Gluten determination by gliadin enzyme-linked immunosorbent assay kit: interlaboratory study.
An interlaboratory study with 10 participants was performed to obtain validation and performance data for an enzyme-linked immunosorbent assay (ELISA) kit developed for quantitative gluten determination in foods. The ELISA kit used for this study is based on 2 monoclonal and 1 polyclonal antibody developed by Immunotech, a Beckman Coulter Co. This kit did not show any false positive results or cross-reactivity with oat, rice, maize, and buckwheat. The gliadin standard from the Working Group on Prolamin Analysis and Toxicity was included in the kit as reference material for calibration. All participants obtained a gliadin ELISA kit with Standard Operational Procedure and a form for recording test results. The study included 13 samples labeled as "gluten-free" and 2 samples spiked by wheat flour. Seven samples had gliadin content below the limit of quantitation (LOQ) of the method, and 1 sample exceeded the highest calibration level. Gliadin content in the range from 10 to 157 mg/kg (1st day) and from 11 to 183 mg/kg (2nd day) was found in 7 samples (including 2 spiked samples). Results of these samples were used for further statistical analysis and evaluation. The Cochran, Dixon, and Mandel statistical tests were applied for detection of outliers. The LOQ of the kit was estimated.
Gabrovská
Dana
D
Food Research Institute Prague, Radiová 7, 102 31 Praha 10, Czech Republic. d.gabrovska@vupp.cz
Rysová
Jana
J
Filová
Vanda
V
Plicka
Jan
J
Cuhra
Petr
P
Kubík
Martin
M
Barsová
Sona
S
D018554:Avena / Q000378:metabolism; D002138:Calibration; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002523:Edible Grain / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D019612:Fagopyrum / Q000378:metabolism; D005504:Food Analysis / Q000379:methods*; D005903:Gliadin / Q000737:chemistry*; D005983:Glutens / Q000032:analysis*; D012275:Oryza / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism; D055315:Prolamins; D015203:Reproducibility of Results; D003313:Zea mays / Q000378:metabolism
D010940:Plant Proteins; D055315:Prolamins; D005983:Glutens; D005903:Gliadin