title: Improved Quantitation of Gluten in Wheat Starch for Celiac Disease Patients by Gel-Permeation High-Performance Liquid Chromatography with Fluorescence Detection (GP-HPLC-FLD).
abstract: Purified wheat starch (WSt) is commonly used in gluten-free products for celiac disease (CD) patients. It is mostly well-tolerated, but doubts about its safety for CD patients persist. One reason may be that most ELISA kits primarily recognize the alcohol-soluble gliadin fraction of gluten, but insufficiently target the alcohol-insoluble glutenin fraction. To address this problem, a new sensitive method based on the sequential extraction of gliadins, glutenins, and gluten from WSt followed by gel-permeation high-performance liquid chromatography with fluorescence detection (GP-HPLC-FLD) was developed. It revealed that considerable amounts of glutenins were present in most WSt. The gluten contents quantitated by GP-HPLC-FLD as sum of gliadins and glutenins were higher than those by R5 ELISA (gluten as gliadin content multiplied by a factor of 2) in 19 out of 26 WSt. Despite its limited selectivity, GP-HPLC-FLD may be applied as confirmatory method to ELISA to quantitate gluten in WSt.
authors.affiliation: Deutsche Forschungsanstalt für Lebensmittelchemie, Leibniz Institut , Lise-Meitner-Straße 34, 85354 Freising, Germany.
authors.forename: Katharina Anne
authors.initials: KA
authors.lastname: Scherf
authors.affiliation: Deutsche Forschungsanstalt für Lebensmittelchemie, Leibniz Institut , Lise-Meitner-Straße 34, 85354 Freising, Germany.
authors.forename: Herbert
authors.initials: H
authors.lastname: Wieser
authors.affiliation: Deutsche Forschungsanstalt für Lebensmittelchemie, Leibniz Institut , Lise-Meitner-Straße 34, 85354 Freising, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Koehler
mesh_terms: D002138:Calibration; D002446:Celiac Disease / Q000178:diet therapy; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D055050:Diet, Gluten-Free; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005453:Fluorescence; D005504:Food Analysis / Q000379:methods*; D005903:Gliadin / Q000032:analysis; D005983:Glutens / Q000032:analysis*; D006801:Humans; D057230:Limit of Detection; D013213:Starch / Q000032:analysis*; D014908:Triticum / Q000737:chemistry*
chemical_list: D005983:Glutens; D013213:Starch; D005903:Gliadin; C037302:glutenin
keywords: celiac disease; enzyme-linked immunosorbent assay (ELISA); gel-permeation HPLC fluorescence detection; gliadin; gluten analysis; glutenin; wheat starch