Pharmaceutical Chemistry Journal
1, 2008
MEDICINAL PLANTS
ANTIOXIDANT PROPERTIES OF MEDICINAL PLANTS
V. F. Gromovaya,1 G. S. Shapoval,1 I. E. Mironyuk,1 and N. V. Nestyuk1
Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 42, No. 1, pp. 26 – 29, January, 2008.
Original article submitted May 15, 2006.
Impulse voltamperometry was used to study the antioxidant activity of a series of tinctures from medicinal
plants. Effective concentrations were determined and a series of comparative evaluations of their antioxidant
and antiradical activities was constructed; these were maximal in St. John’s wort, ginseng, rosewort, and
Eleutherococcus. A possible mechanism of the antioxidant action of these plants in the body is suggested.
Interest in studies of the processes of free-radical oxida-
tion and, thus, in agents able to influence the intensity of
these processes, has increased significantly in recent years.
This is because in conditions of a worsening ecological envi-
ronment, various extreme factors produce significant in-
creases in the concentrations of active oxygen species (AOS,
including O-·, ·OH, H2O2, and others) in the body, able to
damage protein and nucleic acid molecules, inactivate en-
zymes, and damage cell membranes, which in turn leads to
the development of various pathological states in the body.
The question of the possible blockade of these free-radical
processes initiated by AOS at the early stages is therefore
particularly acute [1 – 5].
Free-radical processes in the body are controlled using
biologically active substances (BAS) with antioxidant prop-
erties. These include agents of both synthetic and natural ori-
gin; an important place among these is held by plant prepara-
tions containing bioflavonoids, carotenoids, essential fatty
acids, etc. The use of preparations of this type has enormous
potential - as natural antioxidants, they have easy and unlim-
ited access to metabolic processes in the body and produce
virtually none of the side effects associated with synthetic
agents [6 – 9]. Medicinal plants are widely used in medicine,
though as antioxidants they have received only fragmentary
investigation. Thus, studies of medicinal plants as potential
antioxidants and modeling of the possible mechanisms of
their inhibitory actions at defined stages of the reduction of
oxygen in the body are of significant interest.
The question of selecting effective antioxidants and iden-
tifying their antioxidant activity is a quite complex task.
1
Institute of Bioorganic Chemistry and Oil Chemistry, Ukrainian National
Academy of Sciences, Kiev.
25
Vol. 42, No.
Contemporary in vitro assessment methods are time-consum-
ing and laborious and do not allow evaluation of the
efficacies of agents in relation to their influences at defined
stages of free-radical processes [10 – 14]. Potential ap-
proaches to such studies include our previously developed
methodological approaches to modeling elementary initia-
tory redox reactions involving oxygen, as well as the impulse
voltamperometry method and its derivative express method,
which allows simultaneous evaluation of the antioxidant and
antiradical activity of compounds of interest [15 – 16].
The aim of the present work was to investigate the anti-
oxidant activities of a wide range of tinctures of medicinal
plants and to model the possible mechanisms of their action
at defined stages of the reduction of oxygen in the body.
METHODS
Studies were performed using pharmaceutical tinctures
of motherwort, rosewort, Eleutherococcus, ginseng,
Echinacea, and bearberry, as well as laboratory-prepared (as
described in [17]) water-alcohol tinctures of St. John’s wort,
plantain, mayweed, and sage. Working solutions were pre-
pared in 0.1 M sodium chloride.
Electrochemical studies were performed as described in
[18, 19], using a PU-1 universal polarigraph in the differen-
tial impulse voltamperometry regime with a three-electrode
set-up. The working electrode was made from high-purity
copper. Potentials were measured relative to a silver chloride
reference electrode, and the accessory electrode was a plati-
num coil. Electrochemical studies and processing of results
were performed using a specially developed computer pro-
gram. All studies were performed in sodium chloride solu-
0091-150X/08/4201-0025 © 2008 Springer Science+Business Media, Inc.
26 V. F. Gromovaya et al.

dI/dE, arbitrary units a) dI/dE, arbitrary units b)

1 I
I
0.30 1 0.30
2 2
3
0.25 0.25 3
4

III 4 III
0.20 0.20
5
5
6
0.15 II 0.15 6
II
0.10 0.10

0.05 0.05

0 0
0 0.2 0.4 0.6 0.8 1.0 1.2 –E, V 0 0.2 0.4 0.6 0.8 1.0 1.2 –E, V
Fig. 1. Voltamperometric curves of the reduction of oxygen in the presence of different concentrations of: a) rosewort: 1) baseline 0.1 M NaCl;
2 – 6) 0.2, 0.63, 0.92, 1.11, and 1.16 g/liter; b) Eleutherococcus: 1) baseline 0.1 M NaCl; 2 – 6) 0.523, 0.814, 1.677, 2.532, and 3.142 g/liter.

tion, whose concentration corresponded to physiological sa- BAS and h is the height of the corresponding wave in the
line (0.9%). The oxygen concentration was determined by presence of the study BAS.
equilibrium at atmospheric pressure and a temperature of
20 ± 1°C. RESULTS AND DISCUSSION
Antiradical and antioxidant activity of BAS were as-
sessed in terms of changes in the relative height (h/h0) of Figure 1 shows voltamperometric curves for the reduc-
waves of hydroxyl radicals (I) and hydrogen peroxide (III) tion of oxygen obtained in the copper electrode impulse po-
respectively during the electrochemical reduction of oxygen larization regime in baseline 0.1 M sodium chloride solution.
in the presence of study compounds, where h0 is the height These methodological approaches resulted in recording of
(mm) of the wave of reduction of hydroxyl radicals, molecu- three stages of the reduction of oxygen, characterized by the
lar oxygen, or hydrogen peroxide in the absence of study following waves:
·
Wave I OH + e- > OH-; (1)

Wave II O2 + 2e- + 2H+ > H2O2; (2)

TABLE 1. Comparison of the Antioxidant Activities of Medicinal
Plant Tinctures. Wave III H2O2 + 2e- + 2H+ > 2H2O; (3)
Maximum change in the height
of waves of oxygen reduction in the H2O2 + e- > H2O2- > -OH + ·OH (4)
Maximally presence of study preparations
Preparation effective compared with baseline (h/h0) The ability to define this stepwise reduction of oxygen,
studied concentration, which may serve as a model of the elementary initiory redox
Wave
g/dm3
hydroxyl molecular hydrogen
processes occurring in the body in both two- and one-elec-
radicals oxygen peroxide tron reduction of oxygen, allowed the influences of various
BAS on the individual stages of the reduction of oxygen to
Ginseng 0.8 0.38 1.05 0.36
be studied.
St. John’s wort 1.13 0.68 1.12 0.46
Thus, addition of the study medicinal plant tinctures to
Rosewort 1.11 0.58 1.71 0.34
the baseline solution showed that all induced changes in the
Eleutherococcus 2.7 0.62 1.04 0.48
voltamperometric curves of the reduction of oxygen, though
Mayweed 1.32 0.63 1.09 0.56
these changes were of different natures. Addition of rosewort
Motherwort 0.21 0.73 0.96 0.77
to the oxygen-containing baseline solution induced a signifi-
Bearberry 0.7 0.69 0.93 0.69
cant increase in the wave of reduction of molecular oxygen
Sage 0.88 0.86 1.00 0.96
(wave II) and shifted its potential in the positive direction,
Plantain 1.13 0.69 0.85 0.82
providing evidence that the process of reduction of molecu-
Echinacea 1.02 0.81 0.91 0.98
lar oxygen was facilitated in the presence of this tincture
Antioxidant Properties of Medicinal Plants
h/h0 a)
1.1
0.9
0.7
0.5
0.3
0.1
0 0.4 0.8 1.2 C, g/liter
Fig. 2. Relationship between relative decreases in the heights of waves of oxygen reduction at the copper electrode: wave I (Z); wave II (<);
wave III (D) and concentrations of: a) St. John’s wort; b) ginseng.
(Fig. 1a ). There were significant decreases in the waves cor-
responding to hydroxyl radicals (I) and hydrogen peroxide
(III). This effect increased with increases in the rosewort
concentration. The most effective concentration was ~1.1
g/dm3. A similar effect on hydroxyl radicals and hydrogen
peroxide was seen with Eleutherococcus, though the wave of
reduction of molecular oxygen (wave II) remained at essen-
tially the same level as the Eleutherococcus concentration in-
creased and there was no shift in the potential (Fig. 1b ).
St. John’s wort (Fig. 2a ) and motherwort increased the
magnitude of waves of reduction of molecular oxygen, facili-
tating the process of reduction and significantly suppressing
the waves of reduction of hydroxyl radical and hydrogen per-
oxide. St. John’s wort was rather more effective than
motherwort. Although the maximum effective concentration
for motherwort was lower, the changes in the heights of the
hydroxyl radical and hydrogen peroxide waves seen with St.
John’s wort could not be obtained even using significantly
increased concentrations.
If we take the interaction with peroxides as a measure of
antioxidant activity and the interaction with hydroxyl radi-
cals as a measure of antiradical activity, then the study agent
with the most marked antioxidant and antiradical activities
was ginseng, probably because of the inhibitory action in re-
lation to hydroxyl radicals formed by one-electron reduction
of peroxides (reaction 4), producing a significant decrease in
waves I and III on the voltamperometric oxygen reduction
curve (Fig. 2b ). Bearberry had marked antioxidant and
marked antiradical properties but had virtually no effect on
the reduction of molecular oxygen. The results obtained from
our studies of tinctures of medicinal plants are presented in
Table 1.
The present studies identified effective concentrations of
medicinal plant extracts and tinctures and allowed construc-
tion of a series of comparative evaluations of their antioxi-
dant activity:
27
h/h0 b)
1.2
1.0
0.8
0.6
0.4
0.2
0 0.4 0.8 1.2 1.6 2.0 C, g/liter
ginseng > rosewort > St. John’s wort > Eleutheroco-
ccus > mayweed > bearberry > motherwort > plantain >
Echinacea > sage;
and antiradical activity:
ginseng > rosewort > Eleutherococcus > mayweed > St.
John’s wort > bearberry = plantain > Echinacea > mother-
wort > sage.
Thus, we believe that the voltamperometric characteris-
tics of the potentially antioxidant BAS studied here allow not
only assessment of the antioxidant activity of these agents
overall, as described in [20], but also separation of the effects
associated with their interactions with hydrogen peroxide,
the initial molecular oxygen, and, most importantly, with
hydroxyl radicals, which are the most reactive intermediate
in the reduction of oxygen.
These studies established that the most effective of the
medicinal plant extracts studied here were St. John’s wort,
ginseng, rosewort, and Eleutherococcus and that they may
find application in antioxidant treatment as agents able to re-
duce the toxic effects of free-radical products of the reduc-
tion of oxygen in the body.
Extrapolating these data to biological systems suggests
that these agents have antiradical activity, decreasing levels
of hydroxyl radicals in the study system; they also have anti-
oxidant activity, blocking lipid peroxidation processes. At
the same time, they facilitate the process of the reduction of
molecular oxygen.
This study was performed with financial support from
the Ukrainian Science and Technology Center, Project No.
2238.
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