1. WP7 - RECENT DEVELOPMENTS IN ANALYSIS FOR STARCH, FIBER AND β -GLUCAN AVEQ Second meeting Quedlinburg, 2-4 March 2009

2. STATE OF ART Genotype & environment (year or location): The variability of the results from statistical analyses and the significance of the correlations among physical and nutritional grain traits might depend on both the genotypes and the experimental design used. Climatic conditions: it was reported that dry conditions before harvest result in higher β -glucan contents (Brennan & Cleary, 2005; Grebruers et al., 2008) AVEQ Second meeting, Quedlinburg, 2-4 March 2009

3. THE HEALTHGRAIN CEREAL DIVERSITY SCREEN: CONCEPT, RESULTS AND PROSPECTS J.L. WARD, K. POUTANEN, K. GEBRUERS, V. PIIRONEN, A-M. LAMPI, L. NYSTRÖM, A.A. ANDERSSON, P. AMAN, D. BOROS, M. RAKSZEGI, Z. BEDŐ, P. SHEWRY J. Agric. Food Chem., 2008, 56: 9699-9709 AVEQ Second meeting, Quedlinburg, 2-4 March 2009

4. WARD et al. - INTRODUCTION HEALTHGRAIN is an integrated project in the 6th Framework Programme of European Union, started in 2005 with the aim of improving the well-being and reducing the risk of metabolic diseases of European consumers by increasing the intake of protective grain factors through improved availability of health-promoting and safe ceral-based foods and ingredients of high eating quality. As part of this research program it was initially determined the extent of variation of bioactive compounds in cereals: bread and durum wheat, spelt, monococcum, dicoccum, barley, oat, rye. The target compounds were chosen on the basis of the hypotheses in the literature about their protective properties. AVEQ Second meeting, Quedlinburg, 2-4 March 2009

5. WARD et al. - 2008

6. WARD et al. - CONCLUSIONS The HEALTHGRAIN diversity screen has generated the most extensive database on bioactive components in wheat and other cereals currently available. Detailed comparison of the bread wheat lines show that it is possible to identify lines in which high levels of phytochemicals and dietary fiber components are combined with good yield and processing quality. This means that commercially competitive lines with high levels of bioactive components are a realistic goal for plant breeders. AVEQ Second meeting, Quedlinburg, 2-4 March 2009

7. PHYTOCHEMICAL AND FIBER COMPONENTS IN OAT VARIETIES IN THE HEALTHGRAIN DIVERSITY SCREEN P. SHEWRY, V. PIIRONEN, A-M. LAMPI, L. NYSTRÖM, L. LI, M. RAKSZEGI, A. FRÁS, D. BOROS, K. GEBRUERS, C.M. COURTIN, J.A. DELCOUR, A.A. ANDERSSON, L. DIMBERG, Z. BEDŐ, J.L. WARD J. Agric. Food Chem., 2008, 56: 9777-9784 AVEQ Second meeting, Quedlinburg, 2-4 March 2009

8. SHEWRY et al. - GRAIN COMPOUNDS PROXIMATES: moisture, crude protein, ash content, lipid, available starch, free sugars FIBER COMPONENTS: β-glucan (total and MW distribution), arabinoxylan (total and WE), estimated dietary fiber, Klason lignin PHYTOCHEMICALS: sterols, folates, phenolics, avenanthramides, tocols. AVEQ Second meeting, Quedlinburg, 2-4 March 2009

9. SHEWRY et al. - RESULTS Ash content: 2.1 – 2.6% Protein (wholemeal): 16.9 – 18.7% Lipid: 5.8 – 9.6% Available starch: 47.0 – 57.3% Dietary fiber (est): 18.5 – 23.4%; 10.6% (naked) Klason lignin: 4.2 – 5.9%; 2.6% (naked) β -glucan (wholemeal): 4.5 – 5.6% Total arabinoxylan (flour): 0.97 – 1.26% Folates: 495 – 604 ng/g Tocols: 16.1 – 36.1 μg/g Phenolics: 351 – 874 μg/g Avenanthramides: 42 – 91 μg/g AVEQ Second meeting, Quedlinburg, 2-4 March 2009

10. SHEWRY et al. - CONCLUSIONS Substantial variation was demonstrated in the contents of phytochemicals and more limited variation in the contents of fibre components. The fact that the varieties were grown in adjacent sites reduces the differences due to environment, and indicates that at least some of the differences were genetically determined. In conclusion, there is sufficient genetic variation in the contents of bioactive components to be exploited by breeders to develop varieties with enhanced quality for human nutrition. AVEQ Second meeting, Quedlinburg, 2-4 March 2009

11. NAKED OATS FOR IMPROVING HUMAN NUTRITION. GENETIC AND AGRONOMIC VARIABILITY OF GRAIN BIOACTIVE COMPONENTS Redaelli R., Sgrulletta D., Scalfati G., De Stefanis E., Cacciatori P., Crop Science, in press. Seven naked oat genotypes and two husked cultivars (testers) Two years: 2001-02 and 2002-03 Two locations: Rome and Bergamo Two preceding crops: cereal or legume Two seeding seasons: winter and spring Bioactive compounds: protein, β -glucan, TDF AVEQ Second meeting, Quedlinburg, 2-4 March 2009

12. REDAELLI et al. - RESULTS PROTEIN β -GLUCANTDF ABEL18.25 c4.26 a12.75 b BIKINI18.83 b3.78 c HJA72095N19.22 a3.73 c12.80 b KONRADIN17.52 d3.41 d15.23 a KYNON18.37 c3.14 e NAVE18.46 c3.93 b12.10 cd RHEA x PADARN 54218.48 c3.48 d11.56 d DONATA (H)16.43 e4.31 a12.42 bc FLAVIA (H)19.01 ab3.92 b12.62 bc MEAN18.293.7712.78

13. REDAELLI et al. - CONCLUSIONS Protein content was strongly influenced by environment, for β - glucan a significant effect was observed for rotation and genotype, and for their interaction with the other factors. TDF content was largely determined by genotype, but the interaction with the environment was always significant. A significant correlation (r = 0.56*) was found between β -glucan and yield. On a de-hulled basis, husked cultivars contained more β -glucan than the naked cultivars. AVEQ Second meeting, Quedlinburg, 2-4 March 2009

14. IN VITRO DETERMINATION OF THE INDIGESTIBLE FRACTION IN FOODS: AN ALTERNATIVE TO DIETARY FIBER ANALYSIS AVEQ Second meeting, Quedlinburg, 2-4 March 2009 Saura-Calixto F., Garcia-Alonso A., Goi I., Bravo L., J. Agric. Food Chem., 2000, 48: 3342-3347 AOAC method for DF determination aims to measure the content of the fraction composed by NSP (non starch polysaccharides) plus lignin. However, a significant part of the starch remains in foods (resistant starch, RS) and is not digested in the small intestine, along with other substances such as proteins, oligosaccharides, polyphenolic compounds, etc.

15. FOODS ENZYMATIC TREATMENTS CENTRIFUGATION RESISTANT STARCH RESISTANT PROTEIN CONDENSED TANNINS LIGNIN MINERALS INSOLUBLE NSP OTHER COMPOUNDS RESIDUE = INSOLUBLE IF SUPERNATANT - Oligosaccharides SOLUBLE NSP Neutral sugars + uronic acids DIALYSIS Solvent removal SOLUBLE IF

16. SAURA-CALIXTO et al. –CONCLUSIONS   The method for IF determination combines in a single procedure the methodologies for DF and resistant starch analysis. In this method, samples are analyzed as eaten (fresh, boiled or fried), and the analytical conditions are close to the physiological ones.   In DF analysis starch digestibility is altered artificially, whereas IF can remove most digestible starch, avoiding alterations of naturally occurring RS. IF values could be more useful than DF values from a nutritional point of view and could be a valid alternative to DF for food labeling and food composition tables.

17. WORKPACKAGES 7 & 8 – DECISIONS   Total B-glucan analysis will be carried out on the samples from the trials held in Estonia, Bulgaria and Poland in 2008. The samples from Estonia will be the first set to be ready; Matthias is going to de-hull samples from Bulgaria, whereas samples from Poland should be sent to Alf for de-hulling.   Alf will analyse all the samples by NIRS to make a screening for the amount of total B-glucan, and to check if the ranking of the genotypes is the same in all environments. Then 70 accessions will be chosen, that represent a large variability for B-glucan content. These 210 samples (70 genotypes x 3 environments) will be analysed chemically for: total B-glucan (Alf), soluble B-glucan (Rita), starch and fibre (Malgorzata); antioxidants (tocols by Rita and AVAs by Lena) will be determined on about 100 samples (more samples if possible).

18. SAURA-CALIXTO et al. –CONCLUSIONS   The method for IF determination combines in a single procedure the methodologies for DF and resistant starch analysis. In this method, samples are analyzed as eaten (fresh, boiled or fried), and the analytical conditions are close to the physiological ones.   In DF analysis starch digestibility is altered artificially, whereas IF can remove most digestible starch, avoiding alterations of naturally occurring RS. IF values could be more useful than DF values from a nutritional point of view and could be a valid alternative to DF for food labeling and food composition tables.

19. SAURA-CALIXTO et al. –CONCLUSIONS   The method for IF determination combines in a single procedure the methodologies for DF and resistant starch analysis. In this method, samples are analyzed as eaten (fresh, boiled or fried), and the analytical conditions are close to the physiological ones.   In DF analysis starch digestibility is altered artificially, whereas IF can remove most digestible starch, avoiding alterations of naturally occurring RS. IF values could be more useful than DF values from a nutritional point of view and could be a valid alternative to DF for food labeling and food composition tables.