AOAC ERP Gluten Assays - Dec 2018

OMAMAN‐47: Quantification of Wheat, Rye, and Barley Gluten in Oat and Oats Products SUMMARY OF COMMENTS  quantitative analysis of wheat, rye, and barley gluten in oat flour, groats, oat flakes, and oat  cereals. The sandwich ELISA method detects intact gliadins and related prolamins from rye and barley, high‐molecular weight (HMW)  gluteninsubunits. (GS) from wheat, HMW‐secalins from rye and low‐molecular‐weight  (LMW)–GS from wheat. It does not detect D‐hordeins from barley. The method scope has been precisely defined and the materials used for the collaborative  study fall within the scope of the method. The number of laboratories submitting valid  datasets exceeds the number set for collaborative studies under AOAC/IUPAC harmonized  protocol.  Pros/Strengths of the Manuscript: There has been a significant amount of work done in this study and I think the methodology is  strong.  The manuscript is well organized and readable to target audients.  Data are clearly tabulated,  or presented in figures for effective visual.  Discussions and conclusions are clear and precise.   This is not surprising, giving the authors have long history of successful credentials in allergen  and gluten analysis expertise and publication records. This is a very well‐written and organized manuscript. The collaborative study was conducted  appropriately with excellent participation and performance from the outside laboratories. The  sample preparation methods were easy to understand, and the study data is shown in a clear  and concise manner. The SLV was well thought out and provides extensive information about the performance of  the kit. The kit instructions are excellent, and provide good reference material for the end user. Given  the make‐up of the standard materials, it is good that the kit is recommended only for intact  gluten (though I may suggest using un‐fractionated instead of non‐fragmented). The data generated and yet to be generated through a collaborative study will help to better  define the recovery, repeatability and reproducibility of a method for the quantitative analysis  of gluten in oats for the first time. The method uses four monoclonal antibodies including R5 and can detect all gluten fractions  from wheat, rye and barley, except D‐hordein. The calculation of the method has been simplified to estimate gluten from the standard curve  and the use of conversion factor of 2 is removed. The method estimates gluten, rather than  gliadin.

ER 7

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Good data, more than enough labs covering the globe. 

Good and comprehensive validation study.

The manuscript is well written, the procedures clearly described: The data provided in the  manuscript allow the evaluation of the method against the SMPR 2017.021 Cons/Weaknesses of the Manuscript:

ER 8

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