1. Lyophilized Beer: A Banquet of Bioactive Compounds for Human Health Jairam Vanamala, Ph.D. Assistant Professor Dept. of Food Science and Human Nutrition Colorado State University

2. Ancient Wisdom – Beer & Health Mouthwash Treatment of wounds Enema Sanitation http://philly.thedrinknation.com/articles/read/5586-Ancient-Egyptians-First-Medicinal-Use-of-Beer-#

3. What is in the Beer Ethanol Bioactive compounds –Malt – derived –Hop - derived

4. Beer - Protection Against Mortality 10.14 oz. or 0.3 L/day - reduces mortality for all causes of death Moderate beer consumption - Beneficial European Journal of Cancer 41(13), 2005

5. Beer – Bioactive Compounds Malt-derived compounds –Benzoic acid derivatives –Cinnamic acid derivatives –Catechins –Proanthocyanidins –Flavones

6. Biological Activities Anti-oxidant Anti-microbial Anti-viral Anti-proliferative Anti-platelet Anti-hyperlipidemic Anti-hypersensitive Anti-allergenic Anti-ulcerogenic Anti-neoplastic Anti-inflammatory Anti-atherogenic Anti-hepatotoxic Pro-apoptotic Vasodilator

7. Beer – Bioactive Compounds Hop-derived compounds –Alpha acids –Essential oils –Prenylflavonoids Xanthohumol Isoxanthohumol 8-prenylnarigenin

8. (I) Xanthohumol and (II) Isoxanthohumol Xanthohumol and Isoxanthohumol

9. Xanthohumol Anti- inflammatory activity Anticancer activity Antioxidant activity Anti-obesity activity Anti- microbial activity Estrogenic activity

10. Is There a Synergy Xanthohumol + Hop- and malt-derived bioactive compounds

11. How We Test it Colon cancer cells: HCT - 116 –Proliferation –Apoptosis

12. Why Colon Cancer  World: 1,000,000  US: 1,50,000  80% Advanced  2 yr survival  80% Diet Related

13. Cell proliferation

14. Apoptosis * * * *

15. Xanthohumol & Isoxanthohumol – UPLC/MS/MS 246810121416 Isoxanthohumol Xanthohumol

16. American IPAs Ranger Prefiltered (RGP) T-90 pellets and CO2 extract Ranger Filtered (RGF) DE filtration IPA Prefiltered (OOP) Dry hopped: 0.055 oz. for 0.3 L of beer IPA Filtered (OOF) DE filtration Dark IPA Prefiltered (ODP) 8.7% darker malts compared with OOP/OOF Dark IPA Filtered (ODF) DE filtration Zenith Original Recipe (ZOR) 2 oz. pelleted Centennial hops – at the beginning Zenith New Recipe (ZNR) Dry hopped: 0.016 oz. per 0.3 L of beer Punjabi Pale Ale (PPA) Dry hopped: 0.014 oz. per 0.3 L of beer German Beers Paulaner Pils (PPI) Pilsner malts and German hop varieties

17. Xanthohumol Content - IPAs c d d d d b a a

18. Critical Steps to Increase Beneficial Compounds in Beer XAN Technology –Process –Ingredients XN-Enriched Hop Products Roasted Malts Xan Technology Summary Mol Nutr Food Res 49(9): 874-881 Wort Boiling Boiling and cooling time Addition of hops, roasted malts and XN-enriched hop products Fermentation Appropriate yeast Maturation Addition of hops Filtration DE filtration

19. Addition of Roasted Malts - Xanthohumol 8.7% Darker Malts 9.94 25 % b a

20. b a Xanthohumol - Dry Hopping 66 %

21. aa b c d e Xanthohumol - DE filtration 34.3% 11.1%.5% 8.7% Darker Malts

22. Correlation Between Apoptosis & XN R 2 =0.97 P = 0.0001

23. Future Opportunities Lyophilized Beer –Breads –Soups –Ice creams –Protein Bars –Milk Shakes

24. Acknowledgements Twila Henley Lavanya Reddivari Jack Avens Marissa Bunning Martha Stone Jeff Miller Breweries –Odell –New Belgium –Equinox –CopperSmith

25. Thank You

27. Dry hopping: improve sensory characteristics and bioactive content. –Lyophilized beers Xan Wellness & Xan Tea Future development of XN-enriched products could utilize the dry hopping technique. Future Direction

28. a a a ab abc abcd bcd cd d IPAs - Antioxidant Capacity

29. IPAs - Phenolic Compounds Compared to Pilsners IPAs contain: more hops > variation in malts Roasted malts could lessen removal of phenolic compounds a ab bc c cd d d e