Research Article
Zehra Tekin*
Zehra Tekin*
Corresponding Author
Adıyaman University, Faculty of Pharmacy, Department of Basic Pharmaceutical Sciences, Adıyaman, Türkiy
E-mail: ztekin@adiyaman.edu.tr
Fatümetüzzehra Küçükbay
Fatümetüzzehra Küçükbay
İnönü
University, Faculty of Pharmacy, Department of Basic Pharmaceutical Sciences,
Malatya, Türkiye
Abstract
In this article, we have studied the
changes in phytochemical contents and antioxidant properties of blackberry (Rubus caesius L.) fruits naturally grown
in Battalgazi district of Malatya province during the ripening process. It was determined the total phenolic (23.82-151.36 mg GAE/g extract), total flavonoid (5.65-25.72
mg RE/g extract), and total flavonol (1.30-5.40 mg QUE/g extract) contents of
twelve extracts prepared by six different solvents. According to these results, it was concluded that
phytochemical contents were higher in unripe fruit extracts than the ripe
fruits.
The reducing power, chelating capacity
of metal ions and antioxidant activity of each extract were determined using
the DPPH free radical scavenging method. The working conditions in which the
antioxidant activity values were best obtained by the DPPH method were
explained according to each extract extraction method and it was seen that the
best result was the acidified water extract of the unripe fruit (IC50
27.24 ± 0.00 µg/mL).
The results showed that R. caesius L. can be consumed as a
natural antioxidant due to its antioxidant properties. Supporting these results with in vivo studies will
also clarify the health promoting properties of antioxidant activity of R.
caesisus fruit.
Keywords
Dewberry, Rubus caesius L., antioxidant activity, phenolic, flavonoid,
flavonol, Phytochemical.
1. Introduction
The results
of many epidemiological studies conducted to date have shown that long-term and
regular consumption of foods rich in plant polyphenols can protect against the
formation of acute and chronic diseases, especially cancer [1–3]. It has been understood that the
endogenous defense system of the body should be supported by antioxidant
compounds to be taken with a regular and balanced diet [4,
5]. For this reason, it is important to intake antioxidant rich diet. On
the other hand, synthetic polyphenolic antioxidants which butylated hydroxytoluene,
octyl gallate, dodecyl gallate, propyl gallate, tert-butyl hydroquinone,
butylated hydroxyanisole, alpha-, gamma- and delta-tocopherol have been widely
used as food additives [6, 7] but several
reports have indicated high doses of synthetic antioxidant exert negative
health influences such as skin allergies, DNA damage, DNA cleavage,
gastrointestinal tract problems [6, 8], so
the research of natural antioxidants from fruit and vegetable has increasing
attention. Fruits such as berries
contain a variety of compounds with antioxidant activity, including phenolic
acids, flavonoids, vitamin C, anthocyanins, carotenoids, tannins, flavonols and
flavanols [9].
The R.
caesius, Europan dewberry together comprising approximately 700 species of
the Rubus genus belonging to the Rosaceae family [10]. The dewberry is a
perennial shrub reaching between 50-200 cm in height [10-12]
and this species grows on basic soils [31]. R.
caesius L. is native to Europe but is also widely distributed in many
countries worldwide such as Russia, the United States, Turkey, Italy, Spain [11-14].
It was noted
that Rubus species are widely used in traditional and folk medicine in
different regions of the world, as they show a wide distribution area in the
world. In many parts of the world, the use of roots, leaves and fruits of Rubus
species as antispasmodic, reducing menstrual and labor pains, healing wounds,
diabetes, asthma, hemorrhoids, cholagogue, depurative, constipation and
astringent, diarrhea, tonsillitis, allergic rhinitis is recorded in the
literature. [10, 15-17]. Studies have shown
that Rubus species are rich in antioxidant compounds. [10,17]
and this plant has shown a variety of pharmacological activities like
antioxidant [18, 19], anticancer,
anti-inflammatory, antimicrobial, antibacterial [12],
antidiabetic [11], antitumor [16]. According to the survey of the literature
present a lot of data about antioxidant activities and chemical composition of
the genus Rubus, while there are only a few data of the Rubus
caesius. In addition, no research has been reported on the antioxidant
activity and phytochemical content of unripe blackberries in the literature.
This study was carried out to investigate the changes in the phytochemical and
antioxidant properties of Rubus caesius L. fruits grown naturally in
Battalgazi district of Malatya (Turkey) during the ripening process and to form
a source for future biological, nanotechnological, phytochemical and breeding
studies.
2.
Materials and methods
Fresh
dewberry fruits were harvested in 2020 at different dates during ripening (July
of unripe stage and August of the ripe stage) from Malatya, in Turkey.
Dewberries were classified based on the color green fruit (unripe) red-purple
fruit (ripe). The plant was identified at the University of İnönü by Professor
Arabacı and voucher specimen (ZT1002) was prepared and put in the Pharmacy
Faculty herbarium of the University. All samples were dried in air at room
temperature. After drying, the dried samples were ground powder.
2.1.
Preparation of the extracts
Fifteen grams of each powdered unripe and ripe
Rubus caesius L. were extracted with 250 mL of methanol, ethanol, water,
0.1% acidified methanol, 0.1% acidified ethanol and 0.1% acidified water by
using Soxhlet apparatus for 3 hours. Here, twelve kinds of dewberry extract
were obtained as follows:
1.
Methanol
extract of unripe fruit (UDM 1)
2.
Ethanol
extract of unripe fruit (UDE 1)
3.
Water
extract of unripe fruit (UDW 1)
4.
Acidified
methanol extract of unripe fruit (UDM 2)
5.
Acidified
ethanol extract of unripe fruit (UDE 2)
6.
Acidified
water extract of unripe fruit (UDW 2)
7.
Methanol
extract of ripe fruit (RDM 1)
8.
Ethanol
extract of ripe fruit RDE 1)
9.
Water
extract of ripe fruit (RDW 1)
10.
Acidified
methanol extract of ripe fruit (RDM 2)
11.
Acidified
ethanol extract of ripe fruit (RDE 2)
12. Acidified water extract of ripe fruit (RDW 2)
The solvents
of the extracts were evaporated at 40 °C in the evaporator, (Heidolph, Germany)
and stock solutions were prepared by weighing the remaining solid matter. These
stock solutions were stored in the refrigerator at +4 °C until analysis.
2.2.
Determination of total phenolic content
Determination of total phenols
(TPC) in the extracts was done using Folin-Ciocalteu assay [20]. 50 µL (1 mg/mL) extract was added to a test
tube with 450 µL distilled water. Folin reagent (1.0 N, 250 µL) was added to
the mixture. After five minutes at ambient temperature, 1250 µL of sodium
carbonate (7.5%, w/v) was added and shaken vigorously. Further, the mixture was
permitted to incubate for 120 min and the absorbance was measured using a
Spectrophotometer at 765 nm. After then, the sample concentration was
calculated from the gallic acid standard curve equation (y=0.0095x + 0133, R2=0.9988)
and the result was expressed as mg gallic acid equivalents per gram of extract
(mg GAE/g extract).
2.3.
Determination of total flavonoid content
The number of total flavonoids
(TFC) was measured using aluminum chloride colorimetric assay [21]. 500 µL extract solution (1 mg/mL) was mixed
in a test tube and 4500 µL distilled water and 300 µL of 5.0% sodium nitrite
solution were added. Incubation at room temperature was done for 5 min, and
then 300 µL aluminum chloride (10.0%, w/v) was added followed by 2000 µL NaOH
(1.0 M) after a further 6 min. Finally, 2400 µL of distilled water was added.
Rutin was used to prepare a standard calibration curve (y=0.0065x + 0.0272, R2=0.9970),
where y is the absorbance at 510 nm and x is the sample concentration in µg/mL.
The content of total flavonoid in the extract was expressed as milligram rutin
per gram of extract (mg RE/g extract).
2.4.
Determination of total flavonol content
Total flavonol content (TFLC) was
determined by applying slight modification to the method previously described
by Kumaran and Karunakaran [22]. According
to the method, 1 mL of extract (1mg/mL) was taken into different test tubes
separately. 1.0 mL of 2.0% AlCl3 and 3 mL of 5.0% C2H3NaO2
solution were added to the test tubes. After gentle mixing, all test
tubes were incubated at room temperature for 30 minutes. The absorbance was
then determined using a spectrophotometer at a wavelength of 440 nm. The measurement was compared to a
calibration curve of quercetin (y=0.0161x + 0.049, R2= 0.9976) and
final results were given as milligrams of quercetin equivalents (mg QUE/g
extract).
2.5.
Determination of DPPH radical scavenging activity
The free radical scavenging activities of the extracts were
determined using the DPPH free radical [23]. For
the DPPH• assay, 3.0 mL sample extract or standard (BHA, BHT and α-tocopherol)
with varying concentrations (12.5, 25.0, 37.5, 62.50 and 125 µg/mL) were added
to the 1 mL of DPPH• ethanolic solution (0.10 mM) in a test tube. The mixture
was mixed thoroughly. After 30 minutes of incubation in the dark and at ambient
temperature, the absorbance of the samples was measured at 517 nm. The analyses
were performed in triplicate and the inhibition percentage of DPPH•
discoloration was calculated using the following Eq.
% Inhibition= [(Acontrol-Asample)/Acontrol]x100
2.6. Reducing
power
The method described by Oyaizu [24]
was performed in the present study to determine the reducing power of
extract. 1 mL of plant extract, 1 mL of 0.2 M phosphate buffer solution (pH
6.6) and 2.5 mL of 1% K3[Fe(CN)6 solution were added to a
test tube. Shake vigorously and incubate at 50 °C for 20 minutes. At the end of
the period, 2.5 mL of 10% trichloroacetic acid solution was added and
centrifuged at 6000 rpm for 10 min. After taking 1.25 mL from the solution,
1.25 mL of distilled water and 0.5 mL of 0.1% FeCl3 were added. Finally, absorbance values were read at 700
nm.
2.7. Metal
chelating activity
The chelating capacity of the extracts for Fe2+
ions was determined according to the method specified by Dinis et al. [25]. 3750 µL of extract at different
concentrations (12.5-125 µg/mL) and 50 µL of 2.0 mM FeCl2 were
mixed. It was allowed to incubate for 10 min and then 200 µL of 5.0 mM
ferrosine was added to start the reaction mixture. After mixing the solution,
it was incubated for 20 minutes at room temperature and absorbance was measured
at 562 nm. EDTA was used as a positive control in this assay system. The Fe2+
chelating activity of the dewberry extracts was calculated by the following
equation:
% Chelating Activity =
[(Abscontrol – Abssample)/Abscontrol]x100
3.
Results and discussion
Table 1 shows the percentage yields of extract of unripe and
ripe dewberry fruit. The yield of ripe dewberry fruit extract was found to be
the highest (22.02 %) by methanol extraction, followed by acidified methanol
extraction (20.43 %) whereas, the lowest yield (1.39 %) was obtained from
unripe dewberry fruit of ethanol extraction.
Table 1. Extraction yields (Y) of unripe and
ripe dewberry in
different solvent
Extracts |
Y (%) |
Extracts |
Y (%) |
UDM 1 |
3.49 |
RDM
1 |
22.02 |
UDE 1 |
1.39 |
RDE
1 |
4.87 |
UDW 1 |
2.61 |
RDW
1 |
19.72 |
UDM 2 |
2.66 |
RDM
2 |
20.43 |
UDE 2 |
2.62 |
RDE
2 |
5.14 |
UDW 2 |
2.05 |
RDE
2 |
17.56 |
Phenolic compounds like flavonoids, flavanols, phenolic acids
and anthocyanins are the major determinant of antioxidant potentials in plants
and they could be a natural source of antioxidants, anti-cancer and
anti-mutagenic effects [26, 27]. The
Folin-Ciocalteu method was chosen to the measure total phenolic content of
different dewberry fruit extracts. Generally, the green dewberry fruit (unripe)
had higher total phenolic content compared to the red dewberry fruit (ripe).
Total phenolic content ranged from 23.82 ± 0.00 to 58.72 ± 0.00 mg GAE/g
extract in ripe fruit, from 78.24 ± 0.00 to 151.36 ± 0.00 mg GAE/g extract in
unripe fruit. As can be seen from Table 2, the highest TPC was showed in UDW 2,
while the lowest was observed in RDE 2. Consequently, UDW 2 extract is expected
to display powerful antioxidant activities. Sariburun and his co-workers,
suggested that the phenolic content of the blackberry cultivars ranged from
2279.9 ± 13.0 to 2786.8 ± 21.9 mg GAE/g fresh weight [28]
and Radovanović et al., [29] have
found 7838.26 ± 1.64 mg GAE/kg in acidified methanol extract of wild
blackberry.
Flavonoids comprise a wide range of phenolic compounds with
known properties that include anti-inflammatory, anti-cancer, anti-mutagenic
and antioxidant activities [30, 31]. The
colorimetric method was chosen to the measure total flavonoid content of
dewberry fruit extracts. The result of the total flavonoid content of the
unripe and ripe dewberry fruit extracts is also presented in Table 2. Total
flavonoid content ranged from 7.45 ± 0.02 to 25.75 ± 0.00 mg RE/g extract in
unripe fruit, from 5.65 ± 0.01 to 9.60 ± 0.00 mg RE/g extract in ripe fruit.
The results showed that the highest total flavonoid content was observed in the
methanolic extract of unripe dewberry fruit, while the lowest total flavonoid
content was obtained in the acidified water extract of ripe dewberry fruit.
Caruso et al., [32] reported that the total
flavonoid content of wild blackberry extract ranged between 1136 and 3174 mg
quercetin/100 g fresh weight. Croge et al., [33] reported
that the total flavonoid content of Tupy, Guarani, Xavante and Cherokee
cultivars were between 30.44 and 47.33 mg quercetin/100 g fresh fruit.
Flavonols are a sub-group of flavonoids and the widest flavonols found in food are quercetin, myricetin and kaempferol [34, 35]. Dietary intake of flavonols is associated with many health benefits which include antioxidant, anti-inflammatory effects and reduced risk of vascular disease [35, 36]. The procedure reported by Kumaran and Karunakaran [22] was used to determine the total flavonol in the extracts. The total flavonol content varied from 1.68 ± 0.02 to 5.40 ± 0.00 mg QUE/g extract in unripe fruit, from 1.30 ± 0.01 to 3.42 ± 0.01 mg QUE/g extract in ripe fruit (Table 2). The highest flavonol content was determined in the methanolic extract of unripe dewberry fruit, while the lowest flavonol content was detected in the acidified water extract of the ripe dewberry fruit. Contents of flavonol decreased right along throughout ripening. Similarly, Acosta-Montoya et al., [37] reported a decrease in flavonol values for Rubus adenotrichus Schltdl. during maturing.
Table 2. Total phenol, total flavonoid and total flavonol contents
and antioxidant activity of different unripe and ripe dewberry extracts
Extracts |
TPC (mg GAE/g extract) |
TFC (mg RE/g extract) |
TFLC (mg QUE/g extract) |
UDM 1 |
109.80 ±
0.01 |
25.75 ±
0.00 |
5.40 ±
0.00 |
UDE 1 |
103.94 ±
0.00 |
22.60 ±
0.00 |
4.91 ±
0.00 |
UDW 1 |
78.24 ±
0.00 |
9.92 ±
0.01 |
1.86 ±
0.00 |
UDM 2 |
105.88 ±
0.01 |
21.90 ±
0.00 |
4.66 ±
0.01 |
UDE 2 |
100.78 ±
0.02 |
18.45 ±
0.01 |
3.73 ±
0.00 |
UDW 2 |
151.36 ±
0.00 |
7.45 ±
0.02 |
1.68 ±
0.02 |
RDM 1 |
37.74 ±
0.00 |
9.60 ±
0.01 |
3.42 ±
0.01 |
RDE 1 |
25.20 ±
0.01 |
9.30 ±
0.00 |
2.42 ±
0.00 |
RDW 1 |
47.22 ±
0.00 |
6.60 ±
0.00 |
1.49 ±
0.01 |
RDM 2 |
28.22 ±
0.03 |
8.85 ±
0.01 |
3.29 ±
0.01 |
RDE 2 |
23.82 ±
0.00 |
8.00 ±
0.02 |
2.04 ±
0.01 |
RDW 2 |
58.72 ±
0.00 |
5.65 ±
0.01 |
1.30 ±
0.01 |
The antioxidant capacity of the plant extract is affected by a wide range of factors such as the composition of the extract and solvent type, and cannot be entirely utilized by one single method [38]. Owing to, in the present study, antioxidant activity was assessed by three assays which are propped up various mechanisms of antioxidant action. Fig. 1 exhibits variation in DPPH• scavenging activity of the dewberry extracts which ranged from 3.09 ± 0.00 to 83.16 ± 0.00%.
Figure. 1. DPPH free radical scavenging activity of blackberry extracts (%)
UDW 2 exhibited the highest (83.16 ± 0.00%) activity followed by RDW 1 and RDW 2 (78.69 ± 0.00%) and UDM 1 (78.35 ± 0.00%), whereas RDE 2 showed the lowest activity (20.61 ± 0.00%) at the same concentration of 125 µg/mL. To evaluate the antioxidant activity else, the IC50 for every extract was calculated (Table 3) and described as the concentration of extract inducing 50 % inhibition of absorbance, so a lower value would represent the greater antioxidant activity of the extracts. The results indicated that the highest active DPPH• scavenging activities of the acidified water extract were unripe dewberry (IC50 27.24 ± 0.00 µg/mL), which less than that of BHA (21.74 ± 0.01 µg/mL), but higher than that of BHT (33.45 ± 0.01 µg/mL).
Table 3. Antioxidant activity of unripe and ripe Rubus caesiıs L. different extract
Extracts/ Standards | IC50 (μg/mL) | Extracts/ Standards | IC50 (μg/mL) |
UDM 1 | 52.05 ± 0.01 | RDM 1 | 175.48 ± 0.01 |
UDE 1 | 58.32 ± 0.00 | RDE 1 | 194.44 ± 0.00 |
UDW 1 | 77.05 ± 0.00 | RDW 1 | 70.24 ± 0.01 |
UDM 2 | 87.66 ± 0.01 | RDM 2 | 182.73 ± 0.01 |
UDE 2 | 116.15 ± 0.01 | RDE 2 | 296.18 ± 0.00 |
UDW 2 | 27.24 ± 0.01 | RDE 2 | 67.58 ± 0.01 |
BHA | 21.74 ± 0.01 | α-tocopherol | 1.55 ± 0.01 |
BHT | 33.45 ± 0.01 |
|
|
A comparative analysis on free radical scavenging activity of cultivated varieties of blackberry (Xavante and Cherokee) made by Denardin et al. [39] showed that DPPH• scavenging activity was found IC50 44.70 mg/L and IC50 78.25 mg/L, respectively. Moreover, the DPPH radical scavenging activity of dewberry in this study was much higher than that found by Keser et al. [40]. The antioxidant activities of unripe water and ethanolic blackberry fruit extracts on DPPH radicals were 38.13% and 49.82% at 100 µg/mL, respectively.
Ferric reducing the capacity of the plant extracts is usually related to the existence of reductones (antioxidant agents) that have been indicated to exert antioxidant activity of hydrogen donating and chain-breaking free radicals [41]. The data obtained (Table 4) in this study showed that both unripe and ripe extracts exhibit moderate reducing power capacity as compared with the standards. Keser et al. [40] indicated alterations in the phytochemical composition and antioxidant activity of Rubus discolor fruits for ripening. According to their results, unripe fruit extracts showed higher antioxidant activity using a reducing power assay than ripe fruit.
Table 4. Reducing power ability of different unripe and ripe dewberry extract
Extracts/ Standards | Reducing Power Ability (µg/mL, 700 nm) | |||
5.88 | 14.70 | 29.41 | 44.11 | |
A | A | A | A | |
UDM 1 | 0.130±0.000 | 0.205±0.000 | 0.330±0.000 | 0.429±0.000 |
UDE 1 | 0.126±0.000 | 0.189±0.000 | 0.298±0.000 | 0.408±0.000 |
UDW 1 | 0.124±0.001 | 0.158±0.000 | 0.237±0.000 | 0.328±0.000 |
UDM 2 | 0.135±0.000 | 0.196±0.000 | 0.312±0.000 | 0.405±0.000 |
UDE 2 | 0.118±0.000 | 0.155±0.000 | 0.219±0.000 | 0.296±0.000 |
UDW 2 | 0.184±0.000 | 0.302±0.000 | 0.512±0.000 | 0.701±0.001 |
RDM 1 | 0.123±0.000 | 0.141±0.000 | 0.195±0.000 | 0.259±0.000 |
RDE 1 | 0.113±0.001 | 0.122±0.001 | 0.159±0.000 | 0.192±0.000 |
RDW 1 | 0.131±0.000 | 0.175±0.001 | 0.273±0.000 | 0.377±0.000 |
RDM 2 | 0.114±0.000 | 0.136±0.000 | 0.179±0.001 | 0.230±0.000 |
RDE 2 | 0.113±0.001 | 0.120±0.000 | 0.152±0.001 | 0.197±0.000 |
RDW 2 | 0.139±0.000 | 0.186±0.000 | 0.251±0.000 | 0.331±0.000 |
BHA | 0.601±0.000 | 1.145±0.001 | 2.111±0.000 | 3.356±0.000 |
BHT | 0.348±0.001 | 0.675±0.002 | 1.093±0.000 | 1.816±0.000 |
α-toco-pherol | 0.268±0.000 | 0.521±0.000 | 0.877±0.000 | 1.029±0.001 |
Iron is an important dietary mineral for normal physiology, but an excess of it can lead to damage on a broad array of cellular structures since free iron can donate or accept and electron from next to molecules to bring about cellular injury and to create free radicals from reactive oxygen species [42, 43]. So, the chelating ability of ferrous ion of the plant extracts is an important mechanism for antioxidant activity. In the present study, we assessed the metal chelating ability of extracts of R. caesius and summarized Table 5.
Table 5. The metal chelating ability of different unripe and ripe dewberry extract
Extracts/ Standards | Metal Chelating Ability, Inhibition % | ||||
12.5 µg/mL | 25.0 µg/mL | 37.5 µg/mL | 62.5 µg/mL | 125.0 µg/mL | |
UDM 1 | 1.36±0.00 | 1.21±0.00 | ---- | ---- | ---- |
UDE 1 | 0.15±0.00 | ---- | ---- | ---- | ---- |
UDW 1 | 0.45±0.01 | 1.66±0.00 | 1.81±0.00 | 1.96±0.00 | 3.02±0.00 |
UDM 2 | 0.15±0.00 | ---- | ---- | ---- | ---- |
UDE 2 | 0.30±0.01 | 0.30±0.01 | ---- | ---- | ---- |
UDW 2 | ---- | ---- | ---- | ---- | ---- |
RDM 1 | 1.81±0.00 | 2.87±0.01 | 2.72±0.01 | ---- | ---- |
RDE 1 | 3.17±0.01 | 1.66±0.00 | 1.21±0.02 | ---- | ---- |
RDW 1 | 0.15±0.01 | ---- | ---- | ---- | ---- |
RDM 2 | 4.23±0.00 | 2.26±0.00 | 0.76±0.00 | ---- | ---- |
RDE 2 | 3.93±0.02 | 2.42±0.01 | 1.21±0.01 | ---- | ---- |
RDW 2 | 2.72±0.01 | 1.51±0.00 | ---- | ---- | ---- |
BHA | ---- | ---- | ---- | ---- | ---- |
BHT | 2.42±0.01 | 2.42±0.01 | 2.36±0.00 | 2.11±0.00 | ---- |
α-tocopherol | ---- | ---- | ---- | ---- | ---- |
EDTA | 10.12±0.01 | 53.32±0.00 | 90.18±0.00 | 91.39±0.01 | 93.95±0.01 |
According to obtained data, RDM 2 and RDE 1 extracts showed maximum metal chelating ability at the concentration of 12.5 µg/mL compared to other extracts in each solvent. In addition, the ferrous chelating ability of acidified methanol extract of ripe dewberry was 1.7 times higher than BHT that using as a standard at the concentration of 12.5 µg/mL. Keser et al. [40] reported that ethanol and water extracts of unripe and ripe blackberry possess the potential metal chelating ability. Previous research on the 50% water-methanol extract from among the 20 plants including ripe R. caesius studied and dewberry showed moderated metal chelating activity (51.36%) [14]. This difference might be due to the climatic and soil differences, ripening period of fruit and used extraction solvent.
4. Conclusions
The results of this study, which investigated the chemical contents and biological activities of Rubus caesius L. fruit at different maturity stages, are considered promising. In later studies, the isolation, purification and structure elucidation of active compounds, especially in immature blackberries with bioactivity can be carried out. Obtaining the predicted results may provide possible economic value and form the basis for the identification or formulation of therapeutic or curative antioxidant preparations.
Authors’ contributions
All authors contributed equally.
Acknowledgement
The authors are thankful to Prof. Dr. Turan Arabacı for identifying the Rubus caesius L.
Funding
This research received no specific grant from any funding agency ‘(the public, commercial, or not-for-profit sectors)’.
Conflicts of interest
The authors have declared that no conflict of interest exists.
References
1.
Cory,
H.; Passarelli, S.; Szeto, J.; Tamez, M.; Mattei, J. The role of polyphenols in
human health and food systems: A mini-review. Front. Nutr. 2018, 5(87), 1-9. https://doi.org/10.3389/fnut.2018.00087.
2.
Pandey,
K.B.; Rizvi, S.I. Plant polyphenols as dietary antioxidants in human health and
disease. Oxid. Med. Cell. Longev.
2009, 2(5), 270-278. https://doi.org/10.4161/oxim.2.5.9498.
3.
Vauzour,
D.; Rodriguez-Mateos, A.; Corona, G.; Oruna-Concha, M.J.; Spencer, J.P.E.
Polyphenols and human health: prevention of disease and mechanisms of action. Nutrients. 2010, 2(11), 1106-1131. https://doi.org/10.3390/nu2111106.
4.
World Health Organization. Diet
Nutrition and the Prevention of Chronic Diseases, Report of a Joint FAO/WHO
Expert Consultation. 2003. Available online: whqlibdoc.who.int/trs/WHO_TRS_916.pdf
(accessed on 20 November 2022).
5.
World Health Organization. Fruit and
Vegetables for Health: report of the Joint FAO/WHO Workshop on Fruit and
Vegetables for Health. 2005. Available online: https://apps.who.int/iris/handle/10665/43143
(accessed on 20 November 2022).
6.
Lourenço, S.C.; Moldão-Martins, M.;
Alves V.D. Antioxidants of natural plant origins: from sources to food industry
applications. Molecules. 2019,
24(22), 4132. https://doi.org/10.3390/molecules24224132.
7.
Nieva-Echevarría, B.; Manzanos, M.J.;
Goicoechea, E.; Guillén, M.D. 2,6-Di-tert-butyl-hydroxytoluene and its
metabolites in foods. Compr. Rev. Food Sci. Food Saf. 2015, 14(1), 67-80. https://doi.org/10.1111/1541-4337.12121.
8.
Dolatabadi, J.E.N.; Kashanian, S. A
review on DNA interaction with synthetic phenolic food additives. Food Res. Int. 2010, 43(5),
1223-1230. https://doi.org/10.1016/j.foodres.2010.03.026.
9.
Skrovankova, S.; Sumczynski, D.; Mlcek,
J.; Jurikova, T.; Sochor, J. Bioactive compounds and antioxidant activity in
different types of berries. Int. J.
Mol. Sci. 2015, 16(10), 24673-24706. https://doi.org/10.3390/ijms161024673.
10.
Grochowski, D.M.; Strawa, J.W.;
Granica. S.; Tomczyk, M. Secondary metabolites of Rubus caesius (Rosaceae).
Biochem. Syst. Ecol. 2020, 92,
10411. https://doi.org/10.1016/j.bse.2020.104111.
11.
Schädler, V.; Dergatschewa, S. Rubus
caesius L. Leaves: pharmacognostic
analysis and the study of hypoglycemic activity. Nat. J. Physiol. Pharm.
Pharmacol. 2017, 7(5), 501-508. https://doi.org/10.5455/njppp.2017.7.1234224012017.
12.
Rejewska, A.M.; Sikora, A.; Tomczykowa,
M.; Tomczyk, M. Dewberry (Rubus caesius
L., Rosaceae). Pharmacogn. Commun. 2013,
3(1), 55-57.
13.
Ioncică, R; Bubulică, M.V.; Chirigiu,
L.; Mogosanu G.D.; Popescu, R.; Popescu, H. Researches upon the heavy metals
content of Rubus caesius L. (Rosaceae). Curr. Health Sci. J. 2010, 36(1), 48-51.
14.
Serteser, A.; Kargıoğlu, M.; Gök, V.;
Bağcı, Y.; Ozcan, M.M; Arslan, D. Determination of antioxidant effects of some
plant species wild growing in Turkey. Int.
J. Food Sci. Nutr. 2008, 59(7-8), 643-651. https://doi.org/10.1080/09637480701602530.
15.
Gudej, J.; Tomzcyk, M. Determination of
flavonoids, tannins and ellagic acid in leaves from Rubus L. Species. Arch. Pharm. Res. 2004, 27(11), 1114-119. https://doi.org/10.1007/bf02975114.
16.
Turker, A.U.; Yildirim, A.B.; Karakas,
F.P. Antibacterial and antitumor activities of some wild fruits grown in Turkey.
Biotechnol. Biotechnol. Equip. 2014,
26(1), 2765-2772. https://doi.org/10.5504/bbeq.2011.0156.
17.
Rocabado, G.O.; Bedoya, L.M.; Abad,
M.J.; Bermejo, P. Rubus- A review of its Phytochemical and
Pharmacological Profile. Nat. Prod.
Commun. 2008, 3(3), 423-436. https://doi.org/10.1177/1934578x0800300319.
18.
Grochowski, D.M.; Paduch, R.; Wiater,
A.; Dudek, A.; Pleszczynska, M.; Tomczykowa, M.; Granica, S.; Polak, P.;
Tomczyk, M. In vitro antiproliferative and antioxidant effects of extracts from
Rubus caesius leaves and their quality evaluation. Evid. Based Complement. Alternat. Med. 2016,
2016, 5698685. https://doi.org/10.1155/2016/5698685.
19.
Veličković, I.; Grujić, S.; Džamić, A.;
Krivošej, Z.; Marin, P. D. In vitro antioxidant activity of dewberry (Rubus
caesius L. var. Aquáticus Weihe. & Ness.) leaf extracts. Arch. Biol. Sci. 2015, 67(4),
1323-1330. https://doi.org/10.2298/abs150414109v.
20.
Singleton, V.L.; Rossi, J.A.
Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid
reagents. Am. J. Enol. Vitic. 1965,
16, 144-158. https://doi.org/10.5344/ajev.1965.16.3.144.
21.
Zhishen, J.; Mengcheng, T.; Jianming,
W. The determination of flavonoid contents in mulberry and their scavenging
effects on superoxide radicals. Food Chem. 1999, 64(4), 555-559. https://doi.org/10.1016/s0308-8146(98)00102-2.
22.
Kumaran, A.; Karunakaran, R.J. In vitro antioxidant activities of
methanol extracts of five Phyllanthus species
from India. LWT 2007, 40,
344-352.
https://doi.org/10.1016/j.lwt.2005.09.011.
23.
Blois, M.S. Antioxidant determinations
by the use of a stable free radical. Nature
1958, 181, 1199-1200. https://doi.org/10.1038/1811199a0.
24.
Oyaizu, M. Studies on product of
browning reaction. Antioxidant activities of products of browning reaction
prepared from glucosamine. Japan. J.
Nutr. Diet. 1986, 44, 307-315. https://doi.org/10.5264/eiyogakuzashi.44.307.
25.
Dinis, T.C.P.; Madeira, V.M.C.;
Almeida, L.M. The action of phenolic derivatives (acetaminophen, salicylate,
and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as
peroxyl radical scavengers. Arch.
Biochem. Biophys. 1994, 315(1), 161-169. https://doi.org/10.1006/abbi.1994.1485.
26.
Kumar, N.; Goel, N. Phenolic acids; natural
versatile molecules with promising therapeutic applications. Biotechnol. Rep. 2019, 24, e00370. https://doi.org/10.1016/j.btre.2019.e00370.
27.
Srivastava, A.; Greenspan, P. Hartle,
D.K.; Hargrove, J.L.; Amarowicz, R.; Pegg, R.B. Antioxidant and anti-inflammatory
activities of polyphenolics from Southeastern U.S. range blackberry cultivars. J. Agric. Food Chem. 2010,
58(10), 6102-6109. https://doi.org/10.1021/jf1004836.
28.
Sariburun, E.; Şahin, S.; Demir, C.;
Türkben, C.; Uylaşer, V. Phenolic content and antioxidant activity of raspberry
and blackberry cultivars. J. Food Sci.
2010, 75(4), 328-335. https://doi.org/10.1111/j.1750-3841.2010.01571.x
29.
Radovanović, B.C.; Anđelković, S.M.;
Radovanović, A.B.; Anđelković, M.Z. Antioxidant and antimicrobial activity of
polyphenol extracts from wild berry fruits grown in Southeast Serbia. Trop. J. Pharm. Res. 2013, 12(5),
813-819.
https://doi.org/10.4314/tjpr.v12i5.23.
30.
Li, A.N.; Li, S.; Zhang, Y.J.; Xu,
X.R.; Chen, Y.M.; Li, H.B.; Resources and biological activities of natural
polyphenols. Nutrients. 2014,
6(12), 6020-6047. https://doi.org/10.3390/nu6126020.
31.
Karak, P. Biological activities of
flavonoids: An overview. Int. J. Pharm. Sci. Res. 2019, 10(4), 1567-1574.
32.
Caruso, M.C.; Galgano, F.; Grippo, A.;
Condelli, N.; Cairano, M.D.; Tolve, R. Assay of healthful properties of wild
blackberry and elderberry fruits grown in Mediterranean area. J. Food Meas. Charact. 2019, 13,
1591-1598. https://doi.org/10.1007/s11694-019-00075-x.
33.
Croge, C.P.; Cuquel, F.L.; Pintro,
P.T.M.; Biasi, L.A.; Bona, M.D.C. Antioxidant capacity and polyphenolic
compounds of blackberries produced in different climates. Hort. Sci. 2019, 54(12), 2209-2213. https://doi.org/10.21273/hortsci14377-19.
34.
Lavefve, L.; Howard, L.R.; Carbonero,
F.; Berry polyphenols metabolism and impact on human gut microbiota and health.
Food Funct. 2020, 11,45-65. https://doi.org/10.1039/c9fo01634a.
35.
Dabeek, W.M.; Marra, V.M. Dietary quercetin
and kaempferol: bioavailability and potential cardiovascular-related bioactivity
in humans. Nutrients. 2019,
11(10), 2288. https://doi.org/10.3390/nu11102288.
36.
Panche, A.N.; Diwan, A.D.; Chandra,
S.R. Flavonoids: An overwiev. J. Nutr.
Sci. 2016, 5: e47. https://doi.org/10.1017/jns.2016.41.
37.
Acosta-Montoya, Ó.; Vaillant, F.;
Cozzano, S.; Mertz, C.; Perez, A.M.; Castro, M.V. Phenolic content and
antioxidant capacity of tropical highland blackberry (Rubus adenotrichus Schltdl.) during three edible maturity stages. Food Chem. 2010, 119(4), 1497-1501. https://doi.org/10.1016/j.foodchem.2009.09.032.
38.
El Abed, N.; Guesmi, F.; Mejri, M.;
Marzouki, M.N. (2014). Phytochemical screening and assessment of antioxidant,
antibacterial and cytotoxicity activities of five Tunisian medicinal plants. Int. J. Pharma. Res. Bio Sci. 2014,
3(4), 770-789.
39.
Denardin, C.C.; Hirsch, G.E.; da Rocha,
R.F.; Vizzotto, M.; Henriques, A.T.; Moreira, J.C.F.; Guma, F.T.C.R.; Emanuelli
T. Antioxidant capacity and bioactive compounds of four Brazilian native
fruits. J. Food Drug Anal.
2015, 23(3), 387-398. https://doi.org/10.1016/j.jfda.2015.01.006.
40.
Keser, S.; Celik, S.; Turkoglu, S.;
Yılmaz, Ö.; Turkoglu, I. Antioxidant properties of Rubus discolor L., extracts and protective effects of its flower
extracts against hydrogen peroxide-induced oxidative stress in wistar rats. Turk. J. Pharm. Sci. 2015, 12(2),
187-206. https://doi.org/10.5505/tjps.2015.57966
41.
Abdolahi, N.; Soltani, A.; Mirzaali,
A.; Soltani, S.; Balakheyli H.; Aghaei M. Antibacterial and antioxidant
activities and phytochemical properties of Punica granatum flowers in
Iran. Iran. J. Sci. Technol. Trans.
A: Sci. 42: 1105-1110. https://doi.org/10.1007/s40995-017-0413-8.
42.
Sowndhararajan, K.; Kang, S.C. Free
radical scavenging activity from different extracts of leaves of Bauhinia
vahlii Wight & Arn. Saudi J.
Biol. Sci. 2013, 20(4), 319-325. https://doi.org/10.1016/j.sjbs.2012.12.005.
43. Eid, R.; Arab, N.T.T.; Greenwood, M.T. Iron mediated toxicity and programmed cell death: A review and a re-examination of existing paradigms. Biochim. Biophys. Acta (BBA) Mol. Cell Res. 2017, 1864(2), 399-430. https://doi.org/10.1016/j.bbamcr.2016.12.002

This work is licensed under the
Creative Commons Attribution
4.0
License (CC BY-NC 4.0).
Abstract
In this article, we have studied the
changes in phytochemical contents and antioxidant properties of blackberry (Rubus caesius L.) fruits naturally grown
in Battalgazi district of Malatya province during the ripening process. It was determined the total phenolic (23.82-151.36 mg GAE/g extract), total flavonoid (5.65-25.72
mg RE/g extract), and total flavonol (1.30-5.40 mg QUE/g extract) contents of
twelve extracts prepared by six different solvents. According to these results, it was concluded that
phytochemical contents were higher in unripe fruit extracts than the ripe
fruits.
The reducing power, chelating capacity
of metal ions and antioxidant activity of each extract were determined using
the DPPH free radical scavenging method. The working conditions in which the
antioxidant activity values were best obtained by the DPPH method were
explained according to each extract extraction method and it was seen that the
best result was the acidified water extract of the unripe fruit (IC50
27.24 ± 0.00 µg/mL).
The results showed that R. caesius L. can be consumed as a
natural antioxidant due to its antioxidant properties. Supporting these results with in vivo studies will
also clarify the health promoting properties of antioxidant activity of R.
caesisus fruit.
Abstract Keywords
Dewberry, Rubus caesius L., antioxidant activity, phenolic, flavonoid,
flavonol, Phytochemical.

This work is licensed under the
Creative Commons Attribution
4.0
License (CC BY-NC 4.0).
Editor-in-Chief

This work is licensed under the
Creative Commons Attribution 4.0
License.(CC BY-NC 4.0).