Investigation of catalytic, anti-bacterial, anti-oxidant, and DNA cleavage properties of bimetallic and trimetallic magnetic nanoalloys base on cupper

Article abstract of DOI:10.1080/24701556.2019.1574818

Bimetallic and trimetallic magnetic nanoalloys base on cupper were easily prepared from their starting materials. These nanoparticles (NPs) were used for the synthesis of biscoumarines by two-component one-pot domino Knoevenagel-type condensation/Michael

Full text of DOI:10.1080/24701556.2019.1574818

Inorganic and Nano-Metal Chemistry
ISSN: 2470-1556 (Print) 2470-1564 (Online) Journal homepage: https://www.tandfonline.com/loi/lsrt21
Investigation of catalytic, anti-bacterial, anti-
oxidant, and DNA cleavage properties of bimetallic
and trimetallic magnetic nanoalloys base on
cupper
Kaveh Parvanak Boroujeni, Ahmad Farokhnia, Mansooreh Shahrokh &
Mohsen Mobini
To cite this article: Kaveh Parvanak Boroujeni, Ahmad Farokhnia, Mansooreh Shahrokh &
Mohsen Mobini (2019): Investigation of catalytic, anti-bacterial, anti-oxidant, and DNA cleavage
properties of bimetallic and trimetallic magnetic nanoalloys base on cupper, Inorganic and Nano-
Metal Chemistry, DOI: 10.1080/24701556.2019.1574818
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INORGANIC AND NANO-METAL CHEMISTRY
https://doi.org/10.1080/24701556.2019.1574818
Investigation of catalytic, anti-bacterial, anti-oxidant, and DNA cleavage
properties of bimetallic and trimetallic magnetic nanoalloys base on cupper
Kaveh Parvanak Boroujenia, Ahmad Farokhniaa, Mansooreh Shahrokha, and Mohsen Mobinib
b
aDepartment of Chemistry, Shahrekord University, Shahrekord, Iran; Department of Genetics, Shahrekord University, Shahrekord, Iran
ABSTRACT
ARTICLE HISTORY
Received 24 February 2018
Accepted 2 January 2019
Bimetallic and trimetallic magnetic nanoalloys base on cupper were easily prepared from their
starting materials. These nanoparticles (NPs) were used for the synthesis of biscoumarines by two-
component one-pot domino Knoevenagel-type condensation/Michael reaction between aldehydes
and 4-hydroxycoumarin. Also, anti-bacterial, anti-oxidant, and DNA cleavage properties of these
NPs were investigated.
KEYWORDS
Magnetic nanoalloys;
nanocatalyst; anti-bacterial
activity; anti-oxidant
activity; DNA
cleavage properties
Introduction
industry due to their strong antibacterial properties and bio-
compatibility.[12] For example, Ag–Cu and Ni–Ag, Cu–Ni,
Ag–Au, and Ni–Co NPs exhibit antibacterial properties,[13,14]
with widespread applications in dental and hyperthermia,[15,16]
the biological activities toward human mesenchymal stem cells
(hMSC),[17] and non-enzymatic glucose sensor applications,[18]
respectively.
One of the most important aspects of nanotechnology is
alloying two or more elements to make nanoalloys with
modified properties including high strength, ductility, corro-
sion resistance, moderate strength, magnetic, high tough-
ness, and electrical and thermal conductivity properties.
Among these properties, magnetic behavior is considered as
the most important feature of the magnetic nanoalloys.[1]
For example, Cu and Ag do not have magnetic properties,
but by mixing them with Co or Ni and converting them
into bimetallic or trimetallic alloys, they show magnetic
properties.[2] On the other hand, sometimes, some of the
heterogeneous alloys made of nonmagnetic metals like
Cu–Co or Cu–Ni that are used in the structure of sensors,
exhibit enhanced magnetoresistance in contrast to the
homogeneous single metals like Co or Ni.[3]
Nowadays, the magnetic nanoalloys have been received
considerable attention as catalyst for synthetic applications
due to their high surface-to-volume ratio and easy separ-
ation from reaction vessel via an external magnet and main-
taining their catalytic activity after several reaction cycles.[4]
For instance, several magnetic catalysts based on Cu alloys
have been used as efficient catalysts in some of the import-
ant heterogeneous reactions such as the generation of car-
bon nanotubes by catalytic cracking of methane,[5] synthesis
of vertically aligned carbon nanofibers,[6] methane decom-
position,[7] high-temperature water-gas shift reaction,[8] syn-
thesis of higher alcohols from syngas,[9] the hydrogenation
of p-chloronitrobenzene,[10] and selective oxidation of
ethyl benzene.[11]
Heterocyclic systems are common structural motifs in
many biologically active compounds and therefore intense
effort has been devoted by researchers constantly to intro-
duce newer and efficient protocols for their synthesis.
Coumarin and its derivatives are a large group of hetero-
cycles with a wide variety of biological and therapeutic
properties such as anti-HIV,[19] cytotoxicity and enzyme
inhibitory,[20] anti-fungal,[21] anti-bacterial,[22] anti-oxi-
dant,[23] anti-cancer,[24] and anti-coagulant properties.[25]
Among coumarin derivatives, biscoumarins become more
popular and are usually prepared by the condensation reac-
tion of aldehydes with 4-hydroxycoumarin. This reaction can
be catalyzed by several types of catalysts such as piperi-
dine,[26] I2,[27] sodium dodecyl sulfate,[28] [poly(4-vinylpyri-
dine) BuSO3H]Cl.xAlCl3,[29] MW irradiation,[30,31] [poly(4-
vinylpyridine)–BuSO3H]HSO4,[32] indion 190 resin,[33] choline
hydroxide,[34] poly(AMPS-co-AA)@Fe3O4,[35] Zn(OAc)2,[36]
and triethylammonium hydrogen sulfate.[37] However, these
methods are plagued by the limitation of prolonged reaction
times, low yields, tedious work up, inefficiency of method
when aliphatic aldehydes are used in the reaction, and the
use of expensive, hazardous, difficult to handle or unreus-
able catalysts.
Various synthetic techniques have been reported for pre-
Over the past decade, inorganic material–nanoparticles
have gained increasing attention in the pharmaceutical paring nanoalloys such as reduction method, interaction
CONTACT Kaveh Parvanak Boroujeni
parvanak-ka@sci.sku.ac.ir
Department of Chemistry, Shahrekord University, Shahrekord, (115), Iran.
Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/lsrt.
ß 2019 Taylor & Francis Group, LLC

2
K. PARVANAK BOROUJENI ET AL.
method, vapor quenching, sputtering and gas condensa- Preparation of 3,3’-(4-bromobenzylidene)-bis-(4-
tion.[38,39] Among them, the reduction method is relatively the
hydroxycoumarin) as a typical procedure for the
synthesis of biscoumarins
simplest one. During the reduction process, if the two metals
in a complex have different redox potential, they will form a
core-shell structure, otherwise they generate bimetallic
alloy.[40,41] Along this line and in continuation of our works
on the synthesis and applications of magnetic catalysts, nano-
alloys, and heterogeneous catalysts[35,39,42,43] we now wish to
introduce bimetallic and trimetallic magnetic nanoalloys base
on cupper as new magnetic catalysts for the synthesis of bis-
coumarins. Also, anti-bacterial, anti-oxidant, and DNA cleav-
age properties of these NPs were investigated.
To a solution of 4-bromobenzaldehyde (1 mmol), 4-hydroxy-
coumarin (2 mmol), and toluene (3 mL), Cu–Ni (0.05 g) or
Cu–Ni–Ag (0.04 g) was added at 90 ꢀC. After the completion
of the reaction (monitored by TLC), the catalyst was
removed by an external magnet and washed with toluene
(2 ꢁ 5 mL) and the filtrate was concentrated on a rotary
evaporator under reduced pressure. Whenever required, the
crude product was recrystallized from ethanol to give the
pure product.
Antibacterial test performance
Experimental
At first, E.coli and S.aureus cells were separately transferred
from pure bacterial stocks to nutrient broth media, and incu-
bated at 37 ꢀC until reaching to 0.5 McFarland standard. The
diffusion method is provided by the application of standard
bacterial inoculums to the surface of Muller-Hinton agar
plates (100 mm diameter) using sterile swabs. After incuba-
tion of plates at 37ꢀC for 24h and bacterial growth, distinct
equal wells have been created in the agar plates for antibac-
terial diffusion assay. Then, dispersed solutions of Cu–Ni and
Cu–Ni–Ag NPs were individually poured into the wells with
distinct concentrations (0.25, 0.5, 1 or 2 mg/mL). After incu-
bation at 37ꢀC for 18–24hours, the growth inhibition zone
diameters (in millimeter scale) was determined. Standard
antibiotic discs and sterile distilled water has been used as
positive and negative controls, respectively.
Materials and methods
All chemicals were either prepared in our laboratory or were
purchased from Merck and Fluka. Two bacterial strains
were used: E. coli (ATCC 35218) and S. aureus (ATCC
6538). Reaction monitoring and purity determination of the
products were accomplished by GLC or TLC on silica-gel
polygram SILG/UV254 plates. Gas chromatography was
recorded on Shimadzu GC 14–A. IR spectra were obtained
by a Shimadzu model 8300 FT–IR spectrophotometer. 1H
NMR spectra were recorded on 400 MHz spectrometer in
CDCl3. Melting points were determined on a Fisher–Jones
melting–point apparatus. XRD patterns were recorded by a
Phillips, X–ray diffractometer using graphite monochromat-
ized Cu Ka radiation. A morphological study of the synthe-
sized products was carried out directly by a Hitachi S4160
field emission scanning electron microscope (FE–SEM).
Room temperature magnetic properties were investigated by
Lakeshore device in an applied magnetic field sweeping
between 8000 Oe.
DPPH radical–scavenging activity
The 0.1 mM DPPH in methanol solution was prepared.
450 lL of Tris–HCl buffer (pH= 7.4) and 1 mL of metha-
nolic DPPH solution were added to 50 lL of Cu–Ni and
Cu–Ni–Ag NPs with distinct concentrations (2.5, 5, 10 or
20 lg/mL). The mixtures were left for 30 min at room tem-
perature in the dark and then their absorbance was meas-
ured at 517 nm. The methanol was applied as a blank
solution. The same experiment was performed with BHT as
a positive control. The DPPH free radical scavenging activity
was calculated by the following equation: %DPPH radical
Preparation of Cu–Ni NPs[39]
0.1 g of [Cu(NH3)4][Ni(C2O4)2] complex (0.27 mmol) was
dissolved in 20 mL of a mixture of water-ethanol solution
(50:50). Then, to stirred solution, 5 mL of hydrazine and
10 mL of NaOH (4 M) were gradually added at 70–80 ꢀC.
During the reduction reaction the color of the solution was
turned to red and then after 1 h the black Cu–Ni NPs were
observed. The resulting black NPs were carefully decanted,
washed repeatedly with doubly distilled water, and dried at
room temperature for 24 h.
scavenging ¼ Absorbance
ꢂ
Absorbance(sample)/
(blank)
Absorbance(blank) ꢁ 100.
DNA cleavage assays
The cleavage of pET-28 plasmid DNA by Cu–Ni and
Cu–Ni–Ag NPs was identified using agarose gel electrophor-
esis. Each 40 lL of Cu–Ni and Cu–Ni–Ag NPs with distinct
concentrations (0.25, 0.5, 1 or 2 mg/mL) was mixed with
5 lL of plasmid DNA solution (0.25 lg/mL) in sterile 1.5 mL
micro tubes and incubated at 37 ꢀC for 24 h. Then 10 lL of
the resulting mixture was mixed with 1 lL of gel loading
dye (G2526-5ML Sigma-Aldrich) and loaded into the 1%
agarose gel (W/V) wells. The agarose gel was prepared with
Preparation of Cu–Ni–Ag NPs
In the preparation of the above Cu-Ni nanoalloy after add-
ition of hydrazine and NaOH solution at 70–80 ꢀC, a solu-
tion of silver nitrate (0.1 M, 2.7 mL) and then hydrazine
(2 mL) were gradually added to the reaction solution to yield
Cu–Ni–Ag NPs.

INORGANIC AND NANO-METAL CHEMISTRY
3
TBE 1X buffer (Tris-HCl (0.07 M, pH= 7.4), EDTA (4 mL,
0.5 M, pH= 8.0), and boric acid (5.5 g)) in distilled water
(1 L). After performing electrophoresis (40 mA and 80 volt
for 45 min), the final agarose gel was observed in the gel
duct device (UVItec Limited BTS-20M) under UV rays.
Results and discussion
Synthesis and characterization of Cu–Ni and
Cu–Ni–Ag NPs
In our previous work, the bimetallic nanoalloys Cu–Ni was
produced from the by chemical reduction of the
[Cu(NH3)4][Ni(C2O4)2] using hydrazine.[39] In this work the
reduction of this complex along with silver nitrate (AgNO3)
by excess hydrazine lead to formation of Cu–Ni–Ag NPs.
During the reduction process, the whole reaction proceeds
through co-reduction of Cu2þ, Ni2þ and Agþ so solid solu-
tion Cu–Ni–Ag NPs is formed. The reactions can take place
at 70–80 ꢀC in water-ethanol solution via the following
equations.
Figure 1. X-Ray diffraction patterns for (a) Cu–Ni and (b) Cu–Ni–Ag NPs.
Â
ÃÂ
Ã
CuðNH3Þ4 NiðC2O4Þ2 þ N2H4 þ 4NaOH ! Cu–Ni
þ N2 þ 4NH3 þ 2Na2C2O4 þ 4H2O
Â
Ã
Â
Ã
CuðNH3Þ4 NiðC2O4Þ2 þ AgNO3 þ 5=4N2H4
þ 5NaOH ! Cu–Ni–Ag þ 5=4N2 þ 4NH3
þ 2Na2C2O4 þ NaNO3 þ 5H2O
Figure 2. FT–IR spectra of (a) Cu–Ni and (b) Cu–Ni–Ag NPs.
The crystal structure of these Cu–Ni bimetallic and
Cu–Ni–Ag trimetallic NPs was studied by powder X–ray dif-
fraction (XRD). The powder diffraction pattern of Cu–Ni
bimetallic have been studied completely (Figure 1a).[39] The
diffraction peaks for Cu–Ni–Ag NPs shows that this nanoal-
loy is in the face-center cubic (fcc) phase and also confirm
that these compounds can be considered as Cu/Ni/Ag trime-
tallic NPs. In Figure 1b three distinct diffraction peaks are
clearly observed at 2h values of 43.50, 50.64 and 74.32ꢀ that
could be assigned to 111, 200 and 220 crystal planes of the
metallic Cu, respectively. The results show that diffraction
peaks are in good agreement with the standard values for
Cu (with card no. 04-0836), nickel (with card no. 87-0712)
and silver (with card no. 01-1164). In most studies of the
grain size of nanocrystalline materials, X–ray line-broaden-
ing analysis is used. The crystallite size is estimated from the
full width at half maximum (FWHM) of the diffraction
peaks by the Scherrer formula. The crystal siza obtained for
Figure 3. The magnetization curves and saturation magnetizations of (a)
Cu–Ni[39] and (b) Cu–Ni–Ag NPs.
Magnetic properties of bimetallic and trimetallic NPs
Cu–Ni and Cu–Ni–Ag NPs, relationship at the sharpest were also studied. As show in Figure 3, NPs are characteris-
peak are 29 and 44 nm, respectively.
tic of ferromagnetic material. The saturation magnetization
(Ms) values of Cu–Ni and Cu–Ni–Ag NPs are 13 and
5 emu/g, at 300 K, respectively, that confirm trimetallic alloy
NPs have Ms less than the bimetallic alloy and the nickel
NPs have a Ms less than that of the bulk nickel (about
FT–IR of Cu–Ni[39] and Cu–Ni–Ag NPs are shown in
Figure 2. In the conversion of complex into NPs, the
absorption bands related to the ligand groups in complex
were disappeared and the metals had no absorption bands
in the medium IR region. However, the weak absorption 55 emu/g at 300 K[44]). Also, the magnetic properties of NPs
bands due to the water in the alloy NPs are observed in the show that the Ms decreases with increasing Cu and Ag con-
infrared spectrum.
centration. The reason of the decrease in Ms of Cu–Ni and

4
K. PARVANAK BOROUJENI ET AL.
Cu–Ni–Ag NPs is due to the presence of dissolved Ni in the samples (Figure 4b) show agglomeration of particles with
Cu and Ag matrix (Figure 3b).
different size in spherical shapes. The particle size distribu-
tion graphs for Cu–Ni–Ag samples obtained from FE–SEM
analysis, according to which the average diameter of max-
imum number of particles lies in the range of 30–40 nm.
The different dispersion of the formed nanoalloy phases
can be also argued by considering energy-dispersive X–ray
(EDX) data in SEM micrographs. EDX analysis shows that
these prepared NPs are pure nanoalloys (Figure 5).
The FE–SEM studies from Figure 4a show monodisperse,
spherical and uniform particles of Cu–Ni by average diam-
eter size 15 nm.[39] The images of SEM revealed that uni-
form particles morphology in Cu–Ni changed by increasing
the presence of silver, therefore images of Cu–Ni–Ag
Catalytic application of Cu–Ni and Cu–Ni–Ag NPs in the
synthesis of biscoumarins
The catalytic performance of NPs was investigated in the
condensation reaction of aldehydes with 4-hydroxycou-
marin. For this purpose, as a model reaction, reaction of 4-
bromobenzaldehyde (1 mmol) with 4-hydroxycoumarin
(2 mmol) in the presence of different amounts of NPs in
various solvents and also under solvent free conditions were
studied. We observed that this reaction was performed
smoothly in toluene at 90 ꢀC in the presence of 0.05 g of
Cu–Ni or 0.04 g of Cu–Ni–Ag NPs and the desired product
3,3’-(4-bromobenzylidene)-bis-(4-hydroxycoumarin)
was
obtained in excellent yield (Table 1, entry 4). Afterwards,
under the optimized conditions, various aromatic (entries
1–10), heteroaromatic (entries 11,12), and aliphatic alde-
hydes (entries 13–15) were reacted with 4-hydroxycoumarin
and the corresponding products were obtained in excellent
yields. As the results in Table 1 show, the better yields were
obtained with aromatic aldehyde containing electron with-
drawing groups (entries 9,10). This can be explained by the
reaction mechanism in Scheme 1. Aldehyde forms an oxy-
gen-bonded complex through NPs to give activated alde-
hyde, which is attacked by of 4-hydroxycoumarin to afford
the intermediate I. Then, the intermediate II is produced by
dehydration of the intermediate I. The 1,4-nucleophilic add-
ition of a second molecule of 4-hydroxycoumarin on acti-
vated intermediate II, in the Michael addition fashion,
affords the synthesis of biscoumarin product. The electron
withdrawing groups substituted on aromatic aldehyde in
intermediate II increase the rate of 1,4-nucleophilic addition
reaction because of the alkene LUMO is at lower energy in
their presence compared with electron donating groups.[29]
Figure 4. SEM images of (a) Cu–Ni and (b) Cu–Ni–Ag NPs.
Figure 5. EDX spectrum of (a) Cu–Ni[39] and (b) Cu–Ni–Ag NPs.

INORGANIC AND NANO-METAL CHEMISTRY
5
Table 1. Synthesis of biscoumarin derivatives catalyzed by Cu–Ni and
Cu–Ni–Ag NPs.
Finally, we present a comparison between selected previ-
ously known catalysts and Cu–Ni and Cu–Ni–Ag NPs
employed for the synthesis of biscoumarins. As shown in
Table 2, in comparison with many of catalysts Cu–Ni and
Cu–Ni–Ag NPs have good catalytic performance in terms of
high yields at short reaction times, reusability, easy separ-
ation, and mild reaction conditions.
OH
Cu-Ni (0.05 g) or
Cu-Ni-Ag (0.04 g)
R
OH
OH
2
+ RCHO
Toluene / 90oC
O
O
O
O
O
O
M.P. (oC)
Entry
Aldehyde
Time (min.)
Yield (%)a,b
Found
Lit.
1
2
3
4
5
6
7
8
C6H5
201,152
221,162
251,202
201,172
201,162
201,162
231,202
251,202
151,102
161,102
201,182
201,172
291,242
251,222
301,232
971,982
951,962
961,972
971,982
941,952
941,952
951,962
951,962
971,982
961,972
931,952
941,962
941,942
931,922
941,932
229–231
260–262
242–244
264–266
256–257
220–222
227–229
213-214
233–234
238–240
214–216
201–203
200-202
189–191
125–126
226–228[31]
265–266[30]
244–246[31]
268–270[30]
254–256[31]
215[26]
4-CH3C6H4
4-CH3OC6H4
4-BrC6H4
4-ClC6H4
3-ClC6H4
4-HOC6H4
3-HOC6H4
4-NO2C6H4
3-NO2C6H4
2-Thienyl
The investigation of biological activity of Cu–Ni and
Cu–Ni–Ag NPs
220–224[30]
210.5[26]
The main objective of antibacterial tests is the evaluation of
the resistance of bacteria to chosen empirical antimicrobial
agents.[45] For this purpose, one of the simplest tests
includes the manual method which itself is classified in two
different methods containing disk and gradient diffusion.
The first one has several advantages such as its simplicity,
requirement of simple equipment, flexibility in choice of
disks, and relatively lower costs.[45]
Encouraged by the results obtained from the catalytic
activity of Cu–Ni and Cu–Ni–Ag NPs in the synthesis of
biscoumarins and xanthenes derivatives, we also investigated
the anti-bacterial properties of these NPs on viable typical
microorganisms. Antibacterial activity of NPs was studied
against gram positive and gram negative bacteria using disk
diffusion method and the zone of growth inhibition was
measured quantitatively (Figure 8a–d, Table 3). As the
results show, Cu–Ni–Ag NPs have stronger degree of anti-
bacterial activity than Cu–Ni NPs. Cu–Ni and Cu–Ni–Ag
NPs both exhibit antibacterial properties against E.coli cell.
Cu–Ni NPs do not show antibacterial activity against
S.aureus cell, while Cu–Ni–Ag NPs have an efficient antibac-
terial effect on it, which can be attributed to the presence of
Ag in this triple alloy and its antibacterial features.[13] The
growth inhibition zone of two bacteria increased with incre-
ment of the concentration of NPs.
9
236–237[31]
233–234[30]
213-214[30]
204-205[30]
196[26]
10
11
12
13
14
15
2-Furanyl
C6H5CH ¼ CH
C6H5CH2CH2
CH3(CH2)2-
190-193[32]
123[26]
aIsolated yields.
bAll products are known compounds and were identified by comparison of
their physical and spectral data with those of the authentic samples.
Oxidation is a chemical reaction that can produce free
radicals, leading to chain reactions that may promote some
of illnesses such as cancer. Antioxidants agents are able to
terminate these chain reactions and hence limit harmful
effects of cellular oxidants. Reactive oxygen species (ROS)
and reactive nitrogen species (RNS) are two types of cellular
oxidants which are produced in living cells.[46] Recently,
nanotechnology is focusing on synthesis of NPs having
improved antioxidant properties which can be used for pre-
vention or treatment of diseases. A convenient method for
the determination of antioxidant activity of NPs is radical
scavenging method which in 2,2-diphenyl-2-picrylhydracyl
hydrate (DPPH) is used as a stable compound which accepts
electrons from antioxidant donors.[47]
Scheme 1. Suggested mechanism for the preparation of biscoumarins cata-
lyzed by Cu–Ni and Cu–Ni–Ag NPs.
1H NMR spectra of the 3,3’-(4-bromobenzylidene)-bis-
(4-hydroxycoumarin) is shown in Figure 6.
Recently, we have reported carbon nanotube-supported
butyl 1-sulfonic acid groups and cellulose aluminum oxide
composite-supported imidazolium chloroaluminate ionic
liquid as heterogeneous catalysts for the synthesis of xan-
thene deravatives.[42,43] Along this line and based on the
above results, we found that Cu–Ni and Cu–Ni–Ag NPs can
be used as magnetic catalysts for the synthesis of 1,8-dioxo-
octahydroxanthenes through condensation of aromatic alde-
hydes with two equivalents of dimedone in ethanol at room
temperature (Scheme 2).
Following the obtained results of antibacterial tests, we
tried to evaluate antioxidant activity of these NPs with dis-
tinct concentrations (2.5, 5, 10, and 20 mg/mL) using DPPH.
As shown in Figure 9, effective free radical inhibition was
observed for Cu–Ni and Cu–Ni–Ag NPs. The antioxidant
The stability and reusability of a catalyst are very import-
ant factors for industrial use. Thus, the recovery and reus-
ability of Cu–Ni and Cu–Ni–Ag NPs were investigated and
it was found that the catalysts could be completely recovered
and used again at least six times without any noticeable loss activity of the Cu–Ni NPs were lower than butyl hydroxyto-
of catalytic activity (Figure 7). luene (BHT) and the antioxidant activity of the Cu–Ni–Ag

6
K. PARVANAK BOROUJENI ET AL.
Figure 6. 1H NMR spectra of the 3,3’-(4-bromobenzylidene)-bis-(4-hydroxycoumarin).
Table 2. Comparison of the catalytic efficiency of Cu–Ni and Cu–Ni–Ag NPs
with selected previously catalysts in the condensation reaction of benzalde-
hyde with two equivalents of 4-hydroxycoumarin.
Cu-Ni (0.05 g)1 or
Cu-Ni-Ag (0.04 g)2
EtOH, r.t.
O
O
O
Ar
O
Time
(min.)
Yield
(%)a
2
+
Entry
Reaction conditions
ArCHO
O
1
2
3
4
Piperidine, EtOH, r.t.
240
25
138
36
92[26]
97[27]
90[28]
95[29]
I2, H2O, 100 oC
Ar : C6H5
15 min, 96%1
Sodium dodecyl sulfate, H2O, 60 oC
[Poly(4-vinylpyridine)-BuSO3H]Cl-xAlCl3,
toluene, 90 oC
4-NO2C6H4
C6H5
4-NO2C6H4
10 min, 97%1
13 min, 97%2
8 min, 98%2
5
6
7
Catalyst-free, solvent-free, MW Irradiation, r.t.
Catalyst-free, H2O, MW irradiation, r.t.
[Poly(4-vinylpyridine)-BuSO3H]HSO4,
toluene, 90 oC
3.5
48
5
91[30]
93[31]
98[32]
Scheme 2. Preparation of 1,8-dioxo-octahydroxanthenes using Cu–Ni and
Cu–Ni–Ag NPs.
8
9
Indion 190 resin, toluene, 100 oC
Aaqueous solution of choline
hydroxide, 50 oC
30
60
90[33]
99[34]
10
11
12
Poly(AMPS-co-AA)@Fe3O4, toluene, 90 oC
15
25
30
97[35]
98[36]
88[37]
Zn(OAc)2, H2O, 100 oC
Triethylammonium hydrogen sulfate,
EtOH, 80 oC
13
14
Cu-Ni NPs, toluene, 90 oC
20
15
97
98
Cu-Ni-Ag NPs, toluene, 90 oC
aIsolated yields.
NPs was higher than Cu–Ni NPs. The IC50 value of the
Cu–Ni and Cu–Ni–Ag NPs has been shown in Table 4.
The damage of genomic DNA can be caused by oxygen
and nitrogen free radicals.[48] It can lead to chromosomal
abnormalities such as chromatin fractures that promote cell
death or uncontrolled cell proliferation. Thus, studies on the
Figure 7. Recyclability of Cu–Ni (0.05 g) and Cu–Ni–Ag (0.04 g) NPs in the
reaction of 4-nitrobenzaldehyde (1 mmol) with 4-hydroxycoumarin (2 mmol) in
toluene at 90 oC after 15 and 10 min, respectively.

INORGANIC AND NANO-METAL CHEMISTRY
7
Table 4. The IC50 values of the BHT, Cu–Ni, and Cu–Ni–Ag NPs.
Sample
IC50 (lg/mL)
BHT
Cu–Ni NPs
Cu–Ni–Ag NPs
12.53 2.43
19.12 2.89
9.62 2.09
Figure 8. Evaluation of the antimicrobial activity of a) Cu–Ni NPs against E.coli;
b) Cu–Ni NPs against S.aureus; c) Cu–Ni–Ag NPs against E.coli; d) Cu–Ni–Ag NPs
against S.aureus.
Table 3 Antibacterial activity of Cu–Ni and Cu–Ni–Ag NPs
Zone of growth inhibitiona
(mm)/ Concentration
(mg/mL)
Zone of growth inhibitionb
(mm)/ Concentration
(mg/mL)
Species
0.25
0.5
1
2
0.25
0.5
1
2
Figure 10. Effect of the distinct concentrations of Cu–Ni and Cu–Ni–Ag NPs
(0.25, 0.5, 1, and 2 mg/mL) on the DNA cleavage.
E.coli
S.aureus
1
–
1
–
5
–
8
–
2
–
6
1
7
8
8
11
aCu–Ni NPs.
bCu–Ni–Ag NPs.
shows gel electrophoretic pattern of DNA after incubation
with distinct concentrations of Cu–Ni and Cu–Ni–Ag NPs
in contrast to plasmid DNA mixed with water as the nega-
tive control. After exposure of plasmid DNA to different
concentrations of the nanomaterial suspensions no destruc-
tive effects were observed on DNA.
Conclusion
In conclusion, we prepared Cu–Ni and Cu–Ni–Ag NPs and
investigated their application as magnetic catalysts for the
synthesis of biscoumarin and xanthene derivatives. The cata-
lysts were easily separated from the reaction media by the
application of an external magnetic and work-up of these
reactions is very easy. The products were obtained in high
yields in short reaction times and the recycled catalysts can
be used for further reactions without any activity loss.
Biological activity of these NPs such as anti-bacterial, anti-
oxidant, and DNA cleavage properties were also investi-
gated. The results showed that both Cu–Ni and Cu–Ni–Ag
NPs have antibacterial and antioxidant properties and they
had no significant effect on DNA cleavage.
Figure 9. The DPPH radical scavenging activity of the BHT, Cu–Ni, and
Cu–Ni–Ag NPs.
DNA cleavage by synthetic materials such as NPs have been
the subject of great interest to researchers.[47]
After evaluation of antibacterial and antioxidant activities
of the Cu–Ni and Cu–Ni–Ag NPs, we also examined DNA
cleavage property of these NPs. The cleavage of plasmid
DNA was revealed by gel electrophoresis. In agarose gel
electrophoresis method DNA molecules migrate in the gel
Acknowledgements
The author thanks the Research Council of Shahrekord University for
texture based on their charge, size, and topology. Figure 10. partial support of this work.

8
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