A Study of Avenanthramides in Oats for Future Applications
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UPTEC X 10 023 Examensarbete 30 hp November 2010 A study of avenanthramides in oats for future applications Eléne Karlberg Molecular Biotechnology Programme Uppsala University School of Engineering UPTEC X 10 023 Date of issue 2010-06 Author Eléne Karlberg Title (English) A study of avenanthramides in oats for future applications Title (Swedish) Abstract Avenanthramides are polyphenols existing exclusively in oats. Studies have shown their antioxidant, anti-inflammatory and anti-proliferatory properties, making avenanthramides interesting therapeutic candidates for treatment of diseases with cardiovascular and dermatological origin. This study has shown a method for enrichment of avenanthramides involving steeping and germination at low pH levels, resulting in comparable amounts to the amounts found physiologically significant in previous studies in mice. Since germination at low pH levels potentially means preservation of other health beneficial compounds such as β- glucan, an application based on this method would not only contain the therapeutic effects from avenanthramides, but also the positive effects originating from β-glucan. Keywords Oats, avenanthramides, antioxidants, anti-inflammatory, anti-proliferatory, cardiovascular disease, dermatological disease, germination, β-glucan. Supervisors Ingmar Börjesson Lantmännen Food R&D Scientific reviewer Torgny Fornstedt Uppsala University Project name Sponsors Language Security English Secret until 2012-11 Classification ISSN 1401-2138 Supplementary bibliographical information Pages 48 Biology Education Centre Biomedical Center Husargatan 3 Uppsala Box 592 S-75124 Uppsala Tel +46 (0)18 4710000 Fax +46 (0)18 471 4687 A study of avenanthramides in oats for future applications Eléne Karlberg Populärvetenskaplig sammanfattning Havre är ett av de vanligaste spannmålen i Sverige och förekommer både som foder åt djur och som matprodukter för människor. Havre innehåller höga nivåer av antioxidanter, vitaminer, mineraler och kostfiber som har visats sig kunna minska skadligt kolesterol i blodet, hjälpa till med insulinreglering och även sänka risken att drabbas av hjärt‐ och kärlsjukdomar. Avenantramider är ämnen i havre som fungerar som antioxidanter och studier har visat att de även besitter förmågor att bland annat minska celldelning, inflammation och klåda. Dessa egenskaper gör avenantramider till intressanta kandidater för behandling av en mängd olika sjukdomar. Det är sedan tidigare känt att halterna avenantramider ökar under en groningsprocess, det vill säga där havrekärnor utsätts för behandlingar som får dem att börja gro. Denna studie visar att pH påverkar anrikningen av avenantramider under groningsprocessen och att de högsta avenantramidhalterna fås under groning vid låga pH:n. Dessa halter är jämförbara med halter som i tidigare studier haft fysiologisk effekt i möss, vilket antyder att denna metod för anrikning av avenantramider är fullgod för skapandet av extrakt med terapeutisk potential. Vidare så medför groning vid låga pH:n lägre enzymaktivitet, vilket potentiellt innebär att andra nyttiga ämnen som annars skulle brutits ned bevaras. Havreextrakt på material som behandlats enligt den föreslagna metoden in denna studie skulle kunna inkluderas i en mängd olika applikationer för behandling av bland annat hjärt‐ och kärlsjukdomar, hudåkommor, allergier eller ingå i produkter med nutritionellt värde. Examensarbete 30 hp Civilingenjörsprogrammet i Molekylär Bioteknik Uppsala Universitet 1 2 Table of contents 1. Introduction ..................................................................................................................... 5 1.1 The oat grain ..................................................................................................................... 5 1.2 Avenanthramides .............................................................................................................. 6 1.3 Aim of project ................................................................................................................... 8 2. Materials and methods .................................................................................................... 9 2.1 Empirics ............................................................................................................................. 9 2.2 Oat materials .................................................................................................................... 9 2.3 Chemicals .......................................................................................................................... 9 2.4 Raw seeds ......................................................................................................................... 9 2.5 Freezing ............................................................................................................................. 9 2.6 Steeping .......................................................................................................................... 10 2.7 Germination .................................................................................................................... 10 2.8 Extraction of avenanthramides ...................................................................................... 10 2.9 High performance liquid chromatography (HPLC) analysis ............................................ 11 2.10 Antioxidant activity assay, DPPH‐radical scavenging system ....................................... 11 2.11 Statistical analysis ......................................................................................................... 11 3. Empirics ......................................................................................................................... 12 3.1 Antioxidant and antigenotoxic effects of avenanthramides .......................................... 12 3.2 Avenanthramides and atherosclerosis ........................................................................... 13 3.3 Avenanthramides and colloidal extracts in dermatology ............................................... 19 3.4 Bioavailability of avenanthramides ................................................................................ 22 3.5 Tranilast .......................................................................................................................... 23 4. Results from experiments .............................................................................................. 25 4.1 Germination frequency................................................................................................... 25 4.2 Influence on drying on avenanthramide amounts ......................................................... 25 4.3 Influence of pH and germination on avenanthramide amounts .................................... 26 4.4 Antioxidant activity of avenanthramides ....................................................................... 31 4.5 Statistical analysis ........................................................................................................... 33 5. Discussion ...................................................................................................................... 35 5.1 Germination frequency................................................................................................... 35 3 5.2 Influence on drying on avenanthramide amounts ......................................................... 35 5.3 Influence of pH and germination on avenanthramide amounts .................................... 36 5.4 Antioxidant activity of avenanthramides ....................................................................... 37 5.5 Physiological aspects ...................................................................................................... 38 5.6 Potential applications ..................................................................................................... 38 6. Conclusions .................................................................................................................... 40 7. Acknowledgements ........................................................................................................ 41 8. References ..................................................................................................................... 42 Appendix ........................................................................................................................... 46 4 1. Introduction Oats have been cultivated for more than a thousand years and are used today not only as feeding crop for animals, but also in a variety of different food products for human consumption. The main producers of oats are Finland, Germany and Sweden, with oats landing a third place as the largest crop produced in Sweden. There are many health benefits from consumption of oat based products. The high nutritional content of oats, with high levels of lipids, proteins, dietary fibers such as β‐glucan, minerals such as calcium and iron, vitamins like Vitamin E and a wide variety of antioxidants, contribute to reduce plasma cholesterol, improve glucose and insulin regulation and lower the risks for coronary heart disease. The amounts of proteins present in oats are not only higher but also of a higher quality compared to other cereals, containing the essential amino acids lysine, threonine and methionine (Bryngelsson 2002, Meydani 2009 and Skoglund 2008b). Furthermore, oats are considered to be safe for adults suffering from celiac disease, since only trace amounts of gluten protein