Authentication of shrimp muscle in complex foodstuff by in-solution digestion and high-resolution mass spectrometry

Abstract

A method for shrimp muscle identification in complex foods is required to safeguard the shrimp-allergic population. This study described a method for authentication of shrimp in complex foodstuffs (fish balls) by liquid chromatography tandem QTOF mass spectrometry (UPLC-QTOF-MS). The proteins in shrimp muscle were extracted using a Tris-HCl solution and then digested using tryptic protease. The main allergen proteins, tropomyosin (TM) and arginine kinase (AK), were characterized using the 'bottom up' MS approach. After analysis of their peptide mass fingerprinting based on the UniProt database, two specific heat-stable peptides, ALSNAEGEVAALNR for TM and VSSTLSSLEGELK for AK, were screened as surrogate (signature) peptides. The detection limit, expressed as shrimp meat per kilogram of food, was 8 g kg À1 (usage of TM) or 5 g kg À1 (usage of AK). The developed method is suitable to screen potential addition of shrimp meat in foodstuffs by detection of allergen proteins.

The use of in-strip digestion for fast proteomic analysis on tear fluid from dry eye patients

“…Mass spectrometry (MS) is a fundamental and versatile technique for analyte test in biological samples due to its speed, specificity, sensitivity and throughput [ 15 ]. The current modes for biomarker quantification based on MS are selected/multiple reaction monitoring (SRM or MRM) performed typically on a triple quadrupole mass spectrometer and parallel reaction monitoring (PRM) performed on a hybrid quadrupole-orbitrap [ 16 ] or a quadrupole time-of-flight [ 17 ] MS. For example, Tong et al [ 18 ] quantified of 47 human tear proteins using high resolution multiple reaction monitoring (HR-MRM) of Triple TOF-MS. You et al [ 19 ] have determined human tear lactoferrin using MRM technique with stable-isotopic labeling. However, it should be noted that unlike clinical immunoassays, MS-based analyses of proteins/peptides from biological fluids usually involve complicated pretreatments, such as analyte extraction and tryptic digestion [ 20 ].…”

Analysis of amino acids in human tears by hydrophilic interaction liquid chromatography and quadrupole orbitrap mass spectrometry

“…Moreover, Q-Orbitrap-MS provides product-ion spectra with accurate mass measurement that permit unequivocal conrmation of compounds of interest. 24,25 Accordingly, it might be a good choice for the AAs analysis in tears with Q-Orbitrap. To achieve the highest selectivity and sensitivity, mass spectrometry parameters including ionization mode, capillary voltage, source temperature, sheath gas ow, and collision energy were optimized using AAs standards.…”

Development of a sandwich ELISA and immunochromatographic strip for the detection of shrimp tropomyosin

“…However, there are disadvantages, such as the complicated operation process, time-consuming analyses, and high false-positive rate, to these methods. High performance liquid chromatography coupled with tandem mass spectrometry has also been developed to accurately quantify food allergens (Chen, Pan, & Huang, 2017;Nagai, Minatani, & Goto, 2015;Rahman, Lopata, Randell, & Helleur, 2010). However, it is difficult to perform on-site detection, or use as a high-throughput screen because of the complicated sample pre-treatment procedures, expensive equipment, and trained personnel needed.…”