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Nutritional evaluation of fresh and dried goji berries cultivated in Italy

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The nutritional profile of fresh and dried goji berries cultivated in Italy was investigated. The obtained data confirm goji berries as a source of nutritional and healthy components, such as vitamin E, minerals and fibre. Taking into account the Recommended Daily Allowance (RDA) for minerals and vitamins established by the Commission of the European Communities, Goji berries provide significant amounts of dietary fibre and zeaxanthin and can be declared on the label as a potential source of vitamins E and C. Moreover, dried goji berries can be declared as a source of K, P, Cu, Fe Mn, Zn.
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Ital. J. Food Sci., vol 29, 2017 - 398
PAPER
NUTRITIONAL EVALUATION OF FRESH
AND DRIED GOJI BERRIES CULTIVATED IN ITALY
S. NIRO, A. FRATIANNI*, G. PANFILI, L. FALASCA, L. CINQUANTA
and MD RIZVI ALAM
Department of Agricultural, Environmental and Food Sciences, University of Molise, Via F. De Sanctis, 86100
Campobasso, Italy
*Corresponding author. fratianni@unimol.it
ABSTRACT
The nutritional profile of fresh and dried goji berries cultivated in Italy was investigated.
The obtained data confirm goji berries as a source of nutritional and healthy components,
such as vitamin E, minerals and fibre. Taking into account the Recommended Daily
Allowance (RDA) for minerals and vitamins established by the Commission of the
European Communities, Goji berries provide significant amounts of dietary fibre and
zeaxanthin and can be declared on the label as a potential source of vitamins E and C.
Moreover, dried goji berries can be declared as a source of K, P, Cu, Fe Mn, Zn.
Keywords: goji berries, Lycium barbarum, superfruit, wolfberries
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Ital. J. Food Sci., vol 29, 2017 - 399
1. INTRODUCTION
Fruits of Lycium barbarum L., belonging to Solanaceae family, commonly known as goji
berries or wolfberries, have been used in Chinese traditional medicine for centuries.
Lycium barbarum grows in China, Tibet and other parts of Asia and its fruits are 1-2 cm-
long, bright orange-red ellipsoid berries. The native area of Lycium is not definitively
established but it is likely found in the Mediterranean Basin (POTTERAT, 2010).
Traditionally, goji berries are collected in summer and autumn. The fruits can be eaten
fresh or dried, and they are also found in conventional food products, such as yoghurt,
fruit juices, bakery foods, chocolate and others (MIKULIC-PETKOVSEK et al., 2012a,
2012b). The drying process is intended to remove water from foodstuff in order to prevent
microbial spoilage and chemical alterations, thus prolonging shelf life, while realizing
space and weight saving (CINQUANTA et al., 2010; CUCCURULLO et al., 2012;
FRATIANNI et al, 2013). Traditionally, the berries are dried in the shade until the skin
shrinks and then exposed to the sun until the outer skin becomes dry and hard but the
pulp is still soft (AMAGASE and FARNSWORTH, 2011). The sun drying method is cheap,
but there is a risk of damage due to dust and insect infestation. An alternative is hot air
drying. Today the goji fruit market is significantly expanding because of an increased
awareness of the possible health benefits, as fruits contain different nutrients, such as
polysaccharide complexes, organic acids, phenolic compounds and antioxidants with high
biological activity. Dietary fibre provides several health benefits, including the reduction
of the risk of coronary heart disease, of diabetes, hypertension, obesity, stroke and some
gastrointestinal disorders (EFSA, 2010). Recent studies indicate that polysaccharides from
Lycium barbarum possess a range of biological activities, including antioxidant properties
(AMAGASE and NANCE, 2008; CHANG and SO, 2008). Goji fruits constitute a variety of
antioxidants such as ascorbic acid, different carotenoids (KULCZYŃSKI and GRAMZA-
MICHAŁOWSKA, 2016) and high levels of phenolic compounds (ZHANG et al., 2016).
Carotenoids are a significant group of biologically-active constituents with health
promoting properties (AMAGASE et al., 2009; DONNO et al., 2014) responsible for the
colour of a wide variety of foods (FRATIANNI et al., 2005).
The reddish-orange colour of L. barbarum fruits derives from a group of carotenoids, which
make up only 0.03–0.5 % of the dried fruit. Zeaxanthin is the major carotenoid found in
goji. This is a yellow pigment, an isomer of lutein and a derivative of β-carotene. When
ingested, zeaxanthin accumulates in fatty tissues, but especially in the macula, a region of
the retina, helping in protecting the macula from degeneration, which can be induced by
excessive sun exposure (UV light) and by other oxidative processes. In goji, zeaxanthin is
present as an ester of dipalmitate. Studies focusing on carotenoid goji berries are few and
mainly aimed at the identification and quantification of ester-form carotenoids. INBARAJ
et al. (2008) and ZHAO et al. (2013), in particular, identified free-forms and ester-forms of
carotenoids. Beta-carotene, neoxanthin, and cryptoxanthin are also present at low
concentrations (PENG et al., 2005; WANG et al., 2010). Regarding other antioxidants,
studies made on Lycium chinense Miller reported high amounts of α-tocopherol, together
with other vitamin E compounds (ISABELLE et al., 2010). Vitamin E is a generic term
indicating structurally related compounds, namely tocols, comprising two groups of
vitamers, i.e. tocopherols and tocotrienols, which occur in eight forms: α-tocopherol (α-T),
β-tocopherol (β-T), γ-tocopherol (γ-T), and δ-tocopherol (δ-T) and α-tocotrienol (α-T3), β-
tocotrienol (β-T3), γ-tocotrienol (γ-T3), and δ-tocotrienol (δ-T3). The potential health
benefits of tocols have been the subject of several reviews (TIWARI and CUMMINS, 2009).
Vegetable oils are the main tocol source; however, substantial amounts of these
compounds are also reported in most cereal grains (FRATIANNI et al., 2013; MIGNOGNA
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et al., 2015; PANFILI et al., 2003). To our knowledge, no literature data are available on the
composition and content of tocols in L. barbarum fruits. Goji is also an extremely rich
source of many essential minerals, which are essential for many actions in the body, like
muscle contraction, normal heart rhythm, nerve impulse conduction, oxygen transport,
oxidative phosphorylation, enzyme activation, immune functions, antioxidant activity,
bone health, and acid-base balance of the blood (WILLIAMS, 2005; SALDAML and
SAĞLAM, 2007). An adequate daily amount of minerals is necessary for an optimal
functioning of the body. For the above reported reasons, goji berries are often proposed as
functional foods and have been included in the novel category of “superfruits” or
“superfoods”.
Superfruits, a subcategory of superfoods, is a relatively recent word and is considered a
new marketing approach to promoting common or rare fruits which can be consumed as
foodstuffs or used as ingredients by manufacturers of functional foods, beverages and
nutraceuticals. Superfruits have a high nutritional value due to their richness in nutrients,
antioxidants, proven or potential health benefits and taste appeal (FELZENSZWALB,
2013). In the functional foods market, the products targeting health and mental well-being
have prompted the food industry to increase the research and the development of these
new foods, outlining a rapid expansion market in several countries (VICENTINI et al.,
2016). In the last years, goji berries have been cultivated in Italy and are available both as
fresh and dried fruits. While several papers on the medical effect of goji berries have been
published, little information is available on the nutritional composition of dried and,
above all, fresh goji berries. The aim of the present study is therefore to determine the
compositional and nutritional value of fresh and dried goji berries cultivated in Italy, with
a particular focus on minerals and some antioxidant compounds, such as carotenoids,
tocols and vitamin C, to increase the awareness about their nutritional profile.
2. MATERIAL AND METHODS
2.1. Sample collection and preparation
Fresh goji berries (L. barbarum L.) were provided by Favella Spa farm (Sibari, Southern
Italy). The farm has 21000 plants in 5 Ha (2.5 m x 1 m), with a drip irrigation system. Goji
berries were cultivated in two consecutively growing seasons (2014 and 2015) and were
collected in July. All harvested fruits were randomly collected in the orchard from
different plants and analysed fresh and air-dried. Fresh goji berries (about 1 cm size) were
subjected to freeze-drying before analyses, as reported by FRATIANNI et al., 2013 (fresh
fruits). One-half of collected goji berries were air-dried in a convective dryer
(B80FCV/E6L3, Termaks, Norway), at 60 °C, with an air velocity of 2.1 m/s, until a
constant weight was reached (dried fruits). The drying time was about 21 h.
Results are reported as the average of the two growing seasons (2014-2015).
2.3 Proximate composition analysis
Fresh and dried fruits were analysed for moisture, ash, fat, and protein (N×6.25) contents,
according to standard methods of AOAC (2000). Dietary fibre content was determined
according to AOAC method 991.43 (1995) and AACC method 32-07 (1995). Total dietary
fibre content was the sum of insoluble and soluble dietary fibre content. Vitamin C was
determined by using an enzymatic kit (Megazyme International, Ireland), following the
manufacturer instructions.
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2.4. Mineral analysis
Ultrapure nitric acid for trace analysis, sulfuric acid (96 %) and standard mono elements in
nitric acid 2 % were purchased from Sigma-Aldrich (20151 Milan, Italy). The
determination of metals (K, Ca, Co, Cu, Fe, Mg, Mn, Mo, Na, P, Se, Zn) in goji samples was
carried out by using the technique of nitric mineralization and the analysis by
spectrophotometry plasma emission (Varian ICP 710, OES Inductively Coupled Plasma-
Optical Emission Spectrometers, Palo Alto, CA 94304-1038). Samples were ground and 0.5
g was digested with 10 ml of nitric acid with a mineralizer (SCP Science DIGIprep, Quebec
H9X 4B6, Canada), with the following instrumental conditions: start at 40 °C for 15
minutes; heating at 60 °C for 15 minutes; stay at 60 °C for 15 minutes; heating to 90 °C for
20 minutes. The digested samples were cooled and brought to a volume of 50 ml with
bidistilled water and analysed with the optical ICP. The precision was calculated as a
mean deviation of three measurements.
2.5. Carotenoid extraction and determination
Carotenoid extraction was carried out using the direct solvent extraction method reported
in FRATIANNI et al. (2013) with slight modifications due to the complex structure of goji
berries. About 0.1 g of milled freeze-dried samples (fresh fruit) and air-dried samples
(dried fruit) was weighed and placed in a screw-capped tube. Then, 5 ml of ethanolic
pyrogallol (60 g/L) was added as an antioxidant. The sample was stirred for 10 minutes.
After that, 2 ml of absolute ethanol was added and the sample was stirred again for a few
minutes. The suspension was then extracted with 15 mL of n-hexane/ethyl acetate (9:1
v/v) and stirred; after that 15 mL of sodium chloride (10 g/L) was added. Further
extractions with n-hexane/ethyl acetate (9:1 v/v) were made until the organic layer was
colourless. Finally, the organic layer was collected and evaporated to dryness, and the dry
residue was dissolved in methanol: MTBE 50:50 (v/v). This sample was used to determine
the free carotenoids not esterified with the lipid components and carotenoids esterified
with fatty acids (unsaponified). A volume of 2 ml of this extract was evaporated to dryness
and subjected to alkaline hydrolysis under a nitrogen flux for 1 minute in a screw-capped
tube with 1 ml of ethanolic pyrogallol (60 g/L), 10 ml of solvent HEAT (hexane: ethanol:
acetone: toluene 10: 6: 7: 7 v/v/v/v), 2 ml of methanolic KOH (40 %) and glass balls. The
tubes were placed in a 56 °C water bath and mixed every 5 to 10 min. After alkaline
digestion at 56 °C for 20 minutes, the tubes were cooled in an ice bath, and 15 mL of
sodium chloride (10 g/L) were added. The suspension was then extracted with 15 mL of
n-hexane/ethyl acetate (9:1 v/v) until the organic layer was colourless. The organic layer
was collected and evaporated to dryness, and the dry residue was dissolved in methanol:
MTBE 50:50 (v/v). This sample was used to determine carotenoids esterified with lipid
components (saponified). An aliquot of the carotenoid extract was separated, as in
MOULY et al. (1999), by a reverse-phase HPLC system. An HPLC Dionex (Sunnyvale, CA)
analytical system, consisting of U3000 pumps, and an injector loop (Rheodyne, Cotati)
were used. Separation was performed as in FRATIANNI et al. (2013) by using a YMC
(Hampsted, NC, USA) stainless steel column (250×4.6 mm i.d.) packed with 5 &m silica
spheres that were chemically bonded with a C30 material at a flow rate of 1 mL/min. The
mobile phase was methanol: MTBE (v/v). The eluted compounds were monitored by a
photo-diode array detector (Dionex, Sunnyvale) set at 430 nm.
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2.6. Carotenoid identification and quantification
Carotenoids were identified on the basis of their diode array spectral characteristics,
retention times, and relative elution order, compared with known commercially available
standards. All-trans-β-carotene and lutein were from Sigma Chemicals (St. Luis, MO,
USA); zeaxanthin and β-cryptoxanthin were obtained from Extrasynthese (Z.I. Lyon-
Nord, Genay, France). Zeaxanthin dipalmitate was identified by means of its spectral
characteristics found in literature (INBARAJ et al., 2008). Compounds were identified by
comparison of their retention times with those of known available standard solutions and
quantified on the basis of the calibration curves of standard solutions. Zeaxanthin
dipalmitate was quantified as zeaxanthin.
2.7. Tocol analysis
Tocols were determined after the saponification method of the extract described for
carotenoids. An aliquot of the carotenoid extract was collected and evaporated to dryness,
and the dry residue was dissolved in 2 ml of isopropyl alcohol (1 %) in n-hexane and was
analysed by HPLC under normal phase conditions, using a 250 x 4.6 mm i.d., 5 mm
particle size, and Kromasil Phenomenex Si column (Torrance, CA, USA) (PANFILI et al.,
2003). Fluorometric detection of all compounds was performed at an excitation
wavelength of 290 nm and an emission wavelength of 330 nm by means of an RF 2000
spectrofluorimeter (Dionex, Sunnyvale, USA). The mobile phase was n-hexane/ethyl
acetate/acetic acid (97.3:1.8:0.9 v/v/v) at a flow rate of 1.6 mL/min (FRATIANNI et al.,
2002; PANFILI et al., 2003). Compounds were identified by comparison of their retention
times with those of known available standard solutions and quantified on the basis of the
calibration curves of standard solutions. The concentration range was 5-25 µg/ml for
every tocol standard. Vitamin E activity was expressed as Tocopherol Equivalent (T.E.)
(mg/100 g product), calculated as reported by SHEPPARD et al. (1993).
3. RESULTS AND DISCUSSION
3.1. Nutritional composition
The nutritional composition of fresh and dried goji berries is shown in Table 1. Fresh goji
berries have 77.4 % moisture, 1.1 % fats, 2.5 % proteins, 15.3 % carbohydrates and 2.9 %
fibre. In dried goji berries, 4.4 % fats, 10.2% proteins, 61.3 % carbohydrates and 11.4 % fibre
were found. Similar results on dried goji were reported by ENDES et al. (2015). Our data
suggest that dried fruits contain notable levels of dietary fibre, either as water-soluble
form (2.6 %) or as insoluble form (8.8 %). The ratio between insoluble and soluble fibre is
about 3:1. Dietary fibre intake recommendation for adults is 25 g/day (LARN, 2014). With
the consumption of a portion of 30 g of dried fruits, dietary fibre intake for the adults is
about 14 % of its daily recommended intake. Taking into account the European law
(Regulation CE 1924/2006), dried goji can be declared in label with the claim “high fibre
content”, since it contains at least 6 g of fibre per 100 g. Finally, fresh and dried goji berries
provide about 87 and 348 kcal/100 g, respectively.
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Table 1. Nutritional composition of fresh and dried goji berries (g/100 g) (mean±standard deviation).
Moisture
Fats
Proteins
Carbohydrates*
Fibre
Ash
Soluble
Total
Fresh
77.4±0.4
1.1±0.02
2.5±0.12
15.3
0.7±0.17
2.9
0.84±0.11
Dried
9.3±0.02
4.4±0.45
10.2±0.22
61.3
2.6±0.06
11.4
3.4±0.16
* Calculated by difference.
3.2. Mineral composition
The content of both macro and microelements in goji berries is reported in Table 2.
Potassium (K) is the predominant element (276.2 mg/100 g and 881.9 mg/100 g for fresh
and dried fruits, respectively), followed by sodium (Na). Potassium and sodium play an
important role in regulating blood pressure and the body’s acid-base balance (CLAUSEN
et al., 2013). Goji could also be a good source of phosphorus (P) and calcium (Ca), with an
appreciable concentration of magnesium (Mg), which is needed to prevent heart disease
and growth retardation (CHATURVEDI et al., 2004). A discrete amount of copper (Cu),
iron (Fe) and manganese (Mn) were also found. BELLAIO et al. (2016), ENDES et al. (2015)
and LLORENT-MARTÍNEZ et al. (2013) reported slightly different results. As in any other
plant food, the mineral content of berries reflects the soil in which they are grown. It is
important to highlight that essential and nonessential element concentration is dependent
on the soil characteristics, the physiology of the plant, the water source composition, and
fertilizers, insecticides, pesticides, and fungicides used in the plantations. Plants can
absorb, carry, and accumulate chemical elements. Each species has its own requirements
and differing levels of tolerance when absorbing and accumulating an element. The
movement of the inorganic constituents is selectively controlled by the plant, with some
being easily absorbed and others impeded to a different degree (NAOZUKA et al., 2011).
Table 2. Average values of mineral elements in fresh and dried goji berries (mg/100 g) (mean±standard
deviation).
Fresh
Dried
Ca
26.6±4.90
101.3±22.60
K
276.2±41.00
881.9±239.70
Mg
12.7±2.80
45.9±9.20
Na
57.3±8.70
209.8±72.30
P
48.4±9.26
174.3±32.10
Co
0.001
0.001
Cu
0.3±0.04
0.8±0.25
Fe
0.9±0.22
3.4±1.57
Mn
0.2±0.03
0.5±0.18
Zn
0.5±0.12
1.5±0.62
Se (µg/100g)
0.1±0.01
0.17±0.03
Mo (µg/100g)
0.00
0.00
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Table 3 reports the percentage contribution to the RDA of 100 g of fresh and dried goji
berries, according to Reg. EU 1169/2011. For dried goji berries, the percentage of RDA per
portion (30 g) is also reported. From data, fresh goji berries can be declared on the label as
a source of Cu; in fact, 100 g of fresh goji berries contributed to about 25% of the RDA. The
contribution of other minerals from fresh goji is low. Dried goji berries can be declared as a
source of K, P, Cu, Fe Mn, Zn. A consumption of 30 g of dried goji per day contributes to
the RDA approximately of 25 % for Cu, 13 % for K and less than 10 % for other elements.
Table 3. Percentage contribution to the RDA of minerals in fresh and dried goji berries.
3.3. Carotenoid, tocol and ascorbic acid amounts
Table 4 shows HPLC carotenoid analysis of fresh and dried fruits. Drying of fruits did not
cause significant changes in carotenoid amounts (data not shown). Unsaponified
carotenoids, determined after solvent extraction, and saponified carotenoids, determined
after saponification of the extract, are reported.
Table 4. Average carotenoid amounts in fresh and dried goji berries (mg/100 g) (mean±standard deviation).
Fresh
Dried
Unsaponified
Saponified
Unsaponified
Saponified
Lutein
0.0
1.1±0.02
0.0
5.7±0.44
Zeaxanthin
0.0
53.8± 0.82
0.0
186.0±3.80
β
-cryptoxanthin
0.0
2.3± 0.25
0.0
6.1±0.14
Zeaxanthin dipalmitate
47.8±2.32
0.0
158.8±1.53
0.0
Esters
8.5±1.24
0.0
24.5±2.30
0.0
β-carotene
0.2±0.01
0.1±0.01
0.9±0.09
1.0±0.20
Total carotenoids
56.4±1.23
57.3±0.80
184.2±1.52
198.8±3.80
Unsaponified carotenoids are characterized by a significant peak, identified as zeaxanthin
dipalmitate, the dominant ester of goji berries (WELLER and BREITHAUPT, 2003;
INBARAJ et al., 2008). Beta-carotene is also present. Before zeaxanthin dipalmitate peak,
other unidentified peaks, probably carotenoid esters (INBARAJ et al., 2008), were also
Reg. RDA
Fresh
Dried
Dried
mg/day
% RDA
% RDA
% RDA x 30g
Ca
800
3
13
4
K
2000
14
44
13
Mg
375
3
12
4
P
700
7
25
7
Cu
1
25
84
25
Fe
14
6
24
7
Mn
2
8
26
8
Zn
10
5
15
5
Se (µg)
55
0
0
0
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Ital. J. Food Sci., vol 29, 2017 - 405
detected. The amount of zeaxanthin dipalmitate in dried fruits is about 159 mg/100 g and
of β-carotene is about 1 mg/100 g. INBARAJ et al. (2008) found values of zeaxanthin
dipalmitate and of β-carotene of 114.3 mg/100 g and 2.4 mg/100 g, respectively. This
difference is probably due to the fact that carotenoid levels can be influenced by different
harvest stage fruits, geographical origin, and seasonality (WEN-PING at al., 2008).
Saponification of the extract is necessary to convert esters to free-compounds and it is
often used to remove chlorophylls, lipids and other analytical interferences (FRATIANNI
et al., 2015). The saponified extract of dried fruits shows high zeaxanthin contents (about
190 mg/100 g) and small lutein and β-cryptoxanthin amounts (about 6 mg/100). Small
amounts of lutein were also found, after saponification, in a work of ZAO et al., 2013. As a
dietary supplement for eye health (CHENG et al., 2005), a dose of 15 g per day was
deemed beneficial in supplying adequate zeaxanthin (estimated at 3 mg/day). Thirty g of
our goji samples provide a zeaxanthin amount of 14 mg/day (fresh fruit) and 48 mg/day
(dried fruit). Table 5 shows the tocol amounts in fresh and dried goji berries. As for
carotenoids, the drying treatment did not cause significant changes in tocol contents (data
not shown).
Table 5. Average tocopherol amounts in fresh and dried goji berries (mg/100 g) (mean±standard deviation).
Fresh
Dried
α-tocopherol
1.4±0.10
5.5±0.48
β -tocopherol
1.0±0.01
4.2±0.04
Total tocopherols
2.4±0.04
9.7±0.20
Tocopherol Equivalent (TE) §
2
8
§ Calculated as in SHEPPARD et al., 1993.
Goji berries were found as a source of α- and β-tocopherol (about 1.4 and 1.0 mg/100 g,
respectively, in fresh fruits, and 5.5 and 4.2 mg/100 g, respectively, in dried fruits). A
paper by ISABELLE et al. (2015) reports, in Lycium chinenses, belonging to the same Lycium
species, the presence of α-tocopherol (3.9 mg/100 g), together with γ-tocopherol (0.46
mg/100 g), δ-tocopherol (0.12 mg/100 g), and traces of α-γ-δ tocotrienol (< 0.1 mg/100 g).
Table 5 also reports values of vitamin E activity provided by 100 g of product, expressed
as Tocopherol Equivalent (TE) (mg/100 g product) (SHEPPARD et al., 1993). Taking into
account the Recommended Daily Allowance (RDA) for vitamin E, which is of 12 mg/day
(Regulation EU 1169/2011), 100 g of fresh goji berries contribute approximately 16 % of
the RDA, while 100 g of dried fruits contribute approximately 66 % of the RDA, so that to
be declared in label as a source of vitamin E. A portion of dried goji berries (30 g)
contributes approximately 20 % of the RDA. The concentration of vitamin C was about 40
mg/100 g in fresh fruits and 38 mg/100 g in dried fruits. DONNO et al. (2015) report an
amount of about 42 mg/100 g in dried goji berries. Taking into account the Recommended
Daily Allowance (RDA) for vitamin C of 80 mg/day (Regulation EU 1169/2011), 100 g of
fresh or dried goji berries contribute approximately 50 % of the RDA, so that they can be
declared on the label as a source of vitamin C. A portion of dried goji berries (30 g)
contributes about 16 % of the RDA.
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Ital. J. Food Sci., vol 29, 2017 - 406
4. CONCLUSIONS
Goji berries cultivated in Italy were confirmed as an important source of healthy
compounds, providing a significant contribution to the diet, in terms of some inorganic
nutrients, and of dietary fibre, zeaxanthin, vitamins E and C. In particular, taking into
account the Recommended Daily Allowance (RDA) for minerals and vitamins established
by the Commission of the European Communities, dried goji berries can be declared as a
source of K, P, Cu, Fe Mn, Zn. Moreover, both fresh and dried berries can be declared on
the label as a potential source of vitamins E and C.
The presented results enhance the knowledge of the composition and the nutritional
characteristics of fresh and dried goji berries cultivated in Italy and will help in verifying
the information reported in the label.
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Paper Received October 12, 2017 Accepted February 26, 2017
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