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Rubus Fruticosus L.: Constituents, Biological Activities and Health Related Uses

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Rubus fruticosus L. is a shrub famous for its fruit called blackberry fruit or more commonly blackberry. The fruit has medicinal, cosmetic and nutritive value. It is a concentrated source of valuable nutrients, as well as bioactive constituents of therapeutic interest highlighting its importance as a functional food. Besides use as a fresh fruit, it is also used as ingredient in cooked dishes, salads and bakery products like jams, snacks, desserts, and fruit preserves. R. fruticosus contains vitamins, steroids and lipids in seed oil and minerals, flavonoids, glycosides, terpenes, acids and tannins in aerial parts that possess diverse pharmacological activities such as antioxidant, anti-carcinogenic, anti-inflammatory, antimicrobial anti-diabetic, anti-diarrheal, and antiviral. Various agrogeoclimatological factors like cultivar, environmental conditions of the area, agronomic practices employed, harvest time, post-harvest storage and processing techniques all influence the nutritional composition of blackberry fruit. This review focuses on the nutrients and chemical constituents as well as medicinal properties of different parts of R. fruticosus. Various cultivars and their physicochemical characteristics, polyphenolic content and ascorbic acid content are also discussed. The information in the present work will serve as baseline data and may lead to new biomedical applications of R. fruticosus as functional food.
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Molecules 2014, 19, 10998-11029; doi:10.3390/molecules190810998
molecules
ISSN 1420-3049
www.mdpi.com/journal/molecules
Review
Rubus Fruticosus L.: Constituents, Biological Activities and
Health Related Uses
Muhammad Zia-Ul-Haq 1,*, Muhammad Riaz 2, Vincenzo De Feo 3, Hawa Z. E. Jaafar 4,* and
Marius Moga 5
1 The Patent Office, Kandawala Building, M.A. Jinnah Road, Karachi-74400, Pakistan
2 Department of Pharmacy, Shaheed Benazir Bhutto University, Sheringal, Dir Upper-2500,
Pakistan; E-Mail: pharmariaz@gmail.com
3 Department of Pharmaceutical and Biomedical Sciences, University of Salerno, Salerno 84100,
Italy; E-Mail: defeo@unisa.it
4 Department of Crop Science, Faculty of Agriculture, University Putra Malaysia, Selangor, 43400,
Malaysia; E-Mail: hawazej@agri.upm.edu.my
5 Department of Medicine, Transilvania University of Brasov, Brasov 500036 Romania;
E-Mail: moga.og@gmail.com
* Authors to whom correspondence should be addressed; E-Mails: ahirzia@gmail.com (M.Z.-U.-H.);
hawazej@agri.upm.edu.my (H.Z.E.J.); Tel.: +92-322-250-6612 (M.Z.-U.-H.);
+6-03-8947-4821 (H.Z.E.J.); Fax: +6-03-8947-4918 (H.Z.E.J.).
Received: 21 April 2014; in revised form: 14 July 2014 / Accepted: 16 July 2014 /
Published: 28 July 2014
Abstract: Rubus fruticosus L. is a shrub famous for its fruit called blackberry fruit or more
commonly blackberry. The fruit has medicinal, cosmetic and nutritive value. It is a
concentrated source of valuable nutrients, as well as bioactive constituents of therapeutic
interest highlighting its importance as a functional food. Besides use as a fresh fruit, it is
also used as ingredient in cooked dishes, salads and bakery products like jams, snacks,
desserts, and fruit preserves. R. fruticosus contains vitamins, steroids and lipids in seed oil
and minerals, flavonoids, glycosides, terpenes, acids and tannins in aerial parts
that possess diverse pharmacological activities such as antioxidant, anti-carcinogenic,
anti-inflammatory, antimicrobial anti-diabetic, anti-diarrheal, and antiviral. Various
agrogeoclimatological factors like cultivar, environmental conditions of the area,
agronomic practices employed, harvest time, post-harvest storage and processing
techniques all influence the nutritional composition of blackberry fruit. This review focuses
on the nutrients and chemical constituents as well as medicinal properties of different parts
OPEN ACCESS
Molecules 2014, 19 10999
of R. fruticosus. Various cultivars and their physicochemical characteristics, polyphenolic
content and ascorbic acid content are also discussed. The information in the present work
will serve as baseline data and may lead to new biomedical applications of R. fruticosus as
functional food.
Keywords: Rubus fruticosus L.; pharmacology; phytochemistry; nutrition
1. Introduction
Plant foods (fruits, herbs, nuts, spices, vegetables, legumes and grains) occupy an important
position in the economic, cultural as well as health systems of both developing and developed
countries due to their proven health-promoting claims and immunity-boosting effects. Regular
consumption of fruits, spices, nuts, legumes, vegetables and grains, is vital for a balanced and
nutritious diet and is associated with reduced risk of various ailments like inflammation, arthritis,
cancer, diabetes, cardiovascular disease, atherosclerosis, cataracts, Parkinson’s disease, Alzheimer’s
disease, and aging. The origin of many remedies, recipes and pharmaceuticals can be been plant food
especially fruits. Nutritional information of fruits and their effects on human health is among the most
frequently referenced and most sought-after items on the internet. Fruits are consumed in various
quantities as concentrated sources of energy, nutrition, vitamins, essential minerals and antioxidants by
people of all ages and income groups globally. Rubus fruticosus L. (Rosaceae) is a shrub famous for its
fruit, called blackberry, which is traded globally due to its delicious taste, pleasant flavor and
nutritional profile. The shrub is believed to have its origin in Armenia, and is now distributed throughout
Europe, Asia, Oceania and North and South America [1–3]. It grows wild in the Northern areas of
Pakistan, like Chitral [4], Dir [5], Mansehra [6], Malakand [7,8] and Kotli [9], where it is known by
local names Karwara [4,7], Ach [8], Akhara [6] and Baganrra [10,11]. Although the fruit has wide
acceptance in Pakistan, it is not cultivated on a commercial scale. The Rosaceae family is the 19th
largest family of plants [12]. The genus Rubus, with almost 700 species, is the largest genus of this
family [13]. Rubus comprises 12 subgenera, with few domesticated species [14]. Members of this genus
have been cultivated for centuries for their fruits. These fruits are consumed fresh or processed to make
food products such as jam, wine, tea, ice cream, desserts, seedless jellies and bakery products. Extracted
pigment from fruits is used as a natural colorant in baked products, jellies, chewing gums, fruit-wines
and beverages [15,16]. Due to increasing awareness about the valuable attributes of functional foods
and optimal nutrition among customers, the global consumption of fruits and fruit-based products has
increased considerably, especially in high-income countries. It is well-known now that healthiest diets
are those loaded with plant foods, especially fruit-based diets. Therefore, health care advisors and
nutrition counselors recommend inclusion of fruits and fruit-based products especially juices in the
diet. Blackberries possess a delicious taste, pleasant flavor, nice appearance and excellent nutritional
profile. Fruit are eaten raw or cooked as well as crushed to make juice. Syrups, jams and other
preserves are prepared from fruit [17]. The cooked root is also used as food [18], while leaves, whither
dried or fresh, are used as a tea [19]. The young shoots are used in salads after peeling [20].
Molecules 2014, 19 11000
The present work is a survey of research carried out on this plant. Various search engines like
SciFinder, PubMed and ScienceDirect were used to search the isolated bioactive constituents and
pharmacological activities exhibited by these compounds as well as by the crude extracts by using the
search-terms Rubus fruticosus, chemical constituents and pharmacological activities as keywords. The
main objective of the present review is to compile a comprehensive report covering medicinal,
phytochemical and nutritional attributes of different parts of the blackberry.
2. Botanical Description
2.1. Description
Rubus fruticosus L. is a semi-prostrate to almost erect, scrambling, perennial deciduous prickly,
shrub with entangling and arching stem growing up to 3 m at a fast rate. It grows in woodland garden
sunny edge, dappled shade, shady edges [21]. This bushy plant is thorny, but some cultivated varieties
are free of thorns. Blackberries are perennial, lasting for three seasons or more [3]. Plants typically
bear biennial stems or semi woody called canes. They vary from sprawling to almost erect, spreading
shrubs with thorn and leaves, the stem grow up to 7 m in length that is greenish, purplish or red in
colour. Every spring buds of the woody root produce juvenile canes which grow at a fast pace of
almost 50–80 mm per day [3]. They are categorized into two groups in terms of branch structure:
generative cane (floricane) and vegetative cane (primocane). Vegetative canes formed during first year
convert into generative canes during the second year [22]. The plant flowers in early summer and late
spring. Diameter of a flower is about 2–3 cm having 5 pale pink or white petals. Flowers have multiple
stamens. After fall of petals, fruit develops an aggregate of drupelets that are green earlier and later turn to
red to black on ripening. The color of fruit and fruit juice is an important parameter from commercial point
of view as consumers rate the product depending upon its visual appearance. The color of blackberry
fruit and its juice depends upon natural pigments present in it which in turn depends upon many factors
like cultivar being analyzed, agronomic practices utilized in cultivation, maturity stage of collection
and geological and climatic conditions of area from where fruit is collected, post-harvest storage
conditions employed and enzymatic activity and microbial contamination. Juice may be extracted from
fresh blackberry as well as from frozen. The color of frozen is much better than fresh one. Flowers and
fruit occur in a panicle-like or raceme [3]. They are formed in clusters at the end of floricanes.
Blackberry fruits twice a year both in spring (floricane) and autumn (primocane) [22]. A dense cluster
of separate units or drupelets forms the fruit which on ripening turn black or dark purple from red [3].
Seeds are light to dark brown in colour, round, 2–3 mm long with irregular and deep pits. The upper side
of leaves is dark green while underside is lighter green. Short prickles cover the stalks and veins of
leaves. Leaves are ternate above, tending to 5 or 7 palmate leaflets towards the base. Adaxial sides of
these leaflets are folded into pleats and glabrate which are dark red-purple in fall, green in summer and
deciduous in winter [3].
2.2. Cultivars
Numerous cultivars of R. fruticosus have been developed by farmers by traditional breeding
methods. These cultivars differ in fruit firmness, shape, size, flavor, color, weight, yield, ripening
Molecules 2014, 19 11001
season, nutritional contents and resistance to pests. The most famous cultivars are Jumbo, Chester,
Bartin, Ness, Bursa 1, Bursa 2, Bursa 3, Arapaho, Navaho, Thornfree, Chester Thornless, Dirksen
Thornless, Cacanska Bestrna, Loch Ness, Cherokee, and Black Satin [23,24].
2.3. Physico-Chemical Characteristics of Fruit and Oil
The increasing awareness of consumers about healthy and functional food has led to increased
consumption of fruit and fruit-based products. Physico-chemical characteristics of fruit are the key
parameters that define quality of fruit and products made there from. A good fruit flavor is due to
higher levels of sugar and organic acids. Various parameters of fruit like fruit dimensions, weight,
titratable acidity (TAc), pH and total soluble solids (TSS) contents of cultivated and wild blackberry
fruits were determined by Yilmaz et al. [23]. Fruit weight ranged from 1.2 g to 5.4 g for Arapaho and
Bursa1 cultivars respectively while it was 0.4 g to 1.2 g for wild genotypes. It indicates that cultivated
genotypes have higher mean fruit weight as compared to wild genotypes; same trend was observed for
length and width of fruit. However TSS was less in cultivated genotypes (8.6%–14.1%) than wild
genotypes (12.9%–22.3%) with overall means of 11.6% vs. 16.2%. The total soluble solid means of
wild genotypes was higher by 20%. The pH means of the wild genotypes were slightly but significantly
higher than the cultivated genotypes.
Table 1. Fruit weight, berry size and berry shape index of blackberry fruits grown
in Serbia [22].
Cultivars Fruit Weight (g) Length (mm) Width (mm) Shape Index
Year 2010 2011 2010 2011 2010 2011 2010 2011
Cacanska Bestrna 7.57 7.61 26.62 27.54 20.31 21.35 1.31 1.29
Black Satin 6.45 7.24 25.96 27.08 20.40 21.28 1.27 1.27
Thornfree 4.65 5.32 21.52 23.69 17.52 19.31 1.23 1.22
Loch Ness 7.76 7.61 28.13 27.15 21.78 20.69 1.30 1.32
Dirksen Thornless 4.54 6.91 27.28 28.10 19.31 20.33 1.41 1.38
Chester Thornless 5.31 6.11 24.13 25.01 19.72 20.82 1.23 1.20
Navaho 5.39 5.90 22.65 23.12 19.46 19.80 1.16 1.17
Mean over years 5.95 6.67 25.18 25.96 19.78 20.51 1.27 1.26
Table 2. Soluble solids, titratable acidity and ripening index of blackberry fruits grown
in Serbia [19].
Cultivar Soluble Solids (°Brix) Titratable Acidity (%) Ripening Index
Year 2010 2011 2010 2011 2010 2011
Thornfree 7.70 8.66 1.72 1.60 4.48 5.41
Cacanska Bestrna 6.40 7.82 1.89 1.64 3.39 4.77
Loch Ness 9.25 9.35 1.56 1.42 5.93 6.58
Dirksen Thornless 6.80 9.76 1.51 1.24 4.50 7.87
Black Satin 6.70 6.89 1.57 1.42 4.27 4.85
Chester Thornless 9.20 9.27 1.44 1.27 6.39 7.30
Navaho 9.35 9.67 1.33 1.08 7.03 8.95
Mean over years 7.91 8.77 1.57 1.38 5.14 6.53
Molecules 2014, 19 11002
Milosevic et al., compared physio-chemical characteristics of fruits of different cultivars blackberry
grown in Serbia in two years. A large variation was observed in parameters investigated. (Tables 1 and
2) [24].
The soluble solids which represent sugar level in fruits and pH and titratable acids which represent
total acids contribute to sweetness and acidity of fruits and products made from them. Blackberry
cultivars grown in different regions of Turkey had total soluble solids (8.98%–20.2%), weight (2.0–6.6 g),
pH (3.3–3.6) and acidity (1.0%–3.1%) for cultivated blackberry while weight (1.5–2.1 g), TSS
(11.3%–13.1%), pH (3.33–3.35) and acidity (0.7%–1.0%) for wild blackberries [25–27]. The fruit
acidity is due to presence of organic acids especially malic acid. The balance between soluble solids
contents and titratable acidity is determined by sugars and organic acids ratio and this determines
flavor of fruit. Fruit parameters like fruit dimensions, weight, titratable acidity (TAc), pH and total
soluble solids (TSS) contents depends upon fruit variety, agronomic practices employed, stage of
collection of fruits and climatic and geological condition of area from where fruits are collected.
Determination of these parameters is of main interest and first step during nutritional evaluation of
fruits and it dictates further studies on components which seem more interesting. Dimića et al.,
reported the technological quality characteristics of dried pomace of blackberry as well as total
carotenoid and chlorophyll contents and physio-chemical characteristics of oil (Tables 3 and 4). Fresh
fruits were frozen for 8 months and then pressed to extract juice. The residue obtained from pressing
fruits (pomace) was collected and dried by two ways:
B1 = pomace dried (22 °C) for 3 days
B2 = pomace dried at 63 ± 2 °C and 103 ± 2 °C for 20 h each
Table 3. Technological quality parameters of blackberry seeds grown in Serbia [28].
Parameter Blackberry Seeds
B1 B2
Water content (%) 6.59 5.24
Oil content (%)
- telquel (as is)
- on dry basis
13.05
13.97
13.59
14.34
Impurities content (%)
Pure seeds content (%)
4.68
95.32
4.36
95.64
Weight of 1000 seeds * (g)
- telquel (as is)
- on dry basis
3.21
3.45
3.32
3.50
Specific weight (g/mL)
- pure seeds *
- telquel seeds (as is)
0.999
0.997
0.997
0.993
Weight per liter (g/L):
- pure seeds *
- telquel seeds (as is)
423.6
384.8
429.2
394.0
*: pure separated seeds from pomace by hand.
Molecules 2014, 19 11003
Table 4. Important quality parameters of blackberry seed oils grown in Serbia [28].
Parameter Blackberry Seed Oil
B1 B2
Acid value (mg KOH/g) 6.85 7.05
FFA (% oleic acid) 3.43 3.53
Peroxide value (mmol/kg) 8.89 11.16
Total carotenoids (mg/kg) 32.30 33.92
Total chlorophyll (mg/kg) Cyclohexane
Chloroform
3049.52
1505.78
3094.98
1583.62
Transparency (%) Cyclohexane
Chloroform
25.84
24.33
19.28
18.87
A brown-greenish oil (due to presence of high chlorophyll contents) was obtained by n-hexane
extraction of this pomace. Various physico-chemical parameters of this oil were studied. Total
chlorophyll content as well as transparency (%) of oil was studied by dissolving oil in two different
solvents (i.e., cyclohexane and chloroform) and results were significantly different for both solvents [28].
3. Phytochemistry
The profile and contents of bioactive contents and constituents, fixed and essential oil, fatty acids,
tocopherol and sterols, minerals, amino acids, vitamins, protein and carbohydrate contents of fruits or
products made from them depends upon fruit variety, agronomic practices utilized in cultivation, stage
of collection of fruit, geological and climatic conditions of area from where fruit is collected and the
method utilized for their determination. Proper identification and quantification of bioactive
constituents is necessary to understand the underlying mechanism of biological and pharmacological
activities of extracts of plants as these properties are due to presence of bioactive constituents.
Blackberry fruit itself, and its products as well as by-products are a rich source of phytochemicals and
natural antioxidants which are being explored for their health promoting activities. Detailed quantitative
data of bioactive components is still needed and their structure activity relationship should be investigated.
3.1. Compositional Studies of Fruit
Due to the proven benefits of regular consumption of fruit and vegetables in promoting health and
combating metabolic disorders and chronic diseases like cancer, diabetes mellitus, hypertension,
cardiovascular diseases, gastrointestinal diseases, atherosclerosis, aging, Parkinson’s and Alzheimer’s
disease in humans, their consumption has increased globally. The health benefits of fruits and fruit
products are due to their low calories, less energy density and low fat contents, higher vitamins,
minerals, fibre and simple sugar contents and presence of various bioactive constituents in them. The
nutritional profile of berry fruit indicates presence of carbohydrates, vitamins, minerals as well as
dietary fibre (Table 5).
This profile of fruit indicates its potential use in diet-based therapies for improving human health.
Due to high water content, carbohydrate content of fruit is less as compared to cereals. Like other
fruits, its fruits also have less quantity of protein and sodium. Usually, protein content of fruits is less
than 3.5% with exceptions. Similarly lipid content of its fruits like other fruits is not greater than 1%.
Molecules 2014, 19 11004
Like most other fruits, it is rich in potassium which may help in reducing risk of developing kidney
stones, bone loss and blood pressure. Milosevic et al., performed a comparative study of sugar and
ascorbic acid contents of fresh fruits of blackberry (Table 6) [29].
Table 5. Blackberries nutritive value per 100 g [30].
Component Nutrient Value Percentage of RDA
Energy 43 Kcal 2%
Carbohydrates 9.61 g 7%
Total Fat 0.49 g 2%
Protein 1.39 g 2%
Dietary Fiber 5.3 g 14%
Cholesterol 0 mg 0%
Folates 25 µg 6%
Pyridoxine 0.030 mg 2%
Niacin 0.646 mg 4%
Pantothenic acid 0.276 mg 5.5%
Thiamin 0.020 IU 2%
Vitamin C 21 mg 35%
Vitamin A 214 IU 7%
Vitamin K 19.8 µg 16.5%
Vitamin E 1.17 mg 8%
Potassium 162 mg 3%
Calcium 29 mg 3%
Sodium 1 mg 0%
Magnesium 20 mg 5%
Copper 165 µg 18%
Iron 0.62 mg 8%
Zinc 0.53 mg 5%
Manganese 0.646 mg 3%
Selenium 0.4 µg 1%
Carotene-α 0 µg --
Carotene-β 128 µg --
Lutein-zeaxanthin 118 µg --
RDA = Recommended daily allowance.
Table 6. Sugar and ascorbic acid contents (FW) in blackberry cultivars grown in Serbia [30].
Cultivars Reducing
Sugars (%) Sucrose (%) Total Sugars (%) Ascorbic Acid
(mg /100 g)
Black Satin 5.65 0.98 6.68 38.72
Dirksen Thornless 7.98 1.00 9.04 35.20
Chester Thornless 8.18 0.89 9.12 36.96
Thornfree 6.12 0.86 7.02 40.48
Čačanska Bestrna 7.36 0.85 8.25 42.24
Loch Ness 9.01 0.90 9.96 44.00
Navaho 9.08 1.08 10.22 35.20
Molecules 2014, 19 11005
Since ascorbic acid is a water-soluble vitamin, it is present in excessive amounts in fruits and
vegetables having water contents more than 50%. It explains higher level of ascorbic acid in
blackberry fruit. The fruit is a rich source of carbohydrates most of which is present as sugars thereby
making fruit a high source of energy. These sugars are also a basis of sweetness of fruit. The fruit may
be included in nutritional support and dieto-therapy programs to prevent lifestyle-related diseases like
diabetes mellitus and cancer due to presence of sufficient amount of ascorbic acid and folic acid.
Fruits and fruit juices are very important in human nutrition as vital source of nutrients, non-nutritive
food constituents and for reduction of various disease risks. Therefore ad commercials and campaigns
to increase their consumption are justified as a policy to decrease burden of diseases. Stajcic et al., reported
chemical composition of two blackberry cultivars i.e., Cacanska bestrna and Thornfree (Table 7) [31]. The
compositional data of fruits is a vital information for food scientists as it is an index of total energy
content, nutrients and calories present in that fruit. This information helps to establish the relationships
between fruit intake and disease in specific population and also helps in formulation of recommended
dietary intakes (RDI) and recommended dietary allowance (RDA) values for that fruit.
Table 7. Chemical composition of two blackberry cultivars grown in Serbia [31].
Parameter (g/100 g FW *) Čačanska Bestrna Thornfree
Total solids 11.96 15.57
Ash 0.29 0.41
Cellulose 2.2 2.97
Pectin 0.29 0.30
Pectic acid 0.1 0.10
Protopectin 0.15 0.17
Acidity 1.36 1.39
Total sugars 5.36 5.98
Reducing sugars 1.46 1.32
Sucrose 3.71 4.43
Proteins 1.4 1.49
* FW: fresh weight of berry fruits.
Various other scientists also reported glucose [32], fructose [33] and sucrose from the fruit [34].
Pectins have also been reported from the fruit of R. fruticosus [35]. Organic acids are primary
metabolites found mostly in fruits. Various organic acids like citric [32] malic [33] and galacturonic
acids [35] have been found in the fruit. Organic acids are usually present in minor concentration in
fruits and are responsible for fruit flavor. They help to stabilize anthocyanins and ascrorbic acid in
fruits and these acids in combination with sugar also impart sensory characteristics to fruits. The
composition of sugars detected in blackberry (fruits) indicates that fructose is predominant, followed
by glucose [36]. Since fructose is sweeter than glucose, its high concentration is a desirable
organoleptic characteristic of fruits. In a recent study, blackberry expressed the lowest values of
fructose, sucrose and glucose contents (64.5 mg/g FW, 76.1 mg/g FW, 3.0 mg/g FW) respectively.
“Thornfree” had highest levels of fructose and glucose. Sucrose was present in much lower quantities
as compared to the other sugars in wild varieties, because it is converted to inverted forms during the
ripening process. Significant differences in malic acid content were observed between wild and
Molecules 2014, 19 11006
cultivated species [37]. Vitamins such as A, C, E, and folic acid were reported in fruit powder of
R. fruticosus during anticancer studies on berries [38].
Stefanut et al., reported macro-mineral and micro-mineral concentration of Zn, Cu, Al, Mn, Co, Fe
as 140, 50, 27, 33, 1, 30 (μg/100 g fruits) in fresh blackberry (fruit) respectively [39]. Radocaj et al.,
reported that pomace, even after extended frozen storage, is a good raw material for oil extraction and
a rich source of functional bioactive constituents (Table 8). The quality characteristics of blackberry
seed oils were studied. The results indicated that prolonged freezing time as well as pomace drying
method did not influence fatty acid profile of oils extracted from pomaces. The results indicated that
best drying regime for blackberry pomace was the two step drying process [40]. Presence of higher
amounts of α-tocopherol in pomace and its known highest biological activities than other tocopherols,
indicates potential use of pomace in food, pharmaceutical and cosmetics industries as value added
natural extract.
Table 8. Chemical composition of oils extracted from blackberry pomaces grown in Serbia [40].
Parameter B0 B1 B2
Water content (%) 6.08 6.55 5.20
FFA (% oleic acid) 1.18 3.44 3.54
PV (mmol/kg) 3.73 8.84 11.14
Induction period (h) at 100 °C 7.50 6.30 6.80
Campesterol 781.7 757.1 771.8
Stigmasterol 1090.4 1052.9 1087.1
β-sitosterol 4370.5 4331.9 4337.9
Total sterols content (mg/kg) 6242.6 6159.8 6196.8
α-tocopherol 133.2 79.1 110.7
β-tocopherol 1097.9 1051.9 1062.5
γ-tocopherol 823.2 565.7 624.5
Total tocopherols (mg/kg) 2054.3 1696.7 1797.7
Σ-SFA 7.53 7.13 7.48
Σ-MUFA 19.97 17.87 19.03
Σ-PUFA 78.56 74.94 75.66
Total phenolics content (mg GAE/kg) 306.5 226.9 256.6
B0: fresh dried at 22 °C/72 h; B1: freeze dried at 22 °C/72 h; B2: freeze dried at 63 °C/20 h
and 103 °C /2 h.
α-tocopherol, γ-tocopherol, δ-tocopherol and γ-tocotrienol were reported in seed oils from Korean
thornless blackberry [41–43]. Mazur and co-workers isolated Δ7-avenasterol, squalene, daidzein,
genistein, secoisolariciresinol and matairesinol from fruits of R. fruticosus. Other sterols in the seed oil
of R. fruticosus include campesterol, Δ5-avenasterol, stigmasterol and β-sitosterol [44]. Both saturated
and unsaturated fatty acids have been observed in seed oil, the major fatty acids being lauric, myristic,
palmitic, stearic, oleic, linoleic, α-linolenic and arachidic acids. Lead was detected in shoots and roots [45]
while rare earth elements, viz La, Lu, Ce, Yb, Sm, Tb, Nd and Eu, were found in leaves of
R. fruticosus [46]. Toth and coworkers reported the presence of minerals, viz chromium, zinc,
manganese, calcium, copper, iron and nickel, in fruit and leaves of R. fruticosus [47].
Molecules 2014, 19 11007
3.2. Phenolic Acids, Flavonoids and Anthocyanins
The health promoting properties and immunity-boosting effects of fruits, vegetables and products
made from them depend on concentration and profile of phenolic acids, flavonoids, carotenoids,
anthocyanians, vitamins and minerals present in them as well as on quantity and frequency of their
daily intake and their bio-avilibity to human physiological system after digestion. Therefore determination
of phenolic acids, flavonoids, carotenoids, anthocyanians, vitamins and minerals is of prime importance in
assessment of nautraceutical values. Total phenolic contents determined in a recent study [31] are from
1.74 mg GAE/g to 1.97 mg GAE/g which are in good agreement with previously published data [48].
These results are slightly lower than those obtained in some thornless blackberry cultivars grown in
Italy [49]. Phenolic acids, like ellagic, gallic, caffeic acid and p-coumaric acids, and flavonoids, such
as quercetin, hyperoside, kaempferol, myricitin, (+)-catechin, (–)-epicatechin, epicatechin gallate,
procyanidin B1 and quercetin-3-D-glucoside, have been identified in fruit and leaves of
R. fruticosus [34,50–55]. Radovanović et al. [50] has reported individual contents of phenolic acids
present in blackberry fruit (Table 9).
Table 9. Phenolic acids profile of blackberry fruit [50].
Phenolic Acid Contents (mg/kg Fresh Weight)
Gallic acid 137.98
t-Caftaric acid 0.99
Caffeic acid 0.33
Syringic acid 3.71
Procyanidin B2 1.49
(+)-Catechin 4.09
()-Epicatechin 3.63
Quercetin-3-Glycoside 3.53
Rutin 22.77
Quercetin 3.79
Sellappan and co-workers compared the chemical composition of wild blackberry and common
commercial cultivars for ellagic acid and flavonols. They found significant differences between the
amounts of individual flavonols and ellagic acid. Wild cultivars of R. fruticosus contained the highest
values of myricetin, kaempferol, and ellagic acid contents. Wild cultivars of R. fruticosus had twice
higher contents of ellagic acid as compared to cultivated genotypes [56].
Anthocyanins are a group of flavonoid derivatives and water soluble natural pigments as they give
color to flowers and fruits. Animal model studies indicate that anthocyanins possess anti-carcinogenic,
anti-inflammatory, and anti-obesity activities besides their role in preventing diabetes mellitus and
cardiovascular diseases.The total anthocyanin content of a blackberry crude extract was 17.1 mg/g of
freeze-dried powder, which was equival to 176 mg/100 g of blackberry [57]. The primary anthocyanin
detected in blackberry is cyanidin-3-O-glucoside. Various other anthocyanins are also detected in
blackberry fruit like cyanidin-3-O-xyloside, cyanidin-3-O-dioxaloylglucoside and cyanidin-3-O-(600-
malonyl)-glucoside [58]. Smaller amounts of other anthocyanins reported in blackberry are
pelargonidin-3-O-glucoside, malvidin-3-O-glucoside, cyanidin-3-O-arabinoside, cyanidin-3-O-xyloside,
Molecules 2014, 19 11008
cyanidin-3-O-rutinoside, cyanidin-3-O-dioxalylglucoside and cyanidin-3-O-glucoside acylated with
malonic acid [59–65]. Cyanidin- 3-O-saccharide was also reported from stems and leaves [66]. A
comparative study of presence of these antioxidant constituents in blackberries is given below (Table 10).
Fruits exhibit different antioxidant capacity due to variations in vitamin C and E contents, phenolic,
flavonoid and anthocyanin contents, solvents used for extraction and method used to assess antioxidant
activity. All these factors make it difficult to announce a definite antioxidant potential of fruits
however it helps to get an idea about average antioxidant activity of fruits.
Table 10. Total polyphenols, total anthocyanins and ascorbic acid in blackberry.
Number of
Cultivars
Total Polyphenols
(mg/100 g)
Total Anthocyanins
(mg/100 g)
Ascorbic Acid
(mg/100 g) Reference
4 2030 134–152 15.22 [36]
7 289.3 88.7 12.9 [67]
27 460 141 NR [37]
2 417–555 110–122 NR [56]
6 NR NR 14.9 [68]
3 226 152.8 NR [69]
1 (Chester) 361 NR NR [70]
5 320 80 20.4 [48]
4 07.5 115 NR [71]
2 NR NR 6 [72]
NR = Not Reported.
3.3. Carotenoids
Carotenoids are an important group of fat-soluble natural pigments and are believed to possess
various immunity-boosting properties and health promoting effects. Marinova and Ribarova isolated
lutein, β-cryptoxanthin, lycopene, zeaxanthin, β-carotene and α-carotene from R. fruticosus fruit [73].
Rutz et al., investigated effect of maturity on lutein, zeaxanthin and β-carotene contents in pulp of
blackberry fruit (Table 11). It was observed that carotene contents decreased with maturity stage of
fruit [74].
Table 11. Carotenoid contents (μg/g) of blackberries (cv. Tupy) at different maturity stages [74].
Maturity Stage Lutein + Zeaxanthin β-Carotene
Immature 0.66 0.400
Intermediate 0.235 0.078
Mature 0.00 0.162
β-carotene has potential of transformation to vitamin A, thereby imparting an important nutritional
role of acting as antioxidant to berry fruit.
3.4. Aromatic Compounds
Aromatic compounds are always present in plants as byproducts. Some important aromatic
compounds isolated from the fruit of R. fruticosus are furans, 5-hydroxymethylfurfural and 2, 3-dihydro-3,
Molecules 2014, 19 11009
5-dihydroxy-6-methyl-4H-pyran-4-one. The aroma of blackberry fruit is due to presence of
5-hydroxymethylfurfural [75].
3.5. Triterpene Acids
Triterpenoids are polycyclic compounds that are derived from linear hydrocarbon squalene and
exert various biological activities due to their unique structure. Triterpene acid such as rubutic acid and
rubinic acid were isolated from the leaves of R. fruticosus. 2-α-Hydroxyursolic acid and β-amyrin were
also reported [76,77]. Figure 1 reports some of the main constituents found in R. fruticosus.
Figure 1. Phytochemicals isolated from R. fruticosus.
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Figure 1. Cont.
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Figure 1. Cont.
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4. Traditional Uses
Fruits and fruit bearing plants are believed to possess various health-promoting effects and
immunity-boosting properties since long ago. Romans treated various diseases through the use of tea
prepared from its leaves [20]. R. fruticosus is known as food form about 8,000 years and as a medicinal
plant soon after the Ice Age [77]. Hippocrates recommended blackberry stems and leaves soaked in
white wine to relieve difficulties in childbirth and as an astringent poultice on wounds [78]. Externally
it is used as a gargle to treat gum inflammations, sore throats and mouth ulcers [79,80]. Decoction of
leaves is used as a gargle or mouthwash and also used to treat thrush [17]. The fruit juice is used to
treat asthma [7]. The leaves of the plant are also used in various respiratory problems [81]. Blackberry
juice is recommended in colitis while tea made from its roots is used for relief in labor pain. Poultice is
leaves are applied in skin ulcers. The fruit and juice is recommended in anemia. R. fruticosus leaves or
maceration of the tops in sunlight is claimed as cicatrizing agent [82]. A methanol extract of the aerial
parts has been used for wound healing, as an antiseptic and a disinfectant and to treat cough [83,84].
R. fruticosus cures skin wounds in cattle [85]. A decoction of the twig tops soothes menstruations and
also is used to treat diarrhoea. Its leaves are chewed to strengthen the gums and to cure thrush. Leaves
are wrapped to stop fungal infection and abscesses on skin [86]. R. fruticosus jams, prepared without
sugar, is prescribed to cure throat ailments in children and as an anti-diarrhea [87].The root-bark and
the leaves are depurative, strongly astringent, tonic, vulnerary and diuretic. It is used as an excellent
remedy against diarrhoea, dysentery, cystitis and haemorrhoids [7,86,87].
5. Pharmacological Actions
Fruits, vegetables, herbs and spices have been used since long to cure various human ailments
besides their nutritional importance. This curative potential has been ascribed to various bioactive
constituents and antioxidant components present in them and their synergistic effects. The most
important activities of blackberry are anti-microbial, antioxidant, anti-inflammatory and anti-cancer.
Several factors such as cultivar, agroclimatological conditions, level of ripening and processing
method affect the profile and intensity of these pharmacological activities. Most activities performed
are on crude extracts without sufficient information on preparation and standardization of extracts so
many times results are non-reproducible. Most of pharmacological activities can be linked to various
phenolic compounds which help in scavenging free radicals which are root cause of various
pathological and metabolic disorders. Although many traditional uses have been verified, however
in vitro as well as in vivo pre-clinical and clinical studies are necessary to assess their safety and efficacy.
5.1. Antimicrobial Activity
Riaz and coworkers studied the possible antibacterial activity of the methanol extracts from various
parts of the plant against eight bacterial strains (Salmonella typhi, Escherichia coli, Streptococcus
aureus, Micrococcus luteus, Proteus mirabilis, Bacillus subtilis Citrobacteri sp., Pseudomonas
aeruginosa). All extracts were found to inhibit growth of bacteria. The order of potency on minimum
inhibitory concentration was stem > root > leaves > fruit. The same authors also screened the methanol
extracts for their antifungal potential against nine pathogenic fungal strains (Yersinia aldovae,
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Aspergillus parasiticus, Candida albicans, Aspergillus niger, Aspergillus effusus, Macrophomina
phaseolina, Fusarium solani, Trichophyton rubrum, Saccharomyces cerevisiae) without recording any
biological activity [88]. Blackberry juice inhibited the growth of Bacillus cereus, Bacillus subtilis,
Streptococcus marcescens and Escherichia coli from 50% to 75%. A methanol extract of aerial parts
of R. fruticosus inhibited Mycobacterium tuberculosis with MIC of 1 mg/mL in agar dilution test [89].
Fruit cordials were reported to be bacteriostatic [90]. Abachi et al., reported that MIC values of aqueous
and ethanolic extracts against Helicobacter pylori were 400 and 450 µg/mL while zone of inhibitions
were 8 and 7.3 mm for same extracts respectively [91]. Radovanović et al., reported that blackberry
extracts exhibited strong antioxidant potential against Gram () bacteria S. enteritidis ATCC13076and
against Gram () bacteria S. aureus ATCC 6538, while weak to moderate activity was observed
against Clostridium perfringens ATCC19404, Bacillus subtilis ATCC 6633, Listeria innocua
ATCC33090, Sarcina lutea ATCC9341, Micrococcus flavus ATCC40240 and against gram negative
bacteria like Escerichia coli ATCC25922, Pseudomonas aeruginosa ATCC9027, Shigella sonnei
ATCC25931, Klbsiella pneumonia ATCC 10031 and Proteus vulgaris ATCC 8427 [37]. Yang et al.,
reported that juice of fruits of R. fruticosus had strong antimicrobial potential against food borne
pathogens like Listeria monocytogenes, Salmonella Typhimurium, Escherichia coli, Lactobacillus
casei, Lactobacillus plantarum and Lactobacillus rhamnosus. The results suggest potential use of juice
as a preservative in food processing industries [92]. Salaheen et al., investigated the effect of extracts
of blackberry pomace on growth and pathogenicity of Campylobacter jejuni. The extracts decreased
the growth, swimming and swarming motility of C. jejuni and changed cell-surface hydrophobicity and
auto-aggregation of these bacteria. The results indicate potential use of pomace extracts to reduce
colonization level of C. jejuni in poultry and in controlling growth of pathogens in meat and meat
products [93].
5.2. Antioxidant and Anticancer Activity
There is no standard method of preparation of extract of a plant part and its antioxidant analysis and
it has led to diverse rather confusing reports when comparing the antioxidant potential of extracts of
the same parts of the same plant, even from same regions with similar agro-geo-climatoligical
conditions. Since different protocols for assessment of antioxidant capacity are based on different
mechanisms, scientists therefore use a battery of assays when analyzing the antioxidant potential of
any plant extract. Blackberries are a rich source of natural antioxidants as they contain high levels of
phenols, flavonols and anthocyanins and are therefore well-reputed scavengers and inhibitors of free
radicals [68]. Anthocyanins of blackberry are predominantly cyanidin based in non-acylated form. An
ethanol extract of leaves was reported for its strong antioxidant potential [94]. Due to its antioxidant
activity, blackberry exhibited chemopreventive effects in rats [38]. The antioxidant activity of fruit was
also evaluated using ORAC method [68]. Blackberry extracts effectively suppressed the production of
intracellular peroxyl free radicals induced by AAPH in human intestinal cell line (INT-407 cells) and
this effect was concentration dependent. The suppression of intracellular oxidation by blackberry
extract occurred at concentrations that were not toxic to the INT-407 cell line [95]. Blackberry
powders were mixed with a synthetic diet (AIN-76), at 5% to 10% concentrations and fed to
rats (Fischer 344) before, during and after treatment with the esophageal carcinogen
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N-nitrosomethylbenzylamine. At 25 week of experiment, all berry types inhibited the number of
esophageal tumors (papillomas) in NMBA-treated animals by 24%–56% as compared to controls. This
inhibition was associated with decrease in the formation of the NMBA-induced O-6-methylguanine
adduct in esophageal DNA, indicating that the berries influenced the metabolism of NMBA leading to
reduced DNA damage and thus preventing esophageal cancer in rats [38].
Cyanidin-3-O-glucoside isolated from blackberry possesses strong antioxidant activity and inhibited
neoplastic transformation, metastasis, neoplastic cell migration and invasion, activation of tumor cell
markers (NF-αB, AP-I, COX-2, TNF-α and MAPK), activation of cell migration markers (JNK, p38,
and ERK), and induces apoptosis in neoplastic cell (HL-60 cells) [96].
Halvorsen and coworkers investigated the total antioxidant capacity of cultivated R. fruticosus
collected at three different locations. The wild blueberry, wild blackberry, crowberry, sour cherry,
black currant, wild strawberry, cultivated blackberry and cowberry/cranberry contained very high
amount of total antioxidant concentration (5.03 to 9.17 mmol/100 g) [97]. Presence of anthocyanin in
general and cyanidin-3-glucoside in particular in blackberry is the source of antioxidant capacity to
repress both peroxyl-radical induced chemically and intracellular oxidation [95]. All conventional
anticancer treatments like chemotherapy, surgery and radiotherapy have some side-effects. So scientists are
looking for alternative anti-cancer remedies. Apoptosis of cancer cell is a unique target for
chemoprevention study. A blackberry extract induced apoptosis in human leukemia HL-60 cells [98].
Some blackberry extracts (Hull Thornless, Chester Thornless, Triple Crown) induced apoptosis in
human leukemia cells (HL-60) in a dose-dependent manner. Induction of apoptosis may be due to
presence of various components in the extract; however the possible role of antioxidant potential of the
extract may not be neglected in enhancement of cancer cells apoptosis. This indicates that there is a
significant relationship between antioxidant activity, antioxidant content and anticancer activity in
blackberries [99].
Wang and coworkers found that the pre-harvest application of methyl jasmonate (MJ) increased
blackberry fruit quality significantly. MJ treated blackberries had low titratable acid content and high
soluble solids as compared to untreated fruits. MJ treatment also significantly increased flavonoid
contents, the antioxidant capacity and the inhibition of proliferation of cell lines (A549, HL-60) and
also induced apoptosis in cell lines (HL-60) [100]. Tate and coworkers studied eight varieties of
R. fruticosus (Arapaho, Chickasaw, Hull, Chester, Choctaw, Navajo, Kiowa, Triple Crown) to
determine the inhibitory effect on UV-C–induced mutagenesis in Salmonella typhimurium TA100.
Chester and Navajo varieties showed significant suppression of mutagenesis [101].
Intake of blackberry juice (BJs) prepared in water (BJW) and defatted milk (BJM) affects the
plasma antioxidant power and non-enzymatic and enzymatic antioxidants. Ascorbic acid content
increased significantly in plasma after intake of both BJs. However α-tocopherol and plasma urate
were not affected. The plasma antioxidant capacity increased only after consumption of BJW. Plasma
antioxidant capacity showed a positive correlation with ascorbic acid and a negative correlation with
urate level. However no correlation was found among antioxidant capacity and total cyanidin or total
ellagic acid contents. Intake of blackberry juice also increased plasma catalase level. A significant
decrease in the urinary antioxidant capacity was observed [67]. Antioxidant profile of various cultivars
is given in Table 12 [49]. Since synergetic effects exist between various bioactive compounds, so
antioxidant capacity may be higher than measured for individual bioactive constituent, and therefore
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aggregate antioxidant potential of fruits should be measured instead of individual bioactive
compounds.
Table 12. Phenolic, anthocyanin and ascorbic acid contents and DPPH radical scavenging
activity of blackberry fruits [49].
Cultivar Total Polyphenols
(mg/100 g)
Total Anthocyanins
(mg/100 g)
Ascorbic Acid
(mg/100 g) EC50 (mg)
Thornless Boy Sembes 329.1 126.9 12.5 5.2
Smoothstem 289 86.8 12.4 4.6
Black Diamond 307.4 119.3 13.1 5.7
Darrow 192.8 67.4 12.9 5.7
Hull Thornless 236.7 69.1 13.0 6.2
Chester 351.7 76.2 13.0 7.6
Black Satin 317.3 75.1 13.1 9.5
Means 289.3 88.7 12.9 6.4
Huang et al., reported that blackberry extracts exhibited a strong DPPH scavenging activity
(95.37%) at 2 mg/mL. Antioxidant activity observed was TEAC was 11.48 mmol Trolox/100 g DW,
EC50 of DPPH was 0.44 mg/mL, TAC was 3.99 mg catechin/g DW,TFC was 11.83 mg rutin/g DW,
TPC were v5.58 mg gallic acid/g DW. Phenolic acids, flavonoids and tannins detected were gallic
acid; gallocatechin; protocatechuic acid; epigallocatechin; catechin;7, p-hydroxybenzoic acid; caffeic
acid; malvidin-3-glucoside; p-coumaric acid; catechin gallate; cyanidin; ellagic acid; quercetrin
(quercetin-3-rhamnoside); cinnamic acid and luteolin [102].
Samec et al., investigated effect of temperature and time on blackberry fruits (Table 13). Storage of
blackberry fruits at refrigerator temperature helped in preservation of fruit qualities by 1.6 to 5.5 fold
as compared to at room temperature. Storage at 25 °C led to spoilage of analyzed fruits while storage
at 4 °C did not adversely affect phytochemicals in analyzed fruits [103].
Table 13. Antioxidant components of blackberry fruits stored at 25 °C and 4 °C [103].
Days Total Phenol Content
(mg GAE/100 g FW)
Total Flavonoid Content
(mg CE/100 g FW)
Total Anthocyanin Content
(mg CGE/100 g FW)
25 °C 4 °C 25 °C 4 °C 25 °C 4 °C
0 364.24 66.13 121.82
2 301.33 347.39 68.27 75.20 117.79 134.67
4 391.76 371.39 69.90 63.89 141.37 145.45
9 391.27 65.20 163.90
14 379.88 73.77 144.22
Stajčić et al., reported chemical composition, total phenolic, flavonoid and monomeric anthocyanin
contents as well as antioxidant activity two blackberry cultivars, i.e., Čačanska bestrna and Thornfree
(Table 14) [31].
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Table 14. Total phenolic, flavonoid and monomeric anthocyanin contents and antioxidant
activity two blackberry cultivars [31].
Parameter Čačanska Bestrna Thornfree
Total Phenolic Contents
(mg GAE/100 g FW) 235.09 270.22
Total Flavonoind contents
(mg RE/100 g FW) 143.33 172.95
Total Monomeric
Anthocyanin Contents
(mg CGE/100 g FW)
50.95 102.31
DPPH radical Scavenging Activity
EC50 (mg FW/mL) 0.8188 0.6691
EC50 (mg extract/mL) 0.0616 0.0646
Radovanović et al., also investigated antioxidant potential of blackberry fruits (Table 15). Phenolic
acids identified were galic acid, caftaric acid, siringic acid, ferulic acid, while flavonoids detected were
catechin, epicatechin, quercetin and quercitin-3-glycoside and rutin. All extracts showed high
scavenging effect on DPPH radical with IC50 values ranging from 22.19 to 31.18 mL/g [40].
Table 15. Antioxidant potential of blackberry fruits [40].
Antioxidant Potential Contents
Total phenols(mg GAE/kg) 7838.26
Total tartaric esters(mg CAEb/kg) 291.91
Total flavonols (mg QEc/ kg) 647.68
Radical scavenging activity(ml/g) 31.18
Ştefănuţ et al., reported the anthocyanins, phenolics and antioxidant activity of fresh fruit of
blackberry as 1,343 mg/L, 3,284 mg GAE/L and 17.3 (μM TE/gFM) respectively [39]. Percentage
compositions of anthocyanins detected are reported in Table 16 [39].
Table 16. Anthocyanin contents of acidified ethanol extract of blackberry [39].
Anthocyanin Type % of Total Anthocyanins
Cyanidin-3-sambubioside 0.84
Cyanidin-3-glucoside 90.72
Cyanidin-3-xyloside 3.44
Cyanidin-3-malonylglucoside 2.97
Cyanidin-3-dioxalylglucoside 2.04
Najda and Labuda reported total phenolic contents, anthocyanin contents and flavonoid contents of
fresh fruits which were 101,947, 38,021 and 4,291 per 100 gram of fruits. Values for antioxidant activity
(µMTE/g of fresh fruits) were 1,293, 971 and 517 respectively for DPPH, FRAP and ABTS [104].
Salehi et al., determined effect of solvent on phenolic contents and antiradical activity of blackberry
extracts (Table 17). Methanolic and n-hexane extracts contained highest and lowest amounts of
phenolic contents respectively. Same trend was observed for DPPH radical scavenging assay [105].
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Table 17. Total phenolic content and antiradical activity of blackberry extracts [104].
Extracts Total Phenolic Content
(mg GAE/g of Extract)
DPPH Radical Scavenging
Activity (IC50 μg/mL)
n-Hexane 12 76.5
Dichloromethane 8.9 30.1
Chloroform 27 54.8
Ethylacetate 77.9 35.5
Methanol 79.1 15.2
Ivanovic et al., studied effect of sonication time and temperature on yield, anthocyanin (cyanidin)
contents and antioxidant potential of ultrasound-assisted extracts of blackberry. It was observed that
increase of sonication time as well as temperature increased yield, anthocyanin contents as well as
antioxidant potential of extracts. The results suggest use of ultrasound-assisted extraction technique for
better isolation of anthocyanins from blackberry extracts [100]. Total phenolic contents and
antioxidant activity of liqueurs made from different fruits was comparatively measured with regard to
storage temperature and time (Table 18). In blackberry liqueur, the phenolic compounds, flavonols and
anthocyanins decreased during storage. It is well-known that food commodities and plant parts like
fruits and seeds undergo transformations during storage. Contents and composition of phenolic
compounds present in them also change with the passage of time depending upon storage conditions.
Anthocyanins are degraded because they are prone to oxidation and this process is sped up in the
presence of vitamin C or its products. Similarly degradation process of phenolic compounds is initiated
by various enzymes present in liqueur [106].
Table 18. Phenolic compound contents in liqueurs made from blackberry fruit at various
time and storage intervals [106].
Month
Anthocyanin
(mg cy-3-glu/100 mL)
Flavonols
(mg Quercetin/100 mL)
Sum of Phenolic
compounds (mg/100 mL)
15 ns 15 s 30 ns 30 s 15 ns 15 s 30 ns 30 s 15 ns 15 s 30 ns 30 s
0 26.6 22.4 22.1 22.6 1.9 1.6 1.4 1.5 37.4 33.4 33.0 33.6
3 14.7 15.4 0.2 0.4 0.9 1.0 0.0 0.0 23.6 26.1 8.8 10.6
6 8.8 9.7 0.0 0.0 0.4 0.6 0.0 0.0 16.6 19.0 8.8 10.6
15 ns-liqueurs without sugar stored in 15 C; 15 s-liqueurs with sugar stored in 15 C; 30 ns-liqueurs without
sugar stored in 30 C; 30 s-liqueurs with sugar stored in 30 C.
Saponjac et al., investigared anthocyanin contents and biological activities of two blackberry
cultivars Thornfree (BT) and Cacanska bsetrna (BC). Cyanidin-3-O-glucoside was present in highest
concentration being 1397.7 mg/Kg and 1360.6 mg/Kg in BT and BC respectively. Antioxidant activity
determined via ABTS assay indicated EC50 of 0.007 and 0.06 g/L respectively for BC and BT
respectively [107].
5.3. Anti-Inflammatory Activity
There is convincing evidence that increasing consumption of fruits reduces risk of inflammation.
Fruit were found to be anti-inflammatory in murine model in vivo, with anthocyanins being responsible
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for this activity [108]. A water extract of fruits showed stronger anti-inflammatory activity even from
aspirin by inhibiting hyaluronidase enzyme in vitro [109] thereby confirming traditional use of fruits as
anti-inflammatory remedy. In another study inhibition of hyaluronidase enzyme was linked to GOD-type
tannin [110].
An herbal composition for modulating cytokines in the regulation of inflammatory or immune
diseases includes a blackberry extract [111]. Cyanidin-3-O-glucoside present in blackberry extract
suppresses NO production which leads to anti- inflammatory effects. The mechanism of this inhibition
may be due to an action on the expression/activity of the enzyme. Especially, the protein expression
was inhibited by the attenuation of NF-κB and/or MAPK activation. The NF-κB activation is managed
by mitogen-activated protein kinases (MAPK) [112]. It is used practically in the prevention and
treatment of immune, inflammatory and metabolic diseases [113]. Sangiovanni et al., investigated
effects of allagitannin enriched extracts (ETs) of R. fruticossus for the control of gastric
inflammation by in vitro and in vivo models. ETs inhibited TNFα-induced NF-κB driven transcription
(IC50: 0.67–1.73 mg/mL) and IL-8 secretion (IC50: 0.7–4 mg/mL). Major ETs detected were sanguiin
H-6 and lambertianin C which when tested decreased ulcer index by 88% and 75% respectively. The
results confirm the protective effects of ETs in gastric inflammation [114].
5.4. Antidiabetic Activity
Diabetes mellitus (DM) is an endocrine and metabolic disorder characterized by dyslipidemia,
hyperglycemia and protein metabolism that result from malfunction in regulating either insulin
secretion or insulin action. Persons suffering from DM are more prone to risk of coronary heart
diseases and therosclerosis. Despite the availability of modern hypoglycemic agents, ideal treatment of
diabetes is still to be achieved, so scientists are searching for treatments from natural sources for
diabetes mellitus. An aqueous tea prepared from balckberry fruit was evaluated by an in vitro glucose
diffusion model but no anti-diabetic effect was recorded [115]. The water and butanol fractions of a
R. fruticosus leaves 70% alcoholic extract were active in the treatment and prevention of noninsulin
dependent diabetes. Water and butanol extracts from leaves of R. fruticosus were reported to be active
in non-insulin dependent diabetes [116]. An aqueous extract of leaves was investigated for its possible
anti-diabetic activity in rats. The hypoglycaemic effect demonstrated in normal rats indicates that it is
active because counter-regulatory mechanisms cannot normalize rapidly blood glucose levels [117].
Chromium (Cr3+) and zinc (Zn2+) supplementation alleviates hyperglycemia and tea made from
R. fruticosus leaves decreased diabetic symptoms associated with these metals dependent diabetes [118].
The leaves of R. fruticosus are advised practically to manage diabetes mellitus. Studies in
streptozotocin (STZ)-diabetic mice have evaluated the anti-hyperglycaemic efficacy of RF previously
as a dietary supplement. Blackberry fruit was found to exhibit no effect on glucose homeostasis in
mice [9]. Leaves at daily administration of 5 g/kg of the infusion decreased 50% glucose-induced
hyperglycemia in alloxan-diabetic rabbits [119,120]. Ştefănuţ reported that administration of blackberry
extracts to diabetic rats in drinking water for 5 weeks significantly decreased glucose level from 360 to
270 mg/dL [39].
The general accepted therapeutic strategy for control of postprandial hyperglycemia is by inhibition
of α-glucosidase and α-amylase enzymes. This leads to significant delay of carbohydrate breakdown to
Molecules 2014, 19 11019
monosccharides. Salehi et al., reported that n-hexane and chloroform extract of blackberry exhibited
IC50 value of 0.5 and 6.2 in α-glucosidase inhibition activity while α-amylase inhibition potential
of n-hexane and methanol extract was 53.7 thus indicating that extract may be used as potential
anti-diabetic remedy [104]. Pressed residue of two blackberry cultivars Thornfree and Cacanska bsetrna
exhibited stronger α-glucosidase inhibitory activity even at the lowest concentration, i.e., 0.02 mg/mL,
while complete inhibition was achieved at 0.63–2.50 mg/mL [107]. Collectively, the inhibition of
intestinal α-glucosidase and pancreatic α-amylase activities as well as rich profile of antioxidant
bioactive constituents indicate berry fruit as a promising dietic therapy for DM. Controlled clinical
trials, however, are desirable for well-characterized and standardized blackberry extracts to corroborate
its beneficial effects in diabetic patients. Similarly, traditional claimed use of its fruit to control
hypertension and obesity should also be investigated in future studies.
5.5. Antiviral Activity
Globally viral diseases are increasing and simultaneously research to find new antiviral agents that
are non-toxic and safe for human consumption is also increasing. The berry fruits are an ideal
candidate for this search as these are non-toxic and may be recommended for human trials at lower
costs. R. fruticosus is used in the treatment of influenza in combination with other medicinal plants [121]
the role to control influenza virus may to the presence of polyphenols [122]. Antiviral activity data
shows that very little work has been done on this aspect.
5.6. Neuropharmacological Activity
Our research group evaluated various activities on mice which are grouped as neuropharmacological
activities. R. fruticosus L. fruit, leaves, root, and stem methanolic extracts were administered to mice at
doses of 100, 300, and 500 mg/kg. The order of CNS depressant effect for various parts was
fruit > root > leaves > stem. All extracts were found to be anxiolytic in nature, while no muscle
relaxing activity or sedative effect was observed. The order of central nervous system (CNS)
depressant effect for various parts of R. fruticosus was fruit > root > leaves > stem [123].
5.7. Toxicity Studies and Smooth Muscle Activities
Ali et al., reported that LD50 of acute toxicity studies of crude methanolic extract of blackberry
fruits was 887.75 ± 9.22 mg/mL while CC50 of same extract was 13.28 ± 2.47 μg/mL in brine shrimp
cytotoxic studies. Excellent anthelmintic activity was exhibited by 20 mg/mL of extract against
Raillientina spiralis and Ascaridia galli which was 1.37 times higher than albendazole. The extract
although toxic is safe at 100 mg/kg. EC50 for spontaneous relaxant activity and for 80 mM KCl-induced
contractions was 7.96 ± 0.1 and 6.45 ± 0.29 mg/mL respectively. The extract relaxed the spontaneous
contractions in a concentration dependent manner on jejunum preparations. The results indicated that
smooth muscle activity was mediated via inhibition of voltage gated channels [124].
Molecules 2014, 19 11020
5.8. Nutraceutical Usage
Blackberry juices, prepared with defatted milk and water, increased the ascorbic acid content in the
plasma [27]. Health granules and health beverages are prepared from R. fruticosus and other plants
used as dietary supplement and as immunity enhancer [125,126].
5.9. Miscellaneous Actions and Patents
Blackberry extract exhibited strong inhibitory action against monoamine oxidase B (MAO-B) and
the inhibitory concentration, IC50, was found to be between 4 and 7 mg/mL [127]. Blackberry and its
antioxidant components especially phenolics contribute positively to skin health by inhibiting the
oxidative damage linked with the formation of wrinkles and other skin-disorders like
hyperproliferation and skin dryness. It is used in cosmetic industry due to its specific scent and its
antioxidant potential. It is frequently used in the formulation of skin care products, for facial cleansing,
hair care products, to treat oily skin, acne as well as boils, skin eruptions and burns. Extracts of leaves
are used for skin aging and deodorant composition [128–132].
A water extract of leaves is reported for its angiogenic properties [133]. Extracts of whole plant are
used to prevent and cure inflammatory, immune and metabolic diseases and also as anti-influenza
remedy [113,121]. The whole plant extract possesses diuretic and hypoazotemic activities [134].
An oral pharmaceutical formulation is prepared from Gleditshcia triacanthos powder, powdered
leaves of R. fructicosus, pectin and corn starch and is used for the treatment of digestive disorders in
calves and piglets [135]. A toothpaste containing R. fruticosus as active principle is used for dental
caries, treating gums and cleaning teeth [136]. Leaves and fruits of R. fruticosus are consumed as
traditional foodstuff in normal diet to maintain immune health [80,130]. Powdered fruit is also used as
nutritional supplement [125].
5.10. Acute Toxicity
The lethality of water extract in mice was recorded for 1, 2, 3, 4 and 8 days after oral administration
of various doses of R. fruticosus. The extract did not cause any death or significant changes in general
behavior in mice at low and moderate doses (0–6 g/kg), but resulted in piloerection, rapid respiration
and diuresis at higher doses (>6 g/kg).The LD50 value for the aqueous extract of leaves was 8.1 g/kg
body weight [117].
6. Conclusions
R. fruticosus fruit is packed with numerous plant nutrients such as vitamins, minerals, anti-oxidants,
and dietary fibers that are essential for human health and fitness. These compounds protect from
cancer, aging, inflammation, and neurological diseases. With an increased awareness associated with
potential health benefits of consuming fruits, efforts are being made to enhance fruit quality and color
for consumers. It can be concluded that the wild growing blackberry fruits have a great future potential
to meet nutritional demands of indigenous communities besides their therapeutic efficacy. More work
is needed in identification, quantification and deciphering the bioactive constituents of fruit, seeds,
flesh and peel of these berries and their impact on human health needs to be explored. Community-
Molecules 2014, 19 11021
based trials should be conducted to validate its nutraceutical claims. The consumption of fruits,
vegetables, spices, legumes and grains in Pakistan is insufficient and may be supplemented by
indigenous cost-effective sources like berries. Although not a famous fruit for commercial production,
it has a great production, expansion and consumption potential in Pakistan, if premium prices are paid
and fruits are exploited economically. Even though various types of chemical compounds have been
isolated and characterized, research reports on the bioactivity and the mechanism of action of the
isolated compounds under in vivo conditions are limited. Additionally the effects of these compounds
on other ailments like cancer, HIV, blood pressure, cardio-vascular disease and others, need to be
investigated in detail.
Acknowledgments
The anonymous referee’s comments are highly acknowledged that helped to fine tune the manuscript.
Author Contributions
MZUH and MR initiated and designed the study. VDF, HZEJ and MM, contributed to literatures
collection. MZUH and MR drafted the manuscript. All authors read and approved the final manuscript.
Conflicts of Interest
The authors declare no conflicts of interest.
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... Boysenberry, which is included in the order Rosales, the Rosaceae family, and the Rubus genus, is a shrub-shaped plant with high economic value that is traded all over the world due to its fruit's pleasant taste and high nutritional value [1][2][3]. The cultivation of Boysenberry, which was obtained by crossing Rubus baileyanus and Rubus loganobaccus in the USA between 1921 and 1923, is increasing rapidly due to its suitability for fresh consumption and processing in the food industry, as well as its higher fruit quality compared to other blackberry varieties [4]. ...
... SiNPs were used as a dry white powder (silica nanopowder) with a particle size of 10-20 nm, surface area of 590-690 m 2 /g, and purity of 99.8%. FeNPs (iron nanopowder) were used as black-colored and high-purity iron powders with a particle size of 35-45 nm, a bulk density of 0.5 g/cm 3 , and a true density of 7.9 g/cm 3 . AgNPs (silver nanopowder) with an average particle size of <10 nm, an average surface area of 10.0-15.0 ...
... SiNPs were used as a dry white powder (silica nanopowder) with a particle size of 10-20 nm, surface area of 590-690 m 2 /g, and purity of 99.8%. FeNPs (iron nanopowder) were used as black-colored and high-purity iron powders with a particle size of 35-45 nm, a bulk density of 0.5 g/cm 3 , and a true density of 7.9 g/cm 3 . AgNPs (silver nanopowder) with an average particle size of <10 nm, an average surface area of 10.0-15.0 ...
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Salinity is one of the most important abiotic stress factors that affect plant growth and limit agricultural productivity. In this study, the effects of iron (FeNP), silver (AgNP), and silicon dioxide (SiNP) nanoparticles on the morphological and physiological parameters of in vitro boysenberry plants grown under salinity stress (NaCl) were investigated. According to our study results, higher values were obtained from SiNP application in terms of shoot development parameters; FeNP application was found to be more successful for root development; AgNP application was effective in terms of SPAD, leaf relative water content (LRWC), and relative growth rate (RGR); and FeNP application increased superoxide dismutase (SOD) and catalase (CAT) enzyme activities. Salt stress significantly affected root development, SPAD values, LRWC and RGR, and SOD and CAT enzyme activities. As a result, under salt stress conditions, SiNP, FeNP, and AgNP applications can significantly reduce the negative effects of stress and promote the vegetative development of the plant compared to control conditions.
... According to Table 1, the pH values found for R. ulmifolius, M. nigra and R. fruticosus were 5.59, 3.93 and 3.39, respectively (Table 1). These values are consistent with previous research (Zia-Ul-Haq et al., 2014). In addition, our findings are consistent with previous research suggesting that wild blackberries with higher pH values may taste sweeter than those obtained from cultivated blackberries. ...
... On the other hand, in M. nigra, six anthocyanins were found, namely (1, 2) cyanidin 3-O-glucoside derivatives 1 and 2, respectively, (3) cyanidin 3-O-rutinoside, (4) pelargonidin glucoside, (5) pelargonidin deoxyhexoside-hexoside and (6) cyanidin arabinose/xyloside. The detection of these compounds based on their m/z and fragment ions is in accordance with other studies (Migues et al., 2018;Zia-Ul-Haq et al., 2014). ...
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Berries have gained popularity recently due to the health benefits associated with their bioactive compounds. This work aimed to evaluate the physicochemical characteristics, colour, phytochemicals content and mineral content of three common berries, two types of blackberries (Rubus fruticosus and R. ulmifolius) and one mulberry (Morus nigra), collected in the Covilh˜ a region, Portugal. R. fruticosus presented the greatest length, weight, moisture, firmness and darker colour, while R. ulmifolius showed the greatest width. On the other hand, M. nigra showed the highest ash content (2.49±0.40 %). A total of 20 phenolics were identified and quantified by chromatographic techniques. Using ICP-MS and FAAS, 13 essential and 14 non-essential elements were quantified. Additionally, 54 volatiles were detected through HS-SPME/GC-MS, several compounds were herein reported for the first time in these berries. Our results show that these berries contain a high concentration of highvalue bioactive compounds, significantly influencing their nutritional value, aroma and appeal to consumers, providing data to make it possible to choose more attractive and superior blackberry and mulberry varieties that meet consumer demands.
... Externally, decoctions or infusions of leaves are used as a gargle for gum inflammations, throat cankers, and mouth ulcers. [9][10][11] Rubus ulmifolius Schott is a perennial bush referred to as wild blackberry, capable of reaching heights of 2-3 meters. R. ulmifolius is distributed in North Africa, Asia and Europe. ...
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Rubus ulmifolius Schott commonly known as wild blackberry, is a perennial shrub native to North Africa and Europe. It has been used for both medicinal and food purposes. Different parts of the plant have been used in herbal medicine for a long time to treat numerous diseases such as diabetes, hypertension, gastro-intestinal disorders, mouth ulcers, heart ailments, menstrual pain, hemorrhoids, tonsillitis, skin diseases, etc. Its fruits are consumed fresh or in jams, juices, and marmalades. The shoots are used in omelets and soups. The present review aims to summarize data on ethnomedicinal uses, phytochemistry, pharmacological effects, botanical description, geographical distribution, and toxicity studies of Rubus ulmifolius over a period covering the last 23 years using online scientific databases such as Science direct, Google Scholar, Scopus, PubMed and Springer. Various studies have shown that Rubus ulmifolius possesses different biological activities in vitro and in vivo, including antipyretic, anti-inflammatory, anthelmintic, anticoagulant, antiproliferative, antioxidant, wound healing, amoebicidal, antifungal, antiviral, antibacterial, antihyperglycemic, and antihyperlipidemic properties. Phytochemical studies of Rubus ulmifolius have revealed the presence of various constituents belonging to different classes, such as flavonoids, tannins, terpenoids, and phenolic acids. We hope that our review will serve as an important source of information that can trigger further scientific research on this species.
... Boysenberry, which is included in order Rosales, Rosaceae family, Rubus genus, is a shrub-shaped plant with high economic value traded all over the world with its pleasant taste, high nutritional value fruit [1][2][3]. The cultivation of Boysenberry, which was obtained by crossing Rubus baileyanus and Rubus loganobaccus in the USA between 1921-23, is increasing rapidly due to its suitability for fresh consumption and processing in the food industry, and higher fruit quality compared to other blackberry varieties [4]. ...
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Salinity, one of the most important abiotic stress factors, affects plant growth and limits agricultural productivity. In the study, the effects of iron (FeNP; 0.025 and 0.05 mM), silver (AgNP; 0,2 ve 0,4 mg L-1) and silicon dioxide (SiNP;7,5 ve 15 mg L-1) nanoparticles on morphological and physiological parameters of in vitro blackberry plants grown under salinity stress (NaCl; 15 mM ve 35 mM) were investigated. According to our study results, it was determined that higher values obtained from SiNP application in terms of shoot development parameters, FeNP application found more succesfull for root development, AgNP application was effective in SPAD, leaf relative water content (LRWC) and relative growth rate (RGR), and FeNP application was increased superoxide dismutase (SOD) and catalase (CAT) enzyme activities. Salt stress was significantly affected the root development, SPAD value, LRWC and RGR, SOD and CAT enzyme activities. As a result, under salt stress conditions, SiNP, FeNP and AgNP applications can significantly reduce the negative effects of stress and promote vegetative development of the plant compared to control conditions.
... The wild edible fruits and berries have provided nutrients, minerals, and energy to humans since time immemorial and are considered healthy food resources (1,2). These fruits are also beneficial to health and are used in the pharmaceutical and cosmetic industries for their healing and protective qualities (1,(3)(4)(5). In addition, fruits also offer taste and nutrients for a wide range of culinary uses, making them indispensable. ...
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Rubus ellipticus Smith is an evergreen shrub in the Rosaceae family, commonly known as yellow Himalayan raspberry. The objective of this study is to determine the morphological analysis, minerals, proximate, ascorbic acid, anthocyanins, and carotenoids content in R. ellipticus fruits. The fruit samples were collected from four different sites with different altitudes [500 m (District Bilaspur), 1,000 m (District Hamirpur), 1,500 m (District Solan) and 2,000 m (District Shimla)] of Himachal Pradesh for the two consecutive years (2018 and 2019). The fruit morphological investigation revealed that the maximum length (7.71 ± 0.08 mm), width (8.71 ± 0.03 mm), and weight (0.80 ± 0.01 g) of fruits is achieved at higher altitudes (2,000 m) in the year 2018 as compared to 2019. The mineral content (0.05–36.6 mg/g DW), ascorbic acid content (14.59–23.64 mg/g DW), proteins (95.20–131 mg/g DW), and crude fibers (5.6–11.5%) were also higher in fruits grown at 2,000 m altitude, whereas carbohydrates (210–398 mg/g DW), crude fat (2.4–4.1%), and anthocyanins (0.42–1.35 mg/100 g FW) contents were higher in fruits collected from 500 m altitude. According to the results, R. ellipticus fruits were rich in crude fiber, moisture, carbohydrates, protein, ash, and crude fat, as well as in micronutrients, and displayed significant variation with altitude in nutrient content. This could be due to the different environmental, geographical, and weather conditions. The high nutrient content of R. ellipticus suggests its future potential applications for the food and pharmaceutical industry.
... The major health benefits reported for Rubus fruticosus fruit are anti-oxidant activity, anti-inflammatory activity, antimicrobial activity, anti-diabetic activity and anti-cancer activity (Verma et al. 2014, Schulz et al. 2019, Amini et al. 2021. Preparations made of blackberry juices with milk and water elevate plasma ascorbic acid content and other supplements help in improving immunity (Zia-Ul- Haq et al. 2014). ...
... 18 Besides their nutritional value, fruits, vegetables, herbs, and spices have been traditionally used to cure various human ailments. 19 Dietary guidelines emphasize fruit and vegetable consumption for health benefits, while antibiotics' resistance makes natural products a promising alternative for controlling food-borne infections. 9,20 Several studies have investigated the protective effects of BBE, fruit or leaf and reported that it significantly reduced liver and kidney damage markers and improved their function in injured rats. ...
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The Himalayas, globally acknowledged as one of the four biodiversity hotspots, underscore their ecological significance, boasting abundant flora and fauna. Among these, a diverse array of wild fruits such as Aegle marmelos, Artocarpus lakoocha, Baccaurea spp., Carissa spp., and others provide essential nutrition for local populations. These fruits, rich in bioactive compounds, offer nutraceutical potential, contributing to health aspects like antidiabetic, anti-inflammatory, and anticancer properties. The integration of Himalayan wild fruits into circular practices supports sustainable livelihoods. The responsible harvesting, efficient processing, and value addition of these fruits align with circular principles, striking a balance between conservation and progress. Technologies such as anaerobic digestion, waste-to-energy conversion, and composting can harness waste generated during cultivation and processing, contributing to a circular economy and rural Himalayan community development. Preserving, accessing, and commercializing underutilized fruits can significantly enhance economic prosperity and the quality of life for inhabitants. However, integrating these fruits into agriculture faces multifaceted challenges, spanning social, economic, environmental, agronomic, and political dimensions. Addressing these challenges is crucial for sustainable development, aiming to eradicate poverty, malnutrition, and hidden hunger. Moreover, addressing these challenges is not only vital for sustainable development in the Himalayan region but also for mitigating carbon footprints and tackling issues like poverty, malnutrition, hidden hunger, and climate change. The exploration of these concepts within the Himalayan context holds immense promise for sustainable development and ecological conservation. Graphical Abstract
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The present invention concerns the field of plant extracts and their uses, in particular for cosmetic, oral hygiene and pharmaceutical purposes. In particular the invention concerns blackberry leaf extracts and preparations and medicaments containing them, as well as their use to inhibit irritating and inflammatory skin conditions.
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We studied the element composition of wild-grown Rosa canina L. and Rubus fruticosus L. fruits at a derelict lead-zinc ore mine in North-Hungary. Whole rosehips and Rubus berries were collected from seriously contaminated vs. background spots and analyzed according to standard lab procedures. The species differed in all focal elements except Ba and S. Rubus berries in general were richer in most macro- and microelements, particularly of Mn, Ca, Cu, Sri, Zn, Fe and Ni. The sampling site had a significant effect on all elements except Ca, Cu and Se. Rosehips had very low Cd (< 0.08), As (< 0.12), Pb (< 1.63) concentrations (mg kg(-1) dw). Ba, Cd, Mn, Ni and Sri were found to accumulate consistently in both species' fruits at the contaminated sites. The element composition of entire fruits was shown to be an inadequate indicator of elevated heavy metal levels in the soil in a situation where marked acidification associated with pyrite oxidization is an important process.