Synthesis, Antioxidant and Toxicity Activity of Compounds

(E)-1-(3-bromophenyl)-3-p tolylprop-2-en-1-on

Eti Meirina Brahmana

1,4

, Jamaran Kaban

, Ginda Haro

, Juliati Br Tarigan

, Basuki Wirjosentono

Tamrin

and Mimpin Ginting

Postgraduate Chemistry Study Program, Faculty of Mathematics and Natural Sciences, Universitas

Sumatera Utara, Jl. Bioteknologi No. 1 Kampus USU, Medan, Indonesia

Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara,

Jl. Bioteknologi No. 1 Kampus USU, Medan, Indonesia

Faculty of Pharmacy, Universitas Sumatera Utara, Jl. Tri Dharma Kampus USU, Medan,

North Sumatera, Indonesia

Biology Study Program, Faculty of Teaching Training and Education, Universitas Pasir Pengaraian, Indonesia

thamrinsumut@gmail.com, mimpinginting@usu.ac.id

Keywords: Antioxidant, Chalcone, Synthesis, Toxicity.

Abstract: Halogen substituted analog compound chalcone (E)-1-(3-bromophenyl)-3-p-tolylprop-2-en-1-on was

synthesized from 4-metylbenzaldehyde as aldehydes with 3- bromoacetophenone, as ketones by using aldol

condensation reaction. The compound resulted rendement with value of 62,38% and characterized by using

UV, IR, MS, and 1HNMR. Test of antioxidant activity using DPPH method showed that those compounds

have low potency as antioxidant agent LC

with value 571, 7903 ppm. Toxicity tests using Brine Shrimp

Lethality Test (BSLT) showed that those compounds have a potency as anticancer agent with LC

value

7,94 μg/mL.

1 INTRODUCTION

Chalcones (α, β-unsaturated aromatic ketones) are

medicinally important compounds (Zhuang, 2017).

Chalcones have been targeted by several researchers

in recent years due to their wide biological potentials

(Espinoza, 2016). Various natural and synthetic

chalcones have been shown to display biological

properties to act as potential hits for anticancer (Park

et al, 2018; Michelini, 2018), Antitumor (Fouad,

2018), Antioxidant (Jawad, 2018).

Antioxidants are compounds that have the ability

to ward off free radicals that can cause various

dangerous diseases such as cancer, cardiovascular

diseases and degenerative diseases (Barhe, 2014).

Antioxidant testing in this study used the DPPH

(1,1-diphenyl-2-picrylhydrazyl) method by

spectrophotometry, because this method has the

advantages of being simple, easy, fast, sensitive and

requires only a few samples in testing. The

parameters used in this method are IC

, which is the

sample concentration needed to capture DPPH

radicals by 50% (Polo, 2019).

Cancer is a large group of heterogeneous

diseases characterized by abnormal division and

spread of cells (Mansoori et al, 2017). 2,2-dimethyl-

2,3-dihydro-4(1H)-quinolinone were screened

against the NCI-N87 and DLD-1 cancer cell lines,

with most compounds showing low micromolar

cytotoxic activity (Jean, 2018).

Chalcone compounds can be synthesized through

Claisen-Schmidt condensation from an aldehyde and

aromatic ketone with an acidic or basic catalyst. The

basic catalysts commonly used are NaOH (Suwinto,

2014) and KOH (Brahmana, 2015; Riaz, 2019).

Whereas commonly used acid catalysts are HCl

(Wang, 2019), H

(Dong, 2018) and HClO

-SiO

(Siddiqui, 2015).

Chalcon synthesis in this study used the

Microwave Assisted Organic Synthesis (MAOS)

method. Microwave-induced organic reaction

enhancement (MORE) chemistry is gaining

popularity as a non-conventional technique for rapid

organic synthesis. Important features of this

technique are easy access to very high temperature,

good control over energy input in a reaction, higher

Brahmana, E., Kaban, J., Haro, G., Br Tarigan, J., Wirjosentono, B., Tamrin, . and Ginting, M.

Synthesis, Antioxidant and Toxicity Activity of Compounds (E)-1-(3-bromophenyl)-3-p tolylprop-2-en-1-on.

DOI: 10.5220/0008836400050009

In Proceedings of the 1st International Conference on Chemical Science and Technology Innovation (ICOCSTI 2019), pages 5-9

ISBN: 978-989-758-415-2

 2020 by SCITEPRESS – Science and Technology Publications, Lda. All r ights reserved

yields and rapid synthesis of organic compounds

(Ahmad, 2016). Microwave-assisted organic

synthesis (MAOS) has been risen as new strategy in

the syntheses of dyes with luminescence properties.

Scientists have turned from conventional heating to

microwave assisted chemistry as an astounding and

powerful strategy for their researches in modern

organic synthesis. The technique offers a lot of

advantages as it is simple, rapid, economic, and

efficient. Dyes are effective molecules have pivotal

effects in various fields (Elgemeie, 2018).

However, if viewed from its biogenetic origin,

halogen-substituted chalcone is not possible in

nature. Therefore, to obtain halogen substituted

chalcons, synthesis is carried out. Based on the

description above, it can be seen that chalcone has

been widely used for various medical purposes. The

chalcon compounds to be synthesized are chalcone

substituted with halogen (E) -1- (3-bromophenyl) -3-

p-tolylprop-2-en-1-on. The compounds were

characterized by UV, IR, MS, and

H-NMR

analysis. The compounds were tested for their

cytotoxic activity and antioxidant activities by

standard methods.

2 MATERIALS AND METHODS

2.1 General Information

The chemical used in the research were pro analysis

grade. Melting points were measured with a Fisher

Johns melting point apparatus (SMP 11-Stuart®).

The purity of synthesized compounds was checked

by thin layer chromatography on silica gel GF254

plates, the eluent was mixture of n-hexane/ethyl

acetate in 9:1 ratio and the spot were identified by

UV (Camag®) (254 nm) . The mass spectra was

recorded by MS Water LCT Premier XE

spectrometers, NMR spectra was recorded in CDCl3

on a NMR (AGILENT 500 MHz). The IR spectra

was recorded on a spectrum FTIR (Shimadzu, IR

Prestige-21) spectrophotometer. The UV-Vis spectra

was measured on UV-Vis (Genesys 10S®)

spectrophotometer. Chalcone synthesis used oven

microwave Samsung ME 109 F.

2.2 General Procedure for Chalcone

Synthesis

A mixture of 3- bromoacetophenone (5 mmol) was

dissolved in ethanol (5 mL), then KOH 6 N (2 mL)

was added dropwise. The mixture was stirred for 5

minutes and 4-metylbenzaldehyde (5 mmol) added

to the mixture. The mixture is irradiated using

microwave for 2-5 minutes, with an interval of 30

seconds. After that, the mixture is left for 20 hours

to maximize the results of the reaction (sediment)

obtained. A total of 15 mL of cold distilled water

was added to the mixture and the pH of the mixture

was neutralized with HCl. The precipitate formed is

then filtered with a Buchner funnel, washed with

cold n-hexane, and vacuumed to dry. Stages of

reaction were observed with TLC. The product

obtained was tested for its purity by TLC test and

melting point. The pure product obtained was then

determined by UV, IR, MS and

H-NMR

spectroscopy.

2.3 Antioxidant Activity

DPPH (1,1–diphenyl-2-picrylhydrazyl) radical

scavenging activity was measured by the method of

Lamaison et al. The reaction mixture contained 1.5 ×

−7

M methanolic solution of DPPH and various

concentrations of the test substances and was kept in

the dark for 50 min. Optical density (OD) of samples

was measured at 517 nm against a blank, and IC50

values were calculated (using linear regression

analysis) by plotting a graph, taking concentration

on the X-axis and percentage inhibition on the Y-

axis, at 50% of the percentage inhibition the line was

drawn from Y-axis and aligned with the

concentration on X-axis then the IC

values were

obtained.

2.4 Brine Shrimp Lethality Bioassay

(BSLT)

Brine shrimps (Artemia salina) was hatched using

brine shrimp eggs in a conical flask (1 L), filled with

sterile artificial sea water under constant aeration for

48 h. After hatching, active nauplii free from egg

shells were collected from the brighter portion of the

chamber and used for the assay. Ten nauplii were

drawn through a glass capillary and placed in each

vial containing 5 mL of brine solution. In each

experiment, test substances whose activities are to

be checked were added to the vial according to their

concentrations and maintained at room temperature

for 24 h under light and the surviving larvae were

counted. Experiments were conducted along with

control (vehicle treated), different concentrations

(10, 100 and 1.000 μg/mL) of the test substances in a

set of three tubes per dose. Replicas should be

maintained to get accurate results.

ICOCSTI 2019 - International Conference on Chemical Science and Technology Innovation

3 RESULTS AND DISCUSSIONS

(E)-1-(3-bromophenyl)-3-p-tolylprop-2-en-1-on (1) :

yellow crystals (0.9358 g ; 62.38%), m.p 131-

1320C, Rf = 0,79 (n-hexane/ethyl acetate: 9:1 );

Mass spectrum (HR-MS) m/z: 300,0152 with

formula C16H13Obr; IR (KBr, cm-1): 792 (C-Br),

1512 (C = C of benzene), 1604 (C = O of ketone),

2362 (C-Br), 2960 (-CH3), 3061 (CH from benzene)

and 3468 (Overtone from C = O); 1H-NMR (CDCl3,

500 MHz) δH 8.13 ppm (s; 1H); 7.93 ppm (d: 7.5;

1H); 7.81 ppm (d: 15.5; 1H); 7.70 ppm (dd: 8; 1;

1H); 7.55 ppm (d: 8; 1H); 7.42 ppm (d: 15.5; 1H);

7.38 ppm (t: 8; 1H); 7.24 ppm (d: 7.5; 1H); 2.40

ppm (s; 3H).

Figure 1: Chalcon Structure.

Compound is synthesized via an aldol

condensation reaction, where a new carbon-carbon

bond is formed between α carbon atoms from one

carbonyl and another carbonyl carbon atom. The

acidity of the hydrogen atom α from the carbonyl

compound allows the carbonyl compound to react

with the others to produce a combined product of

both. This reaction is catalyzed by a base (KOH).

Figure 2: Synthesis reaction of chalcone compounds (E)-

1-(3-bromophenyl)-3-p-tolylprop-2-en-1-on.

DPPH is a stable free radical and is used to

evaluate the reduction of free radicals that have a

principle that DPPH will be reduced by the donation

process of hydrogen or electron so that the color will

change from violet to yellow with changes in color

intensity proportional to the number of electron

donations followed by a decrease in DPPH

absorbance (Dris and Jain, 2004). Where, the greater

the decrease in absorbance of DPPH, the stronger

the antioxidant activity.

The antioxidant value of the extract was

determined based on IC

value, namely the

concentration of chalcone which caused a reduction

of DPPH activity by 50%. The smaller the IC

value, the more active the sample is in capturing

DPPH radicals or better antioxidant activity

(Morales, 2013).

The percentage of inhibition was determined by

comparing the DPPH absorbance purely with the

absorbance of DPPH plus chalcone at a wavelength

of 516.5 nm. The results showed that DPPH

inhibition by chalcons was directly proportional to

the concentration of chalcone, which meant that the

greater the concentration of chalcone, the higher the

inhibition percentage. Chalcone compounds work as

antioxidants by breaking radical chain reactions and

donating hydrogen atoms to produce more stable

free radicals (Nimse and Pal, 2015). The antioxidant

ability of chalcone has an IC

value of 571, 7903

ppm and is classified as a weak antioxidant (Jacoeb

et al, 2011).

Toxicity activity tests were carried out on

A.salina larvae using the Brine Shrimp Lethality

Test (BSLT). The selection of this method as an

initial screening in an effort to search for anticancer

compounds because of the low cost of the

experiment, the process is fast and simple. In

addition, these larvae have several advantages

including easy to obtain, easy to breed and can live

in a high range of salinity. The larvae are obtained

by hatching eggs for 48 hours. Hatched larvae will

swim to a bright place. This will make it easier for

the separation and retrieval of these animals that

have become larvae.

Each of the compounds to be tested was made in

concentrations of 1000, 100 and 10 µg/mL in

seawater for 24 hours of testing. The difference in

concentration is intended to determine the level of

activity of each compound against the death of these

larvae. Making test solutions using ethyl acetate

solvents because the analogue compound chalcone

tested was dissolved in ethyl acetate. The solvent is

left until it evaporates perfectly so as not to interfere

with the toxicity tests carried out. Before adding sea

water, dimethyl sulfoxide (DMSO) is added to help

dissolve the test compound in seawater so that the

compounds can be distributed evenly. The amount

of DMSO added is 50 µL, because if more than 50

µL can cause death in the larvae. In this test DMSO

was used as a control, which was not too toxic as the

reason for choosing DMSO to help dissolve

compounds in seawater.

Halogen-substituted chalcone compounds have

significant cytotoxic activity with an LC

value of

7.94 µg /mL. The analogue compounds of chalcone

tested showed toxic effects on the death of A.salina

larvae and showed potential toxicity. Because a

compound is said to be active if the LC

≤ 250





Synthesis, Antioxidant and Toxicity Activity of Compounds (E)-1-(3-bromophenyl)-3-p tolylprop-2-en-1-on

µg/mL and a maximum of 500 µg/mL (Meyer,

1982).

The biological activity of chalcone compounds in

this study, may be influenced by the presence of α,

β-unsaturated carbonyl groups and substituents

bound to the aromatic ring contained in the

compound. (Suvitha, 2012) argue that the possibility

of chalcon compounds can induce cell death by

interfering with mitochondrial function as cell

respiration. Cell respiration is the oxidation process

of food molecules, for example glucose to CO

and

O which form energy in the form of ATP

(Adenosine Tripospat) which is useful in supporting

cell activity that requires energy. If mitochondrial

damage occurs in the cell, it will cause interference

with mitochondrial function in ATP synthesis so that

the cell will die. Cell death will later cause the death

of the larvae themselves.

4 CONCLUSIONS

In conclusion, we have designed and synthesized in

good Halogen substituted analog compound

chalcone (E)-1-(3-bromophenyl)-3-p-tolylprop-2-en-

1-on using aldol condensation reaction reactions

with yield 62,38%. Test of antioxidant activity using

DPPH method showed that those compounds have

low potency as antioxidant agent LC

with value

571, 7903 ppm. Toxicity tests using Brine Shrimp

Lethality Test (BSLT) showed that those compounds

have a potency as anticancer agent with LC

value

7,94 μg/mL.

ACKNOWLEDGEMENTS

The author would like to thank RistekDIKTI the

financial support to complete this study and all

lecturer in the Department of Chemistry Universitas

Sumatra Utara.

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Synthesis, Antioxidant and Toxicity Activity of Compounds (E)-1-(3-bromophenyl)-3-p tolylprop-2-en-1-on