International Journal of Food Science and Technology 2015 1

Original article
Evaluation of colouring ability of main European elderberry
(Sambucus nigra L.) varieties as potential resources of natural
food colourants

Lilla Szal o,1,2 M

oki-Dork ate1 & Laszlo Abranko2*
onika Steger-M
 t, Budapest 1118, Hungary
1 Department of Food Preservation, Faculty of Food Science, Corvinus University of Budapest, 29-33 Villanyi u
t, Budapest 1118, Hungary
2 Department of Applied Chemistry, Faculty of Food Science, Corvinus University of Budapest, 29-33 Villanyi u

(Received 14 November 2014; Accepted in revised form 18 January 2015)

Summary Proling and quantitative analysis of anthocyanins in ve elderberry (Sambucus nigra L.) varieties, namely
Haschberg, Samocco, Samyl, Samident and Sampo, were performed in six dierent maturity stages
from two consecutive years (2012 and 2013) and from two growing areas in Hungary. Cyanidin-3-O-sam-
bubioside-5-O-glucoside, cyanidin-3-O-sambubioside and cyanidin-3-O-glucoside were found and identi-
ed by HPLC-Q/TOF-MS as major anthocyanins and were quantied by HPLC-UV/Vis. In optimum
maturity stage, Samocco showed the highest anthocyanin content with an average of 1237 mg per 100 g
dry weight in both growing areas and vintages. The dominant anthocyanin component of Samocco vari-
ety was cyanidin-3-O-sambubioside, which is according to literature more stable against technology pro-
cessing than cyanidin-3-O-glucoside found in the other four investigated elderberry varieties in the highest
concentration. Samocco, if grown under the climatic conditions of the Carpathian basin, might be a
promising alternative variety for growing as raw material for natural food colourant processing industry.
Keywords Anthocyanin prole, elderberry, food colouring, ripening, Sambucus nigra.

colouration. The only permitted natural food colou-

Introduction
Colours are important quality indicators that may
remarkably contribute to the consumer preference and
acceptance of foods (Downham & Collins, 2000). In
recent days, market for the application of synthetic col-
ourants has decreased in favour of natural colourants,
especially since the human safety of synthetic food dyes
has been legally questioned. The regulation of the
European Commission 1333/2008/EC states that foods
containing certain articial colouring material, namely
Sunset Yellow (E110), Quinoline yellow (E104), Car-
moisine (E122), Allura Red (E129), Tartrazine (E102)
and Ponceau 4R (E124), must be labelled with the fol-
lowing phrase: may have an adverse eect on activity
and attention in children (Ocial Journal of the Euro-
pean Union, 2008). Based on the above regulation
along with the strengthening consumer demand, food
producers tend to make eorts to replace their syn-
thetic food colouring agents with natural food colou-
rants. There are a number of natural colour additives
derived from natural sources, which are legally permit-
ted in the European Union and thus are used for food
*Correspondent: E-mail: laszlo.abranko@uni-corvinus.hu
rants of animal origin are carminic acid and carmines
often referred to as cochineal. However, allergy-like
reactions such as IgE-mediated reactions linked to co-
chineal have been reported (Lucas et al., 2001; Green-
hawt & Baldwin, 2009). Therefore, plants are
considered as primary sources of natural food colou-
rants, yielding anthocyanins, carotenoids and chloro-
phylls as the major groupings (Scotter, 2003). Among
them, anthocyanins demonstrate a high potential due
to their attractive red and purple colours and water sol-
ubility that allows their incorporation into aqueous
food system (Kong et al., 2003; Lee & Finn, 2007; Del
Caro & Piga, 2008). Anthocyanin colourants are suit-
able for food coloration with a low pH level such as
soft drinks, water ice cream, sauce, confectionery, table
jellies and conserves because anthocyanin molecules are
more stable in acidic medium (Bridle & Timberlake,
1997; Cooper-Driver, 2001; Bakowska-Barczak, 2005).
Elderberry (Sambucus nigra L.) may be an excellent
and preferred source of natural food colourant in food
industry because of its high anthocyanin content (Bron-
numhansen et al., 1985; Jakobek et al., 2007; Veberic
et al., 2009). Elderberry pigments are almost exclusively
cyanidin glycosides, from which cyanidin-3-O-glucoside

doi:10.1111/ijfs.12773
2015 Institute of Food Science and Technology
2  ki-Dorko
Coloring potential of elderberry L. Szalo  et al.
and cyanidin-3-O-sambubioside are the major ones
(Dawidowicz et al., 2006). In addition, cyanidin-3-O-
sambubioside-5-O-glucoside and cyanidin-3,5-O-dig-
lucoside were detected as minor compounds (Hong &
Wrolstad, 1990; Kaack et al., 2008; Veberic et al.,
2009). Wu et al. (2004) and more recently, Mikulic-Pet-
kovsek et al. (2014) have reported additional minor
components in S. nigra (cyanidin-3-O-rutinoside, pe-
largonidin-3-O-glucoside, pelargonidin-3-O-sambubio-
side). By nature, chemical forms and the amount of
pigments present in the natural food materials depend
on several factors such as the plant cultivar, the
geographical origin and the time of harvest (Kaack
et al., 2008).
Many cultivars of elderberries are planted in Europe,
the most common is the Haschberg cultivar with sev-
eral selections, which are mainly grown for their fruit
(Veberic et al., 2009). Globalisation of the market for
juice or concentrate resulted that new genotypes with
dierent quality characteristics have appeared over the
last few years, but their colouring properties are less
known. Up until now, only a few studies are available
focusing on the anthocyanin content in elderberry
genotypes at the molecular level (Kaack & Austed,
1998; Wu et al., 2004). In addition, major dierences
occur in these results (e.g. minimum value of cyanidin-
3-O-glucoside is 361 mg per 100 g fresh weight (FW)
and maximum is 1266 mg per 100 g FW) in terms of
the individual anthocyanin content. This remarkable
diversity may be attributed to the dierence in climatic
conditions, geographical areas and applied horticultural
technology in the case of the investigated elderberry
genotypes. In order to be able to adequately evaluate
and assess the colouring potential of elderberry varieties
whose importance is steadily increasing as sources of
natural food colourants, it is essential to carry out sys-
tematic experiments and gain more detailed data on
anthocyanin proles and contents of these cultivars.
The objective of this study was to characterise
anthocyanin types and contents and thus to evaluate
colouring potential of ve elderberry varieties grown
in Hungary (Carpathian Basin climate) that are to be
used as natural food colourant for the food industry.
Samples of this study were harvested in two dierent
growing areas in two consecutive years (2012 and
2013). Eects of vintage and growing area on anthocy-
anins proles along with the total soluble solid con-
tents during ripening were also considered and
evaluated to estimate optimal conditions of harvest.
Materials and methods
Materials
Elderberry fruit [S. nigra (L.) subsp. nigra, Adoxaceae]
samples were harvested in 2012 and 2013 from mid-
International Journal of Food Science and Technology 2015
July till the end of August. Five dierent elderberry
varieties, namely Samocco, Samident, Samyl,
Sampo, Haschberg, were sampled in two dierent
growing locations in Hungary: Val (47210 N, 18400 E)
and Nagyvenyim (46570 N, 18510 E). Harvest started
with fruit clusters containing mostly green berries in
July (1st maturity stage), then progressively darker
berries were collected every week (2nd to 5th maturity
stage). Finally, clusters containing black, fully ripe ber-
ries were harvested (6th maturity stage) in August.
Representative samples of each cultivar were collected
from several plants, and approximately 1.5 kg of elder-
berry berries (with stalks) per sample was packed in
plastic bags and stored at 18 C until further pro-
cessing. The initially picked samples were stored maxi-
mum for 6 weeks, and the fruits collected at the last
harvest date were stored only for some days. Samples
were classied in dierent maturity status based on
visual judgment and harvest dates.
The HPLC-grade acetonitrile (Chromasolv Gradi-
ent) and methanol (Chromasolv for HPLC) were
purchased from Sigma-Aldrich (St. Louis, MO, USA).
Formic acid was obtained from Fluka (Buch, Switzer-
land). Cyanidin-3-O-glucoside and cyanidin-3-O-sam-
bubioside were purchased from Extrasynthese (Genay,
France).
Methods
Sample preparation
Samples were lyophilised prior to extraction using a
Scanvac, CoolSafe freeze dryer (Labogene, Lynge,
Denmark). Each batches of samples was weighed at
0.01 g accuracy before and after freeze drying and its
water content was calculated based on the obtained
weight dierence between original (i.e. before freeze
drying started) and dried conditions. The extraction
procedure used in this study was adopted from Lin &
Harnly (2007) and applied with slight modications.
Briey, 200 mg of lyophilised and homogenised sample
was weighed into a 15-mL plastic centrifuge tube and
10 mL 60% (v/v) aqueous methanol containing 1% (v/
v) formic acid was added. Samples were extracted for
30 min in an ultrasonic bath (<35 C at the end). After
extraction, samples were centrifuged at 4180 g for
5 min at room temperature. Next, the supernatant was
removed and ltered through a 0.2-lm PTFE mem-
brane lter before injecting 10 lL to the HPLC sys-
tem. Sample preparations were carried out in ve
replicates (n = 5).
HPLC-UV/Vis and HPLC-MS analysis
Chromatographic separation of anthocyanin compo-
nents was performed with an Agilent 1200 HPLC sys-
tem (Agilent Technologies, Waldbronn, Germany)
using a 125 9 4 mm, C18, 3 lm particle size column
2015 Institute of Food Science and Technology

(Dr. Maisch GmbH, Ammerbuch-Entringen, Ger-
many). For the elution, 0.5% (v/v) formic acid in high
purity water (mobile phase A) and 0.5% formic acid
in acetonitrile (mobile phase B) were used as solvents
at a ow rate of 0.5 mL min1. The total gradient
program was 25 min and started at 5% solvent B. Sol-
vent B was increased linearly to 25% in 5 min and
from 5 to 10 min solvent B increased to 100%.
Finally, 100% solvent B was held constant for another
5 min then it was immediately reduced to 5% and the
system was running for an additional 10 min at the
initial solvent composition.
The HPLC system, including a diode array detector,
detector was coupled to an Agilent 6530 quadrupole/
time-of-ight mass spectrometer (Q/TOF-MS) instru-
ment equipped with an electrospray ion source (ESI),
which was used in positive mode with the following
instrument parameters: capillary voltage, 4000 V; neb-
ulizer pressure, 40 psig; drying gas ow rate,
13 L min1; gas temperature, 350 C; fragmentor volt-
age, 140 V. Mass spectra were acquired in the range of
m/z 501100. Mass accuracy <2 ppm was achieved in
TOF-only mode and <20 ppm in Q/TOF mode. Quan-
titation of anthocyanins was carried out using an Agi-
lent 1200 HPLC system including a variable
wavelength UV/Vis detector. Samples were analysed at
520 nm.
Measurement of soluble solids
Soluble solid content was determined from thawed
fruits and expressed as Brix with ATAGO PR-301
digital refractometer (ATAGO USA Inc., Belleuve,
WA, USA).
Statistical analysis
T-test was used for the analysis of the eect of vin-
tages on anthocyanin content at optimum ripening
stage. Signicant dierences between 2 years were con-
sidered when P value was <0.05.
Results and discussion
Anthocyanin identification
Based on the UV/Vis chromatogram of elderberry
extracts, three anthocyanin compounds could be
observed in all investigated varieties. Two of three
peaks could be identied with reference standards as
cyanidin-3-O-sambubioside and cyanidin-3-O-gluco-
side. These results are in accordance with litera-
ture data (Hong & Wrolstad, 1990; Wu et al., 2004;
Veberic et al., 2009). In the case of the third com-
pound, neither of our reference standards matched its
retention time; therefore, this compound was subjected
to a targeted Q/TOF-MS experiment in order to iden-
tify it. As a result, the monoisotopic mass of this com-
2015 Institute of Food Science and Technology
 ki-Dorko
Coloring potential of elderberry L. Szalo  et al. 3
pound was determined as m/z 743.2041, and its
elemental formula could be elucidated as C32H39O20.
This compound was further investigated in a tandem
Q/TOFMS experiment. In the product ion spectrum of
this compound shown in Fig. 1, in addition to the pre-
cursor ion at m/z 743.2051, three further fragments
could be observed. The fragment with the experimental
monoisotopic mass m/z 287.0561 was identied as
C15H11O6. The other two fragments (m/z 449.1095 and
581.1516) have been annotated with the following
elemental formulae: C21H21O11 and C26H29O15, respec-
tively. These ion series indicating that m/z 287.0561
corresponds to a cyanidin aglycone (Cy), whereas
m/z 449.1095 can be annotated as [Cy + hexosyl]+,
m/z 581.1516 as [Cy + hexosyl  xylosyl]+ and m/z
743.2041 as [Cy + hexosyl + xylosyl + hexosyl]+.
Based on the acquired MS and MS/MS mass spectra
in combination with the obtained UV/Vis signal at
520 nm, this anthocyanin trisaccharide eluting at
7.1 min was identied as cyanidin-3-O-sambubioside-
5-O-glucoside, which is in accordance with other litera-
ture data (Hong & Wrolstad, 1990; Kaack et al., 2008;
Veberic et al., 2009).
Changes in anthocyanins during ripening
Sampling of elderberries was performed at multiple
occasions to investigate any dierences in the distribu-
tion of individual anthocyanins during ripening as
described in Materials and methods (see Materials).
Quantitative anthocyanin measurements were per-
formed with a HPLC-UV/Vis system applying the
same chromatographic separation method as was used
in the HPLC-Q/TOF-MS system. Measurements were
carried out at 520 nm, and external calibration was
performed using cyanidin-3-O-glucoside reference stan-
dard. Concentrations of each anthocyanin compounds
were assessed from peak areas and are given as cyani-
din-3-O-glucoside equivalent (CGE) per 100 g of dry
weight (DW) elderberry fruit. The amount of total an-
thocyanins (TA) was expressed as sum of individual
anthocyanin concentrations.
It is well known that maturity stage and therefore
harvest time may aect the concentration of phenolic
compounds in berries and other fruits. Awad et al.
(2001) showed on the example of some apple varieties
that concentration of cyanidin-3-O-galactoside was rel-
atively high in the early season, and then it showed
decreasing and increasing trends during ripening.
Other studies presented increase of anthocyanin con-
tent in investigated sweet cherries (Serrano et al., 2005;
Goncalves et al., 2007) and strawberries (Pineli et al.,
2011; Mazur et al., 2014) during ripening in dierent
maturity stages. Up until now, there are only a few
studies investigated anthocyanin changes in elderberry
fruits during ripening (Lee & Finn, 2007; Kaack et al.,
International Journal of Food Science and Technology 2015
4  ki-Dorko
Coloring potential of elderberry L. Szalo  et al.

Figure 1 Full-scan Q/TOF-MS product ion

spectrum of m/z 743, which ion mass corre-
sponds to the compound eluted at 7.1 min.
Spectrum was acquired in the extract of a
Samocco sample, collected in the
Nagyvenyim region in 2012.

1990), and to our best knowledge, it is the rst system-

atic report that considers eects of vintage and grow-
ing area. Our results show that elderberry cultivars
have a maximum point in their anthocyanin contents
during ripening. This was observed for all investigated
samples; however, the maximum point was reached at
a dierent date for each variety. For instance in 2012,
when Sampo, the earliest maturing variety reached
the 5th maturity stage, Haschberg was only in its 3rd
maturity stage. In 2012, all the samples reached the
maximum anthocyanin concentration within the inves-
tigated timeframe except Samyl collected from either
growing areas. This latter observation might be
explained with the very late maturing characteristic of
Samyl in the areas studied. A typical example of
observed anthocyanin concentration curves is shown
in Fig. 2. In 2013, maximum points in anthocyanin Figure 2 Typical anthocyanin concentration curve during ripening
concentration curves were reached at the same matu- in the case of Sampo (Nagyvenyim) in 2012. TA: total anthocyanin
content, Cy3G: cyanidin-3-O-sambubioside-5-O-glucoside, Cy2G: cy-
rity stage for each variety; however, certain maturity
anidin-3-O-sambubioside, CyG: cyanidin-3-O-glucoside.
stage was achieved in both growing regions several
weeks later than in 2012. This suggests that the opti-
mum maturity stage in terms of anthocyanin content stage dependent. Figure 3 shows an example where
is considerably inuenced by climatic conditions, and anthocyanin proles of Haschberg in the 2nd matu-
optimum harvest date can be shifted by several weeks rity stage and Sampo in the 2nd and the 3rd maturity
between vintages. Therefore, acquiring objective data stages are presented. As can be seen, cyanidin-3-O-
such as total anthocyanin content is essential to appro- sambubioside (eluting at 8.9 min) is the dominant
priately schedule optimum harvest date. For example, anthocyanin in Haschberg in the 2nd maturity stage
if Haschberg in Nagyvenyim had been harvested in (chromatogram A in Fig. 3), whereas in Sampo in
2013 at the date this variety reached the maximum the same 2nd maturity stage, these two compounds are
anthocyanin content during ripening in 2012, pigment present in almost equal amounts (chromatogram C in
concentration would have been ca. 25% lower than in Fig. 3). However, this ratio changes in Sampo during
the previous year. Based on similar considerations, our ripening and in the following (3rd) maturity stage,
results suggest that Samident is the most robust and cyanidin-3-O-glucoside becomes more abundant than
Samyl was the most sensitive to the harvest date in cyanidin-3-O-sambubioside (chromatogram B in
both growing areas. Finally, it should be mentioned Fig. 3).
that there were no dierences observed in the kinetics
of concentration changes for the particular anthocya-
Anthocyanin contents at optimum maturity stage
nin compounds; during ripening, each compound has
reached its maximum at the same maturity stage. The most abundant anthocyanin molecule in most
However, in some cases, the distribution pattern of samples at full maturity stage was cyanidin-3-O-gluco-
anthocyanin compounds was variety and maturity side (CyG) that accounted for approximately the half

International Journal of Food Science and Technology 2015 2015 Institute of Food Science and Technology
  ki-Dorko
Coloring potential of elderberry L. Szalo  et al. 5

P value

0.012*
0.000*
0.013*
0.002*

0.000*
0.024*

0.001*
0.361

0.469
0.344
2013

5373
2641
3699
4168
1880
3282
2943
3516
1981
1826

Represents significant differences in anthocyanin content between the two vintages (P < 0.05). Average values of five replicates (n = 5)  standard errors are presented.
2012

6979
7013
2711
4809
2809
4720
3365
4577
2621
4159

Cy3G, cyanidin-3-O-sambubioside-5-O-glucoside; Cy2G, cyanidin-3-O-sambubioside; CyG, cyanidin-3-O-glucoside; NV, Nagyvenyim; TA, total anthocyanin; V, Va l.
TA
Table 1 Anthocyanin concentration in optimum maturity stages of ve elderberry varieties from dierent growing areas in mg CGE per 100 g DW

P value

0.039*
0.000*
0.045*
0.808*
0.001*
0.009*

0.001*

0.000*
0.570

0.288
47.0
248
222
359
355
158

281
478
336
183










2013

1801
877
1863
1676
991
1553
1395
2065
925
747
Figure 3 Three dierent anthocyanin proles of elderberry varieties
from the same growing area of Val in the harvest year of 2012.
Chromatogram A shows the 2nd maturity stage of Haschberg

81.4
355
429
224
122
217
222

202
468
252
variety, whereas chromatograms B and C show the 3rd and 2nd











maturity stages of Sampo, respectively.

2012

2432
2645
1263
2457
1107
2378
1759
2242
1184
1513
CyG
of all determined anthocyanins. The only exception
was Samocco where cyanidin-3-O-sambubioside

P value

0.020*
0.046*

0.000*
0.000*

0.001*

0.001*
(Cy2G) accounted for more than 50% of all analysed

0.850
0.054

0.675

0.137
anthocyanins. This fact gains importance as the nd-
ings of Drdak & Daucik (1990) is considered, who

61.0
reported that Cy2G proved to be more stable during

439
339
309
435
119

254
274
311
218
food processing than CyG. It means that besides the










quantitative aspects of the anthocyanin content of
2013

3118
1372
1590
2102
747
1679
1280
1203
871
886
Samocco, its special anthocyanin prole makes it
more suitable to be used as a raw material for produc-

47.6
ing elderberry concentrate. The third identied
606
609
206
119
265
188

176
464
379
pigment molecule, cyanidin-3-O-sambubioside-5-O-glu-










Cy2G

coside (Cy3G) presented in the smallest concentration

2012

3942
3833
1196
2051
1369
1984
1348
1933
1155
2282
in the elderberry fruits that is consistent with the
ndings of another study on European elderberries
P value

(Veberic et al., 2009).

0.024*
0.000*
0.033*

0.010*
0.000*

0.031*

0.000*
0.156

0.156

0.290
Evaluation of colouring potential of the investigated
elderberry varieties was based upon the measured total
anthocyanin contents in the optimum maturity stage
64.8

50.3
85.4
23.8
11.6
52.9
54.2
67.5
47.5
105

(maturity stage with the highest TA value). TA concen-












trations of each studied varieties ranged from 2621 mg

2013

455
393
246
390
142
249
269
248
185
193

CGE per 100 g DW to 7013 mg CGE per 100 g DW

in 2012 and from 1826 mg CGE per 100 g DW to
95.4
86.8
42.6
23.4
63.8
35.2
10.4
37.2

61.8

5373 mg CGE per 100 g DW in 2013 (Table 1). These

113

results are in accordance with previous investigations











Cy3G

2012

(Bronnumhansen & Flink, 1986; Kaack & Austed,

604
535
252
300
333
358
258
402
282
364

1998; Kaack et al., 2008; Veberic et al., 2009).

Samocco had the highest TA concentration in our
Growing

study in both harvest seasons and both growing areas

area

as well. Maximum concentration reached at the 5th

NV

NV

NV

NV

NV
V

maturity stage in both growing areas in 2012 was

6979 mg CGE per 100 g DW from V al and 7013 mg
Haschberg
Samident
Samocco

CGE per 100 g DW from Nagyvenyim. In 2013, Samo-

Variety

Sampo

Samyl

cco contained considerably (2845%) less anthocyanins

than in 2012; nevertheless, Samocco was still the rst
*

2015 Institute of Food Science and Technology International Journal of Food Science and Technology 2015
6  ki-Dorko
Coloring potential of elderberry L. Szalo  et al.
variety in terms of TA content. If only the TA values
are compared with respect to growing areas, Samocco
from Nagyvenyim contains nearly 1.59 more TA, and
Samocco from V al, nearly 2.59 more than the other
varieties in 2012 and 2013 (Table 1). The variety con-
taining the lowest total pigment content was Samyl
with 4159 mg CGE per 100 g DW and 2621 mg CGE
per 100 g DW in the year of 2012 and 1826 mg CGE
per 100 g DW and 1981 mg CGE per 100 g DW in
2013. Haschberg that is currently considered as lead-
ing variety in Europe, contained only an average of
3173 mg CGE per 100 g DW of anthocyanins that
equals 787 mg CGE per 100 g FW of pigments. This
observed anthocyanin content was in good agreement
with results in literature, for example 737 mg CGE per
100 g FW was reported by Veberic et al. (2009). Simi-
larly, in a study by Kaack et al. (2008) comparing
Samocco, Haschberg and Sampo, Sampo showed in all studied samples and in all maturity stages. For
the highest anthocyanin content (1900 mg per 100 g
DW) and Samocco the lowest (1480 mg per 100 g
DW) in their maximum maturity stages. Haschberg
had also lower value (1770 mg per 100 DW) than that
obtained in our study. Though, it was also reported that
anthocyanin concentration of Haschberg planted in
the USA at the US Department of Agriculture Agricul-
tural Research Service (USDA-ARS) was much lower
(219 mg CGE per 100 g FW) (Lee & Finn, 2007).
In our study, considerable dierence (1966%) in
TA content between the studied 2 years hindered the
possibility to set up an unequivocal ranking of the
remaining varieties, based on maximum TA concen-
trations. These results indicate that growing areas
and climatic conditions such as number of sunny
days, number of rainy days, etc. may have tremen-
dous inuence on TA content of the investigated
elderberries.
With respect to the vintage eect, signicant dier- having the most anthocyanins relative to the other
ences (P < 0.05) were observed for most elderberry
varieties between the two studied years (Table 1.). The
highest dissimilarity between two harvest years was
observed for Samocco and Samyl from Nagyvenyim
growing area. The average TA content in 2013 was
4007 mg CGE per 100 g DW in Samocco and
1906 mg CGE per 100 g DW in Samyl that are less
than the two-third of the amount observed in 2012.
The slightest eect of vintages was found in the case
of Sampo (V al 19%; Nagyvenyim 31%), whereas the
greatest in the case of Samyl (V al 35%; Nagyvenyim
65%). Furthermore, dierences within varieties in total
pigment content from dierent growing areas were
observed in the case of some varieties in both harvest
seasons. In general, two times higher TA concentration
was recorded in elderberries planted in Nagyvenyim
than in V al especially in the harvest date of 2012. The
most remarkable dierence was observed in the case of
Samyl in 2012 because almost two times higher TA
International Journal of Food Science and Technology 2015
concentration was measured in elderberry fruit from
Nagyvenyim than from Val.
Total soluble solid content
In food industry, concentrated elderberry juice is
mostly used for colouring. Concentrate production is
based on the use of cultivars with a high content of
total soluble solids and anthocyanins (Kaack, 1997).
The higher the total soluble solid content of the fresh
fruit, the less water should be evaporated to reach the
targeted concentrate. Therefore, in addition to antho-
cyanin content estimated on a dry weigh basis, water
content of the harvested fruit samples has to be consid-
ered when the potential of these varieties as source of
natural food colourants are evaluated. Therefore, total
soluble solid contents of samples were also determined
each investigated elderberry varieties, Brix values
(determined with a refractometer) increased during rip-
ening. The soluble solid content of raw elderberry fruits
ranged from 11.7 to 17.9 Brix in 2012 and from 7.7 to
14.1 Brix in 2013 in optimum maturity stages, which
results are consistent with literature data (Lee & Finn,
2007; Kaack et al., 2008; Casati et al., 2012). Except
for Haschberg, as this variety had extremely high,
17.9 Brix value from Val in 2012 in our investigation.
As it was previously observed, Samocco contained the
highest amounts of TA in this study. In order to be
able to realistically assess the usability of this variety as
raw material for processing colouring concentrate, this
variety was compared in more details to the most
widely grown Haschberg variety. Besides comparing
measured TA contents of these two varieties, TA con-
tent was normalised to total soluble solids (Table S1).
In terms of TA/Brix value still Samocco comes out as
elderberry varieties in the optimum maturity stage.
Conclusions
Elderberry fruit is used for natural food colourant in
the food industry because of high anthocyanin concen-
tration. According to our results, Samocco variety
had the highest anthocyanin content and high total
soluble solid content, while Samyl showed the lowest
values in both investigated growing areas and seasons.
Furthermore, cyanidin-3-O-sambubioside was the
dominant anthocyanin in Samocco, which proved to
be more stable during food processing technology
according to a previous study than cyanidin-3-O-glu-
coside found the highest concentration in the other
investigated elderberry variety. These characteristics
make Samocco a promising alternative for producing
elderberry concentrate if grown under the climatic con-
ditions of the Carpathian basin. However, it should be
2015 Institute of Food Science and Technology

noted that in addition to anthocyanin content and
total soluble solids of the elderberry fruit, other
aspects such as production yield, resistance, etc. should
also be considered when the overall suitability of a
particular elderberry variety is evaluated as a potential
alternative to currently grown ones.
Acknowledgments
The authors wish to thank Elderberry Grower Cooper-
ative (V
al, Fejer, Hungary) for providing elderberry
genotypes. The nancial support of the OTKA-PD
100506 grant is kindly acknowledged.
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Supporting Information
Additional Supporting Information may be found in
the online version of this article:
Table S1. Comparing the total anthocyanin content
(mg CGE per 100 g DW) and the Brix value of elder-
berry varieties to Samocco in optimum maturity stage
in two harvest years.
International Journal of Food Science and Technology 2015