Scientia Horticulturae 226 (2017) 293–301

Contents lists available at ScienceDirect

Scientia Horticulturae
journal homepage: www.elsevier.com/locate/scihorti

Andean blackberries (Rubus glaucus Benth) quality as affected by harvest MARK

maturity and storage conditions
⁎
Sandra Horvitza, , Diana Chanaguanoa, Iñigo Arozarenab
a
Food Science and Engineering Faculty, Universidad Técnica de Ambato, Campus Huachi, Av. Los Chasquis y Rio Payamino, Ambato, Ecuador
b
Food Technology Department, Universidad Pública de Navarra, Campus Arrosadía s/n 31006, Pamplona, Spain

A R T I C L E I N F O A B S T R A C T

Keywords: Maturity stage at harvest and storage conditions are critical factors determining fruit postharvest quality.
Antioxidant activity Physicochemical (fruit size, mass loss, color, firmness, pH, total soluble solids, titratable acidity), microbiological
Blackberries (total aerobic mesophiles, psychrotrophes, and yeasts and molds) and sensory quality of Andean blackberries
Cold storage harvested at two maturity stages and stored under room (18 ± 2 °C) and cold storage (8 ± 1 °C) was studied.
Polyphenols
Total phenolic compounds, anthocyanins and organic acids content, and antioxidant activity were also eval-
Postharvest quality
uated. The more mature fruit was classified as “big”, according to the Ecuadorian Standard and showed lower
acidity and higher total soluble solids, anthocyanins content and sensory scores compared with the fruit har-
vested earlier, whilst maturity at harvest did not affect the microbial counts of any of the groups studied. Cold
storage was effective in delaying weight loss, softening and microbial growth and also in maintaining a better
sensory quality of the blackberries. What’s more, under refrigeration it was possible to extend the shelf-life of the
fruit to up to 8 d. The main limiting factors for shelf-life were microbial growth and loss of firmness at room
storage and cold storage, respectively. Based on these results, it would be advisable to harvest the fruit at
maturity stage 5 in order to achieve an appropriate fruit size, a high anthocyanin concentration, a better sugars/
acids equilibrium, and a better sensory quality and the fruit should be maintained under refrigerated storage.

1. Introduction chemical changes like accumulation of sugars, reduction in organic

Andean blackberries (Rubus glaucus Benth) are native to the Andes
and grow year round, mainly in cold and temperate climates in South
America (Martínez et al., 2007). The fruit are consumed fresh or pro-
cessed as pulp, juices, jams or desserts and are appreciated for their
color, flavor, bioactive compounds, and nutritional attributes, due to
their high content in polyphenols, anthocyanins, vitamins, and minerals
(Garzón et al., 2009; Junqueira-Gonçalves et al., 2016).
Blackberries are non-climateric fruit and though, they must be
harvested at full maturity, usually when they present a bright, dark
purple/black color and optimum firmness (Bejarano, 1992). At this
stage, also the best flavor quality is obtained (Skrovankova et al., 2015).
Maturity at harvest is one of the main contributing and determining
factors to small fruit quality during transport and commercialization,
due to its direct relation with postharvest shelf-life (Perkins-Veazie and
Collins, 2002). Immature fruit will not reach appropriate organoleptic
characteristics whilst shelf-life of over-mature fruit is generally very
short as the susceptibility to decay also increases (García, 2001).
During ripening, berries soften and become darker in color, and also
acids, and an increase in anthocyanins content occur (Mikulic-
Petkovsek et al., 2015). Different maturity indexes (color, firmness,
total soluble solids, and days after full bloom, among others) are
commonly used in order to determine the optimum maturity stage for
fruit and vegetables harvest. In the case of blackberries, the Ecuadorian
Quality Standard NTE-2427 (INEN, 2010) establishes that harvest can
be done once the fruit reaches, at minimum, the maturity stage 3, de-
fined as light red. However, Ayala-Sánchez et al. (2013) concluded that
the berries should be harvested at stage 5, as at that stage, the fruit
presented an adequate size, the characteristic shape, and appropriate
acids and soluble solids content. In Ecuador, there are no accurate
methods for determining when to pick the berry fruit and this can result
in inappropriate quality of marketable product (Mikulic-Petkovsek
et al., 2015). What’s more, due to agro-climatic conditions, fruit quality
may differ in fruit harvested in different locations, even if their external
color is similar.
In addition, most of the research done on this fruit is focused on
yield and quality comparisons among cultivars, and on fruit quality and
determination of chemical compounds and antioxidant activity under

⁎
Corresponding author.
E-mail address: sandra.horvitz@unavarra.es (S. Horvitz).

http://dx.doi.org/10.1016/j.scienta.2017.09.002
Received 2 March 2017; Received in revised form 30 August 2017; Accepted 1 September 2017
0304-4238/ © 2017 Elsevier B.V. All rights reserved.
S. Horvitz et al.
different storage conditions whilst less attention has been paid to ma-
turity stage at harvest and its relation with quality and commercial
requirements (Ayala-Sánchez et al., 2013).
Another problem with blackberries is that they are highly perishable
fruits, with a usual shelf-life that does not exceed 2–3 d at refrigerated
temperatures, what implies great limitations for fresh market and sto-
rage. In effect, blackberries have a very thin and fragile skin and thus,
are very susceptible to water loss, softening, mechanical injuries and
postharvest diseases like gray mold, caused by Botrytis cinerea, and
Rhizopus rot (Junqueira-Gonçalves et al., 2016).
Hence, the use of techniques that can improve shelf-life duration
without altering the physical, sensorial, and nutritional characteristics
of the fruit is of vital importance to reduce the postharvest losses, which
in some cases can be as high as 60% of the production (Sora et al.,
2006). In this sense, one of the main factors affecting the storage shelf-
life and quality of fruit and vegetables is temperature, as it regulates the
rate of all the metabolic processes that occur in these products. Low
temperatures slow down fungal growth and at the same time, reduce
respiration rate and water loss and therefore, delay ripening and se-
nescence processes (Oliveira et al., 2013a, 2013b). As blackberries are
insensitive to chilling injury, the recommended optimum temperature
for this fruit is at 0–5 °C (Joo et al., 2011).
The aim of this work was to characterize the physicochemical, mi-
crobiological and sensory quality, and the postharvest performance of
Andean blackberries, harvested at two maturity stages and stored under
room temperature and refrigerated storage. Bioactive compounds (total
polyphenols, individual anthocyanins), organic acids and antioxidant
activity were also studied.
2. Materials and methods
2.1. Plant material
Andean blackberries (Rubus glaucus Benth) were hand-harvested in
Tungurahua Province, Ecuador at maturity stages 3 (light red) and 5
(dark purple) in January and July 2016. Maturity stages were defined
by the external color of the fruit, according to the Ecuadorian Standard
(NTE-INEN 2427 (INEN, 2010)). Immediately after harvest the fruit
were transported to the Technical University of Ambato for analyses.
Fruit of uniform size and color, and free from damage and injuries were
selected for the study.
2.2. Packing and storage
200 ± 10 g of blackberries were packaged in transparent poly-
ethylene terephthalate (PET) plastic containers and stored at
18 ± 2 °C (RT) and under refrigerated storage, at 8 ± 1 °C.
2.3. Quality attribute analyses
Quality parameters were evaluated on the harvest day and every 3 d
during the storage period, with the exception of fruit size that was
determined only on day 0.
2.3.1. Physicochemical attributes
2.3.1.1. Fruit size and mass loss. The individual weight (g), the diameter
(mm), and the length (mm) of 30 fruit were measured using an
analytical balance (Radwag AS 310.R2, Poland) and a caliper,
respectively. For mass loss, three trays were randomly selected and
individually weighed at the beginning of the experiment, and on a daily
basis during the storage period, until decay symptoms were visually
observed. Results were expressed as percentage of mass loss relative to
the initial mass.
2.3.1.2. Color and firmness. Color was determined with a HunterLab
spectrophotometer (MINISCAN EZ 4500S, HunterLab, USA) calibrated
294
Scientia Horticulturae 226 (2017) 293–301
to a standard white tile (X 79.80, Y 84.97, Z 90.74) and using the
CIELab color space (CIE, 1976), the illuminant D65 and the 10°
observer as a reference. The measurements were done on two
opposite points of the equatorial zone and on the basis of each fruit
and the hue angle was calculated. Flesh firmness was measured with a
Texture Analyzer (Model CT3, Brookfield Engineering Labs, Inc., USA)
and the TexturePro CT V1.2 Build Software. Each fruit was punctured
once in its equatorial zone with a stainless steel, flat plunger (3 mm
diameter) and the results were expressed as the maximum force (N)
necessary to break the flesh in each fruit.
2.3.1.3. pH, total soluble solids (TSS) and titratable acidity (TA). These
parameters were determined on the juice obtained from each sample.
The pH of the blackberries was measured with a pH-meter (ORION
VersaStar, Thermo Scientific, Denmark). TSS were determined with a
hand refractometer (Atago, Japan) and titratable acidity was
determined by titration with 0.1 N NaOH to pH 8.1 (AOAC, 2000)
and expressed as g citric acid 100 g−1 fruit (fresh weight).
2.3.2. Extraction and determination of organic acids
A 2-g sample of lyophilized fruit was homogenized with 20 mL of
distilled water and shaken for 30 min at 100 rpm in a bench-top orbital
shaker (Infors HT, Switzerland). Afterwards, the solution was cen-
trifuged (15 min, 2100g) at room temperature using a Medifriger BL-S
centrifuge (J.P. Selecta, Spain). The extraction process was repeated
twice, the supernatants were combined and the volume was made up to
50 mL with distilled water. Citric, malic, and ascorbic acids were de-
termined by HPLC, following the method described by Scherer et al.
(2012), with some modifications. For this purpose, a liquid chromato-
graph Waters 2695 coupled to a Waters 996 PDA detector and a
LiChroCART Purospher RP-18 column (250 mm × 4 mm, 5 μm, Merck,
Germany), with a Purospher precolumn (4 mm × 4 mm, 5 μm) were
employed.
The extracts (20 μL) were injected into the chromatograph after
filtration with 0.45 μm PVDF filters (Análisis Vínicos S.L., Spain). An
isocratic elution procedure was used with a 0.01 M KH2PO4 buffer so-
lution, at a flow rate of 0.8 mL per minute, with the column kept at
25 °C. The peaks were identified by comparison of their retention times
and UV-VIS spectra with those of standards. An external standard curve
of each organic acid was used for quantification, at 250 nm for ascorbic
acid, and at 210 nm for malic and citric acids.
2.3.3. Extraction and determination of anthocyanins, total polyphenols and
antioxidant activity
2.3.3.1. Extraction. 1-g of lyophilized fruit was homogenized with
20 mL of ethanol:distilled water:formic acid (50:48:2; v:v:v) and
shaken for 30 min at 100 rpm in a bench-top orbital shaker (Infors
HT, Switzerland). Afterwards, the solution was centrifuged (15 min,
2100g) at room temperature using a Medifriger BL-S centrifuge (J.P.
Selecta, Spain). The extraction process was repeated twice, the
supernatants were combined and the volume was made up to 50 mL
with ethanol:distilled water (v:v).
2.3.3.2. Anthocyanins. Anthocyanins were determined by HPLC
following the method of Vasco et al. (2009), with some
modifications. The equipment and column were the same previously
described for the analysis of organic acids. Mobile phases were 5%
formic acid in water (A) and methanol (B). The flow rate was
0.8 mL min−1. The column was kept at 40 °C. The proportion of
mobile phase B was progressively incremented through the following
elution gradient: 5–10% in 5 min, 10% during 10 min, 10–42% in
20 min, and return to the initial conditions in 5 min. The extracts
(20 μL) were injected into the chromatograph after filtration with
0.45 μm PVDF filters (Análisis Vínicos S.L., Spain). Detection was
made within 250 and 550 nm. The different anthocyanin compounds
detected were quantified using an external standard curve with
S. Horvitz et al.
cyanidin-3-rutinoside (Extrasynthese, France), which is the main
anthocyanin compound in Rubus glaucus blackberries.
2.3.3.3. Total phenolic content (TPC). TPC was determined with the
Folin–Ciocalteu colorimetric method modified for measuring in 96-well
microplates (Bobo-García et al., 2015). Absorbance of the samples was
measured at 750 nm, using a Multiskan GO spectrophotometer (Thermo
Fisher Scientific, Denmark) and the results were expressed as mg gallic
acid equivalent 100 g−1 on a fresh weight basis.
2.3.3.4. Antioxidant activity. Antioxidant activity was evaluated by the
2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, following the method
described by Bobo-García et al. (2015) for microplates. The
absorbance of the samples was measured at 515 nm, using a
Multiskan GO spectrophotometer (Thermo Fisher Scientific, Denmark)
and the results were expressed as μmol Trolox equivalent 100 g−1 on a
fresh weight basis.
2.3.4. Microbiological analyses
Microbiological analyses were performed on days 0, 3, 6, and 9 of
storage. Also, the containers were visually controlled daily in order to
detect symptoms of microbial development. At each evaluation date, 3
samples/treatment were analyzed for total aerobic mesophiles, psy-
chrotrophes, and yeasts and molds. For the analyses, 10 g of fruit were
aseptically transferred to a filter stomacher bag and homogenized in
90 mL sterile buffered peptone water (Difco, USA) for 120 s at 200 rpm,
using a Stomacher 400 circulator (Seward, UK). Serial decimal dilutions
of each homogenized sample were made in peptone water. From each
dilution, 1-mL aliquots were aseptically pour-plated for mesophiles and
psychrothrophes and 0.1 mL was surface-plated for molds and yeasts
analyses. The following media and culture conditions were used: (1)
Plate count agar (PCA, Difco, USA) incubated at 35 ± 2 °C for 48 h and
at 7 °C for 7 days, for total mesophilic (FDA, 1995) and psychrotrophic
(ICMSF, 1982) microorganisms, respectively and (2) Sabouraud-glu-
cose-agar plus chloramphenicol media (Acumedia, USA) incubated at
25 °C ± 2 for 5 days for yeasts and molds. All the samples were ana-
lyzed in duplicate, and microbial counts were expressed as log10
(cfu g−1) of tissue.
2.3.5. Sensory analyses
During storage, three samples per treatment were evaluated on days
0, 3, 6, and 9 by a sensory panel (5 males and 5 females) using a de-
scriptive test. Before the experiments, the panelists were familiarized
with the product and scoring methods, in training sessions in which
appropriate scores for each parameter were agreed. The analyses were
carried out in individual booths, and the samples from the different
treatments (blackberries harvested at different maturity stages and
stored at different temperatures) were presented in groups, coded with
random numbers. Visual quality (color uniformity, injuries and general
appearance) and overall impression were rated from 1 (worst) to 7 (best
quality). Firmness was evaluated after softly pressuring the fruit be-
tween the thumb and the index fingers, from 1 (very soft) to 7 (very
firm). For characteristic aroma, the scale ranged from 1 (none) to 7
(fully typical aroma) and for fruitś color evaluation a color chart (NTE
2427, INEN, 2010) that classifies the fruit into 7 classes, from 0 (light
green) to 6 (dark purple/black) was used. Panelists were also asked to
characterize off-odors, in case they detected any, and scores below 4 in
any of the attributes indicated the rejection of the product.
The shelf-life of the fruit was established as 1 d before the appear-
ance of symptoms of microbial growth, sensorial rejection or a combi-
nation of both.
2.3.6. Statistical analyses
The experiment was repeated twice (6 months apart between har-
vests) and the analyses were done in triplicate, considering each con-
tainer as the experimental unit. For color and firmness, 10 subsamples
295
Scientia Horticulturae 226 (2017) 293–301
were taken, totaling 30 measurements for each of these parameters.
Data were subjected to a one-way analysis of variance (α = 0.05) using
the IBM SPSS Statistics Version 21 software (IBM Corporation, USA).
When significant differences were observed, mean treatments were
compared using Tukey’s test.
3. Results and discussion
As no significant differences were observed between the two har-
vests (January and July 2016) for any of the parameters studied, the
data were pooled. The evaluations were continued until symptoms of
decay were detected on the fruit. So, storage period was of 3 and 9 d, at
18 and 8 °C, respectively.
3.1. Physicochemical attributes
3.1.1. Fruit size
Fruit size increased with ripening. The fruit harvested at stage 5
reached an average weight of 6.25 ± 1.49 g, and 21.8 ± 0.22 and
26.6 ± 0.32 mm diameter and length, respectively, which corre-
sponded to the category “big”, according to the Ecuadorian Standard.
The fruit harvested at maturity stage 3 was classified as “medium”, with
an average weight of 4.46 ± 1.22 g and a diameter and length of
18.9 ± 0.23 and 23.5 ± 0.28 mm, respectively. Consumers associate
larger fruit with better quality and are usually willing to pay more for
this kind of fruit. So, by harvesting when the fruit reaches stage 3, the
fruit obtained could be unacceptable for the market.
3.1.2. Mass loss
Mass loss increased constantly during storage in the blackberries
from both maturity stages, with maximum values of around 9% after 10
d of refrigerated storage and 4–5% after 3 d at RT. No significant dif-
ferences were observed between the mass loss of the more mature and
the more immature blackberries but, at every evaluation date, the mass
loss was lower in the fruit stored at 8 °C in comparison with the fruit
stored at RT. These results are similar to those reported by Ayala-
Sánchez et al. (2013) and Kim et al. (2015), who attributed the greater
mass loss at RT to higher respiration and transpiration rates of the fruit
under these conditions. The maximum admissible mass loss for black-
berries marketing has been reported as 5% (Salgado and Clark, 2016).
This limit was exceeded after 3 d at RT and after 6 d under refrigerated
storage. In effect, the use of low temperatures reduces metabolic pro-
cesses and retards microbial growth which results in better fruit quality
maintenance and longer shelf-life of the fruit (Joo et al., 2011).
3.1.3. Color and firmness
Regardless of storage temperature and maturity stage at harvest, the
°hue of the blackberries remained stable, with slight but no significant
decreases throughout the storage period. The hue of the fruit harvested
at maturity stages 3 and 5 was of 23.75 ± 5.01 (light red) and
11.67 ± 6.20 (dark red), respectively. In this sense, it should be con-
sidered that a great variability in the superficial color of the black-
berries was found, which in turn, may mask differences among the
evaluation dates and storage temperatures. The luminosity (L*) was
significantly higher in the more immature fruit (25.02 ± 2.89) in
comparison with the blackberries harvested at a more advanced ma-
turity stage (17.62 ± 2.17). This parameter remained unchanged in all
the treatments, except the most mature fruit, which showed a sig-
nificant decrease in the L* value after 9 days of cold storage, which
indicates a loss of brightness in the fruit, associated with senescence.
At each evaluation date, the fruit harvested at maturity stage 3 was
firmer than the blackberries harvested at maturity stage 5. During the
storage period, a significant loss of firmness was registered in all the
samples and for both maturity stages (Fig. 1A and B), the softening was
greater in the blackberries stored at RT in comparison with cold sto-
rage.
S. Horvitz et al. Scientia Horticulturae 226 (2017) 293–301

Fig. 1. Firmness evolution of blackberries harvested at maturity

stages 3 (A) and 5 (B) during 3 and 9 days of storage at 18 ± 2 °C and
8 ± 1 °C, respectively. Values represent the mean of 60 measure-
ments for each storage temperature and evaluation date. Error bars
represent the confidence interval (95%) for the mean.

Our results are similar to those obtained by Kim et al. (2015) and
Oliveira et al. (2014). These authors reported a decrease in firmness
with maturation process and attributed the softening to the physiolo-
gical and biochemical changes that occur during ripening like starch to
sugars conversion, biosynthesis of volatiles responsible for odor and
taste, and changes in cell wall structure, due to the breakdown of cel-
lular substances such as pectin, cellulose, hemicellulose, and other
polysaccharides through hydration.
3.1.4. pH, total soluble solids and titratable acidity
The pH, total soluble solids content, and titratable acidity of the
blackberries harvested at two maturity stages and stored at room
temperature and cold storage are shown in Table 1. The more mature
fruit presented higher TSS, lower acidity and similar pH than the fruit
from stage 3 and these differences could be attributed to acid to sugar
conversion during ripening (Ayala-Sánchez et al., 2013). The three
parameters remained unchanged during storage, either at RT or re-
frigerated storage and the values were similar to those reported by
Reyes-Carmona et al. (2005) and Carvalho and Betancur (2015) for
different blackberries cultivars harvested in different locations and at
different maturity stages.
3.2. Organic acids
Together with sugars, organic acids are responsible for blackberries’
flavor and they also have positive effects on human diet and health due
to their antioxidant activity against free radicals (Mikulic-Petkovsek
et al., 2015). Citric acid was the predominant acid found in the fruit,
representing nearly 90% of total organic acids, followed by malic and
ascorbic acids (Table 2). These results are similar to those reported by
Mikulic-Petkovsek et al. (2015) in different berry species. On the con-
trary, Kafkas et al. (2006) found malic as the main acid, followed by
ascorbic whilst citric acid was not detected in any of the cultivars stu-
died. In this sense, the main organic acid and the vitamin C content in
these fruit varies considerably among cultivars, ripeness, growing
conditions and light intensity, and day/night temperatures (Van de
Velde et al., 2016). The concentration of organic acids was higher than
those previously reported by Kafkas et al. (2006) and Guedes et al.
(2013) in different blackberry cultivars and are among the ranges of
5–30 mg 100 g−1 FW for ascorbic acid (Skrovankova et al., 2015) and
87.5–603 and 569–1892 mg 100 g−1 FW for malic and citric acids,
respectively (Fan-Chiang, 1999; Gazioglu-Sensoy et al., 2015). What’s
more, to the best of our knowledge, there are no previous reports on
organic acids content in Andean blackberries.
During storage, some differences between the behaviour of the in-
dividual acids studied were observed. Ascorbic acid content decreased,
regardless of maturity stage at harvest and storage temperature.
However, the losses were greater in the more mature fruit and in the
blackberries stored at RT. After 9 d of cold storage, the citric acid
presented and increase of 28.5% in the fruit harvested at stage maturity
3 and of 114.51% in the fruit harvested at stage maturity 5. In the
blackberries stored at room temperature, there was also an increase of

Table 1
pH, soluble solids content and titratable acidity evolution of blackberries harvested at maturity stages 3 and 5 and stored at room temperature (18 ± 2 °C) and in cold storage (8 ± 1 °C)
during 3 and 9 d, respectively.

pH Total Soluble Solids (%) Titratable aciditya

T (ªC) Day MATURITY STAGE

3 5 3 5 3 5

8 0 2.29 ± 0.05A 2.79 ± 0.34B 9.63 ± 0.69A 11.00 ± 1.67B 3.80 ± 0.05A 2.78 ± 0.21B
3 2.62 ± 0.38A 2.87 ± 0.42A 9.17 ± 0,23A 10.60 ± 0.88B 3.72 ± 0.34A 2.57 ± 0.22B
6 2.63 ± 0.40A 2.92 ± 0.45A 9.33 ± 0.61A 11.50 ± 0.84B 3.69 ± 0.13A 2.58 ± 0.16B
9 2.68 ± 0.41A 2.92 ± 0.38A 9.22 ± 0.39A 12.78 ± 1.96B 3.64 ± 0.20A 2.58 ± 0.16B
18 0 2.29 ± 0.05A 2.79 ± 0.34B 9.63 ± 0.69A 11.00 ± 1.67B 3.80 ± 0.05A 2.78 ± 0.21B
3 2.66 ± 0.43A 2.90 ± 0.49A 9.70 ± 0.40A 9.93 ± 1.17B 3.63 ± 0.23A 2.59 ± 0.19B

Values are the mean ± standard deviation (n = 6). For each storage temperature and evaluation date, different capital letters indicate significant differences between the maturity stages
(p < 0.05).
a
g citric acid 100 g−1 on a fresh weight basis.

296
S. Horvitz et al. Scientia Horticulturae 226 (2017) 293–301

Table 2
Ascorbic, malic and citric acids content in blackberries harvested at two maturity stages and stored under room temperature (18 ± 2 °C) and refrigerated (8 ± 1 °C) storage during 3
and 9 d, respectively.

ORGANIC ACIDS (mg 100 g−1)a

Ascorbic Malic Citric

T (ªC) Day MATURITY STAGE

3 5 3 5 3 5

8 0 8.24 ± 0.50Aa 11.62 ± 3.65Ba 478.10 ± 35.69Aa 295.73 ± 021.04Ba 1004.26 ± 76.91Aa 1739.52 ± 251.29Ba
3 8.42 ± 1.83Aa 8.25 ± 1.04Aab 385.18 ± 52.31Aab 145.43 ± 030.54Ba 1110.66 ± 273.95Aab 1993.13 ± 156.67Aa
6 6.63 ± 1.51Aab 7.72 ± 1.38Ab 340.55 ± 86.44Ab 180.43 ± 075.09Bab 1278.02 ± 223.21Ab 1040.60 ± 102.13Ba
9 5.78 ± 1.46Ab 5.17 ± 1.79Ab 294.09 ± 72.56Ab 263.44 ± 084.44Ab 1290.27 ± 75.56Ab 1586.40 ± 422.17Ab
18 0 8.24 ± 0.50Aa 11.62 ± 3.65Ba 478.10 ± 35.69Aa 295.73 ± 021.04Ba 1004.26 ± 76.91Aa 1739.52 ± 251.29Ba
3 7.38 ± 1.53Aa 7.33 ± 2.78Ab 187.55 ± 7.92Ab 207.18 ± 108.17Ab 1388.49 ± 18.72Ab 1084.45 ± 170.46Bb

Values are the mean ± standard deviation (n = 6). For each storage temperature and evaluation date, different capital letters indicate significant differences between the maturity stages
(p < 0.05). For each storage temperature and maturity stage, different lower case letters indicate significant differences among evaluation dates (p < 0.05).
a
Fresh weight basis.

this acid, of 38.3% and 46.6% in the more immature and more mature
blackberries, respectively. Finally, regardless of storage temperature, a
significant decrease and increase in malic acid content was observed in
the fruit harvested at stage maturity 3 and 5, respectively. Several
studies were found reporting the organic acids content in different
blackberries cultivars at harvest. However, no references were found
regarding the behaviour of these acids during the storage period of this
fruit.
3.3. Anthocyanins, total polyphenol content and antioxidant activity
HPLC-DAD analysis revealed three major anthocyanin compounds
in the blackberries (peaks 1, 2 and 3, Fig. 2). Most of the anthocyanins
previously found in Andean blackberries were cyanidin glycosides
(Vasco et al., 2009). Peak 3, identified as cyanidin-3-rutinoside was
largely the most important anthocyanin, with a relative content of
62–65%. This is an outstanding characteristic of Rubus glaucus
(Arozarena et al., 2012; Garzón et al., 2009; Mertz et al., 2007) that
distinguishes it from other blackberry cultivars, in which cyanidin-3-
glucoside usually prevails. According to the relative retention times and
the spectral characteristics of peaks 1 and 2, they could presumably
correspond to the cyanidin-3-glucoside and the cyanidin-3-
xilorutinoside, respectively (Arozarena et al., 2012; Garzón et al.,
2009).
Fig. 2 shows how the relative content of the three compounds
changed during maturation, particularly in the case of the peaks 1 and
2. While peak 1 increased from 17 at stage 3–26% at stage 5, peak 2
decreased from 18 to 10%. The relative amount of cyanidin-3-rutino-
side also diminished, but in a lower extent, from 65 to 62%. Besides
this, two additional peaks (4 and 5) were detected in the more mature
blackberries. They accounted only the 2% of the total anthocyanin
content. Both peaks had a maximum absorbance wavelength around
500 and 505 nm and a pronounced shoulder in the 400–450 nm region.
These spectral characteristics have been noted to be typical of pe-
largonidin derivatives, which have been also detected in Andean
blackberries (Garzón et al., 2009).
Although the effect of maturation on the relative composition of
anthocyanins was significant, its effect on the absolute content was
more remarkable (Table 3). Blackberries harvested at stage 5 had a total
anthocyanin content (TAC) around 5 times higher than the blackberries
at stage 3. The concentration of anthocyanins at stage 5 was consistent
with values observed in blackberries from different geographical loca-
tions (Probst, 2015). The production and accumulation of anthocyanins
during ripening have been observed in different berry fruit

Fig. 2. Chromatograms at 520 nm of anthocyanin

extracts of Andean blackberries harvested at ma-
turity stages 3 and 5. Retention times, maximum
absorption wavelengths and relative amounts of the
peaks detected.

297
S. Horvitz et al. Scientia Horticulturae 226 (2017) 293–301

Table 3
Total anthocyanin and cyanidin-3-rutinoside content evolution of blackberries harvested at maturity stages 3 and 5 and stored at room temperature (18 ± 2 °C) and in cold storage
(8 ± 1 °C) during 3 and 9 days, respectively.

Cyanidin 3-rutinoside (mg 100 g−1)a Total Anthocyanins (mg 100 g−1)a

T (°C) Day MATURITY STAGE

3 5 3 5

8 0 20.3 ± 4.4 Aa 100.0 ± 18.5 Ba 30.8 ± 6.3 Aa 175.7 ± 36.9 Ba

3 31.0 ± 2.1 Ab 110.7 ± 10.1 Bab 46.2 ± 2.8 Ab 174.9 ± 14.8 Ba
6 45.5 ± 5.0 Ac 125.1 ± 17.1 Bbc 67.1 ± 6.1 Ac 195.7 ± 27.4 Bab
9 41.9 ± 8.8 Ac 143.9 ± 9.0 Bc 64.5 ± 13.0 Ac 228.0 ± 14.7 Bb
18 0 20.3 ± 4.4 Aa 100.0 ± 18.5 Ba 30.8 ± 6.3 Aa 175.7 ± 36.9 Ba
3 36.9 ± 9.5 Ab 84.8 ± 44.8 Ba 61.2 ± 19.1 Ab 137.1 ± 75.4 Ba

Values are the mean ± standard deviation (n = 6). For each temperature and evaluation date, different capital letters indicate significant differences between the blackberries at the two
maturity stages (p < 0.05). For each maturity stage and storage temperature, different lower case letters indicate significant differences among days (p < 0.05). For each maturity stage
and evaluation date no differences were detected between blackberries at different temperatures.
a
Fresh weight basis.

(Skrovankova et al., 2015). In the case of blackberries, increments of
2–4 (Siriwoharn et al., 2004) to 7-fold (Acosta-Montoya et al., 2010) of
anthocyanin concentration have been reported. During the 9 d of cold
storage, TAC significantly augmented, regardless the maturation stage
of the fruit (Table 3). Previous results regarding the behavior of an-
thocyanin compounds of blackberries during refrigerated storage are
not conclusive. Kim et al. (2015) observed that TAC increased after 15 d
storage at 1 °C, or after 13 d at 1 °C plus 2 d at 20 °C. On the contrary,
Joo et al. (2011) described a decline after 18 d at 3 °C, while Wu et al.
(2010) did not see a clear tendency in the evolution of anthocyanins
during 7 d at 2 °C.
Table 4 shows the total phenolic content (TPC) and antioxidant
activity (AA) of the blackberries. TPC values are comparable to values
previously described for blackberries (Probst, 2015). The comparison of
AA results more difficult due to the great variety of analytical methods
and expression units founded in the literature. TPC and AA evolved in a
similar way, and were significantly correlated (r = 0,891). The ma-
turation process did not have a strong influence on them, although
slightly lower values were observed in the more mature blackberries, in
contrast to what happened with anthocyanins. However, it must be
noted that no significant correlation was observed between TAC and
AC. Although anthocyanins are antioxidants, AA could be more related
to other phenolic compounds. It has been shown that ellagitannins and
ellagic acid derivatives are largely the main phenolic compounds in
Andean blackberries (Rubus glaucus Benth.), followed by anthocyanins
and, in a minor extent by other phenolic families, such as flavonols,
flavanols and phenolic acids (Mertz et al., 2007; Vasco et al., 2009).
Acosta-Montoya et al. (2010) studied the phenolic composition of tro-
pical highland blackberries (Rubus adenotrichus Schltdl.) during three
maturity stages, and observed that the strong increment of the antho-
cyanin content was accompanied by a decline on the levels in ellagi-
tannins and flavonols, with the consequence that the total phenolic
content did not vary in a specific way. So, the small effect of maturation
on the TPC and AA levels in our Andean blackberries could be ex-
plained through the same mechanism. The accumulation of anthocya-
nins during ripening would have been counteracted by the fall in el-
lagitannins.
No changes were observed either on the TPC or the AA levels during
storage (Table 4), regardless the maturity stage of the fruit and the
temperature used. Previous results are inconclusive. Whilst Perkins-
Veazie and Kalt (2002) pointed that storage at 2 °C had no influence on
AA of blackberries, Kim et al. (2015) observed a significant increment
on the phenolic content of blackberries after 15 d at 1 °C, or after 13 d
at 1 °C plus 2 d at 20 °C and Wu et al. (2010) showed different evolution
patterns on the anthocyanin content, TPC and AA depending on the
blackberry cultivar considered. These authors speculated that this could
be related to the different phenolic composition, the polyphenol oxidase
activity, and the ripeness of the fruit.
3.4. Microbiological analyses
Yeasts and moulds were the main microbial group causing fruit
decay. In effect, the high water and sugar content and the low pH of the
fruit may limit the growth of many bacteria and at the same time, en-
hance fungal growth (Oliveira et al., 2013a, 2013b).
The microbial counts observed on day 0 ranged between 4.86 and
5.08 (mesophiles), 0.00 and 1.35 (psychrotrophe), and 4.91 and 4.58
(yeasts and moulds) log (cfu g−1) for the blackberries harvested at

Table 4
Total polyphenol content and antioxidant activity of blackberries harvested at maturity stages 3 and 5 and stored at room temperature (18 ± 2 °C) and in cold storage (8 ± 1 °C) during
3 and 9 days, respectively.

Total polyphenol content (mg galic acid 100 g−1)a Antioxidant activity (μmol Trolox 100 g−1)a

T(°C) Day MATURITY STAGE

3 5 3 5

8 0 560 ± 38 Ba 446 ± 27 Aa 6009 ± 408 Ba 5264 ± 318 Aa

3 517 ± 34 Aa 502 ± 32 Aa 5756 ± 377 Aa 5170 ± 328 Aa
6 618 ± 41 Aa 555 ± 36 Aa 6597 ± 441 Aa 5490 ± 354 Aa
9 736 ± 49 Ba 558 ± 34 Aa 6956 ± 472 Ba 5846 ± 355 Aa
18 0 560 ± 38 Ba 446 ± 27 Aa 6009 ± 408 Ba 5264 ± 318 Aa
3 613 ± 40 Aa 585 ± 37 Aa 6586 ± 435 Ba 5594 ± 355 Aa

Values are the mean ± standard deviation (n = 6). For each temperature and evaluation date, different capital letters indicate significant differences between the blackberries at the two
maturity stages (p < 0.05). For each maturity stage and storage temperature, different lower case letters indicate significant differences among days (p < 0.05). For each maturity stage
and evaluation date no differences were detected between blackberries stored at different temperatures.
a
Fresh weight basis.

298
S. Horvitz et al. Scientia Horticulturae 226 (2017) 293–301

Table 5
Microbial growth on Andean blackberries harvested at two maturity stages and stored at room temperature (18 ± 2 °C) and under refrigeration (8 ± 1 °C), during 3 and 9 days,
respectively.

Mesophiles Psychrotrophes Moulds and yeasts

Maturity stage Day STORAGE TEMPERATURE (°C)

8 18 8 18 8 18

3 0 4.86 ± 0.37 Aa 4.86 ± 0.37 Aa 0.00 ± 0.00 Aa 0.00 ± 0.00 Aa 4.91 ± 0.15 Aa 4.91 ± 0.15 Aa
3 4.14 ± 1.11 Ab 5.59 ± 0.42 Bb 0.00 ± 0.00 Aa 3.62 ± 0.47 Bb 4.94 ± 0.43 Aa 6.27 ± 0.41 Bb
6 3.80 ± 0.54 b 1.94 ± 1.44 b 5.17 ± 0.45 a
9 3.69 ± 0.32 b 3.25 ± 0.42 c 5.17 ± 0.70 a
5 0 5.08 ± 0.16 Aa 5.08 ± 0.16 Aa 1.35 ± 1.46 Aa 1.35 ± 1.46 Aa 4.58 ± 0.59 Aa 4.58 ± 0.59 Aa
3 4.23 ± 0.46 Ab 5.24 ± 0.23 Ba 1.45 ± 1.53 Aa 2.60 ± 1.92 Aa 4.66 ± 2.18 Aa 6.96 ± 0.56 Bb
6 3.85 ± 0.12 c 1.87 ± 1.40 a 4.85 ± 0.13 a
9 2.75 ± 0.22 d 2.06 ± 1.52 a 5.06 ± 0.13 a

Values are the mean ± standard deviation (n = 6). For each maturity stage and evaluation date, different capital letters indicate significant differences between the storage temperatures
(p < 0.05). For each maturity stage and storage temperature, different lower case letters indicate significant differences among evaluation dates (p < 0.05).

maturity stages 3 and 5, respectively (Table 5). Regardless of maturity

stage at harvest, an increase in the microbial counts of all the groups
studied was observed during storage at RT. On the contrary, under
refrigerated storage, mesophiles’ counts decreased, yeasts and moulds
remained unchanged and psychrotrophe increased only in the more
immature fruit after 6 d of storage with no significant changes in the
fruit harvested at maturity stage 5 (Table 5). What’s more, on day 3, all
the microbial counts were higher in the fruit stored at room tempera-
ture in comparison with the fruit stored at 8 °C. These results are similar
to those of Kim et al. (2015) and de Arruda Palharini et al. (2015), who
reported that cooling was an efficient technique to reduce microbial
growth. What’s more, handling the fruit carefully, together with rapid
cooling after harvest is essential to reduce fruit damage and delay the
growth of microorganisms like Botrytis, Cladosporium, Penicillium, Al-
ternaria, and Fusarium (Perkins-Veazie et al., 1999).

3.5. Sensory analyses

On day 0, the fruit harvested at stage maturity 5 presented higher

scores than the more immature fruit in all the sensory parameters
studied, with the exception of firmness (Fig. 3). However, the firmness
of the fruit harvested at stage 3 was described as “too firm” by the
panelists, who preferred softer fruit.
During the storage period and regardless of harvest maturity and
storage temperature, a progressive decrease in the scores of the visual
quality, firmness and global impression was recorded (Figs. 4 A,B; and 5
A, B). On the contrary, color and aroma remained unchanged in the Fig. 4. Sensory quality attributes of blackberries harvested at maturity stages 3 (A) and 5
(B) and stored during 9 days at 8 ± 1 °C.

more mature fruit whilst in the fruit harvested at stage maturity 3, an

increase and decrease were obtained for color and aroma scores, re-
spectively. However, even at day 9, the scores of these parameters were
below 4, the limit of acceptance established for all the sensory attri-
butes.
Finally, the use of cold storage was effective in maintaining fruit
quality, especially for the most mature fruit. In effect, on day 3, the
blackberries stored under 8 °C presented higher scores for the visual
quality, aroma, firmness and global impression than the blackberries
kept at room temperature.

4. Conclusions

The Ecuadorian Quality Standard establishes stage maturity 3 as the

minimum maturity for blackberries harvest. However, at this stage and
Fig. 3. Initial sensory quality attributes of blackberries harvested at maturity stages 3 and according to the sensory panelists, the fruit was still “too firm”, did not
5.
present the expected dark, bright purple color and did not develop its

299
S. Horvitz et al.
Fig. 5. Sensory quality attributes of blackberries harvested at maturity stages 3 (a) and 5
(b) and stored during 3 days at 18 ± 2 °C.
full typical aroma. What’s more, the titratable acidity doubled the
maximum of 1.8% allowed in the same Standard, and the anthocyanin
content was around five-fold lower than at maturity stage 5. Though, as
fruit quality is influenced by agro-climatic conditions and may differ in
fruit harvested in different locations, even if their external color is si-
milar, it would be advisable to revise and adapt the Standard to the
local conditions.
Refrigerated storage was effective in delaying weight loss, softening
and microbial growth and in maintaining better sensory quality, mainly
of the fruit harvested more mature. At the same time, it did not affect
either the total phenolic content or the antioxidant activity of the fruit.
When cold storage was used it was possible to extend the shelf-life of
the blackberries from 3 d at RT to up to 8 d. The main limiting factors
for shelf-life were microbial growth and loss of firmness at RT and cold
storage, respectively.
Based on these results, it would be advisable to harvest the fruit at
maturity stage 5 in order to achieve an appropriate fruit size, a high
anthocyanin concentration, a better sugars/acids equilibrium, and a
better sensory quality and the fruit should be maintained under re-
frigerated storage.
Acknowledgements
Special thanks to the Secretaría de Educación Superior, Ciencia y
Tecnología e Innovación (SENESCYT) of the Republic of Ecuador for the
financial support of Dr. Horvitz as Prometeo Researcher.
300
Scientia Horticulturae 226 (2017) 293–301
References
AOAC, 2000. Official Methods of Analysis, 17th ed. Association of Official Analytical
Chemists, Washington, USA.
Acosta-Montoya, O., Vaillant, F., Cozzano, S., Mertz, C., Pérez, A.M., Castro, M.V., 2010.
Phenolic content and antioxidant capacity of tropical highland blackberry (Rubus
adenotrichus Schltdl.) during three edible maturity stages. Food Chem. 119,
1497–1501.
Arozarena, I., Ortiz, J., Hermosín-Gutiérrez, I., Urretavizcaya, I., Salvatierra, S., Córdova,
I., Marín-Arroyo, M.R., Noriega, M.J., Navarro, M., 2012. Color, ellagitannins, an-
thocyanins, and antioxidant activity of Andean blackberry (Rubus glaucus Benth.)
wines. J. Agric. Food Chem. 60, 7463–7473.
Ayala-Sánchez, L.C., Valenzuela-Real, C.P., Bohórquez-Perez, Y., 2013. Caracterización
físicoquímica de mora de castilla (Rubus glaucus Benth) en seis estados de madurez.
Biotecnol. Sector Agropecuario Agroind. 11, 10–18.
Bejarano, W., 1992. Promoción de exportaciones agrícolas no tradicionales (PROEXANT).
Manual de Mora (Rubus glaucus Benth). Quito, Ecuador.
Bobo-García, G., Davidov-Pardo, G., Arroqui, C., Vírseda, P., Marín-Arroyo, M.R.,
Navarro, M., 2015. Intra-laboratory validation of microplate methods for total phe-
nolic content and antioxidant activity on polyphenolic extracts, and comparison with
conventional spectrophotometric methods. J. Sci. Food. Agric. 95, 204–209.
CIE, 1976. Official Recommendations on Uniform Color Spaces, Color Difference
Equations and Metric Color Term. Supplement N° 2. CIE Publications N° 15.
Colorimetry Commision Internationals de L’eclairage, Paris.
Carvalho, C.P., Betancur, J.A., 2015. Quality characterization of Andean blackberry fruits
(Rubus glaucus Benth) in different maturity stages in Antioquia, Colombia. Agron.
Colombiana 33 (1), 74–83.
de Arruda Palharini, M.C., Fischer, I.H., Vegian, M.R.d.C., Fileti, M.d.S., Montes,
S.M.N.M., 2015. Efeito da temperatura de armazenamento na conservação pós-col-
heita de amora-preta. Pesquisa Agropecuária Trop. 45, 413–419.
FDA, 1995. Bacteriological Analytical Manual for Food, 8th ed. AOAC International,
Gaithersburg, M.D., USA.
Fan-Chiang, H.J., 1999. Anthocyanin Pigments, Nonvolatile Acid and Sugar Composition
of Blackberries MSc. Thesis. Oregon State University, Corvallis, Oregon 90 pages.
García, M., 2001. La agroindustria de la mora: Alternativas viables para los fruticultores.
Tecnol. Para Agro 1, 15–17.
Garzón, G.A., Riedl, K.M., Schwartz, S.J., 2009. Determination of anthocyanins, total
phenolic content and antioxidant activity in Andes Berry (Rubus glaucus Benth). J.
Food Sci. 74 (3), C227–C232.
Gazioglu-Sensoy, R.I., Gundogdu, M., Sensoy, S., Celik, F., Dogan, A., 2015. HPLC ana-
lysis of blackberry fruits for organic acid and sugar contents. Acta Hortic. 1089,
77–82.
Guedes, M.N.S., Abreu, C.M.P.d., Maro, L.A.C., Pio, R., Abreu, J.R.d., Oliveira, J.O.d.,
2013. Chemical characterization and mineral levels in the fruits of blackberry cul-
tivars grown in a tropical climate at an elevation. Acta Sci.-Agron. 35, 191–196.
ICMSF, 1982. Microorganismos de los alimentos. Su significado y métodos de enumer-
ación, vol. 1 Editorial Acribia, Zaragoza, Spain.
INEN, 2010. Instituto Ecuatoriano de Normalización. INEN 2427. Frutas frescas. Mora.
Requisitos.
Joo, M., Lewandowski, N., Auras, R., Harte, J., Almenar, E., 2011. Comparative shelf life
study of blackberry fruit in bio-based and petroleum-based containers under retail
storage conditions. Food Chem. 126, 1734–1740.
Junqueira-Gonçalves, M.P., Alarcón, E., Niranjan, K., 2016. The efficacy of potassium
sorbate-coated packaging to control postharvest gray mold in raspberries, black-
berries and blueberries. Postharvest Biol. Technol. 111, 205–208.
Kafkas, E., Koşar, M., Türemiş, N., Başer, K.H.C., 2006. Analysis of sugars, organic acids
and vitamin C contents of blackberry genotypes from Turkey. Food Chem. 97 (4),
732–736.
Kim, M.J., Perkins-Veazie, P., Ma, G., Fernandez, G., 2015. Shelf life and changes in
phenolic compounds of organically grown blackberries during refrigerated storage.
Postharvest Biol. Technol. 110, 257–263.
Martínez, A., Beltrán, O., Velastegui, G., Ayala, G., Jácome, R., Yánez, M., Luciano, E.,
2007. Manual del cultivo de la mora de Castilla, first ed. Convenio INIAP-UTA,
Ambato, Ecuador, pp. 9–16.
Mertz, C., Cheynier, V., Günata, Z., Brat, P., 2007. Analysis of phenolic compounds in two
blackberry species (Rubus glaucus and Rubus adenotrichus) by high-performance liquid
chromatography with diode array detection and electrospray ion trap mass spectro-
metry. J. Agric. Food Chem. 55, 8616–8624.
Mikulic-Petkovsek, M., Rescic, J., Schmitzer, V., Stampar, F., Slatnar, A., Koron, D.,
Veberic, R., 2015. Changes in fruit quality parameters of four Ribes species during
ripening. Food Chem. 173, 363–374.
Oliveira, D.M., Rosa, C.I.L.F., Kwiatkowski, A., Clemente, E., 2013a. Biodegradable
coatings on the postharvest of blackberry stored under refrigeration. Rev. Ciencia
Agron. 44, 302–309.
Oliveira, M., Pereira, J., Verde, S.C., Lima, M.G., Pinto, P., Oliveira, P.B.d., Junqueira, C.,
Marcos, H., Silva, T., Melo, R., Santos, C.N., Botelho, M.L., 2013b. Evaluation of
potential of gamma radiation as a conservation treatment for blackberry fruits. J.
Berry Res. 3, 93–102.
Oliveira, D.M., Kwiatkowski, A., Rosa, C.I.L.F., Clemente, E., 2014. Refrigeration and
edible coatings in blackberry (Rubus spp.) conservation. J. Food Sci. Technol. 51 (9),
2120–2126.
Perkins-Veazie, P., Collins, J.K., 2002. Quality of erect-type blackberry fruit after short
intervals of controlled atmosphere storage. Postharvest Biol. Technol. 25, 235–239.
Perkins-Veazie, P., Kalt, W., 2002. Postharvest storage of blackberry fruit does not in-
crease antioxidant levels. Acta Hortic. 585, 521–524.
S. Horvitz et al.
Perkins-Veazie, P., Collins, J.K., Clark, J.R., 1999. Shelf-life and quality of ‘Navaho’ and
‘Shawnee’ blackberry fruit stored under retail storage conditions. J. Food Qual. 22,
535–544.
Probst, Y., 2015. A review of the nutrient composition of selected Rubus berries. Nutr.
Food Sci. 45, 242–254.
Reyes-Carmona, J., Yousef, G.G., Martinez-Peniche, R.A., Lila, M.A., 2005. Antioxidant
capacity of fruits extracts of blackberries (Rubus sp.) produced in different climatic
regions. J. Food Sci. 70, s497–s503.
Salgado, A., Clark, J.R., 2016. Extended evaluation of postharvest quality and shelflife
potential of blackberries. Acta Hortic. 1133, 379–382.
Scherer, R., Rybka, A.C.P., Ballus, C.A., Meinhart, A.D., Filho, J.T., Godoy, H.T., 2012.
Validation of a HPLC method for simultaneous determination of main organic acids in
fruits and juices. Food Chem. 135 (1), 150–154.
Siriwoharn, T., Wrolstad, R.E., Finn, C.E., Pereira, C.B., 2004. Influence of cultivar, ma-
turity, and sampling on blackberry (Rubus L. hybrids) anthocyanins polyphenolics,
and antioxidant properties. J. Agric. Food Chem. 52, 8021–8030.
301
Scientia Horticulturae 226 (2017) 293–301
Skrovankova, S., Sumczynski, D., Mlcek, J., Jurikova, T., Sochor, J., 2015. Bioactive
compounds and antioxidant activity in different types of berries. Int. J. Mol. Sci. 16,
24673–24706.
Sora, A.D., Fischer, G., Flórez, R., 2006. Almacenamiento refrigerado de frutos de mora de
Castilla (Rubus glaucus Benth.) en empaques con atmósfera modificada. Agron.
Colombiana 24, 306–316.
Van de Velde, F., Grace, M.H., Esposito, D., Pirovania, M.E., Lila, M.A., 2016.
Quantitative comparison of phytochemical profile, antioxidant, and anti-in-
flammatory properties of blackberry fruits adapted to Argentina. J. Food Compos.
Anal. 47, 82–91.
Vasco, C., Riñen, K., Ruales, J., Kamal-Eldin, A., 2009. Phenolic compounds in Rosaceae
fruits from Ecuador. J. Agric. Food Chem. 57, 1204–1212.
Wu, R., Frei, B., Kennedy, J.A., Zhao, Y., 2010. Effects of refrigerated storage and pro-
cessing technologies on the bioactive compounds and antioxidant capacities of
‘Marion’ and ‘Evergreen’ blackberries. LWT-Food Sci. Technol. 43, 1253–1264.