Barley is the third largest agricultural crop grown in Australia averaging over 9 million tonnes per year. Barley is germinated under controlled conditions in a process known as malting, to allow endogenous enzymes to begin the process of cellular degradation for use in the brewing industry. 30-40% of Australia’s national barley crop is malting grade, generating a price premium to feed grade barley and driving the constant development of new malting barley varieties. The recognition of new malting barley varieties is a long process based on pilot and commercial scale malting taking over at least two growing seasons. Once harvested, barley crops are graded as either malting or feed grade based on total protein content, an important marker for barley breeders, growers, maltsters and brewers. A limited number of protein characteristics are regularly assessed, such as diastatic power, but limited regard is given to the overall composition of the barley proteome. Challenges in the proteomic assessment of barley are due to a large number of storage proteins, known as hordeins, representing around 40% of total barley protein content. Hordeins are largely insoluble and contain large regions of glutamine rich repeats which can reduce their detection using standard SWATH-MS workflows. The use of alternative proteolytic enzymes can be utilised to improve proteomic coverage of hordeins but at the expense the total proteomic coverage. Methods have been developed to combine DIA/SWATH-MS data across samples digested with multiple enzymes to improve the proteomic coverage of barley and malted barley. The application of miniaturised methods previously developed for proteomic assessment of different stages of the brewing process1 have also been combined along with newly developed single seed methodologies for malting and brewing. The increased proteomic coverage of the barley and malted barley proteome has been applied to multiple malting barley varieties across malting and brewing with a goal to be implemented to improve the speed of malting accreditation of new barley varieties.