Armen Mirzoian, TTB International Wine Technical Forum May 6, 2015 Allergen Testing Armen Mirzoian, TTB International Wine Technical Forum May 6, 2015 Prepared for the International Wine Technical Forum and presented for purposes of discussion. This is not an authoritative statement of agency policy or guidance.

Allergen Testing in Wine Advantages of ELISA Sensitive (ppm range) Quantitative or semi-quantitative Measure amount of offending food component (i.e. proteins) Fairly rapid Equipment needs are minor (plate reader) Advantages of MS Absolute identification and quantification of allergens Highly sensitive Excellent confirmatory method MS ELISA Disadvantages of MS Requires high level of expertise High cost of equipment Time consuming Laborious Extraction and cleanup needed Not useful for routine analyses Disadvantages of ELISA Cross-reactivity Values obtained from kits do not agree Lack of reference materials Need to do “in house” validation of ELISA

Allergen Testing in Wine Allergen disclosure on wine labels is currently voluntary in the U.S. TTB’s Scientific Services Division is planning to study feasibility of ELISA techniques for screening allergens in wine: Egg (Ovalbumin) Milk (Casein) Lysozyme

Allergen Testing in Wine TTB is collaborating with E&J Gallo and FARRP on developing a Liquid Chromatography-Mass Spectrometry method (LC-MSMS) for egg and milk screening in wine

Gluten in Distilled Spirits Gluten fragmented/hydrolyzed during fermentation Sandwich ELISA is inadequate for measuring hydrolyzed gluten We are evaluating total protein colorimetric assays (Lowry, BCA, Bradford) for detecting trace level protein in DS.

Questions? Thank you!