DOI: 10.

26524/sa1
FULL LENGTH ARTICLE

Microwave Synthesis and Characterization of Antibacterial Activity

Evaluation of 2-(4, 5-dihydro-5-(4-chlorophenyl)-1H-pyrazol-3-yl)
phenol
L. Ilavarasan a, A. Ravi a*, M. Ganapathi a, E. Subitchayini a, K. Sivasakthi a, R. Sapthagiri a,
J. Vinayagam b, K. Mohanraj c, Tamiloli Devendhiran d, Keerthika Kumarasamy d
a
PG & Research Department of Chemistry, Government Arts College, Tiruvannamalai. 606 603, Tamilnadu, India.
b
Department of Chemistry, National Tsing Hua University, Taiwan 30013, R.O.C.
c
Raman Research Laboratory, PG & Research Department of Physics, Government Arts College, Tiruvannamalai. 606 603,
Tamilnadu, India.
d
Department and Graduate Institute of Applied Chemistry, Chaoyang University of Technology, Taichung City-41349, Taiwan
(R.O.C).

*Corresponding Author ABSTRACT: Chalcone derivatives were synthesized by reaction of some benzaldehyde
draravichemetal@[Link] derivatives with acetophenone, then the products obtained were allowed to react with
(A. Ravi) urea, thiourea and hydroxylamine, to give the heterocyclic derivatives of oxazine,
Tel.: +91 9443103974 thiazine and isoxazole, [Link] this study, a series of chalcones and substituted
pyrazole compounds were synthesized according to green chemistry methods of
Received : 01-08-2018 conventional and microwave irradiation by using substituted acetophenone,
Accepted : 18-10-2018
substituted benzaldehyde, hydrazine hydrate and PEG-400. The synthesized
compounds were characterized by UV-Visible, FT-IR and 1H NMR spectral techniques.
The purity of the synthesized compounds were monitored by TLC and tested for their
antibacterial activity by Minimum Inhibitory Concentration (MIC) method against two
different microorganisms Staphylococcus auras (MTCC3381), and Escherichia coli
(MTCC739).
.
Keywords: Chalcones, Pyrazoles, antibacterial activity, PEG-600, MWI, Synthesis

Introduction
Heterocycles are abundant in nature and are of drug design [8-10]. A survey of the literature revealed that
great significance to life because their structural subunits some natural and synthetic chalcones showed significant
exist in many natural products such as vitamins, ALR2 inhibitory activities and this prompted us to
hormones, antibiotics etc [1-5]. Identification of novel investigate potential ARIs derived from chalcone-based
compounds which treat both infectious and inflammatory compounds [11-15]. Thus, we focused on the compounds
states more effectively and which lack side effects having a carboxylic acid moiety that was incorporated into
associated with current therapies remains a major the chalcone backbone and synthesized these compounds.
challenge in biomedical research [6, 7]. In addition, from
the pharmaco economic cost-effective stand-point and
seeking for a better patient compliance, a dual anti-
inflammatory, antimicrobial agent with minimum adverse
effects and a high safety margin is highly desirable. This
promoted us searching for agents that have a dual effect as
anti -inflammatory, antimicrobial agents. Consequently,
the chalcone backbone could be a versatile scaffold for

South Asian J Eng Technol, 01-07 | 1

Vol. 7 Iss. 3 Year 2018 L. Ilavarasan et al/2018 ISSN 2454-9614

Experimental procedure The same PEG was utilized to synthesize further chalcones
Melting point of the synthesised compounds were (Yield – 85% &melting point: 104-108oC) (Table.1).
determined in open capillary tubes and were found
uncorrected. UV spectra were also recorder using Alpha Method – B (Microwave irradiation Method):
Bruker UV spectrophotometer. FTIR spectra (KBr A mixture of compounds 4-chlorobenzaldehyde
pellets)were measured using Alpha Bruker FTIR (0.01mol) and 2-hydroxyacetophenone (0.01mol) and
instrument scanning with the entire region of 4000 - 400 NaOH (0.02 mol) were grinded in to the mortar. Then it
cm-1 with typical resolution of [Link] NMR spectra of was mixed with 15mL of PEG – 600. The mixed
the compounds have been recorded on Bruker AV400 compounds were taken in a 100mL beaker and it was
spectrometer operating at 400 MHz for recording 1H irradiated in a microwave oven for the 3 minutes at 110 W
spectra in DMSO solvent using TMS as internal standard operating at 2450Hz at 30 seconds of intervals. After
Microwave reactions are carried out commercially completion of reaction as followed by TLC examination,
available IFB domestic microwave oven having a chilled water was added to the reaction mixture and
maximum power output of 110W operating at 2450Hz. neutralized by an acid. The solid product was obtained,
Purity of the compounds is checked by TLC plates (Merck) which was filtered, dried and crystallized from an ethanol.
using hexane and ethyl acetate. Silicagel (column grade) The filtrate was evaporated to dryness to remove water
was purchased from Merck. The solvents were purified as leaving behind PEG-600. (Yield – 85% & melting point:
per the standard procedure reported elsewhere. 106-110oC).

Synthesis of 1-(2-hydroxyphenyl)-3-(4- Synthesis of 2-(4, 5-dihydro-5-(4-

chlorophenyl) prop-2-en-1-one by PEG-600 Chlorophenyl)-1H-pyrazol-3-yl) phenol
as Recyclable Solvent (CH1) (CH2)
Method – A: (Conventional Method): A mixture of chalcone (CH1) (0.01 mol) in 25ml of
A mixture of compounds 4-chloroBenzaldehyde absolute alcohol and hydrazine hydrate (0.01 mol) was
(0.01mol) and 2-hydroxyacetophenone (0.01mol) and refluxed in oil bath at temperature 80-90oC for 6 hours.
NaOH (0.02 mol) was stirred in PEG-600 (20 mL) for 1 Then the reaction mixture poured in to ice. The product
hour at 60oC. After completion of the reaction (monitored was isolated and crystallized from ethanol. (Yield – 78% &
by TLC), the crude mixture was worked up in ice-cold melting point: 106-110oC)
water (100 mL). The product which separated out was
filtered. The filtrate was evaporated to remove water
leaving PEG behind.

Synthetic scheme

South Asian J Eng Technol, 01-07 | 2

Vol. 7 Iss. 3 Year 2018 L. Ilavarasan et al/2018

Green chemistry approach Recently, Poly ethylene glycol (PEG) has been found to be
Chalcones are also key precursors in the synthesis an interesting solvent system.
of many biologically important heterocycles such as
benzothiazepine, pyrazolines, 1, 4-diketones and flavones. In continuation work on chalcones as precursors
Hence, the synthesis of chalcones has generated vast in the synthesis of various heterocycles, I have planned to
interest among organic as well as medicinal chemists. synthesize a series of novel hetero chalcones by applying
Reducing or eliminating the use of volatile organic the principles of green chemistry, using PEG-600. PEG is
solvents can minimize the generation of waste, which is a an environmentally benign reaction solvent; it is non-toxic,
requirement of one of the principles of green chemistry. inexpensive, potentially recyclable and water soluble,
which facilitates its removal from the reaction product.

Table 1 Characterization and data of Compounds

Compound Code. Compound General Molecular Molecular Melting point0C Rf Percentage of
Name formula Weight Value Yield,%
CH1 Chalcone C15H11ClO2 258.7 104-108oC 0.65 85%

CH2 Pyrazole C15H13ClN2O 272.73 106-110oC 0.41 78%

South Asian J Eng Technol, 01-07 | 3

Vol. 7 Iss. 3 Year 2018 L. Ilavarasan et al/2018 ISSN 2454-9614

CH1: (R= Cl):m.p. 104-108oC, yield: 85%, UV (in nm):226 (CH=CH), 366(C=O). IR (KBr in cm-1):1633(Ar C=C str.),
3306(C-H str.), 1791(C=O str.), 1431(C-N str.),:Rf: 0.65: 1H NMR( ppm)(CDCl3): 6.74- 7.01 (8H,m,Ar-H), 632- 6.47
(d,2H,CH2=CH2), 11.10(1H,s,Ar-OH), 3.9 (3H,S,CH3).

CH2: (R= Cl):m.p. 106-110oC, yield: 78%, UV (in nm):247 (CH=CH), 312(C=O). IR (KBr incm-1):1717(Ar C=C str.),
3152 (Ar-C-H str.), 1712 (C=N str.):Rf:0.41: 1H NMR( ppm)(CDCl3): 11.11(1H,s,Ar-OH), 6.7 -7.4(4H,s,Ar-H), 4.83
(d,1H,CH=NH), 3.51- 3.63 (2H,t,CH2).

South Asian J Eng Technol, 01-07 | 4

Vol. 7 Iss. 3 Year 2018 L. Ilavarasan et al/2018 ISSN 2454-9614

Antimicrobial activity Staphylococcus aureus (S. aureus) ATCC 19433 and

The micro dilution susceptibility test in Miller- Bacillussubtilis (B. subtilis) ATCC 1042 as Gram-positive
Hinton Broth (Oxoid) and Sabouraud Liquid Medium bacteria and Candidaalbicans (C. albicans) as a yeast
(Oxoid) were used for the determination of antibacterial fungus. Ampicillin trihydrate and clotrimazole were used
and antifungal activity. Test organisms: Escherichia coli (E. as standard antibacterial and antifungalagents,
coli) ATCC 25922 and Salmonella typhimurium respectively. Solutions of the test compounds, ampicillin
([Link]) ATCC 3311 as Gram-negative bacteria, trihydrate and clotrimazole were prepared in DMSO at

South Asian J Eng Technol, 01-07 | 5

Vol. 7 Iss. 3 Year 2018 L. Ilavarasan et al/2018 ISSN 2454-9614

concentration of 1600 mg/mL. From this stock different Results and Discussions
dilutions of the compounds (800, 400, down to 6.25 The Claisen-Schmidt condensation is an important
mg/mL) were prepared. The microorganism suspensions C-C bond formation for the synthesis of 1, 3-diaryl-2-
at 106 colony forming unit/mL/1c concentration were propen-1-ones (chalcones). The aim of my present study
inoculated to the corresponding wells. Plates were was to develop an efficient protocol using PEG-400 as a
incubated at 368C for 24 to 48 h. The incubation chamber recyclable reaction solvent to obtain 1, 3-diaryl-2-propen-
was kept sufficiently humid. At the end of the incubation 1-ones with excellent yields in a short span of time
period, the minimal inhibitory concentrations (MIC) were without formation of any side product and the same time
determined. Controls with DMSO and uninoculated media the solvent can be reused again and again by adopting the
were also maintained. recycling process.
The yields of the synthesized compounds were
Minimum Inhibiting Concentration found to be excellent. The structure of the synthesized
Comp (g/ml) compounds was identified by IR (Fig.2 and 5) and UV-
ounds Staphylococcus Visible (Fig.1 and 4) spectroscopy and confirmed by
Escherichia coli (E.c) 1HNMR
aureus (S.a) spectroscopy (Fig.3 and 6).
CH1 120.00 250.00
Conclusion
CH2 62.5 62.5 All synthesised compounds were characterized by
UV-Visible, FT-IR and 1HNMR spectroscopy techniques
and were found to be in agreement with the chemical
Bar Graph
structures expected. Antibacterial activity of eight
synthesised compounds tested against Staphylococcus
aureus and Escherichia coli. From the antimicrobial
evaluation of all the newly synthesized compounds it was
concluded that out of two compounds CH1, CH2 showed
excellent antibacterial activities against both
Staphylococcus aureus and Escherichia coli with lowest
value of 13.5(g/mL) and 31.25 ((g/mL) respectively. The
activity data obtained during the study will be certainly
useful to go for further research for drug designing and
synthesizing new chalcone and pyrazole derivatives.
Obviously, the comparative evaluation of active
compounds will require further studies; the data reported
in this article may be helpful guide for the medicinal
chemist who is working in this area.

South Asian J Eng Technol, 01-07 | 6

Vol. 7 Iss. 3 Year 2018 L. Ilavarasan et al/2018

References
1. A. Hasan, L. Rasheed, A. Malik, Synthesis and Characterization of Variably Halogenated Chalcones and Flavonols and
Their Antifungal Activity, Asian Journal of Chemistry, 19 (2007) 937-948.
2. R. Kalirajan, S. U. Sivakumar, S. Jubie, B. Gowramma, B. Suresh, Synthesis and Biological evaluation of some
heterocyclic derivatives of Chalcones, International Journal of Chem Tech Research, 1 (2009) 27-34.
3. R. Kalirajan, M. Palanivelu, V. Rajamanickam and G. Vinothapooshan, K. Anandarajagopal, Synthesis and biological
evaluation of some Chalcones derivatives, International Journal of Chemical Science., 5(2007) 73.
4. R. H. Udupi, R. Bhat and K. Krishna, Synthesis and biological activity of Mannich bases of certain 1,2-pyrazolines,
Indian Journal of Heterocyclic Chemistry, 8 (1998) 143-146.
5. G. Urmila, S. Vineeta, K. Vineeta, C. Sanjana, Synthesis and antifungal activity of new fluorine containing 4-
(substituted phenylazo) pyrazoles and isoxazoles, Indian Journal of Heterocyclic Chemistry, 14(2005) 265–266.
6. V. K. Pandey, V. D. Gupta and D. N. Tiwari, Synthesis of substituted benzoxazines as potential antiviral agents ,Indian
Journal of Heterocyclic Chemistry, 13 (2004) 399- 400.
7. B.A. Bhat, K.L. Dhar, S.C. Puri, A.K. Saxena, M. Shanmugavel, GN. Qazi, Bioorganic & Medicinal Chemistry Letters,
15(2005) 3177-80
8. L. E. Michael, M. S. David and S. S. Parsad, Chalcones: a new class of antimitotic agents, Journal of Medicinal Chemistry,
33 (1990) 1948–1954.
9. Pharmacopoeia of India II, (1996) A-100, A-108.
10. S. Chatterjee and S. N. Dan, Anti-arthritic and anti-inflammatory effect of a poly-herbal drug (EASE)@: Its
mechanism of action, Indian Journal of Pharmacology, 28 (1996) 116.
11. A. Barry, Antibiotic in Laboratory Medicine Corian (Ed.), Williams & Wilkins, Baltimore, 1991,Alves A. J. et al., Rev.
Farm., 34, 77 (1999).
12. H. Matsuda, T. Morikawa, I. Toguchida and M. Yoshikawa; Chem. Pharm. Bull., 50,788-795 (2002).
13. K. Aida, M. Tawata, H. Shindo, T. Onaya, H. Sasaki, T. Yamaguchi, M. Chin and H. Mitsuhashi, Isoliquiritigenin: a new
aldose reductase inhibitor from glycyrrhizae radix, Planta Medica, 56, 254-258 (1990).
14. F. Severi, S. Benvenuti, L. Costantino, G. Vampa, M. Michele and L. Antolini, Synthesis and activity of a new series of
chalcones as aldose reductase inhibitors, European Journal of Medicinal Chemistry, 33 (1998)859-866.
15. K. Kazauki, K. Htayama, S. Yokomor and T. Soki; Chem. Abstr., 85, 5913 (1976).

South Asian J Eng Technol, 01-07 | 7