WP7 - RECENT DEVELOPMENTS
IN ANALYSIS FOR STARCH, FIBER
AND β-GLUCAN
AVEQ Second meeting
Quedlinburg, 2-4 March 2009
STATE OF ART
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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
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
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
WARD et al. - 2008
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
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
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
SHEWRY et al. - RESULTS
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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
SHEWRY et al. - CONCLUSIONS
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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
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
REDAELLI et al. - RESULTS
PROTEIN
β-GLUCAN
TDF
ABEL
18.25 c
4.26 a
12.75 b
BIKINI
18.83 b
3.78 c
HJA72095N
19.22 a
3.73 c
12.80 b
KONRADIN
17.52 d
3.41 d
15.23 a
KYNON
18.37 c
3.14 e
NAVE
18.46 c
3.93 b
12.10 cd
RHEA x PADARN 542
18.48 c
3.48 d
11.56 d
DONATA (H)
16.43 e
4.31 a
12.42 bc
FLAVIA (H)
19.01 ab
3.92 b
12.62 bc
18.29
3.77
12.78
MEAN
REDAELLI et al. - CONCLUSIONS
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Protein content was strongly influenced by environment, for βglucan 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
IN VITRO DETERMINATION OF THE INDIGESTIBLE
FRACTION IN FOODS: AN ALTERNATIVE TO DIETARY
FIBER ANALYSIS
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.
AVEQ Second meeting, Quedlinburg, 2-4 March 2009
FOODS
ENZYMATIC TREATMENTS
CENTRIFUGATION
RESIDUE =
INSOLUBLE IF
SUPERNATANT - Oligosaccharides
DIALYSIS
RESISTANT STARCH
RESISTANT PROTEIN
CONDENSED TANNINS
LIGNIN
MINERALS
INSOLUBLE NSP
OTHER COMPOUNDS
SOLUBLE NSP
Solvent removal
Neutral sugars +
uronic acids
SOLUBLE IF
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.
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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.
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.
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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).
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.
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.