Scientific Research Journal of South-West University, June 2009; 2(1): 41-45 UNIVERSITY PUBLISHING

HOUSE
“NEOFIT RILSKI”
45
REVIEW ARTICLE

Quantum-chemical Approach to the Modeling of Antioxidant Activity

(Theoretical descriptors of antioxidants)

ZHIVKO VELKOV

Department of Chemistry, Faculty of Mathematics and Natural Sciences, South-West University "Neofit Rilski"-
Blagoevgrad, Bulgaria
Address: 66 Ivan Mihajlov Str., Blagoevgrad 2700, Bulgaria, E-mail: jivko_av@abv.bg

Abstract
Quantum-chemically calculated descriptors of scavenging/antioxidant activity are a powerful tool in the modeling of compounds
of this type. The usage of a suitable descriptor could demonstrate the correct reaction mechanism and the structural features
responsible for the activity. In this article a generalization is offered of experience in the field. Moreover, two cases are described
in which the above descriptors are irrelevant.

Keywords: antioxidants, radical scavenging, molecular descriptors.

Introduction ganism levels and plays a major role in the patho-

Although aerobic organisms require oxygen to sur-
vive, normal metabolism oxidation produces reac-
tive species - free radicals, some of which are highly
toxic and deleterious for cells and tissues. The free
radicals with high chemical activity attack cellular
components producing harmful molecular debris and
sometimes causing cellular death (Halliwell &
Gutteridge, 1999). To protect the cells from the dam-
age caused by the free radicals, organisms have built
several antioxidant defense mechanisms for a rapid
and efficient removal of reactive oxygen species from
the intracellular environment. In normal circum-
stances, there is a balance between antioxidants and
oxidants. When the equilibrium between oxidants and
antioxidant defense systems is impaired in favor of
the oxidants, the condition is known as oxidative
stress (Halliwell & Gutteridge, 1999). There is abun-
dant evidence that the oxidative stress triggers vari-
ous undesired processes at cellular, tissue and or-
genesis of many human diseases like ischemia/
reperfusion syndrome, atherosclerosis, chronic re-
nal failure, etc. (Halliwell & Gutteridge, 1999).
Therefore, exogenous antioxidants are commonly
administered as prevention against undesired chemi-
cal damage caused by the oxidative stress (Augustin
et al., 1997).
In this paper a generalization is presented of ex-
perience in using quantum-chemically obtained char-
acteristics (descriptors) of scavenging/antioxidant
activity. In particular, a comment is offered on how
indicative they are for the total antioxidant activity;
how they can be related to different reaction mecha-
nisms (presented in Fig. 1) and what their advan-
tages/advantages and limitations are.
The processes summarized in Figure 1 do not ac-
count for any subsequent transformations of Ao•.
The reaction of radical scavenging by an antioxi-
dant can follow the following general pathways (Fig.
1), according to Evans et al., (1992), Nagaoka et al.,

Correspondence: Zhivko Velkov, Department of Chemistry, Faculty of Mathematics and Natural Sciences, South-West University
"Neofit Rilski"- Blagoevgrad, Bulgaria. E-mail: jivko_av@abv.bg

ISSN 1313-4558
46 Zh. Velkov
- (i) Ao-H + + R
e R
(ii) R
Ao-H -H+ (iii)
Ao + R
(iv) e-
R
Ao-H
Figure 1. Mechanisms of radical scavenging by an antioxidant.
(1992), Mukai et al., (1992) and van Acker et al.,
(1996): (i) electron transfer from the antioxidant to
the active radical produces a cation radical and an
anion. The electron transfer is succeeded by proton
transfer from the cation radical to the anion; (ii) di-
rect hydrogen atom transfer between the antioxi-
dant and the active radical; (iii) deprotonation of the
antioxidant followed by electron transfer from the
resulting anion to the active radical. The next step is
protonation of the anion derived from the active radi-
cal; and (iv) proton-coupled electron-transfer for
phenolic antioxidants as a fourth pathway distin-
guished by some authors (Mayer et al., 2002; DiLabio
& Johnson, 2007; Tishchenko et al., 2008). In the
last mechanism of the phenol oxidation, the radical
(R•) has one or more lone pairs of electrons on the
same atom that bears the unpaired electron; a hy-
drogen-bonded complex is formed between the OH
and a lone pair of R•. The proton is then transferred
to the radical’s lone pair with the accompanying pro-
cess of electron moving from the lone pair of the
phenol’s oxygen atom to the radical’s singly occu-
pied molecular orbital.
Apparently the insight into the correct transfor-
mation mechanism of a given antioxidant is rather
essential when its scavenging capabilities are evalu-
ated. Various approaches can be found in literature:
from general chemical considerations and experi-
mental results to quantitative descriptors obtained
by quantum chemistry calculations: bond-dissocia-
tion enthalpy (BDE), highest occupied molecular
orbital (HOMO) energies, spin density, etc. To the
end of assessing the latter parameters, the hybrid
DFT potential B3LYP has been found to be much
more useful than the HF approach (Velkov Zh. et
al., 2007a; Velkov Zh. et al. 2007b).
Reaction Mechanisms and Descriptors
e- +H+ Ao + R-H
R
The probability of mechanism (i) depends on the re-
duction capability of the antioxidant which is diffi-
cult to deduce from general considerations (Velkov
Zh. et al., 2007c]. The electron ionization energy is a
quantitative descriptor of the ability of a compound
to participate in the scavenging of oxidants based
on the same mechanism. Van Acker et al. (1993)
have found a correlation between the ionization po-
tential and the logarithm of the free-radical scav-
enging activity (log ks) for tocopherolic antioxidants.
With reasonable accuracy, the ionization energy can
be estimated by HOMO energy according to
Koopman’s theorem (Ernzerhof, 2009). If the reac-
tion goes through mechanism (i), the HOMO en-
ergy would be a demonstrative descriptor of scav-
enging/antioxidant activity (see Fig. 1).
Mechanisms (ii) and (iii) are only feasible when a
dissociable hydrogen atom or a proton, respectively,
is present. Moreover, the stability of the resulting
structure (Ao• or Ao:) is determined by the possibil-
ity of delocalization of the electrons remaining after
the dissociation. The mechanism (iii) is expected to
occur in antioxidants with an easy-to-deprotonate
functional group such as a phenolic hydroxyl group.
The HOMO energy is in good correlation with the
results for O-H BDE (Velkov Zh. et al., 2007b), a
descriptor related to the mechanism (ii). The BDE
is one of the most frequently used descriptors of
antioxidant activity, albeit a slightly expensive one in
terms of computational resources. The big advan-
tage of the BDE is that it can be obtained both ex-
perimentally and numerically, which allows direct
comparison between theory and experiment.
Korzekwa et al. (1990) were the first to use BDE
for predicting radical scavenging activity. They have
found a linear relationship between experimentally
obtained stability of radicals and activation energies
of hydrogen abstraction in a series of reactions with
similar substrates. The smaller the values of BDE,
the higher the antioxidant activity of the investigated

H H
H H
N H
0.719
-0.032
O -0.192 -0.052
O O
0.034 0.069
0.394
-0.035 -0.033
N 0.089
0.030
H
Figure 2. Spin density distribution in the most probable melatonin radicals.
compounds is. This relation is valid not only for O-
H, but also for C-H and N-H BDE (Velkov Zh. et
al., 2009).
Another informative quantity which is more
closely associated with the reaction degree rather
than the specific radical scavenging mechanism is
the spin density. The delocalization of the single
electron in the radical obtained in the scavenging
reaction is a straightforward indicator of the reac-
tion degree regardless of the reaction mechanism.
The degree of delocalization of the unpaired elec-
tron in the radicals is evident from the spin density
distribution. Therefore spin density serves as a quan-
tum-chemical estimation of the electron/spin delo-
calization and shows the stability of the first product
of the scavenging reaction. The more stable the radi-
cal, the larger the shift of the reaction equilibrium to
the right (see Fig. 1), i.e. the higher the antioxidant
activity of the scavenging compound. For these rea-
sons, spin density is a descriptor of the radical-trap-
ping activity which is widely used in literature (van
Acker et al., 1996). The stability of a phenoxyl radi-
cal may be described also by the C-O bond length.
The more stable the radical is, the higher bond order
must this bond have. In support of this conclusion
we have found a steady correlation between this
topological descriptors and the radical scavenging
activity against DPPH (N,N-diphenyl-N-
picrylhydrazyl, a free radical for fast scavenging test)
(Velkov et al., 2008).
Moreover, if the spin density of a radical formed
from antioxidants is low, the possibility for participa-
tion in undesired oxidation reactions in the cell is
also low. For example, a recent investigation (Velkov
et al., 2007c) showed that the spin density at the
oxygen atom of dissociating hydroxide group has a
linear correlation with the activity of a series of phe-
nolic antioxidants measured against the DPPH. Since
DPPH has been widely used as a standard test-
radical, such quantitative dependences can serve as
a basis for some general conclusions. Thus the fol-
47
N
-0.040
0.022
0.214
O 0.558 0.032
0.015 -0.089
-0.171
0.038 -0.031
N
0.075 0.361
lowing structural requirements for scavenging ac-
tivity can be derived from the obtained results: (1)
The presence of O–H group in the aromatic sys-
tem. This requirement follows from the strong de-
pendence of the scavenging activity on spin density
delocalization. Nevertheless, undisputable evidence
that the extension of the conjugated system leads to
higher scavenging activity has not been found for
the considered group of compounds. It can be as-
sumed that the presence of a benzene ring is a suf-
ficient condition for the activity; (2) The occurrence
of substituents with positive mesomeric and in-
ductive electronic effects. Such substituents in-
crease the reductive properties of the compounds
and the probability to follow pathway (i) (Fig. 1).
Alkyl- and methoxy-groups exhibit positive induc-
tive and mesomeric effects and can play such ΰ role.
From this point of view, the effect of the carboxyvinyl
substituent in the derivatives of cinnamic acid,
coumaric acids, etc., is probably negative but it should
be outweighed by the positive contribution of the p-
system extension to the scavenging activity. The role
of the second ring in the molecules containing
chromane rings like α-tocopherol and trolox can be
interpreted in a similar way; (3) The presence of
hydrogen bonds involving dissociable hydroxyl
group( DHG) and adjacent functional groups.
Particularly effective is the hydrogen bond of the
DHG oxygen and the hydrogen from a neighboring
hydroxyl group.
Two Exceptions
The descriptors considered so far are not universal.
None of them describes all aspects of the total
chemical reaction between antioxidants and active
free radicals. Furthermore, none of them is con-
nected with the AoO• transformations in the organ-
ism.
Our calculations have shown that some radicals
48 ZH. Velkov
Table 1. Antioxidant/scavenging activity and calculated descriptors for the compounds considered in Spasova et al., (2008).
Descriptors
Antioxidants % RSA
Sinapic acid 26.5 (31.9)
Compound (5)* 12.4 (15.9)
Ferulic acid 21.0 (25.1)
Compound (4)* 8.4 (11.9)
*
the notation is from the cited article.
formed by the endogenic antioxidant melatonin have
a rather low degree of delocalization (Velkov et al.,
2009). In Figure 2 it could be seen he spin density
distribution at the most relevant atoms in radicals
which can be formed by melatonin. It is evident that
the spin density at the carbon atom connected to the
third position in the indole ring (Fig. 2, first struc-
ture) is rather high, 0.719. The spin density of the
spin-bearing atom in the second radical is also high,
0.558. However, it is interesting to note that the un-
coupled electron is localized mainly at the third car-
bon atom of the indole ring rather than at the nitro-
gen atom.
The computed values of spin densities are much
larger than the values calculated for radicals obtained
by phenolic antioxidants (Velkov et al., 2007c). Ac-
cording to this descriptor, the melatonin radicals are
rather active and this is not favorable for melatonin’s
antioxidant activity.
It was mentioned above that the considered
mechanisms do not take into account further trans-
formations of the products of the scavenging reac-
tion. However, if any of the products is directly in-
volved in further biochemical transformations, the
stability of the final radical (Ao•) is not so crucial
for the reaction degree. In order to show such an
example, the spin density is calculated of the thiyl
radical (GS•) of glutathione (GSH). The maximum
value obtained (at the sulfur atom) is 0.843, i.e. much
higher than in the melatonin radicals. In vitro, GSH
inactivates prooxidants like OH•, HOCl, RO•, RO2•,
1
O2, etc., and forms GS•. In the next step, GS• is
quickly transformed, forming glutathione disulphide
and superoxide (O2•-). In vivo glutathione disulphide
is reduced to GSH by the enzyme glutathione re-
ductase (Halliwell & Gutteridge, 1999).
This finding leads to the conclusion that the cou-
pling of radical scavenging with other reactions
makes the spin density in this case a descriptor ir-
relevant to the reaction degree.
Having accepted the possibility that the scaveng-
EHOMO [a.u] BDE [a.u.]
-0.20969 0.618341
-0.20355 0.615813
-0.20934 0.624902
-0.20471 0.624001
ing reaction of melatonin is coupled with other reac-
tions, we should also admit the possibility that the
radical scavenging might occur in ways that are con-
ceptually different from the above considered
mechanisms.
Experimental data for % RSA (percent radical
scavenging activity) of a group of natural and syn-
thetic compounds were recently obtained (Spasova
et al., 2008). It is interesting that compounds with
very close energies of HOMO and enthalpies of O-
H bond dissociations expose fairly different activi-
ties (see Table 1).
The reaction of antioxidants with DPPH passes
through a transition state that would be much more
unfavorable for the compounds containing more at-
oms. The reason might be in the much more disad-
vantageous change of the entropy when heavier mol-
ecules form a transition state, which slows down
the reaction. The descriptors considered so far do
not account for this fact.
Acknowledgements
This study was supported by Grand “DVU 01/
0197” of Ministry of Education and Science .
References
Augustin W., Wiswedel I., Noack H., Reinheckel Th., Reichelt O.
(1997) Role of endogenous and exogenous antioxidants in the
defense against functional damage and lipid peroxidation in
rat liver mitochondria. Molecular and Cellular Biochemistry
174, 1-2, 199-205.
DiLabio, G. A. & Johnson, E. R (2007) Lone pair”ð and ð”ð interactions
play an important role in proton-coupled electron transfer reactions.
Journal of the American Chemical Society 129 (19), 6199–6203.
Ernzerhof M. (2009) Validity of the Extended Koopmans’ Theorem.
Journal of Chemical Theory and Computation 5 (4), 793–797.
Evans C.H., Scaiano J.C., Ingold K.U. (1992) Absolute kinetics of
hydrogen abstraction from alpha-tocopherol by several reactive
species including an alkyl radical. Journal of the American Chemical
Society 114, 4589–4593.
Halliwell B. & Gutteridge J.M.C. (1999) Free Radicals in Biology and
Medicine (3rd edition). Ch. ???, p. ???.Oxford University Press,
Oxford.
Korzekwa K.R., Jones J.P., Gillette J.R. (1990) Theoretical studies on
cytochrome P-450 mediated hydroxylation: a predictive model for

hydrogen atom abstractions. Journal of the American Chemical
Society 112, 7042-7046.
Mayer J. M., Hrovat D. A., Thomas J. L., Borden W. T. (2002) Proton-
Coupled Electron Transfer versus Hydrogen Atom Transfer in Benzyl/
Toluene, Methoxyl/Methanol, and Phenoxyl/Phenol Self-Exchange
Reactions. Journal of the American Chemical Society 124, 11142–
11147.
Mukai K., Uemoto Y., Fukuhara M., Nagoaka S., Ishizu K. (1992)
ENDOR Study of the Cation Radicals of Vitamin E Derivatives.
Relation between Antioxidant Activity and Molecular Structure.
Bulletin of the Chemical Society of Japan 65 (8), 2016-2020.
Nagaoka S.I., Kuranaka A., Tsuboi H., Nagashima U., Mukai K. (1992)
Mechanism of antioxidant reaction of vitamin E: charge transfer and
tunneling effect in proton-transfer reaction. Journal of Physical
Chemistry 96 (6), 2754–2761.
Spasova M. G., Paizs Cs., Stoykova B. M., Milkova Ts. S., Irimie F. D.
(2008) Sulfur-Containing Amino Acid Amides of Phenolic Acids
Peptides 14 (8), 34-35, Proceedings of The Thirtieth European
Peptide Symposium, Hilkka Lankinen.
Tishchenko O., Truhlar D. G., Ceulemans A., Nguyen M. T. (2008) A
Unified Perspective on the Hydrogen Atom Transfer and Proton-
Coupled Electron Transfer Mechanisms in Terms of Topographic
Features of the Ground and Excited Potential Energy Surfaces As
Exemplified by the Reaction between Phenol and Radicals.
Journal of the American Chemical Society 130, 7000–7010.
49
van Acker, S.A.B.E., Koymans L.M.H., Bast A. (1993) Molecular
Pharmacology of Vitamin E: Structural Aspects of Antioxidant
Activity, Free Radical Biology & Medicine 15, 311–328.
van Acker S.A.B.E., de Groot M.J., den Berg D.-J. (1996) A Quantum
Chemical Explanation of the Antioxidant Activity of Flavonoids.
Chemical Research in Toxicology 9, 1305–1312.
Velkov Zh., Velkov Y., Balabanova E., Tadjer A. (2007a) First Principle
Study of the Structure of Conjugated Amides and Thioamides.
International Journal of Quantum Chemistry 107, 1765–1771.
Velkov Zh., Balabanova E., Tadjer A. (2007b) Radical scavenging activity
prediction of o-coumaric acid thioamide. Journal of Molecular
Structure: THEOCHEM 821, 133–138.
Velkov Zh. A., Kolev M. K., Tadjer A. V. (2007c) Modeling and Statistical
Analysis of DPPH Scavenging Activity of Phenolics. Collection of
Czechoslovak Chemical Communications 72 (11), 1461–1471.
Velkov Zh., Kolev M., Tadjer A. (2008) Molecular Modelling and
Statistical Analysis of Scavenging/antioxidant Activity of Phenolic
compounds. Proceedings of the Fourteenth National Conference on
Apllication of Mathematics in Biology and Medicin. pp. 134-139,
Leszno n. Warsaw, Poland.
Velkov Zh., Velkov Y., Galunska B., Paskalev D., Tadjer A. (2009)
Melatonin: Quantum-chemical and biochemical investigation of
antioxidant activity. European Journal of Medicinal Chemistry 44
(7), 2834-2839.