JFS C: Food Chemistry

Characterization of Red Raspberry

(Rubus idaeus L.) Genotypes for Their

C: Food Chemistry
Physicochemical Properties
M. TOSUN, S. ERCISLI, H. KARLIDAG, AND M. SENGUL

ABSTRACT: The worldwide tendency for growing more small fruits, including raspberries, shows permanent in-
crease because this group of fruits has a relatively higher content of bioactive nutrients. To study the health benefits
of red raspberry fruits, 11 preselected wild-grown and 1 well-known cultivar, Heritage, were evaluated for some of
their physicochemical properties such as fruit weight, total antioxidant capacity (measured by -carotene bleach-
ing and FRAP assays), total phenolics, ascorbic acid, soluble solid content (SSC), and acidity. Fruit weight, SSC, and
ascorbic acid contents were between 1.47 and 2.32 g, 10.87% and 13.60%, and 21 and 36 mg/100 g, respectively.
Antioxidant activity and total phenolic content varied among genotypes and the ERZ5 genotype had the highest an-
tioxidant capacity as ascertained by both methods. This genotype also had the highest total phenolic (2031 g GAE/g
FW) content. There are linear relationships between antioxidant capacities and total phenols. The present study
demonstrates the potential of certain wild genotypes, notably ERZ5, for improving the nutritional value through
germplasm enhancement programs.
Keywords: antioxidant capacity, red raspberry, total anthocyanin, total phenolics

Introduction Casals and others 2006), sea buckthorns (Ercisli and others 2007),

T here is an abundance of evidence that regular consumption of

fruits and vegetables is associated with reduced risks of chronic
diseases, such as cancers and cardiovascular disease (Dragsted and
others 1993). Fruits and vegetables are primary food sources pro-
viding essential nutrients for sustaining life; they also contain a va-
riety of phytochemicals such as phenolics and flavonoids, which
provide important health benefits (Wang and others 1996). Free
radical-induced oxidative stress has been associated with several
toxic cellular processes including oxidation damage to protein and
DNA, membrane lipid oxidation, enzyme inactivation, and gene
mutation that may lead to carcinogenesis (Poulsen and others
1998). Intake of sufficient amounts of antioxidants is necessary to
prevent free radical-induced oxidative stress. It has been reported
that most of the antioxidant capacity of fruits and vegetables may
come from total phenolics, anthocyanins, and flavonoids (Eber-
hardt and others 2000).
On the other hand, the bioactive content of fruits varies from
genotype to genotype (Ercisli and Orhan 2007). Therefore, attention
has more recently been focused on assessing the distribution on bi-
ologically active compounds among different cultivars/genotypes
(Pantelidis and others 2007).
In addition, increasing recent interest in nutraceuticals and
functional foods has led plant breeders to initiate selection of crops
with higher-than-normal phenolic antioxidant contents, including
blueberries (Prior and others 1998), plums and peaches (Cavallos-
MS 20090370 Submitted 4/26/2009, Accepted 6/1/2009. Author Tosun is with
Dept. of Food Science, Oltu Vocational School, Ataturk Univ., 25800 Oltu-
Erzurum, Turkey. Author Ercisli is with Dept. of Horticulture, Faculty of
Agriculture, Ataturk Univ., 25240 Erzurum, Turkey. Author Karlidag is with
Dept. of Horticulture, Hamza Polat Vocational School, Ataturk Univ., 25900
Ispir-Erzurum, Turkey. Author Sengul is with Dept. of Food Engineering,
Faculty of Agriculture, Ataturk Univ., 25240 Erzurum, Turkey. Direct in-
quiries to author Ercisli (E-mail: sercisli@hotmail.com).
mulberries (Ercisli and Orhan 2007), and strawberries and apples
(Scalzo and others 2005). All these selection programs aim to set
the baseline for establishing breeding efforts, with the intention
of adding value to fruits with respect to the level and diversity of
health benefit properties that crops could impart.
In this regard, it is important to characterize different types of
fruit for the content of single specific antioxidant compounds. But,
because of the difficulty associated with the measurement of in-
dividual antioxidant components in such complex mixtures, vari-
ous methodologies have been developed for obtaining quantitative
data from whole samples. The 2 quantitative methods used in the
present study, FRAP and -carotene bleaching, are just two of many
such assays. These and other assays have been increasingly used
to investigate fruits and vegetables to obtain their antioxidant ca-
pacities, and red fruits appear to be particularly rich in antioxidant
compounds (Wang and others 1996; Prior and others 1998).
Rubus is one of the most diverse genera in the plant kingdom,
comprising over 400 species subdivided into 12 subgenera. The do-
mesticated subgenera contain the raspberries, blackberries, arctic
fruits, and flowering raspberries, all of which have been utilized
in breeding programs. The most important raspberries are the Eu-
ropean red, R. idaeus L. subsp. idaeus, and the place of origin of
raspberry has been postulated to be the Ida Mountains of Turkey
(Jennings 1988).
Raspberries are very rich sources of bioactive compounds such
as phenolics, anthocyanins, organic acids, minerals, and more. The
genus has also very high amounts of antioxidant compounds. It
has been confirmed that the antioxidant ability of raspberry fruit
is derived from the contribution of phenolic compounds in rasp-
berries (Liu and others 2002). Bioactive compounds of raspberry
fruits, their characterization and utilization in functional foods,
and clinical assessment of antimicrobial properties for human
health are among the major targets of contemporary research. The


C 2009 Institute of Food Technologists
R
Vol. 74, Nr. 7, 2009JOURNAL OF FOOD SCIENCE C575
doi: 10.1111/j.1750-3841.2009.01297.x
Further reproduction without permission is prohibited
 Characterization of red raspberry genotypes . . .
evaluation of raspberry fruit genetic resources for the presence of
bioactive compounds and their properties as natural agents is of
doubtless significance and promises to have great benefit for breed-
ers, and the food and pharmaceutical industries (Badjakov and oth-
ers 2008). In addition, it is important to be able to identify the
metabolic differences between cultivated and wild forms to assess
their quality and to monitor it during the domestication process
C: Food Chemistry
(Spina and others 2008).
Previously, health-promoting components of mainly cultivated
raspberries have been reported (Wang and Lin 2000; Liu and others
2002; Kafkas and others 2008). However, there were a few reports
that dealt with bioactive compounds of wild raspberries, and de-
tailed information about the health-promoting components of wild
raspberry genotypes is needed to better understand their use as
functional foods and as ingredients in pharmaceuticals, nutraceu-
ticals, and in medicine.
Therefore, we propose in this study that wild raspberry selec-
tions, rich in antioxidants and phenolics may yield fruits with en-
hanced functional properties such as antioxidant, colorant, and
antimicrobial properties.
Materials and Methods
Collection and preparation of raspberry fruit samples
Eleven promising red raspberry genotypes, previously selected
from the Erzurum region in Turkey according to high yield and free
of pest and disease characteristics, were used. The well-known cul-
tivar Heritage was also used in this study to compare it with wild
types.
Approximately 1/2 kg raspberry fruits per genotype were har-
vested from shrubs at the ripe stage in the autumns of 2007 and
2008. There were no statistically significant differences in the pro-
files and yields of the different phytochemicals being analyzed be-
tween the 2 y, therefore the data of the different years were pooled
(this could be a consequence of similar growing and climatic con-
ditions in both years in the area where the plants grow). The fully
mature uniform fruits were harvested by hand and transferred to Statistical analysis
the laboratory for physical and phytochemical analysis. They were
frozen in plastic bags until used.
Reagents
FolinCiocalteau reagent was purchased from Merck (Darm- was used to correlate antioxidant activity determination methods
stadt, Germany). Trolox (6-hydroxy-2,5,7,8-tetramethylchromane- and total phenolic content results.
2-carboxylic acid), TPTZ (2,4,6-tripyridyl-s-triazine), FeCl3 6H2 O,
and gallic acid were obtained from Sigma-Aldrich Chemical Co. (St.
Louis, Mo., U.S.A.).
Determination of fruit weight, SSC, vitamin C,
and acidity in raspberry fruits
Twenty fruits from each cultivar/genotype were used for fruit
weight determination. Fruit weight was measured by using a dig-
ital balance with a sensitivity of 0.001 g (Scaltec SPB31). For each
genotype, 50 fruits were thawed at room temperature and homog-
enized in a standard food blender. Homogenates were assayed for
titratable acidity, soluble solids content (SSC), and vitamin C. Sol-
uble solid content (SSC) were determined by a digital refractome-
ter (Model RA-250HE; Kyoto Electronics Manufacturing Co. Ltd.,
Japan) at 22 C. Titratable acidity (TAc) was measured by the titri-
metric method (AOAC 1984). Titratable acidity was expressed as
percent citric acid. Ascorbic acid of samples was quantified with the
reflectometer set of Merck Co. (Merck RQflex), according to their
protocol for the juice of red fruit, and expressed as milligram ascor-
bic acid/100 g fresh sample.
C576 JOURNAL OF FOOD SCIENCEVol. 74, Nr. 7, 2009
Determination of total phenolics and
antioxidant activity in raspberry fruits
For extraction, fruit homogenates obtained with a blender were
extracted with a buffer containing acetone, water, and acetic
acid (70 : 29.5 : 0.5, v/v/v) for 1 h in darkness (Singleton and
Rossi 1965). This extract was filtered and used for phytochemical
analysis.
Total phenolics in the methanol extracts were determined colori-
metrically using FolinCiocalteu reagent as described by Slinkard
and Singleton (1977). Gallic acid was used as the standard and re-
sults were expressed as microgram gallic acid equivalents per gram
fresh weight basis.
Total antioxidant capacity of samples was determined by hy-
drogen atom transfer reactions (-carotene bleaching assay) and
an assay based on electron transfer (ferric reducing antioxidant
power, FRAP assay). In the -carotene bleaching assay, antioxi-
dant capacity is determined by measuring the inhibition of the
volatile organic compounds and the conjugated diene hydroperox-
ides arising from linoleic acid oxidation (Kaur and Kapoor 2002).
Antioxidant capacities of the samples were compared with those of
the synthetic antioxidant butylated hydroxyanisole (BHA) and the
blank.
In the FRAP assay, the method of Benzie and Strain (1996)
was used. The assay was conducted using 3 aqueous stock so-
lutions containing 0.1 mol/L acetate buffer (pH 3.6), 10 mmol/L
TPTZ [2,4,6-tris(2-pyridyl)-1,3,5-triazine] acidified with concen-
trated HCL, and 20 mmol/L ferric chloride. These solutions were
prepared and stored in the dark under refrigeration. Stock solutions
were combined (10 : 1 : 1, v/v/v) to form the FRAP reagent just prior
to analysis. For each assay, laboratory duplicates from each repli-
cate plus 2.97 mL of FRAP reagent and 0.03 mL of sample extract
were mixed. After 30 min, the absorbance of the reaction mixture
at 593 nm was determined on a spectrophotometer (Model Nico-
let 100, Thermo Scientific, London, U.K.). Results are expressed as
microgram Trolox equivalent (TE)/g fw.
The experiment was a completely randomized design with 5
replications. Data were subjected to analysis of variance (ANOVA)
and means were separated by Duncans multiple range test at
P < 0.05 significant level. The regression analysis (Pearsons index)
Results and Discussion
Fruit weight, soluble solids contents,
and acidity of raspberry fruits
Fruit weight, SSC (soluble solids content), and acidity (%) of
raspberry genotypes are shown in Table 1. Great variability existed
among the examined raspberry fruit samples, regarding their fruit
weight, SSC, and acidity (P < 0.05, Table 1).
Average fruit weight values of raspberry genotypes ranged be-
tween 1.47 (ERZ 6) and 2.32 g (Heritage) (Table 1). The highest SSC
content was observed in ERZ 7 genotype (13.60%), followed by ERZ
10 genotype (13.25%) and ERZ 9 genotype (13.10%). The lowest SSC
was recorded in the ERZ 3 genotype (10.87%). Acidity of red rasp-
berry genotypes was from 1.26% (ERZ 11) to 1.82% (ERZ 9 geno-
type) (Table 1).
Previously, fruit weights of raspberry genotypes grown in differ-
ent agro-climatic regions of Turkey were reported to be between
0.50 and 4.17 g (Gercekcioglu and others 2003; Kaplan and oth-
ers 2003; Turemis and others 2006). The SSC and acidity contents
Characterization of red raspberry genotypes . . .

of raspberry genotypes had been reported to be between 8.40%
and 14.70% and 1.27% and 2.78%, respectively (Agaoglu and others
2003; Turemis and others 2006; Orhan and others 2006). Our fruit
weight, SSC, and acidity results are comparable with the data from
these studies. Many factors are known to affect the fruit weight,
SSC, and acidity in fruit species, including cultivar/genotype, alti-
tude, environmental conditions, and more (Hardisson and others
dependent phenolic contents had been determined in raspberries,
strawberries, and blueberries (Kalt and others 1999). It was reported
that wild strawberry and blackberry germplasm was responsible
for a higher total phenolic content than that of cultivated plants
(Reyes-Carmona and others 2005; Scalzo and others 2005). Spina
and others (2008) showed that wild plants had higher total phe-
nolic contents than cultivated ones. In fact, phenolics may play

C: Food Chemistry
2001; Ercisli and others 2007). an important role in an effective defense mechanism that renders
plants resistant to numerous potential pathogens. Moreover, there
Total phenolics content, antioxidant activity, is a complex set of interaction between wild plant growth and abi-
and ascorbic acid in raspberry fruits otic environmental factors: nutrient stress, irrigation or lack of wa-
The differences in total phenolics content, antioxidant activity, ter, as well as other conditions, which may enhance the production
and ascorbic acid among raspberry genotypes were statistically sig- of phenolic (Vaya and others 1997).
nificant (P < 0.05, Table 2). The antioxidant activity results using -carotene bleaching and
The total phenolic content of raspberry genotypes was in the ferric-reducing antioxidant power (FRAP) in raspberry genotypes
range of 1145 to 2031 g GAE/g fresh weight basis. The wild geno- are shown in Table 2.
types, in general, had higher total phenolic contents than the culti- Statistically significant difference (P < 0.05) was found among
vated one (Table 2). Modern consumers are increasingly interested the samples and BHA in the -carotene bleaching assay. All rasp-
in their personal health and expect the foods they purchase to be berry genotypes, except ERZ 10 and ERZ 11, showed higher antiox-
tasty, attractive, and also safe and health promoting. Phenolic com- idant activity than the cultivated one, even sometimes higher than
pounds are one of the most diverse groups of secondary metabo- the reference antioxidant, BHA (88.10%). The highest antioxidant
lites in raspberries (Badjakov and others 2008). Previously, a wide activity was observed in the ERZ 5 genotype as 89.60%, followed by
variation was observed on total phenolic contents in fruits of rasp- ERZ 1 (89.35%) and ERZ 6 genotypes (88.70%) (Table 2).
berries, namely, 1442 to 2146 g gallic acid equivalents per gram Data obtained using the FRAP assay confirmed the patterns
fresh weight basis (Khanizadeh and others 2009). Our total pheno- obtained using the -carotene bleaching assay (Table 2). The
lic results were comparable with those in the literature. Genotype- highest antioxidant capacity, as assessed by both methods, was
that from the ERZ 5 genotype (FRAP: 31.1 mol TE/g fw and
-carotene: 89.60%). Heritage, a domesticated red raspberry cul-
Table 1 --- Fruit weight, soluble solid content, and acidity tivar studied as a comparison, had FRAP = 20.9 mol TE/g fw
of raspberry fruits. and -carotene = 83%. The average FRAP value of 11 wild grown
Fruit weight SSC Acidity genotypes was 24.1 mol TE/g fw. Previously, FRAP values of rasp-
Genotype (g) (%) (%) berry cultivars were found to be 19.4 to 22.2 mol TE/g fw (Ozgen
ERZ 1 1.64bc 12.24ab 1.50ab and others 2006) which supports our findings. The antioxidant ac-
ERZ 2 1.57bc 11.60b 1.45ab tivity may be explained by looking at the combination of different
ERZ 3 2.08ab 10.87c 1.37ab phytochemicals functioning additively or synergistically account-
ERZ 4 1.94ab 12.07ab 1.29b ing for the total antioxidant activity of raspberries. Previously, total
ERZ 5 1.51bc 12.35ab 1.45ab
ERZ 6 1.47c 11.10bc 1.60ab phenolic and flavonoid contents have been reported to contribute
ERZ 7 1.70bc 13.60a 1.64ab significantly to the antioxidant activity of raspberries (Liu and oth-
ERZ 8 1.97ab 11.80ab 1.35ab ers 2002). The cancer-protective ability of colorful fruits, including
ERZ 9 1.54bc 13.10ab 1.82a raspberries, is attributed to the ability of the antioxidants in them
ERZ 10 1.67bc 13.25ab 1.64ab
to scavenge free radicals, thus preventing DNA damage and subse-
ERZ 11 1.92b 11.85ab 1.26ab
Heritage 2.32a 11.40bc 1.55ab quent mutation.
A wide variation was found among raspberry genotypes in terms
Values in the same column with different lowercase letters are significantly differ-
ent at P < 0.05. n = 5. of ascorbic acid content, ranging from 21 to 36 mg/100 g. The

Table 2 --- Total phenolic content, antioxidant activity, and ascorbic acid content of raspberry fruits.
Total phenolics -Carotene-bleaching FRAP (mol Trolox Ascorbic acid
Genotypes (g GAE/g FW) assay (%) equivalent/g FW) (mg/100 g)
ERZ1 2014a 89.3ab 117.7ab 36a
ERZ2 1516d 84.6def 88.0de 27bc
ERZ3 1741bc 87.7abcd 97.7cd 30b
ERZ4 1617cd 86.1cde 90.0de 26bc
ERZ5 2031a 89.6a 124.3a 34a
ERZ6 1926a 88.7abc 107.0bc 31ab
ERZ7 1877ab 87.2abcd 105.0c 29bc
ERZ8 1538d 85.8cde 86.7e 21d
ERZ9 1644cd 86.4bcde 90.7de 25cd
ERZ10 1181e 82.3f 81.3e 22cd
ERZ11 1145e 81.0f 83.0e 26bc
Heritage 1214e 83.0ef 83.7e 24c
BHA 88.1b
Mean 1620 86.0 96.3 28
LSD0.05 168 2.8 10.9 5.9
Values in the same column with different lowercase letters are significantly different at P < 0.05. n = 5.

Vol. 74, Nr. 7, 2009JOURNAL OF FOOD SCIENCE C577

 Characterization of red raspberry genotypes . . .
C: Food Chemistry
Figure 1 --- Regression analyses among the antioxidant ac-
tivity determination methods and total phenolics for 12
raspberry genotypes sampled in Turkey.
genotype ERZ 1 had the highest ascorbic acid content in its fruits
(36 mg/100 g). Ascorbic acid content of raspberries had previously
been reported to be between 17 and 37 mg/100 g (de Ancos and
others 2000; Pantelidis and others 2007) and it was shown that the
total antioxidant activity of raspberries was mainly from the other
phytochemicals in the fruit rather than from vitamin C (Deighton
and others 2000; Liu and others 2002). The wild raspberry samples,
in general, showed higher ascorbic acid contents compared to the
cultivated ones, except ERZ 8 and 10.
The relationships between total phenolics, FRAP, and the -
carotene assays are presented in Figure 1. -Carotene bleaching as-
say and FRAP values were highly correlated with phenol content
(R2 = 0.96 for -carotene and R2 = 0.83 for FRAP (Figure 1). In
the literature, the high correlation between antioxidant activity and
total phenolic content has been reported for different small fruits
including raspberries (Prior and others 1998; Liu and others 2002;
Pantelidis and others 2007).
Conclusions
T his study clearly shows the potential value of wild raspberry
germplasm. Raspberry fruits are a significant source of phe-
nolic compounds and ascorbic acid. Antioxidant activity was high
C578 JOURNAL OF FOOD SCIENCEVol. 74, Nr. 7, 2009
in fruits and varied greatly among the genotypes. It is clear that
raspberries higher in phenolics have higher antioxidant activity.
Therefore raspberries must be considered a good source of natural
antioxidants. In addition, a wide range of agronomic characteris-
tics, such as higher fruit weight and pest and disease resistance, of
selected genotypes could be incorporated into superior raspberry
cultivars.
Acknowledgments
This study was supported by The Scientific and Technological Re-
search Council of Turkey (TUBITAK) (project nr TOVAG-105O354);
we greatly acknowledge the financial support.
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