Escolar Documentos
Profissional Documentos
Cultura Documentos
discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/238376275
CITATIONS
READS
72
275
2 authors, including:
Ilkay Koca
Ondokuz Mays niversitesi
44 PUBLICATIONS 497 CITATIONS
SEE PROFILE
Scientia Horticulturae
journal homepage: www.elsevier.com/locate/scihorti
A R T I C L E I N F O
A B S T R A C T
Article history:
Received 7 October 2008
Received in revised form 4 March 2009
Accepted 6 March 2009
Seven wild and ten cultivated blackberries (Arapaho, Bartin, Black Satin, Bursa 1, Bursa 2, Cherokee,
Chester, Jumbo, Navaho, and Ness), and six lowbush (Vaccinium arctostaphylos) and four highbush
(Vaccinium corymbosum) blueberries fruits (Ivanhoe, Jersey, Northland, and Rekord) were analyzed for
total anthocyanins, total phenolics, and antioxidant activity as ferric reducing antioxidant power (FRAP)
in this study. The respective ranges of total anthocyanin and total phenolic contents of the tested
samples were: blackberries, 0.951.97 and 1.733.79 mg g1 and blueberries, 0.182.94 and 0.77
5.42 mg g1. FRAP values varied from 35.05 to 70.41 mmol g1 for blackberries, 7.41 to 57.92 mmol g1
for blueberries. Wild blackberries had the highest FRAP values while wild blueberries had the highest
total phenolic and total anthocyanin contents. A linear relationship was observed between FRAP values
and total phenolics for blueberries (r = 0.981). The anthocyanin pigments in samples were isolated and
characterized by high-performance liquid chromatography (HPLC) with UVvisible detection. Cyn-3-glu
was the predominant anthocyanin in all blackberry fruits.
2009 Elsevier B.V. All rights reserved.
Keywords:
Anthocyanin
Blackberry
Blueberry
FRAP
HPLC
Phenols
1. Introduction
Blackberries and blueberries are known to contain appreciable
levels of phenolic compounds, including anthocyanins, avonols,
chlorogenic acid and procyanidins, that have high biological
activity and may provide health benets as dietary antioxidants
(Cho et al., 2005). Anthocyanins are probably the largest group of
phenolic compounds in human diet. Anthocyanins have been used
for several therapeutic purposes including the treatment of
diabetic retinopathy, brocystic disease, and vision disorders.
Anthocyanins also have the potential to serve as radiationprotective agents, vasotonic agents and chemoprotective agents
and can act against carbon tetrachloride induced lipoperoxidation
(Veberic et al., 2009; Cacace and Mazza, 2003; Wang et al., 1997).
Phenolic extracts of berries inhibited human low-density lipoprotein (LDL) and liposome oxidation (Heinonen et al., 1998). Berries
have also shown a remarkably high scavenging activity toward
* Corresponding author. Tel.: +90 362 3121919; fax: +90 362 4576035.
E-mail address: itosun@omu.edu.tr (I. Koca).
Abbreviations: Cyn, cyanidin; plg, pelargonidin; peo, peonidin; malv, malvidin;
delp, delphinidin; pet, petunidin; rut, rutinose; glu, glucoside; gal, galactoside; ara,
arabinoside; HPLC, high-performance liquid chromatography; FRAP, ferric reducing
antioxidant power; TACN, total anthocyanin; ORAC, oxygen radical-absorbing
capacity; std dev, standard deviation.
0304-4238/$ see front matter 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.scienta.2009.03.015
448
2.2. Samples
Correlation analyses were carried out with the SPSS 11.0 for
Windows software package.
2.3. Methods
2.3.1. Determination of antioxidant activity
Antioxidant activity was determined by FRAP assay. To
determine the antioxidant activity, an aliquot of the acetone/
methanol/water/formic acid (40:40:20:0.1) extracts of berry
samples was dried at 30 8C vacuum using Buchi rotary evaporator
(Newcastle, Del, USA), and redissolved in same volume of water.
Aqueous samples were mixed with 0.95 mL of ferric-TPTZ reagent
and measured at 593 nm. FeSO4 was used as a standard and
antioxidant power was expressed as mmol g1 FRAP (Tural and
Koca, 2008).
2.3.2. Determination of total phenolics
Total phenolic content was measured using the Folin-Ciocalteau method (Singleton and Rossi, 1965). Results were expressed
as milligrams of gallic acid equivalents per gram of fresh weight.
2.3.3. Determination of total anthocyanins
Total anthocyanin content was determined with the pH
differential method (Wrolstad, 1976), and expressed as milligrams
of cyanidin-3-glucoside equivalents per gram of fresh weight, on
the basis of the molar absorptivity (29,600) of cyn-3-glu for
blackberry. Calculations were based on malvidin-3-glucoside with
molar absorptivity 28,000 for blueberry. All spectrophotometric
analyses were performed using a UVvisible spectrophotometre
(Jasco V-530, Japan).
2.3.4. HPLC anthocyanin analysis
Sample preparation was carried out according to the procedure
described by Tural and Koca (2008). A berry sample was
homogenized with 1% HCl in methanol, then centrifuged at
3000 rpm for 10 min. After appropriate dilution with 1% HCl in
methanol, the sample was ltered through a 0.45 mm lter
(Biocrom MN 718020, Phonex nylon lter 25 mm), seperated and
characterized by HPLC. Samples were analyzed using a Thermoquest (USA) HPLC system equipped with a model P100 pump, a SN
4000 controller unite, a model AS 2000 autosampler and a model
Total phenolics
(mg g1)
Wild blackberries (n = 7)
Ranges
52.8870.41
Means std dev
59.36 5.84
2.643.79
3.26 0.41
1.301.97
1.59 0.22
Cultivated blackberries
Arapaho
Bartin
Black Satin
Bursa 1
Bursa 2
Cherokee
Chester
Jumbo
Navaho
Ness
2.19
1.73
1.76
2.10
2.34
3.05
2.25
2.39
2.10
1.76
1.15
0.96
1.20
1.27
0.95
1.15
1.41
1.07
0.98
1.58
Wild blueberries (n = 6)
Ranges
34.4557.92
Means std dev
43.97 8.93
3.085.42
3.95 0.91
0.592.94
1.50 0.87
Cultivated blueberries
Jersey
7.41
Ivanhoe
12.36
Northland
13.69
Rekord
7.56
8.20
1.40
1.84
0.77
0.25
0.25
0.29
0.18
38.08
35.05
39.16
38.67
40.32
43.44
42.16
38.68
36.32
41.36
Cyn-3-glu
(%)
Cyn-3-rut Plg-3-glu
(%)
(%)
Peo-3-glu
(%)
Wild blackberries (n = 7)
Ranges
0.000.14 80.0682.71
Means std dev 0.10 0.02 81.47 1.10
0.220.48 0.000.06
0.32 0.10 0.05 0.01
Cultivated blackberries
Arapaho
0.31
Bartin
Black Satin
0.12
Bursa 1
0.12
Bursa 2
0.17
Cherokee
0.24
Chester
0.15
Jumbo
0.14
Navaho
0.15
Ness
0.20
0.10
0.33
0.11
0.11
0.17
0.15
0.14
0.06
0.11
0.11
90.42
78.19
77.47
81.05
85.79
84.60
82.14
82.29
80.58
82.81
9.09
0.38
0.11
0.04
0.09
0.12
0.10
0.13
Table 3
Percent contribution of individual anthocyanins to total anthocyanins in blueberries.
Cyn-3-glu (%)
Wild blueberries (n = 6)
Ranges
9.2012.52
Means std dev 10.44 1.53
Cultivated blueberries
Jersey
3.11
Ivanhoe
3.51
Northland
3.30
Rekord
1.79
Cyn-3-rut (%)
Plg-3-glu (%)
Peo-3-glu (%)
8.3118.72
12.31 4.13
1.293.74
2.50 0.89
34.0550.26
44.23 6.68
1.50
4.70
0.90
4.35
36.21
40.00
26.04
35.22
3.87
5.64
2.29
5.45
449
2002; Sellappan et al., 2002; Wada and Ou, 2002; Siriwoharn et al.,
2004).
Cho et al. (2004) determined the total anthocyanins in arapaho
and navaho genotypes as 1.798 and 1.823, respectively. Dai et al.
(2009) determined the total anthocyanins and total phenolics in
Chester purees as 7.95 mg g1 vs. and 25.31 mg g1, respectively
and in Black Satin purees as 7.16 mg g1 vs. and 22.91 mg g1,
respectively. These ndings are generally higher than our results.
Ehlenfeldt and Prior (2001) evaulated antioxidant activity, total
phenolic and total anthocyanin contents in highbush blueberry,
including Ivanhoe, Jersey and Northland varieties. They found that
the respective average values of antioxidant activity, total
anthocyanin and total phenolic of samples were: Ivanhoe, 16.1
ORAC (oxygen radical-absorbing capacity) units, 2.01 and
0.95 mg g1; Jersey, 19.3 ORAC units, 2.05 and 1.06 mg g1 and
Northland, 17.2 ORAC units, 1.29 and 1.37 mg g1. Kalt et al. (1999)
reported that phenolics and anthocyanins values were 27.7 and
4.35 mmol g1 for lowbush blueberry, respectively. Also, they
found that phenolics and anthocyanins values 22.7 and
2.67 mmol g1 for highbush blueberry, respectively. In addition,
Zheng and Wang (2003) found that antioxidant activity (assessed
by ORAC), total anthocyanin and total phenolic contents in
blueberry were 28.9 mmol g1, 1.20 and 4.12 mg g1, respectively.
Total phenolic content of blueberries reported by Ehlenfeldt
and Prior (2001) is generally lower than the value determined for
highbush blueberries in the present study, whereas anthocyanin
values reported by them is higher. The total anthocyanin content of
blueberries was lower than the values (1.438.22 mg g1) reported
by Cho et al. (2004), which may be attributed to differences in
environmental growing conditions and genotypes. Kalt et al.
(2001) also indicated that synthesis of anthocyanins and other
phenolic compounds can be inuenced by various abiotic and
biotic factors, including temperature, irradiation, herbivory, and
pathogenic infection.
Antioxidant activity, total anthocyanin and total phenolic
values of lowbush blueberries were higher than highbush blueberries. On average, lowbush blueberries had higher levels of
anthocyanins, total phenolics, and antioxidant activity than
highbush blueberries; which is consistent with the earlier reports
(Kalt et al., 1999, 2000).
FRAP values in blueberries were highly correlated with phenol
content (r = 0.981), whereas a less linear correlation between total
antioxidant activity and anthocyanin content was recorded
(r = 0.817), implying that the antioxidant activity of blueberries
is largely due to presence of phenolic compounds. Similar results
have been reported by other researchers (Wang and Lin, 2000),
who found a linear correlation between total antioxidant activity
and phenol content in blackberries (r = 0.961). In addition,
Deighton et al. (2000) reported that there were apparent linear
relationships between antioxidant activity (assessed by FRAP) and
total phenols (r = 0.965), whereas anthocyanin content had a
minor inuence on antioxidant activity (r = 0.588) of Rubus juices.
Also, the correlation coefcient for antioxidant activity and total
phenols content (r = 0.798) in blackberries is better than that for
antioxidant activity and anthocyanin content (r = 0.530). Similar
results have been reported by other researchers (Wada and Ou,
2002) for blackberries.
In this study, both wild and cultivated blackberries generally
contained 12 kinds of anthocyanins. Six peaks of anthocyanins
were identied. The amounts of anthocyanins in the blackberries
varied markedly. Cyn-3-glu was the major anthocyanin in blackberries. Cyn-3-glu ranged from 77.47% to 90.42% of peak area
(Table 2). The data were consistent with previous reports (Sapers
et al., 1986; Wada and Ou, 2002; Cho et al., 2004; Fan-Chiang and
Wrolstad, 2005). The other minor anthocyanins identied included
cyn-3,5-di glu, peo-3-glu, plg-3-glu, cyn-3-rut (only in Nessy
450