Available via license: CC BY-NC-ND 3.0
Content may be subject to copyright.
Vol. 61 No. 3 2015
DOI: 10.1515/hepo-2015-0022
From Botanical to Medical Research
Herba Pol 2015; 61(3): 53-66
Antiradical activity and amount of phenolic compounds in extracts
obtained from some plant raw materials containing methylxanthine
alkaloids
ZBIGNIEW SROKA
1*
, MARIA JANIAK
1
, ANDRZEJ DRYŚ
2
1
Department of Pharmacognosy
Faculty of Pharmacy
Wroclaw Medical University
Borowska 211a
50-556 Wroclaw, Poland
2
Department of Physical Chemistry
Faculty of Pharmacy
Wrocław Medical University
Borowska 211a
50-556 Wrocław, Poland
*corresponding author: phone.: +4871 784 0220, fax: +4871 784 0218,
e-mail: zbigniew.sroka@umed.wroc.pl
Summary
Raw materials containing methylxanthine alkaloids such as yerba mate, guaraná, white
and green tea, coffee seeds, chocolate and cacao seed powder and extracts obtained from
these raw materials were investigated for their antioxidant features and the amount of
phenolic compounds. The level of phenolic compounds was measured with the colori-
metric method using Folin-Ciocalteu’s phenol reagent and antioxidant features was deter-
mined with the use of DPPH (2,2-diphenyl-1-picrylhydrazyl radical). Amounts of phenolic
compounds were presented in percentages per mass of extracts and mass of raw materials.
Antiradical potential was shown as the number of TAU
DPPH
units per mg of extracts and g
of raw materials. The highest number of antiradical units TAU
DPPH
as well as the amount of
phenolic compounds was calculated for white tea and its extracts and lowest for choco-
late. The correlation coefficient between the content of phenolics and antiradical features
of raw materials is equal to r=0.994.
Key words: antioxidants, free radical scavengers, plant extracts, plant phenolics, methylxanthine
alkaloid raw materials, antiradical units
EXPERIMENTAL PAPER
Unauthenticated
Download Date | 6/1/16 4:50 AM
54
Z. Sroka, M. Janiak, A. Dryś
Plant raw materials containing methylxanthine alkaloids are rich in phenolic
compounds [1-4]. Also extracts obtained from these sources contain large quan-
tities of phenols with strong antioxidative activity. Tea leaves and extracts from
tea are among the most effective. Among them white and green tea exhibit the
strongest antioxidant or antiradical properties [5].
Guaran
á, Paullinia cupana Kunth, is a member of the climbing plant family of
Sapindaceae. The plant is native to the Amazon basin including Brazil. Guaran
á
seeds are rich in caffeine and other methylxanthine derivatives, also contains
tannins and saponins. This raw material contains about 12% tannins, mainly pro-
anthocyanidins, about 6% (+)-catechin and 3.8% (-)-epicatechin [6]. Guaran
á was
consumed for centuries by Indians, who added it to drinks and foods in order to
decrease fatigue and to increase vigor [4, 7]. Nowadays, guaran
á is used for the
production of soft drinks. Guaran
á powder is used as a nutritional supplement to
enhance vigor, to inhibit the platelet aggregation, protect against gastric injury
caused by ethanol, and for antimicrobial and antioxidant activity [4].
White tea (Camellia sinensis (L.) Kuntze is a less processed kind of tea (steamed,
then dried without withering) and exhibit slightly higher antiradical properties
than green teas [5, 8]. It acts advantageously in the prevention of many diseases,
such as heart and neurodegenerative diseases as well as cancers [8]. White tea is
produced from buds and leaves of Camellia sinensis. The white color comes from
the hair of unopened buds. White tea contains similar amount of catechin deriva-
tive as epigallocatechin gallate as green tea but the amount of epigallocatechin
is higher than in green teas. Main catechins in white tea are catechin, epigallo-
catechin gallate, epigallocatechin, and epicatechin [5, 8].
Yerba mate beverage origins from the leaves of Ilex paraguariensis A. St.-Hil. It
is consumed in South America countries such as Brazil (South part), Argentina,
Uruguay and Paraguay. The raw material to be infused consist of dry leaves of Ilex
paraguariensis belonging to the botanical family Aquifoliaceae. According to the
literature, yerba mate has hypocholesterolemic and hepatoprotective activity, and
also exhibits a stimulating effect on the central nervous system [1, 9]. There was
also observed diuretic and antioxidant activity [1, 10, 11], an advantageous effect
on the cardiovascular system [12, 13], and protection of DNA and LDL against
oxidation [14]. Leaves of Ilex paraguariensis contain polyphenols such as phenolic
acids (chlorogenic, caffeic, dicaffeoylquinic acids), flavonoids (quercetin, kaemp-
ferol, rutin), methylxanthines such as caffeine and smaller quantity of theobro-
mine [9]. The leaves of yerba mate also contain amino acids, some vitamins such
as C, B
1
, B
2
, and mineral components (Fe, P, Ca) [9]. In some cases during use of
yerba mate serious adverse effects have been observed, such as oral, oropharyn-
geal and esophageal cancers [15].
Coffee is the most frequently consumed drink in the world, along with tea. Seeds
of coffee contain numerous compounds with antiradical activity such as chlorogen-
ic, 5-caffeoylquinic, feruloylquinic and caffeic acids, melanoidins and caffeine [16].
Unauthenticated
Download Date | 6/1/16 4:50 AM
55
Antiradical activity and amount of phenolic compounds in extracts obtained from some plant raw materials containing...
Vol. 61 No. 3 2015
Cocoa beans originate from plants of Theobroma cacao L. The amount of total
phenols in cocoa powder is about 6%. There are mainly flavan-3-ols, such as mo-
nomeric catechin or epicatechin, their oligomers (procyanidins) and flavonoids
among other derivatives of quercetin [16, 17].
Chocolate is obtained from cacao. Over many years the use of cacao (Theobroma
cacao) evolved into the product called chocolate. Chocolate is rich in lipids with
high amounts of saturated fatty acids such as stearic (30–33%) and palmitic (28-30%)
[18]. Besides, some amounts of sterols were identified as well as minerals, flavo-
noids, catechin, epicatechin, and procyanidins (in cocoa beans 12
–
48%). Chocolate
has a concentration of flavan-3-ol higher than most plant-based foods [19, 20]. The
polyphenols present in chocolate and cacao beans might exert advantageous vascu-
lar effects [21]. These compounds might reduce cardiovascular morbidity [22], and
diseases with chronic character [23]. According to scientific opinion demonstrated
in EFSA (European Food Safety Authority) monomeric catechins (mainly epicatechin)
and oligomers (procyanidins) maintain ”endothelium-dependent vasodilation which
contributes to healthy blood flow” at a daily dose of 200 mg [24, 25].
It is necessary to mention that chocolate are high processed products and the
amount of phenols and the antioxidant activity varies widely. The strongest anti-
oxidant activity exhibit dark chocolate but weakest milk chocolate [26].
All above described raw materials are known to have high amounts of phenolic
compounds such as tannins, mainly condensed [4, 27], catechins and gallic acid
derivatives (epigallocatechin gallate, EGCG) [28], and other coupling of catechins
such as procyanidins [29], and simple phenolic acids (gallic, caffeic) [30-32]. Other
phenols belong to the very wide group of flavonoids such as, the most common,
derivatives of quercetin, or kaempferol [31, 33].
The demonstration of methylxanthine raw materials’ strong antiradical or anti-
oxidant activity might expand their therapeutic use as a weapon to fight diseases
caused by free radicals and reactive oxygen species.
Free radicals and reactive oxygen species (ROS) are formed in normal physio-
logical processes and in some diseases, especially chronic [34]. The physiological
level of free radicals and ROS is maintained with an enzymatic system (superoxide
dismutase, catalase), and natural antioxidants such as glutathione, or NADPH, and
vitamins E and C can decrease the amount of ROS [35-37]. During chronic inflam-
mation, in diseases such as diabetes, free radicals are formed in excess and cannot
be effectively eliminated from the organism. Then, providing strong antioxidants
or free radical scavengers in diet or as a medicine could be advantageous.
As it was stated above, raw materials containing methylxanthines (caffeine,
theobromine, theophylline) are rich in polyphenols with strong antioxidant prop-
erties. Thanks to the content of phenolic compounds, these raw materials could
have many beneficial effects to health such as antiatherosclerotic effect.
Although methylxanthines exhibit some antioxidant properties, the main
components responsible for antioxidant activity are polyphenols. There were de-
scribed studies [26] that antioxidant activity of different types of chocolate posi-
tively correlated with the content of phenolic compounds.
Unauthenticated
Download Date | 6/1/16 4:50 AM
56
Z. Sroka, M. Janiak, A. Dryś
The aim of this work was to measure the antioxidant activity of these raw ma-
terials and study the correlation between antioxidant activity and the amount of
phenolic compounds in extracts.
White tea leaf from Camellia sinensis (L.) Kuntze (wt) – 44.5 g
Guaraná seeds from Paullinia cupana Kunth (gs) – 48.6 g
Yerba mate leaf from Ilex paraguariensis A. St.-Hil. (ym) – 39.8 g
Green tea leaf (gunpowder) from Camellia sinensis (L.) Kuntze (gt) – 50.0 g
Costa Rica coffee seeds from Coffea arabica L. (cc) – 48.0 g
Arabica coffee seeds from Coffea arabica L. (ca) – 46.0 g
Chocolate (Ristora) (cr) – 48.1 g
Chocolate (van Heuten) (ch) – 50.0 g
Chocolate (Tazza) (ct) – 48.1 g
Cacao (van Heuten) seed powder from Theobroma cacao L. (c) – 50.0 g
All raw materials were of commercial origin, edible quality.
Raw materials were extracted according to the modified method described by
Kasprzyk et al., [38] with methanol (900 ml) at a temperature of 50°C for 48 h.
Twenty percent of methanol extract (180 ml) was separated and concentrated
to dryness under reduced pressure to obtain the WA residue. The remaining
part of methanol extract (720 ml) was concentrated to dryness under reduced
pressure and then dissolved in 600 ml of water at 45°C. Water solution of ex-
tract was stored at 4°C for 48 h. The precipitate was separated with filter paper
(Filtrak, 388, 80 g/m
2
) and then dried under reduced pressure to obtain the WD
residue.
After precipitate separation, aqueous solution was exhaustively extracted with
ethyl acetate. Aqueous remaining and ethyl acetate solution were concentrated
to dryness under reduced pressure to obtain WB and WC residues, respectively.
Extracts from white tea were marked additionally with “wt”, extracts from
guaraná seeds with “gs”, extracts from yerba mate “ym”, extracts from green tea
with “gt”, extracts from Costa Rica coffee “cc”, extracts from Arabica coffee “ca”,
extracts from chocolate (Ristora) with “cr”, extracts from chocolate (van Heuten)
with “ch”, extracts from chocolate “Tazza” with “ct”, and extracts from cacao
(van Heuten) with “c”. For example, extracts obtained from white tea leaves are
marked with WAwt, WBwt, WCwt and WDwt.
Unauthenticated
Download Date | 6/1/16 4:50 AM
57
Antiradical activity and amount of phenolic compounds in extracts obtained from some plant raw materials containing...
Vol. 61 No. 3 2015
During extraction to the ethyl acetate, in most cases the emulsion was formed,
which was individually separated and then condensed to dryness under reduced
pressure to obtain WE extract.
The amount of total phenolic compounds was measured with use the method
of Singleton and Rossi [39]. Seven ml of water, then 0.5 ml of Folin and Ciocalteu’s
phenol reagent (3H
2
O x P
2
O
5
x 13WO
3
x 5MoO
3
x 10H
2
O) and 0.5 ml of methanol
solution of extract was poured into the test tube. After 3 min, 2 ml of 20% aque-
ous solution of sodium carbonate was added. The mixture was heated at 100°C
for 1 min. After cooling, absorbency was measured at 685 nm. The measurement
was repeated for five times. Maximal error was calculated with total differential
method. Phenolic compounds were expressed as gallic acid and calculated in per-
centage per weight of extract.
Antiradical activity of extracts was measured by the method of Brand-Williams
et al. [40]. The decrease of absorbency of DPPH (2.2-diphenyl-1-picrylhydrazyl
radical) solution in methanol (94 µmol/l) at 515 nm is measured in the presence
of a substance with antiradical activity. The rate of the decrease of absorbance is
proportional to antiradical activity of the substance.
DPPH was dissolved in methanol (gradient grade, Merck). The reagent was prepared
a day before the experiment so that absorbency at 515 nm of solution was stable.
2 ml of DPPH solution (94 µmol/l) was placed in glass cuvette with optical path
of 1 cm. 50 µl of methanol solution of extract was added. The absorbance of the
solution was measured at 0 and 60 s.
The antiradical potential of the extract was demonstrated as a number of
antiradical units TAU
DPPH
per mg of extract (TAU
DPPH/mg
) and per g of raw material
(TAU
DPPH/g
) calculated according to the equations (1) and (2), respectively.
One unit of antiradical activity is the quantity of antioxidant that scavenges 1 µmol
of DPPH radical in 1 ml of reaction mixture during 1 minute of reaction at 25°C.
(1)
7990.0
10
c
AA
TAU
ss
DPPH/mg
−
⋅=
(1)
The above equation was derived using absorption attenuation coefficient (ε)
equal to: 1.2509∙10
4
L mol
-1
cm
-1
[40], where TAU
DPPH/mg
is the number of antiradical
units per mg of substance, A
s0
is absorbency of DPPH solution at the beginning of
the reaction, A
s1
is absorbency of DPPH solution after 1 min of reaction, c is con-
centration of substance in reaction mixture [mg/ml].
Unauthenticated
Download Date | 6/1/16 4:50 AM
58
Z. Sroka, M. Janiak, A. Dryś
The number of antiradical units per gram of raw material (TAU
DPPH/g
) was cal-
culated by summing up all TAU
DPPH/mg
units calculated for all extracts multiplied
by the total mass of extracts obtained from given raw material according to the
equation:
/ / / / /
/
( )( )( )( )( )
(2)
DPPH mgWA WA DPPH mgWB WB DPPH mgWC WC DPPH mgWD WD DPPH mgWE WE
DPPH g
R
TAU m TAU m TAU m TAU m TAU m
TAU
W
⋅+ ⋅+ ⋅+ ⋅+ ⋅
=
(2)
where TAU
DPPH/g
is the number of antiradical units calculated per g of raw material;
TAU
DPPH/mgWA,
TAU
DPPH/mgWB,
TAU
DPPH/mgWC,
TAU
DPPH/mgWD,
TAU
DPPH/mgWE
is the number of anti-
radical units per mg of extract WA, WB, WC, WD and WE respectively; m
WA,
m
WB,
m
WC,
m
WD,
m
WE
is whole mass of WA, WB, WC, WD, WE extracts [mg], respectively;
W
R
is mass of raw material [g] taken for extraction.
The number of antiradical units calculated per mg of extracts (TAU
DPPH/mg
) and
amount of phenolic compounds in extracts and raw materials (Ph%) is demonstrat-
ed in table 1, figure 1, and figure 2. The general observation is that the highest
values of TAU
DPPH/mg
were obtained for extracts WC for all investigated raw materi-
als; also the amount of phenolic compounds for this extract was the highest. The
strongest antiradical properties exhibited extracts from white tea, among them
WCwt was the strongest (6.86±0.55) among all investigated extracts. Similar,
in terms of antiradical activity, were extracts obtained from leaves of green tea.
Among them the strongest was WCgt extract (6.73±0.21).
WBc
1670.0 0.061±0.007 3.72±0.17
WCc
349.8 0.37±0.03 9.88±0.52
WDc
580.7 0.082±0.012 1.26±0.18
WEc
4.3 0.078±0.012 2.56±0.19
Figure. 1.
Number of antiradical units (TAU
DPPH/mg
) per mg of extracts obtained from white tea (wt), extracts
from guarana seeds (gs), extracts from yerba mate (ym), extracts from green tea (gt). As a positive
control trolox was used, in figure marked with light gray colour
Figure 1.
Number of antiradical units (TAU
DPPH/mg
) per mg of extracts obtained from white tea (wt), extracts
from guaraná seeds (gs), extracts from yerba mate (ym), extracts from green tea (gt).
Unauthenticated
Download Date | 6/1/16 4:50 AM
59
Antiradical activity and amount of phenolic compounds in extracts obtained from some plant raw materials containing...
Vol. 61 No. 3 2015
Figure 2.
Number of antiradical units (TAU
DPPH/mg
) per mg of extracts obtained from coffee Costa Rica (cc),
coffee Arabica (ca), chocolate Ristora (cr), chocolate van Heuten (ch), chocolate Tazza (ct) and cacao
(c). As a positive control trolox was used, in figure marked with light grey colour
Figure 2.
Number of antiradical units (TAU
DPPH/mg
) per mg of extracts obtained from coffee Costa Rica (cc), coffee
Arabica (ca), chocolate Ristora (cr), chocolate van Heuten (ch), chocolate Tazza (ct) and cacao (c).
The weight of extracts [mg], number of antiradical units per mg of extract (TAU
DPPH/mg
), amount of
phenolic compounds in extracts expressed in percentage, number of antiradical units per g of raw
material (TAU
DPPH/g
), amount of phenolic compounds in raw materials (Ph%) expressed in percentage.
Raw
material
Extract
Weight of
extract [mg]
TAU
DPPH/mg
The amount
of phenolic
compounds in
extracts [w/w]
TAU
DPPH/g
The amount of
phenolic compounds
in raw materials Ph%
[w/w]
2523.9 6.16±0.78 43.2±3.22
1268.6±133.5 12.26±10.57
4606.8 2.55±0.10 21.1±2.25
3160.6 6.86±0.55 65.6±3.24
2616.6 2.66±0.17 49.2±2.23
213.8 2.45±0.11 15.8±0.89
1654.5 3.10±0.31 30.5±2.01
474.8±44.6 4.42±3.93
1899.6 1.98±0.21 18.1±1.29
1518.7 4.12±0.35 37.1±2.55
2226.3 3.55±0.16 33.2±2.25
1701.8 1.17±0.45 23.5±1.46
445.8±62.4 4.21±4.26
4095.4 1.98±0.16 17.6±1.37
1220.8 4.98±0.25 41.3±3.16
555.7 2.76±0.17 9.3±1.42
2984.7 4.45±0.15 30.4±5.07
1184.4±64.4 9.79±0.56
4808.4 1.71±0.12 15.84±2.64
2831.4 6.73±0.21 59.2±9.87
3474.2 5.32±0.11 42.09±7.01
420.5 0.39±0.08 20.17±3.36
Unauthenticated
Download Date | 6/1/16 4:50 AM
60
Z. Sroka, M. Janiak, A. Dryś
Raw
material
Extract
Weight of
extract [mg]
TAU
DPPH/mg
The amount
of phenolic
compounds in
extracts [w/w]
TAU
DPPH/g
The amount of
phenolic compounds
in raw materials Ph%
[w/w]
1168.9 0.57±0.10 10.98±3.85
60.9±6.5 1.36±0.19
1168.3 0.73±0.005 11.79±1.39
831.2 0.71±0.075 24.06±0.83
1245.2 0.32±0.05 9.39±1.25
76.7 0.52±0.04 13.43±0.46
998.0 0.34±0.02 6.14±1.88
41.8±4.2 0.94±0.15
1237.9 0.57±0.02 13.63±0.94
644.8 0.76±0.075 20.11±2.60
1530.4 0.24±0.034 4.63±1.56
36.5 0.44±0.038 12.53±1.0
857.8 0.065±0.008 1.51±0.13
6.3±1.1 0.088±0.004
3238.5 0.055±0.009 0.59±0.10
75.3 0.56±0.067 9.85±1.72
212.3 0.11±0.024 2.25±0.20
67.8 0.06±0.01 1.54±0.19
2033.4 0.068±0.01 1.81±0.20
10.4±3.2 0.260±0.052
7757.0 0.031±0.01 0.77±0.20
131.6 0.56±0.065 16.52±0.68
348.6 0.10±0.007 1.23±0.38
194.4 0.17±0.03 3.77±1.13
671.4 0.05±0.006 1.37±0.30
3.0±0.44 0.067±0.02
2405.9 0.029±0.003 0.51±0.29
86.2 0.29±0.025 8.54±0.58
207.1 0.05±0.015 1.0±0.17
92.4 0.04±0.006 0.82±0.08
646.2 0.28±0.008 5.73±0.55
9.2±0.87 0.282±0.019
1670.0 0.061±0.007 3.72±0.17
349.8 0.37±0.03 9.88±0.52
580.7 0.082±0.012 1.26±0.18
4.3 0.078±0.012 2.56±0.19
WA - methanol extract, WB - water remaining extract, WC - ethyl acetate extract, WD - precipitate,
WE - emulsion (layer formed between the water and ethyl acetate), “wt” white tea, “gs” guaraná
seeds, “ym” yerba mate, “gt” green tea, “cc” coffee seeds (Costa Rica), “ca” coffee seeds (Arabica),
“cr” chocolate Ristora, “ch” chocolate van Heuten, “ct” chocolate Tazza, “c” cacao. Value TAU
DPPH/mg
for trolox is 3.22 ± 0.06.
Unauthenticated
Download Date | 6/1/16 4:50 AM
61
Antiradical activity and amount of phenolic compounds in extracts obtained from some plant raw materials containing...
Vol. 61 No. 3 2015
White and green tea extracts appeared to be the strongest antiradicals, and
white tea was even more active than green tea. Our previous research exhibited
strong antiradical features of green tea leaves [41]. Green and white teas do not
undergo a fermentation process. These two kinds of teas contain phenolic com-
pounds such as gallocatechin and gallocatechin gallate [5], which appeared to be
very effective antiradical scavengers.
The raw materials such as yerba mate (ym) and guaraná seeds (gs) appeared to
have lower antiradical properties than those of white and green teas (tab. 1, fig. 3).
Yerba mate contains many classes of caffeoyl derivatives (caffeic, chlorogenic and
5-caffeoylquinic acids and flavonoids, such as rutin [9]. Phenolic compounds present
in yerba mate leaves exhibit the antioxidant activity in vitro and in vivo, and have the
ability to scavenge free radicals and reactive oxygen species [10, 11].
Figure. 3.
Number of antiradical units per g of raw material (TAU
DPPH/g
) for white tea (wt), guarana seeds (gs),
yerba mate (ym), green tea (gt)
Figure 3.
Number of antiradical units per g of raw material (TAU
DPPH/g
) for white tea (wt), guaraná seeds (gs),
yerba mate (ym), green tea (gt)
Guaraná seeds is a raw material rich in methylxanthine derivatives, and contain
more caffeine than other raw materials [42]. This raw material contains polyphe-
nols with strong antioxidant activity such as tannins, especially condensed tannins
[4]. It is believed that condensed and hydrolysable tannins apart from antioxidant
activity have other therapeutic properties [43, 44].
Our investigation showed that extracts obtained from coffee seeds (cc, ca) have
average antioxidative activity (tab. 1, fig. 2). The antioxidant features are posi-
tively correlated with amount of phenolic compounds in these extracts (tab. 1).
Correlation coefficient (r) between amount of phenolic compounds in extracts
and number of antiradical activity TAU
DPPH/mg
was equal to 0.91 (fig. 4). Correlation
coefficient (r) between amount of phenols and number of activity units in raw
materials (TAU
DPPH/g
) was equal to 0.99.
Unauthenticated
Download Date | 6/1/16 4:50 AM
62
Z. Sroka, M. Janiak, A. Dryś
Figure 4.
Correlation coefficient (r) between number of antiradical units per mg of extract (TAU
515/mg
) and
amount of phenolic compounds in extracts (% w/w)
Seeds of coffee are known to contain effective antiradical phenolic compounds
such as caffeic and chlorogenic acids [45]. The chlorogenic acid, apart from an-
tioxidant activity, also exhibits some astringency and bitterness. Other phenolic
compounds present in coffee seeds are ferulic, p-coumaric, vanillic, and syringic
acids [46], which are rather weak antioxidants. Phenolic compounds play an im-
portant role in the quality and flavor of coffee seeds. Other compounds present
in coffee seeds with antioxidant activity are melanoidins [47] and caffeine [48].
Our investigation showed that chocolates (cr, ch, ct) appeared to be the weak-
est antioxidants among all raw materials investigated in this work (tab. 1, fig. 5),
which correlated positively with small amount of phenolic compounds. Choco-
late has variable amounts of phenolic compounds. White chocolate contains
low amounts of phenols, dark chocolate is rich in phenols [45]. The amount of
phenolic compounds correlated positively with antioxidant activity in dark, milk
and white chocolate [49]. According to Miller et al. [19] the antiradical activity
varies from 66.7±8.7 (µmol TE/g) for chocolate syrup to 499±35.5 (µmol TE/g)
for baking chocolate and even to 816±37.6 (µmol TE/g) for cacao powder. The
main phenolic compounds present in chocolate are epicatechin, catechin, pro-
cyanidin B2, procyanidin C1, and procyanidin B5 which are antioxidants, valu-
able for health [50].
In our research, cacao powder and extracts from this raw material were found
to be weak antioxidants (tab. 1, fig. 5). The literature describes rather strong an-
tiradical activity of cocoa [19, 51] due to presence of high amounts of phenolic
compounds.
Unauthenticated
Download Date | 6/1/16 4:50 AM
63
Antiradical activity and amount of phenolic compounds in extracts obtained from some plant raw materials containing...
Vol. 61 No. 3 2015
Figure. 5.
Number of antiradical units per g of raw material (TAU
DPPH/g
) for (Costa Rica) coffee (cc), (Arabica)
coffee (ca), (Ristora) chocolate (cr), (van Heuten) chocolate (ch), (Tazza) chocolate (ct), cacao (c)
Figure 5.
Number of antiradical units per g of raw material (TAU
DPPH/g
) for Costa Rica coffee (cc), Arabica coffee
(ca), Ristora chocolate (cr), van Heuten chocolate (ch), Tazza chocolate (ct), cacao (c)
1. It can be concluded that the extracts of non-fermented, white and green teas
exhibited very high antioxidant features. Much lower activity was observed for
guaraná seeds and yerba mate leaves, and chocolates and cocoa powder ap-
peared to be the weakest antioxidants.
2. The antioxidant properties highly positively correlated with the amount of
phenolic compounds.
The authors would like to thank the technical staff of the Department of Phar-
macognosy, Wrocław Medical University for the help in the research.
The study was financially supported by the Wroclaw Medical University, grant
No. ST-527.
1. Berté KAS, Beux MR, Spada PKWDS, Salvador M, Hoffman-Ribani R. Chemical composition and
antioxidant activity of Yerba-Mate (Ilex paraguariensis A. St.-Hil., Aquifoliaceae) extracts as obtained by
spray drying. J Agric Food Chem 2011; 59:5523-5527.
Unauthenticated
Download Date | 6/1/16 4:50 AM
64
Z. Sroka, M. Janiak, A. Dryś
2. Horžić D, Komes D, Belščak A, Ganić K, Ivecović D, Karlović D. The composition of polyphenols and
methylxanthines in teas and herbal infusions. Food Chem 2009; 115:441-448.
3. Nsor-Atindana J, Zhong F, Mothibe KJ, Bangoura ML, Lagnika C. Quantification of total polyphenolic
content and antimicrobial activity of cocoa (Theobroma cacao L.) Bean Shells. Pakistan J Nutr 2012;
11:574-579.
4. Yamaguti-Sasaki E, Ito LA, Canteli VCD, Ushirobira TMA, Ueda-Nakamura T, Dias-Filho BP. et al.
Antioxidant capacity and in vitro prevention of dental plaque formation by extracts and condensed
tannins of Paullinia cupana. Molecules 2007;12:1950-1963.
5. Unachukwu UJ, Ahmed S, Kavalier A, Lyles JT, Kennelly EJ. White and green teas (Camellia sinensis var.
sinensis): variation in phenolic, methylxanthine, and antioxidant profiles. J Food Sci 2010;75:541-548.
6. Friedhelm M. Analysis of guaraná seeds II. Studies on the composition of the tannin fraction. Z Lebensm
Unters 1990; 190:429-431.
7. Sombra LL, Gómez MR, Olsina R, Martinez LD, Silva MF. Comparative study between capillary
electrophoresis and high performance liquid chromatography in „guaraná ” based phytopharmaceuticals.
J Pharmaceut Biomed 2005;36:989-994.
8. Espinosa C, López-Jiménez JA, Cabrera L, Larqué E, Almajano MP, Arnao MB. et al. Protective effect of
white tea extract against acute oxidative injury caused by adriamycin in different tissues. Food Chem
2012; 134:1780-1785.
9. Heck CI, De Mejia EG. Yerba mate tea (Ilex paraguariensis): a comprehensive review on chemistry, health
implications, and technologica considerations. J Food Sci 2007; 72:R138-R151.
10. Anesini C, Turner S, Cogoi L, Filip R. Study of the participation of caffeine and polyphenols on overall
antioxidant activity of mate (Ilex paraguariensis). LWT - Food Sci Technol 2012; 45:299-304.
11. Valerga J, Reta M, Lanari MC. Polyphenol input to the antioxidant activity of yerba mate (Ilex
paraguariensis) extracts. LWT - Food Sci Technol 2012;45:28-35.
12. Schinella G., Fantinelli JC, Mosca SM. Cardioprotective effects of Ilex paraguariensis extract: evidence for
a nitric oxide-dependent mechanism. Clin Nutr 2005;24:360-366.
13. Schinella G, Fantinelli JC, Tournier H, Prieto JM, Spegazzini E, Debenedetti S. et al. Antioxidant and
cardioprotective effects of Ilex brasiliensis: A comparative study with Ilex paraguariensis (yerba mate).
Food Res Int 2009; 42:1403-1409.
14. Bracesco N, Sanchez AG, Contreras V, Menini T, Gugliucci A. Recent advances on Ilex paraguariensis
research: Minireview. J Ethnopharmacol 2011; 136:378-384.
15. Goldenberg D, Golz A, Joachims HZ. The beverage mate: a risk factor for cancer of the head and neck.
Head Neck 2003; 25:595-601.
16. Rawel HM, Kulling SE. Nutritional contribution of coffee, cacao and tea phenolics to human health. J
Verbrauch Lebensm 2007;2:399-406.
17. Subagio A, Sari P, Morita N. Simultaneous determination of (+)-catechin and (-)-epicatechin in cacao and
its products by high-performance liquid chromatography with electrochemical detection. Phytochem
Analysis 2001;12:271-276.
18. Marques AJV, Coelho JAP. Determination of fat contents with supercritical CO
2
extraction in two commercial
powder chocolate products: comparison with NP-1719. J Food Process Eng 2011; 34:1597-1608.
19. Miller KB, Stuart DA, Smith NL, Lee CY, Michale NL, Flanagan JA et al. Antioxidant activity and polyphenol
and procyanidin contents of selected commercially available cocoa-containing and chocolate products
in the Unites States. J Agric Food Chem 2006; 54:4062-4068.
20. Steinberg FM, Bearden MM, Keen CL. Cocoa and chocolate flavonoids: Implications for cardiovascular
health. J Am Diet Assoc 2003;103:215-223.
21. Ding EL, Huftfless SM, Ding X, Girotra S. Chocolate and prevention of cardiovascular disease:
A systematic review. Nutr Metab 2006; 3:1-12.
22. Galleano M, Oteiza PI, Fraga CG. Cocoa, chocolate and cardiovascular disease. J Cardiovasc Pharmacol
2009; 54:483-490.
23. Grassi D, Desideri G, Ferri C. Blood pressure and cardiovascular risk: What about cocoa and chocolate?
Arch Biochem Biophys 2010; 501:112-115.
24. Scientific Opinion on the substantation of a health claim related to cocoa flavonols amd maintenance
of normal endothelium-dependent vasodilation pursuant to Article 13 (5) of Regulation (EC) No
1924/2006. EFSA J 2012; 10:2809.
Unauthenticated
Download Date | 6/1/16 4:50 AM
65
Antiradical activity and amount of phenolic compounds in extracts obtained from some plant raw materials containing...
Vol. 61 No. 3 2015
25. Scientific Opinion on the modification of the authorization of a health claim related to cocoa flavonols
and maintenance of normal endothelium-dependent vasodilation pursuant to Article 13 (5) of
Regulation (EC) No 1924/2006
1
following a request in accordance with Article 19 of Regulation (EC) No
1824/2006. EFSA J 2014; 12:3654.
26. Todorovic V, Redovnikovic IR, Todorovic Z, Jankovic G, Dodevska M, Sobajic S. Polyphenols,
methylxanthines, and antioxidant capacity of chocolates produced in Serbia. J Food Compos Anal 2015;
41:137-143.
27. Vieira MA, Maraschin M, Pagliosa CM, Podesta R, De Simas KN, Rockenbach II et al. Phenolic acids and
methylxanthines composition and antioxidant properties of mate (Ilex paraguariensis) residue. J Food
Sci 2010;75:C280-C285.
28. Wheeler DS, Wheeler WJ. The medicinal chemistry of tea. Drug Develop Res 2004; 61:45-65.
29. Molan AL, De S, Meagher L. Antioxidant activity and polyphenol content of green tea flavan-3-ols and
oligomeric proanthocyanidins. Int J Food Sci Nutr 2009; 60:497-506.
30. Clifford MN. Chlorogenic acids and other cinnamates – nature, occurence and dietary burden. J Sci
Food Agr 1999; 79:362-372.
31. Isolabella S, Cogoi L, López P, Anesini C, Ferraro G, Filip R. Study of the bioactive compounds variation
during yerba mate (Ilex paraguariensis) processing. Food Chem 2010; 122:695-699.
32. Lin JK, Lin CL, Liang YC, Lin-Shiau SY, Juan IM. Survey of catechins, gallic acid, and methylxanthines in
green, oolong, pu-erh, and black teas. J Agric Food Chem 1998; 46:3635-3642.
33. Karakaya S, Nehir ELS. Quercetin, luteolin, apigenin and kaempferol contents of some foods. Food
Chem 1999; 66:289-292.
34. Burek CL, Rose NR. Autoimmune thyroiditis and ROS. Autoimmun Rev 2008; 7:530-537.
35. Majima HJ, Indo HP, Suenaga S, Matsui H, Yen HC, Ozawa T. Mitochondria as possible pharmaceutical
targets for the effect of vitamin E and its homologues in oxidative stress-related diseases. Curr Pharm
Design 2011; 17:2190-2195.
36. Mari M, Morales A, Colell A, Garcia-Ruiz C, Fernandez-Checa JC. Mitochondrial glutathione, a key
survival antioxidant. Antioxid Redox Sign 2009; 11:2685-2700.
37. Peng Y, Kwok KHH, Yang PH, Ng SSM, Liu J, Wong OG et al. Ascorbic acid inhibits ROS production, NF-κB
activation and prevents ethanol-induced growth retardation and microencephaly. Neuropharmacology
2005; 48:426-434.
38. Kasprzyk A, Żbikowska B, Sroka Z, Gamian A. The antiradical activity of some plant raw materials and
extracts obtained from these raw materials. Postepy Hig Med Dosw 2012; 66:146-152.
39. Singleton VL, Rossi JA. Colorimetry of total phenolics with phosphomolybdic-phosphotungustic acid
reagents. Am J Enol Viticult1965; 16:144-158.
40. Brand-Williams W, Cuvelier ME, Berset C. Use of free radical method to evaluate antioxidant activity.
Lebensm Wiss Technol 1995; 28:25-30.
41. Wojciechowski D, Sroka Z, Gamian A. Investigation of antiradical potential of different kinds of teas and
extracts from these teas using antiradical activity units (TAU). Postepy Hig Med Dosw 2011;65:796-803.
42. Mehr CB, Biswal RN, Collins JL, Cohran HD. Supercritical carbon dioxide extraction of caffeine from
Guaraná . J Supercritic Fluids 1996; 9:185-191.
43. Piluzza G, Sulas L, Bullitta S. Tannins in forage plants and their role in animal husbandry and
environmental sustainability: a review. Grass Forage Sci 2013; 69:32-48.
44. Iqbal Z, Mufti KA, Khan MN. Anthelmintic effects of condensed tannins. Int J Agric Biol 2002; 4:438-
440.
45. Ludwig IA, Sanchez L, Caemmerer B, Kroh LW, De Pena MP, Cid C. Extraction of coffee antioxidants:
Impact of brewing time and method. Food Res Int 2012; 48:57-64.
46. Somporn C, Kamtuo A, Theerakulpisut P, Siriamornpun S. Effect of shading on yield, sugar content,
phenolic acids and antioxidant property of coffee beans (Coffea arabica cv. Catimor) harvested from
north-eastern Thailand. J Sci Food Agric 2012;92:1956-1963.
47. Cämmerer B, Kroh LW. Antioxidant activity of coffee brews. Eur Food Res Technol 2006; 223:469-474.
48. Devasagayam TPA, Kamat JP, Mohan H, Kesavan PC. Caffeine as an antioxidant: Inhibition of lipid
peroxidation induced by reactive oxygen species. BBA-Biomembranes 1996; 1282:63-70.
49. Meng CC, Jalil AM, Ismail A. Phenolic and theobromine contents of commercial dark, milk and white
chocolate on the Malaysian market. Molecules 2009; 14:200-209.
Unauthenticated
Download Date | 6/1/16 4:50 AM
66
Z. Sroka, M. Janiak, A. Dryś
50. Mcshea A, Ramiro-Puig E, Munro SB, Casadesus G, Castell M, Smith MA. Clinical benefit and preservation
of flavonols in dark chocolate manufacturing. Nutr Rev 2008; 66:630-641.
51. Afify AEMMR, Shalaby EA, El-Beltagi HS. Antioxidant activity of aqueous extracts of different caffeine
products. Not Bot Horti Agr 2011; 39:117-123.
AKTYWNOŚĆ PRZECIWRODNIKOWA ORAZ ZAWARTOŚĆ ZWIĄZKÓW FENOLOWYCH
W WYCIĄGACH OTRZYMANYCH Z SUROWCÓW ROŚLINNYCH ZAWIERAJĄCYCH ALKALOIDY
METYLOKSANTYNOWE
ZBIGNIEW SROKA
1*
, MARIA JANIAK
1
, ANDRZEJ DRYŚ
2
1
Katedra i Zakład Farmakognozji
Wydział Farmaceutyczny z Oddz. Analityki Medycznej
Uniwersytet Medyczny we Wrocławiu
ul. Borowska 211a, 50-556 Wrocław
2
Katedra i Zakład Chemii Fizycznej
Wydział Farmaceutyczny z Oddz. Analityki Medycznej
Uniwersytet Medyczny we Wrocławiu
ul. Borowska 211a, 50-556 Wrocław
*autor, do którego należy kierować korespondencję: tel.: +4871 784 0220,
faks: +4871 784 0218, e-mail: zbigniew.sroka@umed.wroc.pl
Streszczenie
Surowce takie jak yerba mate, guaraná , biała i zielona herbata, nasiona kawy, czekolada
i kakao zawierające alkaloidy metyloksantynowe były badane na zawartość związków fe-
nolowych i aktywność przeciwutleniającą. Poziom związków fenolowych określono meto-
dą kolorymetryczną za pomocą odczynnika Folin-Ciocalteu, a właściwości przeciwutlenia-
jące mierzono stosując rodnik DPPH (difenylo-pikrylohydrazylowy). Ilość związków feno-
lowych wyrażono w procentach masy wyciągów i surowców. Potencjał przeciwrodnikowy
określono jako liczbę jednostek TAU
DPPH
na mg wyciągów i g surowców. Największą liczbę
jednostek przeciwrodnikowych TAU
DPPH
oraz najwyższą zawartość związków fenolowych
wykazano dla białej herbaty i wyciągów otrzymanych z tego surowca, a najniższe wartości
uzyskano dla czekolady. Współczynnik korelacji pomiędzy ilością związków fenolowych
i aktywnością przeciwrodnikową surowców wynosił r=0,994.
przeciwutleniacze, zmiatacze wolnych rodników, wyciągi roślinne, fenole roślin-
ne, surowce zawierające alkaloidy metyloksantynowe, jednostki przeciwrodnikowe
Unauthenticated
Download Date | 6/1/16 4:50 AM
