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Wheat gluten imparts “unique” functional characteristics to dough which allow it to make bread. While High Molecular Weight Glutenin Subunits (HMW-GS) are partially responsible for gluten strength Canadian wheat varieties in the premium CWRS class have the same HMW-GS composition yet there is a wide range in their gluten strength. Low molecular weight subunits (LMW-GS) play a key role in gluten polymer size and strength yet limited investigation of Canadian wheat has been done. Four varieties of CWRS, Glenn, Viewfield, Carberry and Unity, representing a wide range in gluten strength were grown at six sites across the western prairies. LMW-GS were prepared via sequential extraction, precipitation, alkylated with 4-vinyl pyridine and analyzed of an ESI-ToF mass spectrometer to characterize differences in these key proteins relative to gluten strength due to different growing environments. Molecular weight accuracy was 6 Da or an RSD of 0.02% with a precision RSD of 0.0013%. The strongest varieties; Glenn, Viewfield and Carberry displayed similar LMW-GS protein patterns above 42,000 Da while the weakest, Unity, exhibited no proteins in this range. The two strongest varieties, Glenn and Viewfield shared a common, most abundant, protein of 42,927 Da however Carberry’s most abundant protein exhibited a mass of 33,553 Da. Unity, the weakest, had its most abundant protein at 40,343 Da. Only 3 LMWs were present in all varieties at all sites: 40,587, 33,278, and 32,444 Da. The 3 strongest varieties only had 8 proteins in common above 30,000 Da, with 6 of these above 40,000 Da, suggesting they are serine-B-type LMWs. All varieties exhibited the presence/absence of different proteins depending upon their growing environment. Only 3 proteins, found in all 4 varieties, displayed absence/presence of the protein due to growing environment; 40,424, 33,296 and 32,382 Da.