Novel liquid chromatography–mass spectrometry method for sensitive determination of the mustard allergen Sin a 1 in food

Abstract

Mustard is a condiment added to a variety of foodstuffs and a frequent cause of food allergy. A new strategy for the detection of mustard allergen in food products is presented. The methodology is based on the analysis by liquid chromatography coupled to mass spectrometry. Mustard allergen Sin a 1 was purified from yellow mustard seeds. Sin a 1 was detected with a total of five peptides showing a linear response (lowest LOD was 5 ng). Sin a 1 was detected in mustard sauces and salty biscuit (19±3mg/Kg) where mustard content is not specified. Sin a 1, as an internal standard, allowed quantification of this mustard allergen in foods. A novel LC/MS/MS SRM-based method has been developed to detect and quantify the presence of mustard, a principal allergen in foods. This method could help to detect mustard allergen Sin a 1 in processed foods and protect mustard allergic consumers.Dear editor, I send you the revised version of our manuscript "Novel liquid chromatography-mass spectrometry method for the sensitive determination of mustard allergen Sin a 1 in food."We have corrected the text according to the reviewers recommendations and added additional data requested by the reviewers. Below you can find the response, point by point, to the issues indicated by the reviewers. All the modifications are included in the revised version. We expect that the revised version is now suitable for publication in Food Chemistry Journal. Looking forward to hearing from you, Carlos Pastor-VargasCover Letter Reviewer #1The manuscript "Novel high-throughput methodology for the sensitive and quantification of mustard allergens in foods" presents an interesting study because it covers an interesting issue within food safety "allergens" and develop an analytical method to determine these allergens that uses currently used techniques. The study is very innovative Importantly, the study offers solutions for the problem of those food that do not declare in the label the addition of mustard. Because all these reasons, the manuscript is clearly worthy of publication in Food Chemistry. From the experimental point of view, the design of the experiments has been properly performed in order to ensure the reliability of the results as well as to avoid possible interferences and to eliminate false negatives. From the formal point of view, however, the manuscript raises some concerns: (i) the excessive use of the first plural person (try to be more impersonal)The sentences with the use of the first plural person have been changed as suggested by the reviewer (ii) the inappropriate use of some chromatographic terminology (see recommendations below).Following your suggestions, chromatographic terminology has been changed (iii) lack of discussion in some parts.New paragraphs in discussion section have been added as suggested by the reviewer Then, the manuscript should be revised. Some comments and questions that need to be addressed are:1.-The title is little descriptive and does not give information on the analytical techniques used. A more a...

Food Allergy

“…Target specificity, selectivity, sensitivity, quantification, and environmental factors that influence the target molecular properties are the major challenges in the selection of the appropriate LC detector. The LC-MS method has therefore received more favorable consideration (van Hengel 2007, Kirsch et al 2009, Monaci & Visconti 2009, Faeste et al 2011, Cucu et al 2013, Posada-Ayala et al 2015.…”

Development of a real-time quantitative PCR assay using a TaqMan minor groove binder probe for the detection of α-lactalbumin in food

“…Enzyme-linked immunosorbent assay and PCR are most commonly used methods for the detection of food allergens (Stephan and Vieths, 2004;. Recently, more novel methods were developed for food allergens detections such as chromatography, mass spectrometry, and magnetoimmunosensing (Posada-Ayala et al, 2015;Ruiz-Valdepeñas Montiel et al, 2015). An ELISA is an immunoassay method that is frequently used to detect allergens in food (Koch et al, 2003;López-Calleja et al, 2013).…”

Food Allergen Analysis: Detection, Quantification and Validation by Mass Spectrometry

“…Table 4a. Quantification of food allergens in different food products by mass spectrometry using label-free quantification with an (1) external calibration curve [75][76][77][78][79]88], (2) unlabelled modified synthetic peptide [78], and (3) standard addition [79]. Table 4b.…”

“…It was used by Posada-Ayala et al for the quantification of commercial food products [79]. This approach consists in adding different known quantities of extracted allergen proteins directly to the sample to be analysed before digestion and in quantifying the target allergens with the resulting calibration curve.…”