara h 1	GTGNLELVAVR
80	Milk	Bos d 9	HQGLPQEVLNENLLR
80	Cashew	Ana o 2	LDALEPDNR
80	Almond	Pru du 6	LLSATSPPR
80	Soy	Gly m 6	LSAEFGSLR
80	Milk	Bos d 10	NAVPITPTLNR
80	Peanut	Ara h 1	NNPFYFPSR
80	Pistachio	Pis v 2	NSFDISR
80	Almond	Pru du 6	QQEQLQQER
80	Soy	Gly m 5	QQQEEQPLEVR
80	Peanut	Ara h 3	RPFYSNAPQEIFIQQGR
80	Peanut	Ara h 1	SFNLDEGHALR
80	Milk	Bos d 12	SPAQILQWQVLSNTVPAK
80	Peanut	Ara h 3	SPDIYNPQAGSLK
80	Soy	Gly m 6	SQSDNFEYVSFK
80	Soy	Gly m 5	TISSEDKPFNLR
80	Walnut	Jug r 2	VFSNDILVAALNTPR
80	Soy	Gly m 6	VLIVPQNFVVAAR
80	Milk	Bos d 11	VLPVPQK
80	Hazelnut	Cor a 9	WLQLSAER
80	Milk	Bos d 12	YIPIQYVLSR
80	Milk	Bos d 9	YLGYLEQLLR