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Fruit Composition of Elderberry (Sambucus spp.) Genotypes Grown in Oregon and Missouri, USA.

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TLDR
Of the total antioxidant tests used, total phenolics was the best quick test, as it is the easiest of the assays, showed the least change among years, and correlated highly with FRAP.
Abstract
Twelve Sambucus nigra subsp. canadensis genotypes were grown at two Missouri and one Oregon (USA) locations to characterize fruit composition in 2004, 2005, and 2006. Fruit was also sampled from an additional 10 genotypes of subsp. canadensis and nigra grown in Oregon. Soluble solids content ranged from 8.9 to 12.5 °Brix, and titratable acid (as citric acid) was 0.4 to 1.7 g/100ml. Ferric ion reducing activity potential (FRAP) values were 15.6 to 30.7 μmol/g trolox equivalents. Total anthocyanin content ranged from 85 to 385 mg/100 g cyanidin-3-glucoside equivalents (C3GE) and total phenolic content was 421 to 719 mg/100 g gallic acid equivalents (GAE). Of the genotypes sampled in three locations, six genotypes were high (250-350), and four were very low in total anthocyanin (<150 mg/100 g C3GE), respectively. 'Barn' and 'Scotia' were highest, and 'Nova' lowest in total anthocyanin content among genotypes grown in Oregon. Of the total antioxidant tests used, total phenolics was the best quick test, as it is the easiest of the assays, showed the least change among years, and correlated highly with FRAP.

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Fruit Composition of Elderberry (Sambucus spp.) Genotypes
Grown in Oregon and Missouri, USA
P. Perkins-Veazie
1
, A.L. Thomas
2
, P.L. Byers
3
, and C.E. Finn
4
1
Plants for Human Health Institute, North Carolina State University, Kannapolis, NC, USA
2
University of Missouri, Southwest Research Center, Mt. Vernon, MO, USA
3
University of Missouri, Cooperative Extension Service, Springfield, MO, USA
4
USDA-ARS-HCRU, Corvallis, OR, USA
Abstract
Twelve
Sambucus nigra
subsp.
canadensis
genotypes were grown at two Missouri and one Oregon
(USA) locations to characterize fruit composition in 2004, 2005, and 2006. Fruit was also sampled
from an additional 10 genotypes of subsp.
canadensis
and
nigra
grown in Oregon. Soluble solids
content ranged from 8.9 to 12.5 °Brix, and titratable acid (as citric acid) was 0.4 to 1.7 g/100ml.
Ferric ion reducing activity potential (FRAP) values were 15.6 to 30.7 μmol/g trolox equivalents.
Total anthocyanin content ranged from 85 to 385 mg/100 g cyanidin-3-glucoside equivalents
(C3GE) and total phenolic content was 421 to 719 mg/100 g gallic acid equivalents (GAE). Of the
genotypes sampled in three locations, six genotypes were high (250-350), and four were very low
in total anthocyanin (<150 mg/100 g C3GE), respectively. ‘Barn’ and ‘Scotia’ were highest, and
‘Nova’ lowest in total anthocyanin content among genotypes grown in Oregon. Of the total
antioxidant tests used, total phenolics was the best quick test, as it is the easiest of the assays,
showed the least change among years, and correlated highly with FRAP.
Keywords
anthocyanin; phenolics; antioxidant; acidity; FRAP; trolox
INTRODUCTION
While the European elderberry (
Sambucus nigra
L. subsp.
nigra
) is well known and
cultivated domestically in Europe and elsewhere, the native North American elderberry
[
Sambucus nigra
L. subsp.
canadensis
(L.) Bolli] is a niche crop (Charlebois et al., 2010).
The unique anthocyanins in both subspecies (cyanidin sambubiosides) have generated
interest for use in pharmaceuticals and dietary supplements (Mohebalian et al., 2012).
Elderberries possess anti-inflammatory and anti-viral properties (Zakay-Rones et al., 1995;
Roschek et al., 2009). Additionally, the acylated anthocyanins common in American
elderberry are prized for imparting color stability to food products (Nakatani et al., 1995;
Lee and Finn, 2007). In this paper, we examine the fruit composition characteristics of
elderberries grown in diverse geographic locations.
HHS Public Access
Author manuscript
Acta Hortic
. Author manuscript; available in PMC 2016 May 06.
Published in final edited form as:
Acta Hortic
. 2015 January 12; 1061: 219–224. doi:10.17660/ActaHortic.2015.1061.24.
Author Manuscript Author Manuscript Author Manuscript Author Manuscript

MATERIALS AND METHODS
Plantings of 12 American elderberry genotypes were established in 2003 at three U.S.
locations: Mt. Vernon, Missouri (latitude 37°4’N; longitude 93°53’W), Mountain Grove,
Missouri (latitude 37°13’N; longitude 92°26’W), and Corvallis, Oregon (latitude 44°30’N;
longitude 123°28’W). Orchard and experimental details, including genotypes, sites,
environmental conditions, horticultural methods, harvest protocols, etc. are described in
Thomas and Byers (2000), Finn et al. (2008) and Thomas et al. (2013). Fruit from replicated
plots at the three sites was harvested 2004, 2005, and 2006. An additional 10 genotypes were
sampled from Corvallis in 2004 and 2005. All fruit was harvested at peak ripeness, frozen,
and held at −20°C until analyzed at the former USDA-ARS Wes Watkins Agricultural
Research Laboratory, Lane, OK.
About 50 g berries were destemmed and thawed, pureed with double distilled water (1:1 wt./
volume) using a blender cup, then ground and homogenized. Soluble solids content (SSC;
°Brix) pH, titratable acidity, total anthocyanin and total phenolics content were measured as
outlined in Thomas et al. (2013). Ferric reducing activity potential using trolox substrates
was measured on the same extracts used for total phenolics following methods of Benzie and
Strain (1999).
Data were subjected to analysis of variance (Proc GLM; SAS Institute, Cary, NC) and means
separated by least significant difference test at
p
≤ 0.05. Linear regression was performed
using SAS to determine relationships between compositional assays.
RESULTS AND CONCLUSION
When averaged across locations in all years, genotypes varied in all fruit composition
characteristics evaluated (Table 1). The pH, SSC, and acidity of elderberry genotypes grown
in the three locations averaged 4.92, 10.9 °Brix, and 0.53 g/100 ml citric acid, respectively.
Among genotypes grown at all locations, ‘Highway O’ and ‘Johns’ were highest in pH, but
low to lowest in SSC and acidity. ‘Johns’ and ‘Netzer’ were consistently low in anthocyanin
content, appearing brown in color (Thomas et al., 2013). This effect was also seen in total
phenolics content, where ‘Johns’ and ‘Netzer’ were lowest (439 and 514 mg/100 g,
respectively) while ‘Adams 2was highest (719 mg/100 g). Lee and Finn (2007) also
reported high total anthocyanin and phenolic values for ‘Adams 2. The
canadensis
genotypes used in this study were found to be somewhat lower in total anthocyanins
compared with values reported for
nigra
by Veberic et al. (2009).
Genotypes with the highest FRAP values were ‘Adams 2, ‘Wyldewood’, ‘Bob Gordon’, and
‘Eridu 1. The first three are commercially-available cultivars, and these results support their
value as cultivars for use in dietary supplements and other products. ‘Johns’, another
commercial cultivar, had very low FRAP values. As reported in Thomas et al. (2013),
‘Johns’ productivity was poor at both Missouri sites. The poor productivity plus poor fruit
composition of ‘Johns’ makes it an unsuitable cultivar for the midwestern US.
Of the elderberries grown only in Oregon, pH, SSC, and acidity ranged from 4.03 to 4.75;
8.9 to 12.5 °Brix; and from 0.73 to 1.65 g/100 ml, respectively (Table 2). The genotypes
Perkins-Veazie et al.
Page 2
Acta Hortic
. Author manuscript; available in PMC 2016 May 06.
Author Manuscript Author Manuscript Author Manuscript Author Manuscript

‘Haschberg’, ‘Korsør’, and ‘Golden City’ were low in pH and high in acidity while SSC was
highest in ‘Barn’, ‘Scotia’, and ‘Korsør’. Total anthocyanin content ranged widely, from 85
to 385 mg/100 g. ‘Barn’ was high in total anthocyanin, phenolic, and FRAP values. In
contrast, ‘York’ and ‘Nova’ were low in total anthocyanin and FRAP values relative to the
other genotypes. The representation of
nigra
genotypes in this study was small but indicates
that both subspecies may have similar processing characteristics when cultivated in Oregon.
When total anthocyanin or total phenolic contents were regressed to FRAP values, strong
linear relationships were seen among these variables (Fig. 1). Regression fit was similar
between total anthocyanin and FRAP (r
2
=0.66), total phenolic content and FRAP (r
2
=0.63),
and total anthocyanin and total phenolic content (r
2
=0.61).
Results from this study support earlier findings (Finn et al., 2008; Özgen et al., 2010;
Thomas et al., 2013) that the fruit composition character of elderberry is influenced by both
genetic and environmental factors. Correlations of antioxidant tests and fruit composition
assays indicate that pH, SSC, and titratable acidity cannot be used as simple predictors of
total phenolic, total anthocyanin, or FRAP values (data not shown). Of the total antioxidant
tests used, total phenolics appears to be the best quick test, as it is the easiest of the assays,
showed the least change among years, and correlated highly with FRAP.
ACKNOWLEDGEMENTS
The assistance of Jungmin Lee, Mary Peterson, Ted Mackey, Chris Rennaker, Sheila Magby, and Brian Yorgey is
gratefully acknowledged. This work was partially funded through the Center for Agroforestry, University of
Missouri under cooperative agreements with the USDA-ARS Dale Bumpers Small Farm Research Center,
Booneville, AR. We thank the USDA-Agricultural Research Service (ARS) CRIS numbers 5358-21000-041-00D,
5358-21000-037-00D and Northwest Center for Small Fruits Research for funding as well as for their technical
assistance with this project. This publication was also made possible by Grant Number P50AT006273 from the
National Center for Complementary and Alternative Medicines (NCCAM), the Office of Dietary Supplements
(ODS), and the National Cancer Institute (NCI), and with funds from the USDA Crop Germplasm Committee,
Small Fruits. Its contents are solely the responsibility of the authors and do not necessarily represent the official
views of the NCCAM, ODS, NCI, USDA, or the National Institutes of Health. Mention of trade names or
commercial products in this publication is solely for the purpose of providing specific information and does not
imply recommendation or endorsement by the USDA.
Literature Cited
Benzie IFF, Strain JJ. Ferric reducing/antioxidant power assay: direct measure of total antioxidant
activity of biological fluids and modified version for simultaneous measurement of total antioxidant
power and ascorbic acid concentration. Methods in Enzymology. 1999; 299:15–27. [PubMed:
9916193]
Charlebois, D.; Byers, PL.; Finn, CE.; Thomas, AL. Elderberry: botany, horticulture, potential. In:
Janick, J., editor. Horticulture Reviews. Vol. Vol. 37. Wiley-Blackwell; New Jersey: 2010. p.
213-280.
Finn CE, Thomas AL, Byers PL, Serçe S. Evaluation of American (
Sambucus canadensis
) and
European (
S. nigra
) elderberry genotypes grown in diverse environments and implications for
cultivar development. HortScience. 2008; 43:1385–1391.
Lee J, Finn CE. Anthocyanins and other polyphenolics in American elderberry (
Sambucus canadensis
)
and European elderberry (
S. nigra
) cultivars. J. Sci. Food Agr. 2007; 87:2665–2675. [PubMed:
20836175]
Mohebalian PM, Cernusca MM, Aguilar FF. Discovering niche markets for elderberry juice in the
United States. HortTechnol. 2012; 22:556–566.
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Nakatani N, Kikuzak H, Hikida J, Ohba M, Inami O, Tamura I. Acylated anthocyanins from fruits of
Sambucus canadensis
. Phytochemistry. 1995; 3:755–757. [PubMed: 7766167]
Özgen M, Scheerens JC, Reese RN, Miller RA. Total phenolics, anthocyanin contents and antioxidant
capacity of selected elderberry (
Sambucus canadensis
L.) accessions. Pharmacogn. Mag. 2010;
6:198–203. [PubMed: 20931079]
Roschek B Jr. Fink RC, McMichael MD, Li D, Alberte RS. Elderberry flavonoids bind to prevent
H1N1 infection in vitro. Phytochemistry. 2009; 70:1255–1261. [PubMed: 19682714]
Thomas, AL.; Byers, P. Multi-locational elderberry cultivar and management study. Proc. Missouri
Small Fruit Conf.; Springfield, MO: Southwest Missouri State Univ.; 2000. p. 37-40.
Thomas AL, Perkins-Veazie P, Byers PL, Finn CE, Lee J. A comparison of fruit characteristics among
diverse elderberry genotypes grown in Missouri and Oregon. J. Berry Res. 2013; 3:159–168.
Zakay-Rones Z, Varsano N, Zlotnik M, Manor O, Regev L, Schlesinger M, Mumcuoglu M. Inhibition
of several strains of influenza virus in vitro and reduction of symptoms by an elderberry extract
(
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1995; 1:361–369.
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Fig. 1.
(A) Regression of total monomeric anthocyanin content to trolox values, (B) regression of
total phenolic content to trolox values, (C) regression of total monomeric anthocyanin to
total phenolic content, all across multiple elderberry genotypes and locations.
Perkins-Veazie et al. Page 5
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Author Manuscript Author Manuscript Author Manuscript Author Manuscript

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References
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Ferric reducing/antioxidant power assay: direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration

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TL;DR: Sufficient variability was seen among these genotypes to suggest that a successful breeding program could be carried out to improve levels of the various compounds evaluated in this study.
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Elderberry flavonoids bind to and prevent H1N1 infection in vitro

TL;DR: The Direct Binding Assay established that flavonoids from the elderberry extract bind to H1N1 virions and, when bound, block the ability of the viruses to infect host cells.
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Inhibition of several strains of influenza virus in vitro and reduction of symptoms by an elderberry extract (Sambucus nigra L.) during an outbreak of influenza B Panama

TL;DR: Considering the efficacy of the elderberry extract in vitro on all strains of influenza virus tested, the clinical results, its low cost, and absence of side-effects, this preparation could offer a possibility for safe treatment for influenza A and B.
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Total phenolic, anthocyanin contents and antioxidant capacity of selected elderberry (Sambucus canadensis L.) accessions

TL;DR: Within this limited study of 14 accessions, variability for phytonutrient content and antioxidant capacity suggested the employment of wild germplasm within an elderberry improvement program to incorporate an array of superior horticultural, post-harvest or processing traits into new or existing cultivars with superior phytonUTrient profiles.
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