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Asian Journal of
Pharmaceutical Sciences
and Clinical Research
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Asian Journal of Pharmaceutical Sciences and Clinical Research (AJPSCR) Vol. 1, Issue 1 (2011), 6-12

RESEARCH ARTICLE

ANTIMICROBIAL SCREENING AND CHARACTERIZATION OF SOME NEWLY

SYNTHESIZED MANNICH BASES OF CIPROFLOXACIN: A GREEN CHEMISTRY
APPROACH

Atul R. Bendale*1, Nilam Dhonde2, Sushil P. Narkhede2, Sachin B. Narkhede2, Anil G. Jadhav2,
G. Vidyasagar3
1
Suresh Gyan Vihar Universe, Jaipur, Rajsthan, India
2
Smt. B. N. B. Swaminarayan Pharmacy College, Salvav (Vapi), Gujarat, India
3
Department of Pharmacy, Kutch University, Bhuj, Gujarat, India

Received 14 March 2011, Received in revised form 21 June 2011, Accepted 28 June 2011

Abstract
Herein we profitably joined some aldehydes to the triazole group through the Schiff reaction to give
different Schiff bases. Ciprofloxacin was incorporated to the new series of Schiff bases of 1, 2, 4- triazole
via Mannich reaction. The new compounds have been evaluated in vitro for their antimicrobial activity
against Staphylococcus Aureus and E.coli. All the compounds showed in vitro gram positive and gram
negative activity which was comparable or superior to that of activity of parent drug (ciprofloxacin).

Keywords: Ciprofloxacin, Schiff bases, Mannich bases, antimicrobial

Introduction:
The Mannich reaction is an organic reaction which consists of an amino alkylation of an acidic
proton placed next to a carbonyl functional group with formaldehyde and ammonia or any
primary or secondary amine. The final product is a β-amino-carbonyl compound also known as a
Mannich base. Reactions between aldimines and α-methylene carbonyls are also considered
Mannich reactions because these imines form between amines and aldehydes.
______________________________________________________________________________
* Corresponding author. Tel: +91 8000701337
E-mail address: atulbendale123@gmail.com (Atul R. Bendale)
© Pharmaboon. All rights reserved.

Atul Bendale et al.: Asian Journal of Pharmaceutical Sciences and Clinical Research, Vol.1, Issue1 (2011), 6-12
 Asian Journal of Pharmaceutical Sciences and Clinical Research (AJPSCR) Vol. 1, Issue 1 (2011), 6-12

The reaction is named after chemist Carl Mannich. The essential feature of the reaction is the
replacement the active hydrogen atom by an amino methyl or substitute amino-methyl
group.[1,2] Ciprofloxacin is a quinolone antibiotic drug used mainly to treat the respiratory
infections (pneumoniae, pseudomonas, influenzae), urinary tract infections, the gastrointestinal
surgery, typhoid fever, gonorrhoea (enterotoxigenic strains of E. coli),
and septicaemia. Ciprofloxacin act by inhibiting the bacterial enzymes DNA gyrase [3].

Ciprofloxacin is the most potent first generation of fluoroquinolones. In the 1990s, compounds
with additional flouro and other substitutions have been developed-further extending
antimicrobial activity to gram-positive cocci and anaerobes, and/or conferring metabolic stability
(longer t½), these are referred as ‘second generation’ fluoroquinolones. The inhibition of DNA
gyrase and cell permeability of fluoroquinolones is greatly influenced by nature of C- 7
substituents on the standard structure of 4-Quinolones-3-Carboxylic acid. In addition, bulky
functional groups substitution is permitted on C-7 position.[4-6] Structure activity relationship of
antibacterial fluoroquinolones has been extensively investigated and the substituent at the C-7
position has a great impact modulating potency, spectrum, bio-pharmaceutics and
pharmacokinetics. The piperazine moiety of 7 piperazinyl quinolnes possesses enough structural
flexibility to allow product optimization. In addition, the C-7 substituent affects the interaction
with the target and both the activity spectrum and kinetic profile can be controlled at C-7. [7-11]
Since our strategy is to achieve a better antimicrobial profile of ciprofloxacin, it has been
planned to incorporate Schiff bases of compounds into the piperazine ring of the ciprofloxacin
via Mannich reaction and to screen antimicrobial activity for these compounds.

Materials and methods:

Ciprofloxacin was procured as gift sample from S Kant Health Care Ltd. India. All the chemicals
and solvents used in studies were of GR grade, dried and purified before use. Melting points
were obtained using capillary method in paraffin bath and are uncorrected. The purification of
synthesized compounds was performed by recrystallization with appropriate solvent system.
Infrared spectra were recorded on FTIR spectrophotometer 8400S, Shimadzu corporation, Mass
spectra were recorded in QP-2010 PLUS GC-MS system. Nuclear Magnetic Resonance spectra
were recorded with AVANCE 300 MHz , using CDCl3 and D2O. The purity of the compounds
was checked using TLC technique; spots were developed by exposure to iodine vapors and UV
cabinet.

Experimental:
Scheme I (Synthesis of schiff base)
Synthesis of 2-methoxy-4 {(E)-[(4-methylphenyl)imino]methyl]} phenol :
0.1 Mole of p-toludine dissolved in 5ml methanol and in another beaker 0.1 Mole of vaniline
dissolved in 5ml methanol. Both the contents are mixed in a beaker; a drop of acetic acid is
added as a catalyst and beaker place in sonicator at 45oC for 9-10 min. A pale yellow coloured
product is formed which indicated formation of product. The synthesized product recrystalized
by shock cooling method, using Ethanol as a solvent to get fine crystals of 2-methoxy-6-[ (4-
methylphenyl) imino] methylphenol (2-M-4-MPIMP). The reaction monitored by TLC and
confirmed by IR. [12]
Atul Bendale et al.: Asian Journal of Pharmaceutical Sciences and Clinical Research, Vol.1, Issue1 (2011), 6-12
 Asian Journal of Pharmaceutical Sciences and Clinical Research (AJPSCR) Vol. 1, Issue 1 (2011), 6-12

NH2 CHO H3C O

-H2O
+ HO

O CH3 N

CH3 OH
p-toluidine 4-hydroxy-3-methoxybenzaldehyde
CH3
2-methoxy-4-{(E)-[(4-methylphenyl)imino]methyl}phenol
( Comp:I)
Fig. 1 Synthesis of Schiff Base

Scheme II (a, b, c) (Synthesis of mannich bases)

1. Synthesis of Formaldehyde- ciprofloxacin Mannich base (IIa)
2. Synthesis of Salicylaldehyde- ciprofloxacin Mannich base (IIb)
3. Synthesis of Benzaldehyde- ciprofloxacin Mannich base (IIc)

Procedure: [12]
Equimole quantity of- Schiff base (I), appropriate aldehyde and ciprofloxacin were dissolved
separately in 10 ml of ethanol. Mix well and stirred at room temperature for 6 hr. The resulting
product was concentrated by heating on water bath. Coloured solid precipitate was collected and
recrystallized with hot ethanol. Analytical data of synthesized compounds is summarized in
Table-2
Antimicrobial Evaluation: [13,14]
All the synthesized compounds were screened for their antibacterial activity against
Staphylococcus aureus and E.coli by using disc diffusion method. Bacteria were cultured in
nutrient agar medium and used as inoculum for study. The test compounds were dissolved in
DMF to obtain a solution of 10µg/ml concentration. All the compounds exhibited moderate
activity against bacteria. The data are given in Table-4.

H3C O

HO
N
COMP:1 + H CHO + Ciprofloxacine N
N N
formaldehyde -H2O

F COOH
O
CH3
IIa

Fig. 2 Synthesis of Mannich Base IIa

Atul Bendale et al.: Asian Journal of Pharmaceutical Sciences and Clinical Research, Vol.1, Issue1 (2011), 6-12
 Asian Journal of Pharmaceutical Sciences and Clinical Research (AJPSCR) Vol. 1, Issue 1 (2011), 6-12

H 3C O

CHO OH
HO
OH N
N
COMP: I
+ + Ciprofloxacine N N
- H2O
salicy la ld e h yd e
F COOH
O
CH3 II b
IIb

Fig. 3 Synthesis of Mannich Base IIb

H 3C O

CHO
HO
N
COMP: I + + Ciprofloxacine N
N N

benzaldehyde - H2O
F COOH
O
CH 3
II c
IIc
Fig. 4 Synthesis of Mannich Base IIc

Compound No. Mobile phase

I Benzene : Acetone (4.0 :1.0)
IIa Benzene : Acetone (4.0 :1.0)
IIb Benzene : Acetone (4.0 :1.0)
IIc Benzene : ethyl acetate (4.0 :1.0)
Table 1: Mobile phase used for determination of Rf value of synthesized compounds.

Result:
Mole.
Molecular Melting % C, H, N Analysis:
Comp. Weight Rf Value
Formula Point (0C) Yield
(gm/mol)
I C15H15N1O2 241.16 220-2220C 96 % 0.79 -
IIa C33H37FN4O5 588.27 280-2820C 82 % 0.85 67.33, 6.34, 9.52
IIb C39H41FN4O6 680.76 262-2640C 91 % 0.63 68.81, 6.07, 8.23
IIc C39H41FN4O5 664.31 284-2860C 78 % 0.75 70.46, 6.22, 8.43

Table 2: Analytical Data for compounds

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 Asian Journal of Pharmaceutical Sciences and Clinical Research (AJPSCR) Vol. 1, Issue 1 (2011), 6-12

Spectral data for synthesized coumpound:

Comp IIa:
I.R. (KBr, cm-1) : 3652.68 (Free –OH stretching), 3310 (NH stretching), 1720.35 (C=N
stretching), 1628.75 (-COOH stretching), 1498.66 (Aromatic group), 1388.47 ( -CH2 –Bending),
1125.31 ( -C=O –stretching), 1038.95 (-C-F - stretching), 892.82 (-C-C stretching).
NMR (CDCl3) : 5.35 (aromatic C–OH), 11.0 (1 OH, carboxylic acid), 1.35 (CH, cyclopropane),
3.44 (2 H, methylene), 6.09 (CH, 1- benzene), 4.10 (CH), 3.83 (3 H, methyl).
MS (m/z): 588.27 (100%), 589.28 (36.3%), 590.28 (8.0%), 589.27 (1.5%), 591.28 (1.2%).

Comp IIb:
I.R. (KBr, cm-1): 3753.40 (-OH stretching), 3529.20 (10 –NH stretching), 3384.94 (C=O
stretching), 2619.36 (S-H - stretching), 1711.08 (-C=N - stretching), 1626.98 (-COOH
stretching), 1496.39 (Aromatic group), 1385.45 (-CH2 - stretching), 1217.11 (-OH - Bending),
1026.13 (C-F - stretching), 893.16 (C-C -stretching).
NMR (CDCl3): 5.35 (CH, aromatic C-OH), 11.0 (OH, carbocylic acid), 1.35 (CH,
cyclopropane), 3.13 (2 H, methylene), 6.97 (CH, benzene), 4.10 (CH, methine), 2.34 (3 H,
methyl).
MS (m/z): 680.30 (100%), 681.30 (43.7%), 682.31 (10.2%), 683.31 (1.8%).

Comp IIc:
I.R. (KBr, cm-1) : 3652.68 (Free –OH stretching), 3310 (NH stretching), 1720.35 (C=N
stretching), 1628.75 (-COOH stretching), 1498.66 (Aromatic group), 1388.47 ( -CH2 –Bending),
1125.31 ( -C=O –stretching), 1038.95 (-C-F - stretching), 892.82 (-C-C stretching).
NMR (CDCl3) : 5.35 ( Aromatic C-OH), 11.0 (OH-carboxylic acid), 1.35 (CH-cyclopropane),
0.80 (2H-1 α-N from methane), 3.13 (2H, methylene), 6.09 (6H- benzene), 3.83 (3H, methyl).
MS (m/z): 664.31 (100%), 665.31 (42.8%), 666.31 (10.4%), 665.30 (1.5%), 667.32 (1.2%).

Result of antibacterial activity: (Zone of Inhibition in mm)

Concentr- Staphylococcus aureus E. coli
Sr
ation
No. IIa IIb IIc Ciprofloxacin IIa IIb IIc Ciprofloxacin
(µg/ml)
1 10 29.3 29.5 27.7 27.5 35.3 37.4 35.4 34.8
2 20 30.5 31.7 29.8 29.7 38.7 37.6 36.7 36.2
3 30 31.4 31.9 31.5 30.2 38.1 38.5 37.0 36.9

Table 4. Antibacterial activity of synthesized compounds

Discussion and Conclusion:

Mannich bases of ciprofloxacin were synthesized by modification of Schiff base 2-methoxy-4
{(E)-[(4-methylphenyl)imino]methyl]}phenol by using different aldehydes.

Atul Bendale et al.: Asian Journal of Pharmaceutical Sciences and Clinical Research, Vol.1, Issue1 (2011), 6-12
 Asian Journal of Pharmaceutical Sciences and Clinical Research (AJPSCR) Vol. 1, Issue 1 (2011), 6-12

Herein we referred greener processes of synthesis for Schiff and Mannich bases. From which
compounds IIa, IIb and IIc were synthesized in good percentage yield, i.e. 82 %, 91 % and 78 %
respectively. Among them compound IIb was formed with very good yield. Zone of inhibition of
compound IIb was larger than that of ciprofloxacin as well as than other two compounds IIa and
IIc. It is worthwhile from above discussion, Mannich base of Salicylaldehyde with Ciprofloxacin
have good yield and better antimicrobial activity rather among others.

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 Asian Journal of Pharmaceutical Sciences and Clinical Research (AJPSCR) Vol. 1, Issue 1 (2011), 6-12

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