2011 International Conference on Chemistry and Chemical Process

IPCBEE vol.10 (2011) © (2011) IACSIT Press, Singapore

Synthesis, Characterization and Biological Activity of Schiff Bases

Muhammad Aqeel Ashraf, Karamat Mahmood, Abdul Wajid

Department of Chemistry, Faculty of Science
The Islamia University of Bahawalpur, 63100 Pakistan
Currently studying at Department of chemistry, University of Malaya, 50603 Kuala Lumpur, Malaysia
[email protected]
Mohd. Jamil Maah, Ismail Yusoff
Department of Chemistry, Faculty of Science
Department of Geology, Faculty of Science
University of Malaya 50603, Kuala Lumpur, Malaysia
Abstract. Three new series of biologically active amino substituted Schiff bases with general formula,
R1N=CHR2. Here R1 = 2-amino-benzthiazole, 4-amino-salicylic acid and 4-aminophenol. R2 =4-chloro-
benzaldehyde, 2-chloro-benzaldehyde, salicylaldehyde, vanillin and benzaldehyde were synthesized by the
reaction of three different amino substituted compounds and substituted aldehydes in ethanol. Such
compounds were characterized by different physico-chemical techniques like, melting point, elemental
analysis, multinuclear NMR (1H, 13C). The free ligands and their metal complexes have been screened for
their in vitro biological activities against bacteria, fungi and yeast. The metal complexes show more potent
activities compared with Schiff base ligands.
Keywords: schiff bases; benzthiozol; aminophenol; antibacterial; antifungal.

1 Introduction
Schiff bases are condensation products of primary amines with carbonyl compounds and they were first
reported by Schiff [1] in 1864. The common structural feature of these compounds is the azomethine group
with a general formula RHC=N-R1, where R and R1 are alkyl, aryl, cyclo alkyl or heterocyclic groups which
may be variously substituted. These compounds are also knows as anils, imines or azomethines. Several
studies [2-8] showed that the presence of a lone pair of electrons in an sp2 hybridized orbital of nitrogen atom
of the azomethine group is of considerable chemical and biological importance. Because of the relative
easiness of preparation, synthetic flexibility, and the special property of C=N group, Schiff bases are generally
excellent chelating agents,[6-12] especially when a functional group like –OH or –SH is present close to the
azomethine group so as to form a five or six membered ring with the metal ion. Versatility of Schiff base
ligands and biological, analytical and industrial applications of their complexes make further investigations in
this area highly desirable.
Schiff bases have been known since 1864 when Hugo Schiff reported the condensation of primary
amines with carbonyl compounds (1). Nowadays, the research field dealing with Schiff base coordination
chemistry has expanded enormously. The importance of Schiff base complexes for bioinorganic chemistry,
biomedical applications, supramolecular chemistry, catalysis and material science, separation and
encapsulation processes, and formation of compounds with unusual properties and structures has been well-
recognized and reviewed (2).
Schiff bases resulted from aromatic aldehydes ortho-substituted with a hydroxyl group have initially
arouse the researchers' interest because of their ability to act as bidentate ligands for transitional metal ions
1
[16-20]. Later, in studies concerning quantitative structure-antitumor activity relationship of a series of Schiff
bases derived from variously substituted aromatic amines and aldehydes, it has been shown that azomethines
from salicylaldehydes gave the best correlation [21, 22]. Schiff bases of salicylaldehydes have also been
reported as plant growth regulators [23] and antimicrobian [24] or antimycotic [25] activity. Schiff bases also
show some analytical applications [26]. Schiff Bases are characterized by the -N=CH- (imine) group which
imports in elucidating the mechanism of transamination and rasemination reaction in biological system [27,
28]. Schiff bases are active against a wide range of organisms for example; Candida Albicans, Escherichia
coli Staphylococcus aureus, Bacillus polymxa, Trychophyton gypseum, Mycobacteria, Erysiphe graminis and
Plasmopora viticola.
A large number of different Schiff base ligands have been used as cation carriers in potentiometric sensors
as they have shown excellent selectivity, sensitivity, and stability for specific metal ions such as Ag(II),
Al(III), Co(II), Cu(II), Gd(III), Hg(II), Ni(II), Pb(II), Y(III), and Zn(II) [29-34]. Schiff bases have been
studied for their important properties in catalysis [35]. They show catalytic activity in hydrogenation of olefins
[36]. They find applications in biomimetic catalytic reactions.
An interesting application of Schiff bases is their use as an effective corrosion inhibitor, which is based on
their ability to spontaneously form a monolayer on the surface to be protected. Many commercial inhibitors
include aldehydes or amines, but presumably due to the C=N bond the Schiff bases function more efficiently
in many cases [37]. The principal interaction between the inhibitor and the metal surface is chemisorption [38].
The inhibitor molecule should have centers capable of forming bonds with the metal surface by electron
transfer. In such cases the metal acts as an electrophile and the inhibitor acts as a Lewis base. Nucleophilic
centers, such as oxygen and nitrogen atoms, of the protective compound have free electron pairs which are
readily available for sharing. Together with the atoms of the benzene rings they create multiple absorption
sites for the inhibitor thus enabling stable monolayer formation [39].
Imines also have biological importance. An imine linkage between the aldehyde derived from vitamin A
and the protein opsin in the retina of the eye plays an important role in the chemistry of vision. Vitamins are
also called coenzymes, meaning that they are to the functioning of many enzymes, which are large proteins
that catalyze chemical changes in cell. An example of a biologically important aldehyde is pyridoxal
phosphate, which is the active form of the vitamin B6. Vitamin B6 serves as a coenzyme by forming an imine
with an amino acid grouping an enzyme. The coenzyme, bound to the enzyme, is involved in transamination
reaction, the transfer of the amino group from one amino acid to another, which is important in the
metabolism and the biosynthesis of amino acids. In the last step, enzyme-catalyzed hydrolysis cleaves the
imine to pyridoxal and the modified amino acid.
Schiff bases have been reported in their biological properties, such as, antibacterial, antifungal activities
[40-43]. Their metal complexes have been widely studied because they have anticancer and herbicidal
applications [44, 45]. They serve as modals for biologically important species.
O-phenylenediamine Schiff bases show clinical properties [46]. Isatin Schiff bases were reported to
possess antiviral, anti-HIV, antiprotozoal and anthelmintic activities [47]. They also exhibit significant
anticonvulsant activity, apart from other pharmacological properties [48]. Certain cobalt Schiff base
complexes are potent antiviral agents [49]. Schiff bases derived from 4-dimethylamine benzaldehyde shows
antibacterial activity. In medicines used as antibodies and anti-inflammatory agents [50-54]
This paper presents a series of new Schiff bases with a potential biological activity resulted from the acid-
catalyzed condensation of aryl aldehydes with aromatic and heteroaromatic amines. These compounds could
also act as valuable ligands. The structures of the Schiff bases synthesized from 2-amino-Benzthiazole, 4-
amino-Salicylic acid and 4-aminophenol are shown in scheme 1:
OH

HO COOH
N
R1 : H, OH, Cl
N R2 : H, OCH 3
R3 : H, OH, Cl
N
S N CH CH
CH
R1 R1
R1

R2 R2 R2

R3 R3 R3

2
2 Material and Methods
Schiff bases' melting points were taken on a Stuart Melting point apparatus SMP-3 and are uncorrected.
Elemental analysis was carried out at Fisons EA 1108 CHNSO Micro analyzer, lH and 13C NMR spectra were
determined in DMSO (internal standard TMS) on Bruker spectrometer.
2-amino-benzthiazole, 4-amino-salicylic acid, 4-aminophenol, 4-chloro-benzaldehyde, 2-chloro
benzaldehyde, Salicylaldehyde, Vanillin, Benzaldehyde were purchased from Fluka and used without further
purification. All organic solvents were purchased from Merck.

2.1 Synthesis of 2-amino-benzthiazole Schiff Bases

2g of 2-amino-benzthiazole was mixed with equivalent amount of corresponding aldehyde in 25 ml of
ethanol. The resulting mixture was left under reflux for 2 h and the solid product formed was separated by
filtration, purified by recrystallization from ethanol, washed with ethanol, and then dried.

2.2 Synthesis of 4-amino-salicylic acid Schiff Bases

2g of 4-amino-salicylic acid was mixed with equivalent amount of corresponding aldehyde in 25 ml of
ethanol. The resulting mixture was left under reflux for 2 h and the solid product formed was separated by
filtration, purified by recrystallization from ethanol, washed with ethanol, and then dried.

2.3 Synthesis of 4-aminophenol Schiff Bases

2g of 4-aminophenol was mixed with equivalent amount of corresponding aldehydes in 25 ml of ethanol.
The resulting mixture was left under reflux for 2 h and the solid product formed was separated by filtration,
purified by recrystallization from ethanol, washed with ethanol, and then dried.

2.4 Biological Activity

The synthesized Schiff bases were screened for antibacterial and antifungal activity.

2.5 Antibacterial Testing

The bacterial cultures for B. subtilis, S. aureus, and E. coli were obtained from Department of
Microbiology University of Malaya, Kuala Lumpur, Malaysia. The bacterial cultures were incubated at 30 ±
0.1oC for 24 hours by inoculation into nutrient agar. Schiff bases were stored dry at room temperature and
dissolved 20mg/ml in dimethylsulfoxide (DMSO). Antibacterial activities of each compound were evaluated
by the agar disc-diffusion method. Mueller Hinton Agar Media (15 cm3) kept at 45oC was poured in the petri-
dishes and allowed to solidify. Poured Petri plates (9 cm) were incubated with 50µL of normal saline solution
of above culture media (105-106 bacteria per ml). Discs injected with prepared Schiff bases (50µL) were
applied on the solid agar medium by pressing tightly. The Petri plates were placed at 37oC for 24 hours. At the
end of period the inhibition zones formed on media were measured with a zone reader in millimeters.

Figure 1. showing zone of inhabitation against subtilis, S. aureus, and E. coli

2.6 Antifungal Testing

Pathogenic strains of Asperigillus niger and Chalara corda were obtained from Department of
Microbiology University of Malaya, Kuala Lumpur, Malaysia. Schiff bases were stored dry at room
temperature and dissolved 20mg/ml in dimethylsulfoxide (DMSO). Antifungal activities of each compound
were evaluated by the agar disc-diffusion method. Sabarod’s agar media (15 cm3) kept at 45oC was poured in
the petri-dishes and allowed to solidify. Sterile, filter paper discs of 10mm diameter were impregnated with
prepared Schiff bases (50µL) and were placed on to the media, seeded with fungus. The plates were then
3
incubated at 27oC for 1-7 days. At the end of period the inhibition zones formed on media were measured with
a zone reader in millimeters.

3 Results and Discussion

3.1 Scheme 1
Twelve new Schiff bases have been synthesized from the condensation of 2-amino-Benzthiazole, 4-
amino-Salicylic acid and 4-aminophenol with 4-chloro-benzaldehyde, 2-chloro-benzaldehyde, salicylaldehyde,
vanillin and benzaldehyde (Scheme 2, 3, 4). The analytical and physical data are listed in Table 1.

Scheme 2 Scheme 3
O N
O HO COOH
CH
S N CH CH
R1
N Reflux R1 N
R1 HO COOH
Ethanol Reflux
CH
S NH 2 Ethanol
R2
R2 H2N R2 R1
R3
R3 R3
R1 : H, OH, Cl R1: H, OH, Cl R2
R2 : H, OCH3 R2: H, OCH3
R3 : H, OH, Cl R3: H, OH, Cl R3

Scheme 4
O OH

OH CH
R1 N
Ref lux
CH
Ethanol
R2 R1

NH2 R3
R 1: H, OH, Cl R2
R 2: H, OCH3
R 3: H, OH, Cl R3

1
TABLE I. PHYSICAL DATA OF SCHIFF BASES WITH GENERAL TABLE II. H NMR DATA A-E OF COMPOUNDS WITH GENERAL
FORMULA R1N=CHR2. FORMULA R1N=CHR2

3.2 NMR Spectroscopy

1
H NMR
1
H NMR spectral data in deturated DMSO solution of the synthesized compounds are given in Table 2
The resonance of protons has been assigned on the basis of their integration and multiplicity pattern. The 1H
NMR spectra of the Schiff bases in DMSO exhibits signals at 9.218, 9.50, 9.4, 9.22 and 9.36ppm for
compounds 1, 2, 3, 4, and 5, attributed to CH=N- protons, respectively. The multisignals within the 6.89-
4
8.1ppm range are assigned to the aromatic protons of both rings. The 1H NMR spectra of the Schiff bases
synthesized from Salicylic acid revealed a signal at 10.39 and 10.26ppm due to the -CH=N- group on
compounds 6 and 7 respectively. The OH moiety of Salicylic acid was observed at 9.04 and 8.97ppm. It
should be noted that the phenolic protons have always given a singlet in off-set at high δ values, thus
confirming its involvement in an intramolecular hydrogen bond with the neighboring nitrogen atom [55]. The
free NH2 protons usually show a broad singlet peak in a region at 4-6ppm [56]. This signal is absent in the
observed spectra of Schiff bases which indicates the formation of the Schiff bases.
13
C NMR
The 13C NMR spectra provide further support for the structural characterization of the Schiff bases. 13C
NMR spectral data of compounds (1-12) have been listed in Table 3. The number of signals found
corresponds with the presence of magnetically nonequivalent carbon atoms, which were assigned by
comparison with literature values. The aromatic carbon present in the structures of Schiff bases were assigned
by comparing the experimental chemical shifts with those calculated from the incremental method [57].The
13
C-NMR spectral data of the Schiff bases are in accord with the proposed structures.

3.3 Antibacterial activity

The results of the antibacterial screening of the Schiff bases at a concentration of 20mg/ml against all
bacteria have been found. The inhibition zones were measured in mm and results are shown in Table 4. The
results of antimicrobial screening, indicate that Schiff bases show significant activity against Staphylococcus
aureus, Escherichia coli, Bacillus subtilis than Aspergillus niger and Chalara corda while compound 1, 2 were
found to be more active against all tested bacterial strains because of the presence of chloro group in the
aldehydic group which itself is active against microbes.
Antibacterial activity of these compounds show ascending order. When we increase concentration, area of
inhibited growth also increased.

3.4 Antifungal activity

From the results obtained by the antifungal activity it is found that the benzthiazole Schiff bases are more
active against all tested fungi then the salicylic acid Schiff bases. Compound 1, 2, 3, 4 and 5 are the most
potent candidates against all type of tested fungi. The greater activity of these compounds is probably due to
the presence of benzthiazole moiety. Compound 7 show good activity against all tested fungi as compared to
standard drug. Compound 6 is significantly active against Aspergillus niger. The antifungal activity results are
shown in Table 5.
13
TABLE III. C NMR DATA A-E OF COMPOUNDS WITH GENERAL FORMULA R1N=CHR2

TABLE IV. ANTIBACTERIAL ACTIVITY DATAA-C OF COMPOUNDS WITH GENERAL FORMULA: R1N=CHR2 (IN VITRO)

5
 TABLE V. ANTIFUNGAL ACTIVITY DATAA-D OF COMPOUNDS WITH GENERAL FORMULA: R1N=CHR2 (IN VITRO)

4 Conclusion
Schiff bases of 2-amino-Benzthiazole, 4-amino-Salicylic acid and 4-aminophenol were synthesized and
characterized by analytical and spectral techniques. These compounds exhibited significant activity against all
the tested microorganisms.

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