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This project is part of a larger task pursuing the development of naturally produced, health-contributing, polyphenol-enriched milk
products that are derived from byproduct plant matter from agricultural sources. Apple pomace (AP), an abundant agricultural
byproduct in Canada has high polyophenol content (1000-1415 g Gallic acid equivalent/100 g dry weight). The objective of this
study is to evaluate the effects of in vitro rumen digestion on the total polyphenol content and free radical scavenging activity of AP.
AP will be collected fresh and kept at -20 �°C until analysis. Rumen inoculum will be collected from a mature lactating cow and in
vitro digestion will be performed for 24 and 48 hours of digestion in an ANKOM RF Gas Production System as per manufacturer�s
instructions. After digestion, the samples will be centrifuged at 26.940 g and filtered through 0.2 um polyethersulfone filters to
remove bacterial cells. All samples (treated and untreated) will be freeze-dried and ground, after which polyphenols will be extracted
by solvent extraction (80% methanol at 21 �°C for 1 hour). Fast Blue BB assay for total polyphenol quantification and 2,2-diphenyl-1-
picrylhydrazyl (DPPH) free radical scavenging activity assay will be performed on all samples in triplicate with 3 technical replicates
to assess the extent of polyphenol degradation in the rumen. The fraction of polyphenols remaining after in vitro digestion will be
roughly representative of the amount available for absorption into the cow�s bloodstream and subsequently the mammary gland.