Preparation, spectroscopic, thermal, antihepatotoxicity, hematological parameters and liver antioxidant capacity characterizations of Cd(II), Hg(II), and Pb(II) mononuclear complexes of paracetamol anti-inflammatory drug

Article abstract of DOI:10.1016/j.saa.2014.04.108

Keeping in view that some metal complexes are found to be more potent than their parent drugs, therefore, our present paper aimed to synthesized Cd(II), Hg(II) and Pb(II) complexes of paracetamol (Para) anti-inflammatory drug. Paracetamol complexes with g

Full text of DOI:10.1016/j.saa.2014.04.108

Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 131 (2014) 534–544
Contents lists available at ScienceDirect
Spectrochimica Acta Part A: Molecular and
Biomolecular Spectroscopy
journal homepage: www.elsevier.com/locate/saa
Preparation, spectroscopic, thermal, antihepatotoxicity, hematological
parameters and liver antioxidant capacity characterizations of Cd(II),
Hg(II), and Pb(II) mononuclear complexes of paracetamol
anti-inflammatory drug
Samy M. El-Megharbel a,b, Reham Z. Hamza , Moamen S. Refat
c
a,d,
⇑
a
Department of Chemistry, Faculty of Science, Taif University, Al-Haweiah, P.O. Box 888, Zip Code 21974, Taif, Saudi Arabia
Department of Chemistry, Faculty of Science, Zagazig University, Zagazig, Egypt
Department of Zoology, Faculty of Science, Zagazig University, Zagazig, Egypt
b
c
d
Department of Chemistry, Faculty of Science, Port Said University, Port Said, Egypt
h i g h l i g h t s
g r a p h i c a l a b s t r a c t
ꢀ
The Cd(II), Hg(II), and Pb(II)
complexes of paracetamol have been
synthesized.
ꢀ
ꢀ
ꢀ
The general formula of complexes is
[
M(Para)
2
(H
2
O)
2
]ꢁnH
2
O.
In vivo the antihepatotoxicity effect
were measured.
2
+
The Cd + Para complex has
succeeded in improvement of the
antioxidant capacities.
a r t i c l e i n f o
a b s t r a c t
Article history:
Keeping in view that some metal complexes are found to be more potent than their parent drugs, there-
Received 28 January 2014
Received in revised form 11 April 2014
Accepted 21 April 2014
fore, our present paper aimed to synthesized Cd(II), Hg(II) and Pb(II) complexes of paracetamol (Para)
anti-inflammatory drug. Paracetamol complexes with general formula [M(Para)
2
(H
2
O)
2
]ꢁnH
2
O have been
synthesized and characterized on the basis of elemental analysis, conductivity, IR and thermal (TG/DTG),
Available online 30 April 2014
1
H NMR, electronic spectral studies. The conductivity data of these complexes have non-electrolytic nat-
ure. Comparative antimicrobial (bacteria and fungi) behaviors and molecular weights of paracetamol
with their complexes have been studied. In vivo the antihepatotoxicity effect and some liver function
parameters levels (serum total protein, ALT, AST, and LDH) were measured. Hematological parameters
and liver antioxidant capacities of both Para and their complexes were performed. The Cd2 + Para
complex was recorded amelioration of antioxidant capacities in liver homogenates compared to other
Para complexes treated groups.
Keywords:
Paracetamol
Heavy metal
Spectroscopic
Antihepatotoxicity
Liver function parameters
Antioxidant capacities
+
Ó 2014 Elsevier B.V. All rights reserved.
Introduction
⇑
Corresponding author at: Department of Chemistry, Faculty of Science, Taif
University, Al-Haweiah, P.O. Box 888, Zip Code 21974, Taif, Saudi Arabia. Tel.: +966
51352497.
E-mail address: msrefat@yahoo.com (M.S. Refat).
Paracetamol (Para; Fig. 1) was a widely used as analgesic and
antipyretic drug [1–6], also, it was known to be hepatotoxic in
5
http://dx.doi.org/10.1016/j.saa.2014.04.108
386-1425/Ó 2014 Elsevier B.V. All rights reserved.
1

S.M. El-Megharbel et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 131 (2014) 534–544
535
Physical measurements
OH
O
Carbon, hydrogen and nitrogen contents were determined using
a Perkin–Elmer CHN 2400. The metal content was determined by
atomic absorption spectrometer model PYE-UNICAM SP 1900 and
the corresponding lamps were used for this purpose. Infrared spec-
N
H
tra were recorded on Bruker FTIR Spectrophotometer (4000–
ꢂ1
4
00 cm ) in KBr pellets. The UV–Vis spectra were studied in the
Fig. 1. Paracetamol (Para) drug structure.
ꢂ3
DMSO solvent with a concentration of 1.0 ꢃ 10 M for the Para
and their complexes using Jenway 6405 Spectrophotometer with
1
cm quartz cell, in the range 800–200 nm. Molar conductance’s
man and various experimental animals upon overdose [7–9]. Tak-
ing the presumed molecular mechanisms of analgesic activity as
well as of the hepatotoxicity of paracetamol into consideration,
there have been several efforts to improve its analgesic activity
while preventing its toxicity by modifying its structure [10–13].
In an attempt to improve the analgesic activity of paracetamol by
mono-substitution ortho to the hydroxyl group, Harvison et al.
of the freshly prepared solutions of the Para complexes with
1
ꢂ3
.0 ꢃ 10 M in DMSO were measured using Jenway 4010 conduc-
1
tivity meter. H NMR spectra were recorded on a Varian Gemini
00 MHz spectrometer using DMSO-d as solvent. Thermogravi-
2
6
metric analyses (TG/DTG) were carried out in a dynamic nitrogen
atmosphere (30 mL/min) with a heating rate of 10 °C/min using a
Shimadzu TGA-50H thermal analyzer.
[
11] showed that 3-methyl paracetamol was equipotent to para-
cetamol with respect to analgesic activity in mice. Unfortunately
however, hepatotoxicity was also equal, the hepatotoxicity of
paracetamol was decreased by 2-methyl substitution (meta to
the hydroxyl group), however, the analgesic activity was also
decreased. N-methyl paracetamol was found to be completely
devoid of hepatotoxicity but also of analgesic activity [14]. In
addition to mono-substitution, it has been shown that dialkyl-
substitution at the 3- and 5-positions of the aromatic nucleus of
paracetamol did not reduce the analgesic activity [15]. A toxicolog-
ical study showed that the in vivo hepatotoxicity of the 3,5-dialky-
lated analogs was reduced almost completely [16]. Recently, it was
reported that aromatic ring-substitution by one or two fluorines
decreased the analgesic activity of paracetamolacetamol in mice
Antimicrobial activities
According to Gupta et al. [20], the antimicrobial tests were
done. The investigated isolates of bacteria were seeded in tubes
with nutrient broth (NB). The seeded NB (1 cm ) was homogenized
in the tubes with 9 cm of melted (45 °C) nutrient agar (NA). The
homogeneous suspensions were poured into Petri dishes. The holes
3
3
(
2
diameter, 4 mm) were done in the cool medium. After cooling,
ꢃ 10 dm of the investigated compounds were applied using
ꢂ3
3
a micropipette. After incubation for 24 h in a thermostat at 25–
2
7 °C, the inhibition (sterile) zone diameters (including disc) were
measured and expressed in mm. An inhibition zone diameter of
over 7 mm indicates that the tested compound is active against
the bacteria under investigation. The antibacterial activities of
the investigated compounds were tested against Escherichia coli
[
17]. It was also shown that these modifications decreased the
in vivo toxicity [18]. The formation of complexes of paracetamol
and Zn(II) was studied in aqueous media at pH 7.2 by polarography
and spectroscopy [19]. The stoichiometry of the Zn(II)-paracetamol
complex was 1:1. Analgesic studies on the drug and its metal com-
plex have been performed in albino mice. Revealing the complex to
be more potent in analgesic activity compared to the paracetamol
alone drug.
(
Gram, ꢂve), Bacillus subtilis (Gram, +ve) and antifungal (Asperagil-
lus oryzae, Asperagillus niger, and Asperagillus flavus).
Experimental animals
Antihepatotoxicity effect in male albino rats
The present work was built on the study of the interactions
between paracetamol drug and some of heavy metal ions like
The present study was carried out at Zoology Department, Fac-
ulty of Science-Zagazig University, Egypt using fifty clinically healthy
mature adult male albino rats. The animals were obtained from the
animal House of Faculty of Veterinary Medicine, Zagazig University,
Egypt. Their weights ranged from (200–250 g). The animals were
housed in standard conditions, where the animals were housed in
metal cages and bedded with wood shavings and kept under stan-
dard laboratory conditions of aeration and room temperature at
about 25 °C. The animals were allowed to free access of standard diet
and water ad-libtum, We have followed the European community
Directive (86/609/EEC) and national rules on animal care. The ani-
mals were accommodated to the laboratory conditions for 2 weeks
before being experimented, adaptation and 10 rats were placed into
each cage. Six groups were established in the study as follows.
The present study was undertaken to assess the effects of single
or multiple-dose administration of Para and it is complexes in nor-
mal treated rats. All experiments were performed during the same
time of day, between 10 am and 1 pm to avoid variations due to
diurnal rhythms.
Cd(II), Hg(II), and Pb(II). The elemental analysis, conductivity, IR
and, thermal (TG/DTG), 1H NMR, electronic spectral studies of
these complexes were discussed and deduced the suggested struc-
ture which associated via deprotonation of AOH hydroxyl group.
The antihepatotoxicity, hematological parameters and antioxidant
effects of the Para complexes were investigated on the treated rats
upon the calculations of liver function parameters levels (serum
total protein, ALT, AST, and LDH), SOD, GST and TAC.
Experimental
Materials and preparations
Analytical grade of chemicals used were purchased from
Aldrich and Merck chemical companies. Paracetamol drug was
received from Egyptian International Pharmaceutical Industrial
Company (EIPICo.). The Para complexes were prepared by mixing
twice amount of Para (2 mmol) and 1 mmol of metal(II) nitrates
3
(
Cd(II), Hg(II) and Pb(II)) in MeOH/H
2
O (50/50, w/w; 40 cm )
Experimental design
solvent, then pH of the mixtures was adjusted to 7–8 using 5%
alcoholic ammonia solution. The reaction mixtures were stirred
at 60 °C for 2 h and left to stand overnight. The precipitated
– Test compounds; Para, Cd2+/Para, Hg2+/Para, and Pb2+/Para.
Animal groups
complexes were filtered off, washed with MeOH/H
in vacuum at room temperature over anhydrous CaCl
2
O and dried
Treatment schedule (each group comprises 10 rats): In order to
optimize Para drug absorption, all animals were starved overnight
2
.

5
36
S.M. El-Megharbel et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 131 (2014) 534–544
prior to Para exposure and administration of its complexes. Para-
cetamol and its complexes were administered to rats following
Lipid profile
Triglycerides, cholesterol and high density lipoprotein-choles-
1
2 h of fasting and allowed free access to food and water 1 h after
terol (HDL-c) were determined using the commercial kits. Low
density lipoprotein-cholesterol (LDL-c) levels were calculated by
using the following formula of Muruganandan et al. [26]:
LDL-c = total cholesterol ꢂ (HDL-c + triglycerides)/5. Volatile low
density lipoprotein-cholesterol (VLDL-c) levels were calculated
by using the following formula of Prakasam et al. [27]: VLDL-
c = triglyceride/5. The risk ratio was calculation by dividing the
total cholesterol by HDL-c.
vehicle/Para and its complexes administration.
i. The 1st control group
Animals received 1 ml of distilled water orally daily for 4 weeks,
and served as the control group.
ii. The 2nd Para treated group
Animals were daily received orally Para, the Para was dissolved
in DMSO (650 mg/kg b.w) [21] for 30 successive days using metal-
lic stomach tube.
Tissue homogenate preparation
Liver was immediately removed; weighed and washed using
chilled saline solution. Tissues were minced and homogenized
iii. The 3rd Cd2+/Para treated group
(
10% w/v), separately, in ice-cold sodium, potassium phosphate
Animals were received orally Cd2+/Para (650 mg/kg b.w) for 30
successive days using metallic stomach tube.
iv. The 4th Hg2+/Para treated group
buffer (0.01 M, pH 7.4) containing 1.15% KCl in a Potter–Elvehjem
type homogenizer. The homogenate was centrifuged at 10,000g
for 20 min at 4 °C, and the resultant supernatant was used for
the determination of antioxidant enzyme assays, (liver only).
Animals were received orally Hg2+/Para (650 mg/kg b.w) for 30
successive days using metallic stomach tube.
Assessment of antioxidants capacities
v. The 5th (Pb2+/Para treated group
Animal were given orally Pb2+/Para (650 mg/kg b.w) for 30 suc-
cessive days using metallic stomach tube.
The rats included in the groups that were given Para and its
complexes were weighed daily, and accordingly the dose that
was given per body weight was calculated and administration by
metallic tube was considered to be more accurate and safety.
The principle of SOD activity assay was based on the inhibition
of nitro blue tetrazolium (NBT) reduction. Illumination of ribofla-
vin in the presence of O and electron donor like methionine gen-
2
erates superoxide anions and this has been used as the basis of
assay of SOD. The reduction of NBT by superoxide radicals to blue
colored formazan was followed at 480 nm [28]. Glutathione-
S-transferase (GST) was determined according to the method of
Habig et al. [29]. While Glutathione level (GSH) as non-enzymatic
antioxidant was estimated based on the method of Beutler et al.
Collection and processing of samples
Twenty-four hours after the last administration of the indicated
substances, blood samples were collected from the retro-orbital
vein, which is a simple, convenient and successful procedure that
allows bleeding of the same animal more than one time with min-
imal stress [22]. Individual blood was drawn by orbital puncture
[
30] who reported that the 5,50-dithiobis-(2-nitrobenzoic acid) is
reduced by SH group to form 1 mol of 2-nitro-5-mercaptobenzoic
acid per mole of SH.
Total antioxidant capacity of hepatocytes was determined using
ferric reducing antioxidant power assay according to the method of
Prieto et al. [31]. FRAP reagent (300 mM acetate buffer, pH 3.6,
(
from eye plexus) using microhematocrit capillary tubes into dry
tubes from 10 rats in each group, Serum was harvested from blood
without EDTA. And their Kidney tissue was excised. Blood samples
were centrifuged at 336 rpm for 10 min for the separation of sera.
The serum samples obtained were transferred into eppendorf
tubes and were preserved in a deep freezer at ꢄ80 °C. Part of the
Kidney tissue was transferred into 10% buffered formalin for histo-
pathological examination.
1
0 mM 2,4,6-tri(Z-pyridy)-S(riazine, 99%) (TPTZ) in 40 mM HCl
and 20 mM FeCl O in the ratio of 10:1:1 was prepared.
ꢁ6H
.5 mL FRAP reagent was added to 50 L of homogenized liver
3
2
1
tissues and incubated at 37 °C for exactly 5 min. The change in
absorbance was measured at 593 nm due to the formation of a blue
colored Fe(II)-tripyridyltriazine complex from the colorless oxi-
dized Fe(III) form by the action of electron donating antioxidants.
The absorbance of the sample was read against reagent blank
(1.5 mL FRAP reagent and 50 L distilled water) at 593 nm. The data
was expressed as percentage of the control value.
Determination of key liver functions biochemical markers
Serum total protein, ALT, AST, and LDH levels were measured.
Some Liver function parameters:
Hematological parameters
i. Determination of serum aminotransferase enzymes
activities
Hematological parameters A complete blood count includes five
major measurements were determined using cell counter (Sysmex,
model KX21N). White blood cells (WBC) were measured in thou-
sands per cubic milliliter of blood. Red blood cells (RBCs) were
measured in millions per cubic millimeter of blood. Hemoglobin
Activities of AST and ALT in the serum were determined calori-
metrically by using bio Merieux kit (France), using method
adopted by [23].
ii. Determination of serum total protein concentration
(
Hb) was measured in grams per deciliter (g/dL) of blood. The
Colorimetric determination of the total protein based on the
principle of Biuret reaction (copper salts in an alkaline medium).
Protein in plasma or serum forms a blue colored complex when
treated with cupric icons in alkaline solution. The intensity of the
blue color is proportional to the protein concentration [24].
iii. Determination of LDH level
hematocrit value (PCV) is the percentage of red blood cells in rela-
tion to total blood volume was also determined. Mean cell volume
(MCV), mean cell hemoglobin (MCH) and mean cell hemoglobin
concentration (MCHC) were also calculated. Platelets were mea-
sured in thousands per cubic millimeter of blood.
The reaction velocity is determined by a decrease in absorbance
at 340 nm resulting from the oxidation of NADH. One unit causes
the oxidation of one micromole of NADH per minute at 25 °C and
pH 7.3, under the specified conditions [25] by using Diamond kit.
Preparation of tissues for histopathological examination
Tissue samples were taken from the liver of animals and fixed in
10% formalin neutral buffer solution. The trimmed tissues were

S.M. El-Megharbel et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 131 (2014) 534–544
537
first washed with tap water followed by dehydration through a
graded alcohol series and then passed though xylol and paraffin
series before finally blocked in paraffin. The paraffin blocks were
cut into 5–6 mm sections using a The trimmed tissues were first
washed with tap water followed by dehydration through a graded
alcohol series and then passed through xylol and paraffin series
before finally blocked in paraffin. The paraffin blocks were cut into
5
–6 lm sections using a microtome stained using hematoxylin and
eosin and examined under a light microscope microtome stained
using hematoxylin and eosin and examined under a light micro-
scope [32].
Statistical analysis
Data were collected, arranged and reported as mean ± standard
error of mean (S.E.M) of nine groups (each group was considered as
one experimental unit), summarized and then analyzed using the
computer program SPSS/version 15.0) The statistical method was
one way analyzes of variance ANOVA test (F-test), and if significant
differences between means were found, Duncan’s multiple range
test (whose significant level was defined as P < 0.05) was used
according to [33] to estimate the effect of different treated groups.
Fig. 2. Infrared spectrum of Cd2+/Para complex.
Table 2
ꢂ1
IR frequencies (cm ) of Para and its metal complexes.
Assignments
Compounds
Para
I
II
III
Results and discussion
3
3694
3184
785
3626
3323
3626
3323
1653
1013
1324
611
t
t
(OAH)
(NAH)
3434
3434
3535
3535
Molar conductivities
3
162
t
t
(C@O)
(CAO)
1650
1256
1324
–
1653
1022
1324
666
1653
1047
1324
605
The molar conductance values for the Cd(II), Hg(II) and Pb(II)
ꢂ3
complexes of Para in DMSO solvent (1.00 ꢃ 10 M) were found
t(CAN)
t
(MAO)
ꢂ1
2
ꢂ1
to be in the range 40–52
X
cm mol at 25 °C, suggesting them
511
567
412
to be non-electrolytes (Table 1). These data matched with the cal-
culated elemental analysis.
ꢂ1
shifted to 1047, 1022 and 1013 cm with a marked change of
intensity. If we assume that the bands arise from CAO vibration,
we may conclude that the CAO group is probably involved in coor-
dination, therefore, the metal M(II) chelate was assigned through
hydroxyl group.
Infrared spectra
The infrared spectra of Para free ligand and its complexes were
recorded in Fig. 2 and Table 2. The spectra of Para complexes are
similar; this is due to the same place of coordination toward Cd(II),
Hg(II) and Pb(II) ions. The most interesting feature in the infrared
spectra of these complexes is the shift and decreasing in the inten-
sities of bands in the 3360–3500 cm region. However, the ligand
exhibited four bands at 3785, 3694, 3184 and 3162 cm . This fact
is hard to reconcile with either structure proposed by both Medv-
edovskii [34,35] and Fazakerley et al. [36]. Ionization of the OH
groups with subsequent ligation through oxygen atoms seems a
1
H NMR spectrum of Cd(II) complex
ꢂ1
ꢂ1
1
The H NMR data of Para and its Cd(II) complex, as an example
are listed in Table 3 and shown in Fig. 3. The H NMR spectrum of
Para show the signal at d = 9.27 ppm, which is assigned to the pro-
tons of the amide group which is the same in the Cd(II) complex
and confirm that the amide group does not contribute in the com-
plexation between Para and Cd(II) metal ions. The proton NMR
1
ꢂ1
plausible explanation. Weak bands at 3184 and 3162 cm can
be assigned to NH which is not shifted; we can conclude that the
NH group is not involved in coordination process. The band at
spectrum for [Cd (Para)
3.38 ppm. This singlet peak is not observed in the free Para ligand
spectrum and can be assigned due to protons of H O molecules,
2
(H
2
O)
2
]ꢁ6H
2
O show a singlet at peak at
ꢂ1
1
653 cm , assigned to C@O vibration, is not shifted in the com-
ꢂ1
plex but has decreased intensity. Band at 1324 cm in the free
ligand is not changed in the complexes. Nakamoto [37] assigned
the band at 1324 cm in the metal oxamido complexes to the
2
supporting the complexes formula. The disappearance of the signal
ꢂ1
1
of the proton of the hydroxyl group in the H NMR spectrum of the
ꢂ1
CAN vibration. If we accept that the 1324 cm band is due to
complex confirms that the hydroxyl group contribute in the com-
plexation between Para and Cd(II) ion, therefore, the hydroxyl
group is disappear in the complex of Para moiety. So, the complex-
ation between two molecules of Para with Cd(II) ion has been
CAN vibration, we conclude that the amido group is not involved
in coordination. The free ligand has a band in the CAO region,
ꢂ1
namely at 1256 cm . On complexation the maxima are significant
Table 1
Elemental analysis and physical data for Para complexes.
Complexes
Mwt.
Color
Content (found) calculated
A (S cm2 molꢂ1
)
%
C
% H
% N
% M
[
[
[
Cd(Para)
Hg(Para)
Pb(Para)
2
(H
(H
(H
2
O)
O)
O)
2
]ꢁ6H
]ꢁ4H
]ꢁ4H
2
O (I)
558
611
617
White
White
White
34.40 (34.33)
31.42 (31.53)
31.11 (31.04)
6.09 (6.31)
4.90 (4.87)
4.86 (4.75)
5.01 (4.95)
4.58 (4.70)
4.53 (4.62)
20.16 (20.21)
32.89 (32.76)
33.54 (33.63)
40
47
52
2
2
2
2
O (II)
2
2
2
2
O (III)

5
38
S.M. El-Megharbel et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 131 (2014) 534–544
formed by the chelation of the metal ion with the hydroxyl group
to form the complex 2-Para-Cd(II) including the oxygen metal [38].
within the temperature range 170–290 °C and can assigned to
the loss of C NO (organic moiety) and Hg vapor and the mass
H
5 6
3
loss due to this step was (obs. = 54.17%, calc. = 53.84%). The last
step is occurring at 300–500 °C and corresponding to the evolution
of 3(C H ) (organic moiety) representing a weight loss of 7.93% and
4 2
Electronic absorption spectra
its calculated value is 8.18%. The final products resulted at 800 °C
contain 2 carbon atoms.
The formation of the Cd(II), Hg(II) and Pb(II) complexes of Para
was also confirmed by UV–Vis spectra. The electronic absorption
spectra of the M(II) complexes in DMSO in the 200–600 nm range
were discusses. It can see that free Para has two distinct absorp-
[Pb(Para)
2
(H
2
O)
2
]ꢁ4H
2
O
⁄
tion bands. The first one at 300 nm may be attributed to
p
?
p
The Pb(II) complex is decomposed in three steps. The first step
is occurring at 50–175 °C and corresponding to the evolution of
intra-ligand transition of the aromatic ring. The second band
⁄
observed at 390 nm is attributed to n ?
p
electronic transition.
5(H
culated value is 14.58%. While, the second decomposition step is
occurring at 180–250 °C and corresponding to the loss of H
and C 10N (organic moiety), representing a weight loss of
2
O) molecules, representing a weight loss of 14.38% and its cal-
In the spectra of the M(II) complexes, the two bands are hypso-
chromically affected clearly, suggesting the ligand is deprotonated
and the lone pair of electrons on nitrogen and carbonyl-oxygen
atoms of the amide group is not participated in the complexation.
These results are clearly in accordance with the results of the FTIR
2
O
9
H
25.42% and its calculated value is 25.93%. The horizontal bump in
Fig. 4C, it is meaning to the start of melting point of lead metal
which is completely melting up to 320 °C. This confirms the con-
fused between logically dissociation of paracetamol ligand and
melting point of lead metal. So the non-horizontal bump with mass
loss refers to decomposition of ligand and horizontal bump with
non-mass loss refer to melting of lead metal. The final decomposi-
tion step takes place in the range of 290–430 °C and corresponding
1
and H NMR spectra.
Thermal analysis of Para complexes
The heating rates were controlled at 10 °C/min under nitrogen
atmosphere and the weight loss was measured from ambient tem-
perature up to ffi136 °C. The data are listed in Table 4 and shown in
Fig. 4. The weight losses for each chelat calculated within the cor-
responding temperature ranges.
to the elimination of C
5.94% (calc. = 26.58%). The final product resulted at 800 °C contain
PbO.
7 8 3
H NO with an observed weight loss of
2
Kinetic and thermodynamic parameters
[
Cd(Para)
2
(H
2
O)
2
]ꢁ6H
2
O
The thermal decomposition of [Cd(Para)
2
(H
2
O)
2
]ꢁ6H
2
O complex
The kinetic and thermodynamic parameters were determined
by non-isothermal methods. The non-isothermal kinetic analysis
for the thermal decomposition of all ligands and complexes in this
work was carried out by the application of the Coats–Redfern [39]
method. The kinetic parameters were evaluated for all decomposi-
tion stages. Kinetic studies were applied for the decomposition
stages that occur within a temperature range, resulting in a TG
curve providing enough data to be collected. The kinetic parame-
occurs at three steps. The first degradation step takes place in the
range of 50–240 °C and they corresponds to the elimination of
2
(H
loss of (obs. = 45.92%, calc. = 45.69%). The second step falls in the
range of 250–440 °C which is assigned to the loss of C NO
organic moiety) with a weight loss (obs. = 14.28%, calc. = 14.51%).
The final decomposition step within the temperature range 450–
75 °C is accompanied by mass loss of 16.62% (calc. = 16.84%),
which are assigned to the loss of C 10 (organic moiety). The
CdO is the final product remains stable till 800 °C.
2 5 5 2
O) and C H NO (organic moiety) with an observed weight
4
H
3
(
5
i f a
ters (T , T , Dm, and E ) were calculated according to the Coats–Red-
fern method [39].
7
H
Methods
[
Hg(Para)
The Hg(II) Para complex is decomposed in three steps. The first
step is exhibited at 50–160 °C and corresponding to the loss of
(H O) and C 10NO (organic moiety), representing a weight loss
2
(H
2
O)
2
]ꢁ4H
2
O
From the TGA curves (TG, DTG) recorded for the successive
steps in the decomposition process of ligands and their complexes,
it was possible to determine the following characteristic thermal
parameters for each reaction step as follows.
Initial point temperature of decomposition (T
which DTG curve starts deviating from its base line. Final point
temperature of decomposition (T ): the point at which DTG curve
6
2
5
H
of (obs. = 34.12%, calc. = 34.04%). The second step takes place
i
): the point at
Table 3
1
H NMR spectral data of Para and its Cd(II) complex.
f
Compounds
d (ppm) of hydrogen
returns to its base line. Peak temperature, i.e. temperature of max-
imum rate of mass loss (TDTG): the point obtained from the inter-
section of tangents to the peak of DTG curve. Mass loss at the
H; CH
3
2
H; H O
H; ArH
H; NH
Para
Cd(II) complex
1.96
1.96
–
3.38
6.57–7.28
6.63–7.34
9.57
9.64
decomposition step (
D
m): it is the amount of mass that extends
on the TG curve. The material
from the point T up to the point T
i
f
Fig. 3. 1H NMR spectrum of Cd2+/Para complex.

S.M. El-Megharbel et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 131 (2014) 534–544
539
Table 4
Weight % (%)
Derivative Weight % (%/m)
Thermal data of Para complexes.
1
1
20
00
2.00E+00
Compounds DTG
TG weight
peak/(°C) loss (%)
Assignments
Residue
(A)
0.00E+00
Calc.
Found
-
-
-
-
-
2.00E+00
4.00E+00
6.00E+00
8.00E+00
1.00E+01
I
240
45.69 45.92
14.51 14.28
16.84 16.62
8H
2
O + C
NO
5
H
H
5
NO
2
CdO
2C
8
0
4
5
45
75
C
C
4
H
H
3
7
10
II
III
160
34.04 34.12
53.84 54.17
6H
2
O + C
5
10NO
60
2
5
90
00
C
C
5
H
H
6 3
NO + Hg vabour
8.18
7.73
4
2
4
2
0
0
0
175
14.58 14.36
25.93 25.42
26.58 25.94
5H
O + C
NO
3
2
O
PbO
2
4
40
30
H
2
9 10
H N
C
7
H
8
-1.20E+01
1000
released at each step of the decomposition is identified by attribut-
ing the mass loss ( m) at a given step to the component of similar
0
200
400
600
800
D
weight calculated from the molecular formula of the investigated
compounds, comparing that with literatures of relevant com-
pounds considering their temperature. This may assist identifying
the mechanism of reaction in the decomposition steps taking place
in the compounds under study.
Weight % (%)
Derivative Weight % (%/m)
120
2
0
(B)
100
-2
8
6
4
0
0
0
Coats–Redfern method
-4
-6
a
Activation energy (E ) of the decomposition step: the integral
method used is the Coats–Redfern equation [40] for reaction order
n – 1, which when linearized for a correctly chosen n yields the
activation energy from the slope;
-8
-10
12
h
i
h
i
1
ꢂn
20
0
1
ꢂð1ꢂ
a
Þ
ZR
2RT
E
a
2:303RT
-
log
log
2
¼ log qE 1 ꢂ Ea
ꢂ
for n–1
T
ð1ꢂnÞ
a
-14
600
h
i
h
i
ꢂlogð1ꢂ
aÞ
ZR
a
2RT
E
a
0
100
200
300
400
500
¼ log qE 1 ꢂ Ea
ꢂ
for n ¼ 1
2
2:303RT
T
Weight % (%)
Derivative Weight % (%/m)
where
a
is the fraction of weight loss, T the temperature (K), n the
120
20
order of reaction, Z the pre-exponential factor, R the molar gas con-
stant, E the activation energy and q is the heating rate. The activa-
tion energies (E ) are calculated from the slopes of the best fit
(C)
a
a
100
0
straight lines (r ꢅ 1) obtained when the plots of the Coats–Redfern
equation [40] are used for the best values of reaction order (n).
Order of reaction (n): it is the one for which a plot of the Coats–
Redfern expression gives the best straight line among various trial
values of n that are examined, i.e., by trial and error for various trial
8
6
0
0
-20
-40
-60
-80
-100
values of n, estimated by the Horowitz–Metzger method [40]. The
Ã
40
20
0
thermodynamic parameters: entropy change
(D
S ), enthalpy
Ã
Ã
change (
D
H ) and free energy of activation change (
DG ) were
calculated using the following equations:
D
D
D
Sà ¼ R½lnðZh=KTފ
Ã
0
100
200
300
400
500
600
700
H ¼ E
a
ꢂ RT
s
Fig. 4. TG/DTG curves of (A): Cd2+/Para, (B): Hg2+/Para and (C): Pb2+/Para
complexes.
Ã
Ã
Ã
G ¼
D
H ꢂ T
s
D
S
where Z, K and h are the pre-exponential factor, Boltzman and Plank
constants, respectively [42].
Ã
The negative
D
S values indicate that the activated complexes
may be attributed to the differences of the electronegativities of
the central metal ions and the ionic size. This can be discussed in
terms of the associated repulsion between the non-bonded elec-
trons (on the donating oxygen) and the bonding electron pairs in
the valance shell of the metal ions. The extent of covalent character
in the metal–ligand bond influences the stabilities of the com-
plexes [45]. It is noticeable that as the electronegativities of the
central metal increases the thermal stability increases because
the covalent bond character increases, giving a strong interaction
between the metal and the ligand [46]. The higher stabilities of
have more ordered structure than the reactants and the reactions
are slower than normal [41,42]. The positive values of
the non-spontaneous character for the reactions at the transition-
Ã
D
G indicate
Ã
state. The positive
D
H values show endothermic transition-state
reactions [43]. From the abnormal values of Z, the reactions of
the complexes at the transition-state can be classified as a slow
reaction [44]. It can be concluded that the thermal decomposition
of these three complexes is of a several of degrees of similarity
(
Table 5). The variation in the thermal stability of these complexes

5
40
S.M. El-Megharbel et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 131 (2014) 534–544
Table 5
Kinetic and thermodynamic parameters of the thermal decomposition of Para complexes.
ꢂ1
ꢂ1
⁄
ꢂ1
ꢂ1
⁄
ꢂ1
⁄
ꢂ1
Complexes
r
Z (s
)
T
max
k
E (kJ mol
)
D
S
(J k mol
)
D
H
(kJ mol
)
DG (kJ mol )
9
6
2
I
II
III
0.9950
0.9992
0.9991
1.54 ꢃ 10
1.32 ꢃ 10
7.21 ꢃ 10
240
160
175
72
86
84
ꢂ70
69
83
77
72
121
182
ꢂ130
ꢂ199
r = correlation coefficient of the linear plot, n = order of reaction, Z = pre-exponential factor.
the metal complexes than that of the ligand may be due to the
molecular symmetry [47].
Table 7
Effect of Para, Cd2+/Para, Hg2+/Para, and Pb2+/Para complexes on liver functions in
male albino rats.
Groups
Total
protein
AST
ALT
LDH
310.66 ± 20.63d
Antimicrobial activity
a
c
12.20 ± 0.54d 13.00 ± 0.56
e
Control
8.18 ± 0.07
6.64 ± 0.71
The results of antimicrobial activities (bacteria and fungi)
in vitro of the Para ligand and their Cd(II), Hg(II) and Pb(II) com-
plexes, Table 6, and Fig. 1S, show that, the [Hg(Para)
c
20.40 ± 0.30d
78.80 ± 1.82
91.00 ± 1.18
70.45 ± 1.18
cd
Para
55.40 ± 1.12
87.80 ± 6.02
177.80 ± 7.39
175.80 ± 7.39
345.69 ± 17.33
341.30 ± 23.10
1922.40 ± 25.37
2+
b
d
e
b
a
a
bc
a
cd
b
Cd /Para 7.98 ± 0.41
2+
2
(H O)
2 2
]ꢁ4H
2
O
Hg /Para 3.88 ± 0.20
2+
c
2200.40 ± 25.37a
test complex has high activities against A. flavus > E. coli > A. ory-
zae = A. niger > B. subtilis. On the other hand, the cadmium(II) and
lead(II) Para complexes have antimicrobial activities against
E. coli, A. oryzae and A. niger, A. flavus and B. subtilis, respectively.
These results clearly obviously that, some metal ions after
complexation give the sensitive nature for the ligand against some
bacteria and fungi, also, we found that the antimicrobial activities
increasing with higher molecular weights (Fig. 1S).
Pb /Para 1.88 ± 0.30
Means within the same column in each category carrying different letters are sig-
nificant at (P 6 0.05) using Duncan’s multiple range tests, where the highest mean
value has symbol (a) and decreasing in value were assigned alphabetically.
serum ALT when compared with normal control group but with
significant decrease when compared with other two complexes
2
+
2+
(
Hg or Pb ) (Table 7 and Fig. 3S).
Our results come in parallel with Galal et al. [49] who reported
Hepatotoxicity effect and histopathological changes of liver tissues
that administration of paracetamol to adult rats caused significant
elevation in ALT and AST levels after treatment for 7 successive
days, several researchers have reported significant elevations in
serum ALT and AST following administration of toxic doses of par-
acetamol in rats [50]. Owing to their high concentrations and ease
of liberation from the hepatocyte cytoplasm, ALT and AST are indi-
cators which are very sensitive of necrotic lesions within the liver
[51]. Hence, the marked release of transaminases into the circula-
tion is indicative of severe damage to hepatic tissue membranes
during paracetamol intoxication and this damage was treated par-
Effect on serum total protein
The administration of Para in it is recommended doses for suc-
cessive 30 days into normal rats elicited significant decrease
(P < 0.05) in serum total protein level compared with normal con-
trol group but the effect was less intense as well as group treated
2
+
with Cd /Para. Whereas, a highly significant decrease (P < 0.05)
2+
2+
were observed in the groups treated with Hg /Para and Pb /Para
complexes after the end of the experiment when compared with
normal control group (Table 7 and Fig. 2S).
2
+
tially by Cd /Para and also, approve the deleterious effect of
Pb(Lead) exposure on hepatic functions parameters.
Our results were supported with Ekam and Ebong [48] who
reported significant decrease in serum total protein after treatment
of male rats with paracetamol for 14 days as a result of hepatic
damage induced by paracetamol administration and this assure
our findings that paracetamol cause significant damage in liver
and this level was increased significantly in Cd2 /Para complex
treated group which showed improvement in liver functions
parameters due to complexation between paracetamol and
Cadmium metal ion as compared to Para treated group.
Effect on serum lactic dehydrogenase enzyme (LDH) activity
2
+
Treatment of normal rats with Para and Cd + Para for succes-
sive 30 days in their recommended doses elicited non-significant
changes in serum LDH activity after the end of the study when
compared with normal control group. While the other treatments
+
2
+
2+
with Para complexes (Hg and Pb ) elicited a marked elevation
in serum LDH activity when compared with normal control group
(
Table 7 and Fig. 4S). The significant increases of LDH are mainly
Effect on serum GOT and GPT (AST&ALT)
2+
due to leakage of these enzymes into the blood because of Pb
HG toxicity in liver [52] and we consider the first author to dem-
onstrate these results on Para and these complexes.
,
Serum transferases (AST&ALT) levels were slightly elevated
after 30 days post Para administration to normal rats when com-
pared with normal control group. Meanwhile highly significant
increase (P < 0.05) in serum AST&ALT were recorded in normal rats
2
+
2+
2+
in response to administration of Pb /Para or Hg /Para but the
Effect on lipid profile in male albino rats
2
+
administration of Cd /Para has revealed significant increase in
Normal healthy control was found to be stable in their choles-
terol values (Table 8). Total cholesterol in the Para treated group
was significantly higher when compared with the other groups
except the group treated with Hg + Para as it was increased sig-
nificantly when compared with other groups and control group.
Table 6
2+
Antimicrobial data of Cd+2/Para, Hg+2/Para and Pb+2/Para complexes.
Complexes
Inhibition zone (cm)
2+
The Cd + Para treated group showed non-significant change
E. coli
B. subtilis
Asperagillus
oryzae
Asperagillus
niger
Asperagillus
flavus
when compared with control group while the group treated with
2+
Pb /Para was significantly increased when compared with the
2
+
Control
I
II
III
0
0.7
4
0
0
0
1
1
3.5
0
0
5
2
control group but not as Hg + Para treated group.
1.5
1.3
1.3
1.5
3.5
3.5
Treatment the animals with Para induced highly significant
increase in triglycerides contents when compared with normal con-
4
2
+
trol group while treatment of animals with Cd + Para afforded

S.M. El-Megharbel et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 131 (2014) 534–544
541
Table 8
Effect of Para, Cd2+/Para, Hg2+/Para, and Pb2+/Para complexes on lipid profile in male albino rats.
Groups
Cholesterol (mg/dl)
Triglycerides (mg/dl)
98.99 ± 4.77e
HDL-c (mg/dl)
LDL-c (mg/dl)
VLDL-c (mg/dl)
19.79 ± 2.45d
Risk ratio (%)
e
ab
33.24 ± 2.52e
1.85 ± 0.24e
Control
68.22 ± 4.57
36.78 ± 3.62
25.25 ± 3.24
87.52 ± 3.54c
149.24 ± 9.45
110.23 ± 8.65
173.62 ± 9.54
bc
c
59.21 ± 4.12b
29.84 ± 4.21
b
3.46 ± 0.52c
Para
2
+
de
d
31.20 ± 5.24b
de
c
d
Cd /Para
70.11 ± 2.15
39.20 ± 5.42
49.52 ± 3.52
22.04 ± 3.52
2.24 ± 0.53
2
+
a
a
d
c
a
14.76 ± 2.11a
13.71 ± 1.63b
Hg /Para
255.14 ± 22.1
17.28 ± 3.21
34.72 ± 5.75
2
+
bc
142.32 ± 10.25c
e
a
28.46 ± 6.75b
Pb /Para
99.41 ± 6.75
7.25 ± 0.42
210.23 ± 12.45
Means within the same column in each category carrying different letters are significant at (P 6 0.05) using Duncan’s multiple range tests, where the highest mean value has
symbol (a) and decreasing in value were assigned alphabetically.
Low density lipoprotein cholesterol (LDL-c) = Total cholesterol ꢂ (HDL-c + triglyceride)/5, volatile.
Low density lipoprotein cholesterol (VLDL-c) = Triglycrid/5, risk ratio = total cholesterol/high density lipoprotein cholesterol (HDL-c).
significant decrease in TG when compared with Para group but
with normal control group as it ameliorate the total antioxidant
capacity when compared with Para treated group and other
complexes.
induced significant increase when compared with control group.
2
+
2+
Pb and Hg + Para groups did not improve the TG contents as
compared to the normal control group but the higher increase
LPO is supposed to cause the destruction and damage of cell
membranes leading to changes in membrane permeability and flu-
idity, enhancing the protein degradation in mice [53]. In the pres-
ent study, the levels of LPO were increased indicating an increased
in the generation of free radicals in the liver tissues of Para treated
2
+
was noticed in Hg + Para treated group after 30 days of treatment.
Table 8 shows the levels of HDL-c for all treatment groups with
Para and it is complexes to be lower than control group except the
2
+
Cd + Para treated group showed non-significant change in HDL-c
when compared with normal control group.
2
+
2+
animals and Hg , Pb Para complexes [54]. After administration
2
+
Table 8 and (Fig. 5S) revealed that all groups treated with Para
and it is complexes afforded significant increase in LDL-c except
of Cd complex with Para for 30 days, the LPO were decreased
in the liver tissues (Fig. 6S) indicating decreased in the generation
2
+
2+
Cd + Para which showed non-significant changes when com-
pared with control group but with less increase in LDL-c content,
on the other hand, VLDL-c was significantly increased in group
of free radicals. It is possible that Cd + Para complex act as a scav-
enger of oxyradicals and thus could prevent liver damage. Accord-
2
+
ing to the present data, Cd + Para complex has a protective effect
by decreasing LPO.
2
+
treated with Hg + Para when compared with control group while
2
+
Cd + Para group showed the less increase in VLDL-c when com-
pared with control group and it was the best treated group.
So, as a result of the previous data, the risk factor will be high in
Paracetamol is mainly metabolized in liver to excretable glucu-
ronide and sulfate conjugates. However, the hepatotoxicity of par-
acetamol has been attributed to the formation of toxic metabolites
when a part of paracetamol is activated by hepatic cytochrome
P-450, to a highly reactive metabolite N-acetyl-P-benzoquinonei-
mine (NAPQI). NAPQI which is believed to play an important role
in paracetamol mediated toxicity is initially detoxified by conjuga-
tion with reduced glutathione (GSH) to form mercapturic acid.
However, when the rate of NAPQI formation exceeds the rate of
detoxification by GSH, it oxidizes tissue macromolecules such as
lipid or ASH group of protein and alters the homeostasis of calcium
after depleting GSH [55]. In the present study the Cd2+/Para com-
plex competes with oxygen thus inhibits the generation of the
anions.
2
+
2+
Para treated group and it is complexes with Hg and Pb but the
risk ratio was more less in Cd2 treated group with Para.
+
Effect on SOD, GST and total antioxidant capacities (TAC) in male
albino rats
Table 9 and (Fig. 6S) showed that all groups treated with
Para complexes showed significant decrease in SOD, GSH and
TCA capacities when compared with control group unless the
group treated with Cd2+/Para complex which showed improve-
ment in the results of antioxidant capacities when compared
Table 9
Effect of Para, Cd+2/Para, Hg+2/Para, and Pb+2/Para complexes on SOD, Glutathione–
S-transferase (GST) and total antioxidant capacity (TAC) in male albino rats.
Effect on hematological parameters in male albino rats
Table 10 and (Fig. 7S) represents effect of Para and it is com-
plexes on the hematological parameters in normal and rats during
the 30 days exposure. The level of RBCs, Hb, HCT, MCH and WBCs
decreases in Para complexes treated groups as compared to the
control group. However, the level of RBCs, Hb, HCT, MCH and WBCs
were non-significantly changed when compared with control
group with less decrease in platelets counts. The major pathologi-
cal consequences of free radical induced membrane lipid peroxida-
tion include increased membrane rigidity, decreased cellular
Groups
SOD (U/mg)
1.66 ± 0.20a
GST (U/mg)
97.23 ± 3.11
TAC (%)
a
a
c
Control
100.20 ± 6.32
76.42 ± 4.42
b
a
77.21 ± 5.32c
Para
1.23 ± 0.32
1.53 ± 0.42
2
+
b
83.10 ± 4.65b
Cd /Para
80.32 ± 6.21
2
+
1.10 ± 0.53c
0.89 ± 0.33d
57.25 ± 5.20d
50.02 ± 4.20d
d
Hg /Para
54.21 ± 3.61
45.32 ± 3.54
2
+
e
Pb /Para
Means within the same column in each category carrying different letters are sig-
nificant at (P 6 0.05) using Duncan’s multiple range tests, where the highest mean
value has symbol (a) and decreasing in value were assigned alphabetically.
Table 10
Effect of Para, Cd2+/Para, Hg2+/Para, and Pb2+/Para complexes on the hematological parameters in male albino rats.
6
3
3
3
3
3
Groups
RBCs (10 /mm )
Hb (g/dl)
HCT (%)
WBCs (10 /mm )
Platelets (10 /mm )
a
a
a
c
ab
665.32 ± 7.85a
534.33 ± 9.52c
Control
8.99 ± 1.90
14.43 ± 3.20
47.30 ± 3.65
39.07 ± 3.22
8.52 ± 2.10
c
Para
7.57 ± 1.35c
13.20 ± 2.85b
5.68 ± 2.35
2
+
b
b
44.42 ± 2.85b
34.25 ± 2.75d
33.20 ± 4.21d
7.69 ± 1.98b
2.98 ± 1.77d
3.20 ± 1.65d
b
Cd /Para
8.56 ± 1.75
13.43 ± 3.61
10.47 ± 2.10
9.63 ± 1.95
580.95 ± 15.32
2
+
5.20 ± 1.68d
5.34 ± 1.45d
c
d
Hg /Para
342.35 ± 8.52
336.10 ± 7.12
2
+
c
d
Pb /Para
Means within the same column in each category carrying different letters are significant at (P 6 0.05) using Duncan’s multiple range tests, where the highest mean value has
symbol (a) and decreasing in value were assigned alphabetically.

5
42
S.M. El-Megharbel et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 131 (2014) 534–544
deformability, reduced erythrocyte survival, and lipid fluidity [56]
and thus this explain the significant decrease in the number of
R.B.Cs and other blood parameters in groups treated with Para
– Para group
Microscopically, severe congestion of the hepatic blood vessels
was seen (Fig. 6).
and it is complexes with Hg2 and Pb
+
2+
.
– Cd2+/Para group
Histopathology of liver
–
Control group
Microscopically: Normal liver tissues formed of small central
Liver tissue showing severe fatty change in the hepatocytes
(Fig. 7) and liver tissue showing necrosis in liver cells in the form
of pyknotic nucleic (") and more eosinophilia of the cytoplasm
was seen in (Fig. 8).
vein surrounded by cords of hepatocytes showing central vesicular
nuclei and eosinophilic cytoplasm (Fig. 5).
Fig. 5. Cross section of control rat liver formed of small central vein surrounded by
cords of hepatocytes showing central vesicular nuclei and eosinophilic cytoplasm
Fig. 8. Cross section of rat liver tissues of Cd+2/Para treated group showing some
necrosis in liver cells in the form of pyknotic nuclei (") and more eosinophilia of the
(H and E ꢃ 200) where CV: central vein, EN: eosinophilic nuclei.
cytoplasm (H and E ꢃ 200) (PN: Pyknotic nuclei).
Fig. 9. Cross section of rat liver tissues of Hg+2/Para treated group showing severe
Fig. 6. Cross section of rat liver tissues of Para treated group showing severe
congestion of the hepatic blood vessels (H & E stain ꢃ 150) where S: Severe
congestion.
fatty change of hepatocytes and markedly congested central vein (H
& E
stain ꢃ 400) (F: Fatty change , CCV: Congested central vein).
Fig. 7. Cross section of rat liver tissues of Cd+2/Para treated group showing severe
fatty change in the hepatocytes (H and E ꢃ 200) (SFC: Severe fatty change).
Fig. 10. Cross section of rat liver tissues of Pb+2/Para treated group showing severe
congestion of the hepatic blood vessels) H & E stain ꢃ 400) (SC: Severe congestion).

S.M. El-Megharbel et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 131 (2014) 534–544
543
–
Hg2+/Para group
Microscopically, severe fatty change of hepatocytes and mark-
edly congested central vein was shown (Fig. 9).
–
Pb2+/Para group
Microscopically, variable degrees of congestion of the hepatic
blood vessels, severe congestion were seen (Fig. 10) and also
marked necrosis of hepatocytes with congested blood sinusoids
(
Fig. 11).
Liver tissue showing markedly dilated central vein filled by
large number of red blood cells and surrounded by hepatic cords
Fig. 12).
Our results come in harmony with Galal et al. [49] who reported
(
Fig. 11. Cross section of rat liver tissues of Pb+2/Para treated group showing
marked necrosis of hepatocytes with congested blood sinusoids (H & E stain ꢃ 400)
that histological examinations of liver sections of rats that was
given paracetamol proved the presence of hepatotoxic markers
which revealed degenerative changes that involved the hepato-
cytes and cells that line the blood sinusoids. The damage related
to the majority of the hepatic lobule with marked loss of its normal
pattern. These changes positively correlated with the noted
increases in transaminase activities and this is greatly agreed with
our recorded results.
(
N :Necrosis).
Conclusion
The complexation between metal ions like (Cd(II), Hg(II) and
Pb(II) with Para produced 1:2 M ratio (metal: Para) as a monoden-
tate via hydroxyl group and give general formula: [M(Para)
(
H
2
O)
2
]ꢁnH
2
O, where n = 6 or 4 (Fig. 13). The resulted Para
compounds were assigned by infrared and electronic spectra. Ther-
mogravimetric analyses have proved the thermal stability feature
of Para complexes. The antimicrobial activities of the metal com-
plexes of Para recorded a significant effect against some bacteria
and fungi. The Para/Hg(II) and Para/Pb(II) complexes duplicate
Fig. 12. Cross section of rat liver tissues of Pb+2/Para treated group showing
markedly dilated central vein filled by large number of red blood cells and
surrounded by hepatic cords (DCCV: dilated congested central vein).
Fig. 13. The mode of chelation of the Para complexes where, M = Cd (II), Hg (II) and Pb(II).

5
44
S.M. El-Megharbel et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 131 (2014) 534–544
the highly dangerous effect of paracetamol on liver and showing
atrophy in the liver tissues and make disruption in liver function
parameters, While, Para/Cd (II) complex has greatly succeeded in
alleviating the hepatotoxic effect of Paracetamol and improved
the antioxidant capacities in male rats as compared with paracet-
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