Global Scientific and Academic Research Journal of Multidisciplinary Studies ISSN: 2583-4088 (Online)

Global Scientific and Academic Research Journal of Multidisciplinary Studies

ISSN: 2583-4088 (Online)
Frequency: Monthly
Published By GSAR Publishers
Journal Homepage Link- https://gsarpublishers.com/journals-gsarjebm-home/

SYNTHESIS, CHARACTERIZATION, AND ANTIBACTERIAL ACTIVITY OF Ni (II),

COMPLEX DERIVED FROM CLOXACILLIN AND CEPHALEXIN
BY
B. A. Saje1 S. Suleiman2 and M. N. Yaro3
1
Department of Chemistry, Aminu Saleh College of Education, Azare Bauchi State
2 and 3
Department of Chemistry, Federal University Dutse

Abstract
A new metal complex of Ni (II) with cloxacillin and cephalexin (mixed ligands), [Ni (Clox)
(Cep)Cl2] was synthesized in 1:1:1 ratio. The complex was characterized for its physicochemical
properties and spectral studies. From the IR and electronic spectral studies, the spectra of the
complex were found to be different from that of the ligands suggesting the formation of
Article History coordination compound. The ligands coordinated with the Ni (II) ion through the carbonyl group
Received: 31/07/2023 (C=O) of the carboxylate and the carboxyl groups of the amide in both the cloxacillin and
Accepted: 23/08/2023 cephalexin antibiotics. This has made the complex to assume octahedral geometry. The complex
Published: 25/08/2023
has shown good antibacterial activity. It is therefore recommended among others that;
Vol – 2 Issue – 8 pharmaceutical companies and other researchers should exploit these findings to process the
complex further to produce potent antimicrobial drugs.
PP: - 14-19
Keywords: Ligands, Complex, Synthesis, Antibacterial activity, cloxacillin, cephalexin

Introduction Many approaches (like solution-based and solid-state) were

The ever-increasing spread of bacterial infection with published for the synthesis of ligand-metal complexes and the
antibacterial drug resistance has become a matter of global various spectroscopic analyses such as IR spectroscopy, UV-
concern in the modern times. Therefore, such serious visible spectroscopy, NMR spectroscopy have been used to
infections leading to high mortality rate has necessitated elucidate the properties of complexes [6]. Various transition
search for classes of effective drugs for curing bacteria with metals with drugs molecules complexes show wide range of
minimum toxicity [9]. pharmaceutical activities, that place them in several
biochemical processes and antimicrobial agents.
It was reported that among the synthesized Co (II) and Ni (II)
coordination compounds of thiosemicarbazone has shown In this article, the solution-based synthesis of a new transition
relatively promising results against mycobacterium metal complex of mixed antibiotics (Cloxacillin and
tuberculosis. Also, antimicrobial studies of the ligands and Cephalexin), i.e. [Ni (Clox)(Cep)Cl2] was obtained by
complexes indicate that the activity increases on refluxing the antibiotics and the inorganic salt of NiCl2 and
chelation.[12] also to determine the antimicrobial activity of the complex.

Synthesis and characterization of mixture of cephalexin (cep) MATERIALS AND METHODS

and amoxicillin (amx) in 1:1 mole ratio interacted with All the reagents were of analytical grade and used without any
transition metal ions to give [ M(cep) (amx)]. 3H2O further purification. The active pharmaceutical ingredients
complexes in 1:1:1 mole ratio, where M = Co (II), Ni (II), Zn (API), cephalexin, and cloxacillin as well as Nickel (II)
(II), and Fe (III). The complexes were characterized by chloride were obtained from Sigma Aldrich Nigeria. All the
physicochemical and spectroscopic analysis. The UV/Vis glassware and other apparatus used for the synthesis of the
spectral data suggested an octahedral[13] geometry for Co complex were obtained from Department of Chemistry
(II), Ni (II), and Fe (III) structure, while Zn (II) complex Laboratory, Federal University Dutse.
adopted tetrahedral geometry. The IR spectral data shows that
the ligands coordinated by metal ions through their υ (NH.), SYNTHESIS OF METAL (II) COMPLEXES
υ(COO), and υ (C = O) functional group respectively due to An aqueous (20 ml) solution of the antibiotics (10 mmol of
the similarity in the structures. cloxacillin trihydrate (Clox) and 10 mmol of cephalexin
monohydrate (Ceph) were mixed in 1:1 mole ratio. The

*Corresponding Author: B. A. Saje © Copyright 2023 GSAR Publishers All Rights Reserved Page 14
 Global Scientific and Academic Research Journal of Multidisciplinary Studies ISSN: 2583-4088 (Online)

solution of the mixed antibiotics was further mixed with 10 ml at the National Research Institute for Chemical Technology
aqueous solution of NiCl2.6H2O, in 1:1:1 mole ratio. The Zaria. The IR spectra were recorded on the IR
reaction mixture was refluxed for 5 hours on a hot plate spectrophotometer in solid state [5].
magnetic stirrer at 50oC. The volume of each solution was UV-Vis Spectral studies
concentrated to half of the initial volume. The product
Perkin Elmer Lambda 35 UV-Visible Spectrophotometer of
obtained was allowed to cool, washed with water, diethyl
200-700nm wavelength was used for UV-absorbance
ether, and then dried in a vacuum over CaCl2 [13].
measurement and the results were recorded at the Central
Physical properties of the synthesized complex Laboratory Bayero University Kano. The UV -Vis spectra
Based on the method by [13], the melting point and were determined by UV- visible spectrometer using DMF as
temperature decomposition were carried out using Stuart solvent [2].
Melting Point apparatus (SMP 10), solubility of the ligands
Elemental Analysis (C.H.N.S)
and complex were determined in different solvents ranging
Elemental analysis technique was recorded on Euro EA
from polar to non-polar such as distilled water, methanol,
Elemental Analyser (2000oC) at Central Laboratory Umaru
ethanol, benzene, CCl4, diethyl ether, acetone, dimethyl
Musa Yar’Adua University Katsina. Also, the metals contents
formamide (DMF) and dimethyl sulfoxide (DMSO) and the
of the complexes were measured using flame atomic
molar conductivity was carried out using conductivity meter
absorption spectroscopy at the same laboratory.
of model DDS-307 in 1x10-3 at Central Laboratory Bayero
University Kano. Antimicrobial Activity Method
Sensitivity discs were punched from Whatman no. 1 filter
Magnetic Susceptibility Measurement
paper, sterilized in bijou bottles by autoclaving at 121oC for
15 minutes. Sensitivity discs were prepared by weighing
0.008 mg of the extract or fraction and serial doubling dilution
Where n = number of coordinated ligands
in DMF followed by placing the improvised paper discs in the
= mole fraction of maximum absorbance
solution such that each disc took up 0.01mL to make the disc
Magnetic susceptibility measurement of the complexes was potency of 500 g, 1000 g, 2000 g, and 4000 g.
recorded using Magnetic Susceptibility balance of Sherwood Standardized inoculate of each isolate were swabbed onto the
Scientific Cambridge UK. The prepared metal complex was surface of Mueller Hinton Agar in separate Petri dishes and
introduced into the balance’s capillary tube up to a given mark discs of the extracts and standard antibiotic (Ciprofloxacin
and the reading recorded using the magnetic susceptibility 500 g). The plates were inverted and allowed to stand for 30
balance. The formula below was used to calculate the minutes for the extract to diffuse into the agar after which the
magnetic susceptibility (Xg) [2]. plates were incubated aerobically at 35oC for 18 hours. This
was followed by measurement of zone inhibition formed by
the test organisms around each of the extract and standard
antibiotic discs [15].
Spectral studies
FTIR spectra were recorded using Fourier Transform Infrared
Spectrophotometer of Shimadzu of 4000-400 nm wavelength

RESULTS AND DISCUSSION

Table 1: Physical Properties of the Antibiotics and their Metal (II) Complex
Compound Molecular Formula Molar Colour Melting Point Decomposition
Mass o
( C) Temperature
(g/mol) (o C )

Cloxacin C19H18CIN3O5S 435.9 White 170 ___

Cephalexin C16H17N3O4S 347.4 White 197 ___

[Ni(Cep)(Clox)Cl2] C35H35Cl3N6O9S2Ni 912.19 Pale ___ 262

green

The synthesis of Ni (II) inorganic complex was completed within 1 to 2 hours. The mixed antibiotic ligands (cloxacillin and
cephalexin) produced pale green [Ni (Cep) (Clox) Cl2] complex. The colour is due to d-d transition and reactivity between the ligands
and the metal ion which is similar with results reported by [4].
The complex exhibit higher melting decomposition temperature than the individual free ligands as presented in Table 1. Cloxacillin
melted at 170oC, Cephalexin melted at 197oC while their complex decomposed at 262oC. The higher value of the mixed complex

*Corresponding Author: B. A. Saje © Copyright 2023 GSAR Publishers All Rights Reserved Page 15
 Global Scientific and Academic Research Journal of Multidisciplinary Studies ISSN: 2583-4088 (Online)

provides evidence of complexation of the ligands to the metal ion. It also revealed the more stable nature of the complex. These values
are in agreement with similar metal (II) complexes reported by [13] and [3].

Table 2: Solubility of the Antibiotics and their Metal (II) Complex

Compound Distilled CCl4 Benzene Diethyl Acetone Ethanol Methanol DMF DMSO
Water ether

Cloxacillin S IN IN SS S IN IN S S
Cephalexin S IN IN SS S IN IN S S
[Ni(Cep)(Clox)Cl2] SS IN SS SS SS IN IN S S

As presented in Table 2, cloxacillin and cephalexin are both soluble in distilled water, acetone, DMF, and DMSO, slightly soluble in
diethyl ether while insoluble in CCl4, benzene, ethanol, and methanol respectively. Ni (II) complex prepared is insoluble in CCl4,
ethanol, and methanol, slightly soluble in distilled water, benzene diethyl ether, and acetone, and soluble in DMF and DMSO. It can
be realized that the metal (II) complex is either insoluble or slightly soluble in some organic solvents such as ethanol, methanol,
diethyl ether, and acetone but soluble in DMF and DMSO. The values would suitably be used to conduct conductivity test. This is
because polar compounds are expected to be soluble in polar solvents and vice-versa (like dissolves like) which is in agreement with
the results reported by [15] and [8].

Table 3: The IR Spectra (4000-400 cm-1) of Antibiotics and their Metal (II) Complex
Compound υ(O-H) υ (C = O) υ (C = O) M–O M – Cl
of CO2 of amide

Cloxacillin 3677.85 1689.66 1771.53 -------- ------

Cephalexin 3392.90 1687.77 1757.21 ---------- -------
[Ni(Cep)(Clox)Cl2] 3398.69 1639.55 1755.28 466.79 854.49

The IR spectra data of the free ligands and their complex as presented in Table 3 were discussed and compared with each other to
predict the coordination mode and the expected final shape depending on ligand behaviour and other coordination molecules such as
chloride ions. Based on the spectra, the absorption bands at 3677.85 cm-1 in cloxacillin and 3392.90 cm-1 in cephalexin ligands is
attributed to O–H stretching which is shifted averagely to low frequency of 3398.69 cm-1 in the Ni (II) complex. This is in agreement
with coordination through the oxygen in the OH of the carboxylic group in both the two ligands. The bands at the of 1689.66 cm-1 and
1687.77cm-1 are assigned to the carbonyl group (C=O) of carboxylate groups in cloxacillin and cephalexin ligands respectively which
were shifted to different frequencies of 1639.55cm-1 in [Ni (Cep)(Clox)Cl2], indicating occurrence of complexation through carboxyl
of the carboxylate group with the metal ion. The bands at 1771.53cm-1 and 1757.21cm-1 are assigned to the carboxyl group of amide in
the ligands respectively which clearly differs from 1757.21cm-1 in the complex. The shifting of values in the complex shows the
complexation through the carboxyl of amide with metal ion under investigation as reported by Waziri et al., [12]. The complex [Ni
(Cep)(Clox)Cl2] indicated new bands at 466.79 and 854.49 cm-1 which are assigned to υ (M – O) and υ (M – Cl), which could not be
traced in the free ligands proves the formation of the new compound as reported by [6].

Table 4: Electronic Spectra, Conductance in DMF Solvent and Magnetic Moment (B.M) for Synthesized Metal Complex with
its suggested geometry
Ligands/Complex Electronic spectra Conductance Magnetic Suggested
Wavelength Energy Transition ( cm2mol-1) moment geometry
( ) ( ) (BM)
Cloxacillin 202 48309
207 49505
239 41841 CT band
Cephalexin 220 45455
233 42918
[Ni (Cep)(Clox)Cl2] 221 45249 MLCT 20.00 2.98 Octahedral
3 3
230 43478 A2g T2g
3 3
237 42194 A2g g

*Corresponding Author: B. A. Saje © Copyright 2023 GSAR Publishers All Rights Reserved Page 16
 Global Scientific and Academic Research Journal of Multidisciplinary Studies ISSN: 2583-4088 (Online)

The molar conductance in DMF is presented in Table 4. Conductivity measurements have been used in elucidating the
structure of complexes within the limits of their solubilities in a given solvent as reported [1] which suggested that the values of less
than < 60 in DMF are considered to be non-electrolytes. The Ni (II) complex has molar conductance of 20 which
indicates that it is a non-electrolyte which lies within octahedral geometry in DMF as reported [1] and [6].
The electronic spectra of both the ligands and their complex are presented in Table 4. The free ligand of cloxacillin exhibits three
bands at 202 nm, 207 nm, and 239 nm while the free ligand of cephalexin exhibits two bands of 220 nm and 233 nm respectively.
These were assigned in the assignment of , , CT- band, . These absorption bands also appeared in
the electronic spectra of the complex, but they are shifted to different values which proves the coordination of the ligands to the central
metallic ions [11]. Ni (II) complex shows three bands at the regions (221 nm, 43478 cm-1), (230 nm, 43478 cm-1) and (237 nm, 42194
cm-1) which are attributed to metal to ligand charge transfer (MLCT), 3A2g 3T2g, and 3A2g 3T1g transitions for an octahedral Ni
(II) complex [10].
Table 5: C.H.N.S. Elemental Analysis and Metal Content of the Synthesized Complex
Compounds Molar Mass Elemental and metal analysis found (calculated) %
(g/mol)
C H N S M

Ni(Cep)(Clox)Cl2] C35H35N6S2O9Ni 46.07 3.68 8.96 5.52 8.00

(805.69) (52.13) (4.34) (10.43) (7.9) (7.28)

The result of the elemental analysis of the Ni (II) complex is presented in Table 5. From the result obtained, the percentage of C, H, N,
and S are in conformity with the proposed structure. Based on the evaluation of the elemental analysis data, it shows that the
compound analysed as [Ni((Cep)(Clox)Cl2)] which shows the coordination of the metal to the two ligands as 1:1:1. The percentage of
the metal ion also agree with the proposed structure. The result is in line with that of [7] and [14].

Table 6: Antibacterial test of the Antibiotics and their Metal (II) Complex
Compounds Concentration Staph. Aureus Salmonella typhi E. Coli
( (mm) (mm) (mm)

Cloxacillin 4 000 11 10 10
2 000 10 11 11
1 000 12 8 8
500 9 7 7
Cephalexin 4 000 11 11 10
2 000 9 11 11
1 000 7 9 9
500 00 8 7
[Ni(Cep)(Clox)Cl2] 4 000 16 15 17
2 000 15 14 15
1 000 14 15 13
500 8 00 7
Standard
Ciprofloxacin 500 30 27 25
200 - - -
observed that the nickel (II) complex of the mixed antibiotics
From Table 6, the result of the antibacterial test for the
has shown a better antibacterial activity than the individual
cloxacillin and cephalexin as well as their metal (II) complex
antibiotics (ligands). Similar result was reported by [6].
are presented. The result shows that the free ligands as well as
the mixed complex were active against all the three isolates of Based on the analytical data obtained such as melting point,
bacteria in all concentrations especially at higher ones. UV-spectroscopy, elemental analysis, magnetic susceptibility,
However, at lowest concentration of 500 | agar-well, and FTIR spectral studies, the tentative proposed structure of
inactivity was observed by cephalexin ligand against the metal (II) complex of the mixed antibiotics can be
staphylococcus aureus. Likewise, the complex exhibited illustrated as follow:
inactivity against Salmonella typhi isolate. It can be

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 Global Scientific and Academic Research Journal of Multidisciplinary Studies ISSN: 2583-4088 (Online)

quinolin-2-ol and its Metal Complexes and their

Evaluation for Antibacterial and Antifungal
Activity. Algerian Journal of Engineering and
Technology. 02; 218-231.
5. Jibril, S., Sani, S., Kurawa, M. A., & Shehu, S. M.
(2019). Mechanochemical synthesis,
characterization, and antimicrobial screening of
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Bayero Journal of Pure and Applied Sciences,
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Cloxacillin and cephalexin are among the interesting 6. Jibril S., Baraya K. A., Muhammad S., Musa I.,
antibiotics which are used as ligands in synthesizing metallic Garba A., Adamu S. B., and Bello I. (2021). Solid
complexes to improve their activity by some researchers using Synthesis, Spectral and Biological Studies of Fe (II)
refluxing solution-based synthesis. This work has Complex Derived from Paracetamol. J.Chem. Soc.
demonstrated the use of such method to obtain similar result Nigeria, Vol 46, No. 6, pp 1068-1074.
with effective antibacterial activity. 7. Lawal, A., Ayanwale, P.A, Obaleye J.A, Rajee,
A.O, Babamale, H. F. and Lawal, M. (2017);
From the values obtained in conductivity, magnetic
Synthesis, Characterization and Biological Studies
measurement, and spectral studies, octahedral geometry is
of Mixed Ligand Nicotinomide-Trimethoprim
suggested. In the spectral studies of both IR and UV-Visible
Complexes. International Journal of Chemical
spectroscopy, the ligands (antibiotics) are coordinated to the
material and Environmental Research. 4(1) 97-101
metal ion to form the complex of [Ni (Cep)(Clox)Cl2] through
8. Mustapha, A. N., Ndahi, N. P., Paul B. B., & Fugu
the υ (C = O) carboxylic acid and that of amide giving rise to
M. B. (2014). Synthesis, characterization, and
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Co (II) and Mn (II) Complexes with Ciprofloxacin
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