title: Development of an enzyme-linked immunosorbent assay method to detect mustard protein in mustard seed oil.
abstract: An enzyme-linked immunosorbent assay for the detection of mustard protein was developed. The assay is based on a polyclonal antiserum directed against a mixture of mustard proteins raised in rabbits. The assay has a detection limit of 1.5 ppm (milligrams per kilogram) and is suitable for the detection of traces of mustard protein in mustard seed-derived flavoring ingredients. Limited cross-reactivity testing showed that no other plant proteins reacted significantly. From the animal proteins tested, only milk showed some cross-reactivity. With this sensitive assay, it was shown that refined mustard seed oil produced by steam distillation does not contain detectable amounts of mustard protein. Mustard seed oil is used as a flavoring in very low quantities, typically between 40 and 200 mg/kg. Thus, 100 g of a food product flavored with 200 mg of mustard seed oil per kg containing < 1.5 mg of protein per kg would represent an amount of mustard seed protein of <30 ng. Taking into account the published literature on allergic reactions to the unintended ingestion of mustard, this conservatively low calculated level indicates that it is unlikely that food products containing mustard seed oil as a flavoring ingredient will elicit an allergic reaction in mustard-allergic individuals.
authors.affiliation: TNO Nutrition and Food Research, Zeist, The Netherlands/Kerklaan 53, NL-3731 EG De Bilt, The Netherlands. stefkoppelman@zonnet.nl
authors.forename: Stef J
authors.initials: SJ
authors.lastname: Koppelman
authors.forename: Riek
authors.initials: R
authors.lastname: Vlooswijk
authors.forename: Gina
authors.initials: G
authors.lastname: Bottger
authors.forename: Gert
authors.initials: G
authors.lastname: Van Duijn
authors.forename: Peter
authors.initials: P
authors.lastname: Van der Schaft
authors.forename: Jacco
authors.initials: J
authors.lastname: Dekker
authors.forename: Hans
authors.initials: H
authors.lastname: Van Bergen
mesh_terms: D000485:Allergens / Q000032:analysis*; D000818:Animals; D003257:Consumer Product Safety; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005504:Food Analysis; D005512:Food Hypersensitivity / Q000209:etiology / Q000517:prevention & control*; D006801:Humans; D009149:Mustard Plant / Q000276:immunology*; D010938:Plant Oils; D010940:Plant Proteins / Q000032:analysis / Q000276:immunology*; D011817:Rabbits
chemical_list: D000485:Allergens; D010938:Plant Oils; D010940:Plant Proteins; C027793:mustard oil