Synthesis 2-(3-Phenylallylidenamino) Pentanedioic Acid by

Condensation of Cinnamaldehyde with Glutamic Acid

and the Activity Test as Antibacterial

Mimpin Ginting*, Deni Aprilina Lumban Gaul, Indra Masmur and Andy Candra

Departement of Chemistry,Faculty Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan

Keywords: Antibacterial, Condensation, Cinnamaldehyde, Glutamic Acid, Schiff Base.

Abstract: 2-(3- Phenylallyldenamino) Pentanedioic Acid as Schiff base has been synthesize by condensation between

cinnamaldehyde and glutamic acid as the source of primary amine. Condensation between cinnamaldehyde

and glutamic acid is in reflux condition by using ethanol as a solvent for 5 hours. The yield’s percentage of

reaction is 49.39%. The formed Schiff base is proven by FT-IR spectrum with the appearance of a vibration

peak at wave number in 1627 cm

-1

as the sign of C=N functional group. UV-Vis spectrum showed the change

of cinnamaldehyde maximum wave number in 238 nm become Schiff base in 321 nm. The result of research

also show that the Schiff base has strong antibacterial activity for S. aureus by obstacle zone 13.3 mm and

weak antibacterial activity for E. coli by obstacle zone 6.6 mm.

1 INTRODUCTION

Condensation between primary amine with

carbonyl from ketone and aldehyde in organic

compounds in special condition will produce Schiff

base (Cimerman et al. 1997, da Silva, et al, 2011).

Schiff base is known structurally as azometine (-

CH=N-) and one of organic compound with many

uses like as pigment and dye, catalyst, intermediary

in organic synthesize, and polymer stabilizer (Dhar

and Taploo, 1982). Schiff base also has biology

activity such as antifungal, antibacterial, antimalarial,

antiproliferative, anti-inflammation, antivirus, and

antipyretic (Dhar and Taploo, 1982, Przybylski, et al,

2009). The different use of Schiff base is based on

basic material from aliphatic, aromatic, heterocyclic

of primary amine and the source of carbonyl in use

(da Siva, et al, 2011).

Many researches also prove that Schiff base is

very effective as corrosion inhibitor for metal by form

a layer to protect material environmentally (Li et al.

1999, Munir, at al, 1985). The former researcher also

tested Schiff base from cinnamaldehyde, 2-

aminophenol condensation as corrosion inhibitor for

iron in HCl 0.5 N with inhibitor efficiency 92%

(Qasim, 2011). Schiff base from cinnamaldehyde as

the source of carbonyl and ethylenediamine as the

source of amine also can be used as corrosion

inhibitor in 7000 ppm of zinc in HCl 0.1 N with

efficiency 90.17% (Ginting, et al, 2016). Beside as

corrosion inhibitor, Schiff base also has antimicrobial

properties (Amanullah et al, 2011). Aysen et al (2010)

also synthesize Schiff base by using 4-

benzylloxybenzaldehide with 2-aminophenol and the

result has good antibacterial properties in E. coli,

B.subtilis, S. aureus (Sirumapea et al, 2015). Wang

et al (2015), synthesize Schiff base from

cinnamaldehyde with some of amino acid and found

that it has antibacterial properties in Eschericia coli,

Aspergillus niger, Penicillium citrinum and

Staphylococcus aureus. Cinnamaldehyde is a natural

product kind of phenylpropionate (C6-C3) which is

instead of synthesizing, it is also as the main

component of cinnamon oil (Guenther, 1990). The

utilization of Schiff base should be improved by

changing cinnamaldehyde to its derivative. By its

chemical properties, cinnamaldehyde has benzene

ring, alkene, and aldehyde so it can be transformed to

cinnamaldehyde derivatives (Ngawidiyana et al,

2007). Glutamic acid is an amino acid used by

organism in protein biosynthesis. This kind amino

acid is one of nonessential amino acid for human

which mean it can be synthesized in their body

(IUPAC-IUB, 2008). Glutamic acid as component of

protein contain in food, but only can be tasted in

Ginting, M., Aprilina Lumban Gaul, D., Masmur, I. and Candra, A.

Synthesis 2-(3-Phenylallylidenamino) Pentanedioic Acid by Condensation of Cinnamaldehyde with Glutamic Acid and the Activity Test as Antibact.

DOI: 10.5220/0008919702290233

In Proceedings of the 1st International Conference on Chemical Science and Technology Innovation (ICOCSTI 2019), pages 229-233

ISBN: 978-989-758-415-2

229

original form. Most of glutamic acid is exist in many

foods like cheese and ketchup. Glutamic acid also

used as additive in food, and as flavor in its natrium

salt known as monosodium glutamic (MSG). Based

on the description, the researcher interest to

synthesize Schiff base by using cinnamaldehyde with

glutamic acid and it used as antibacterial.

2 MATERIALS AND METHODS

2.1 Materials and Equipment

The tools used in this study include: two neck flasks,

reflux devices, thermometers, glassware, vacuum

pumps, analytical balance sheets, chromatographic

columns, chambers, UV lamps, petri dishes, osseous

needles, paper discs, incubators, FT-IR

spectrophotometer, UV-Vis spectrophotometer.

While the materials used in this study include:

Cinnamaldehyde, ethanol, glutamic acid, ethyl

acetate, n-hexane, silica gel HF254, silica gel 60 are

all pro-analysis made by E. Merck.

2.2 Synthesis of 2- (3- Phenyl

alylidinamine) Pentanadioate Acids

About 6.6 g of cinnamaldehyde were dissolved in 25

mL of ethanol, then put in a two-volume 250 mL neck

flask. Next, a drop of 5.9 g of glutamic acid was

dissolved with 50 mL ethanol through a dropper

funnel slowly into the mixture. Then reflux for 5

hours while stirring. Furthermore, the ethanol solvent

used was evaporated with rotary evaporator. Excess

Cinamaldehyde is evaporated by vacuum distillation

at 100oC and a pressure of 20mmHg. The residual

weight was obtained, then thin layer chromatography

(TLC) analysis was performed using developer n-

hexane: ethyl acetate (8: 2 V / V), kiesel 60 HF254

adsorbent and UV lamp stain, then purified using

column chromatography. The results obtained were

analyzed by FT-IR and UV-Vis spectroscopy

followed by an antibacterial activity test.

2.3 Schiff Base Analysis Synthesis

The purity of the Schiff base was analyzed using thin

layer chromatography, using several developers and

mixed developers with the stationary phase of silica

gel HF254. The analysis results provide a single stain.

The results obtained were then carried out FTIR

spectroscopic analysis with KBr and UV-Vis pellet

media in ethanol solvents then followed by an

antibacterial activity test.

2.4 Antibacterial Activity Test

2.4.1 Making Nutrient Agar Media

A total of 7 g was dissolved with 250 ml of aquadest

in an Erlenmeyer glass and heated to dissolve and

boil, then sterilized in an autoclave at a temperature

of 121

C for 15 minutes.

2.4.2 Manufacture of Oblique and Stock

Media for Bacterial Culture

In a sterile test tube, 3 ml of sterile NA media are

inserted, left at room temperature until it solidifies at

an angle to form an angle of 30-45

C. Bacterial

culture from the main strain was taken with a sterile

osseous needle and then inoculated on the sloping NA

media surface by scraping, then incubated at 35

C for

18-24 hours.

2.4.3 Making Mueller Hinton Agar Media

(MHA)

As much as 19 g of Mueller Hinton Agar powder, put

in an Erlenmeyer glass and then dissolved in 500 ml

of aquadest and heated until all dissolve and boil.

Then sterilized in an autoclave at 121

C for 15

minutes.

2.4.4 Making Bacterium Inoculum

A total of 3.25 g of nutrient broth was dissolved in

250 ml of aquadest in an Erlenmeyer glass and heated

to all dissolve and boil, then sterilized in an autoclave

at 121

C for 15 minutes and cooled. Then the

microbial colonies were taken from the stock of

culture using sterile osseous needles then suspended

into 10 ml sterile nutrient media in a test tube and

incubated at 35

C for 3 hours, then measured the

wavelength using a UV-Vis spectrophotometer

wavelength 580-600 nm.

2.4.5 Determination of Antibacterial

Activity

Determination of antibacterial activity was obtained

by agar diffusion method where the paper disk (ǿ 6

mm) which had been immersed with Schiff Bases 3%

and in direct contact with the media which had been

inoculated by E. coli and S. aureus, then clear zones

formed after incubation were observed for 24 hours.

The clear zone shows the inhibition that is produced

ICOCSTI 2019 - International Conference on Chemical Science and Technology Innovation

230

from the Schiff Base on E. coli and S. aureus. The

clear zone formed is measured by using the calipers

(accuracy mm).

3 RESULT AND DISCUSSION

3.1 Synthesis of Bases Schiff

The Schiff base is a 2- (3- Phenylalylidinamine)

Pentanadioate which is produced through

condensation between cinnamaldehyde as a source of

aldehyde and glutamic acid as a source of amine in

ethanol solvents under reflux conditions for 5 hours.

The excess of cinnamaldehyde from the reaction

results was evaporated through vacuum distillation

and analyzed by thin layer chromatography using

developer n-hexane: ethyl acetate (8: 2 v / v), the price

of product Rf still mixed was 0.11 and 0.75. From 6.6

g of cinnamaldehyde used the results were as much as

9.47 g (93.20%) yellowish brown solids. The

purification is continued through column

chromatography with eluent n-hexane: ethyl acetate

(8: 2 v / v), where purification results are 0.53 g, so

that the total yield is 49.39%. The results obtained

were analyzed using thin layer chromatography using

developer n-hexane: ethyl acetate (8: 2 v / v) giving a

single stain at an Rf price of 0.67. The physical form

of the Schiff base obtained is a solid form with a

melting point 124-129

C indicating that changes have

been made from the solid base material namely

glutamic acid with a melting point 247- 249

C. The

results of the Schiff base analysis produced using FT-

IR spectroscopy obtained spectra with absorption

vibration peaks at the wave number area 3410 cm

-1

3024 cm

-1

, 2924 cm

-1

, 1720 cm

-1

, 1674 cm

-1

, 1627 cm

-1

, 1450 cm

-1

, 1126 cm

-1

(Figure 1).

Figure 1: FT-IR Spectrum of Schiff Base.

From the results of UV-Vis spectroscopy there was

an increase in wavelength of 238 nm for

cinnamaldehyde (Figure 2) to 321 nm for the Schiff

base (Figure 3). This shows the addition of

conjugated double bonds to the products produced

due to the presence of new functional groups that are

bound to the synthesized part of the molecule.

Figure 2: UV-Vis Spectrum of Cinnamaldehyde.

Figure 3: UV-Vis Spectrum of Schiff Base.

The Schiff base results from a condensation reaction

between cinnamaldehyde and glutamic acid as

evidenced by the support of the FT-IR spectrum

which shows the emergence of a stretch of C = N at

the absorption peak of the wave number 1627 cm

-1

supported by stretching C-N at wave number 1126

-1

. Uptake in the wave number area 3024 cm

-1

shows that C-H aromatic from the benzene ring

supported by stretching absorption of 1674 cm

-1

shows the vibration of C = C of aromatic compounds.

This is also supported by the vibration peak in the

wave number region 2924 cm

-1

which shows the

typical absorption of vibration stretching (C-H) sp

which is supported by bending vibration (C-H) sp

the wave number region of 1450 cm

-1

. Hypothetically

the Schiff base formation reaction is shown in Figure

Synthesis 2-(3-Phenylallylidenamino) Pentanedioic Acid by Condensation of Cinnamaldehyde with Glutamic Acid and the Activity Test as

Antibact

231

Figure 4: Schiff Base Formation Reaction.

3.2 Antibacterial Activity Test

Antibacterial tests on the Schiff base using

Escherichia coli and Staphylococcus aureus can be

seen in Table 1.

Table 1: Antibacterial Activity Test.

Bacteria

Inhibited Zone

Diameter (mm)

Diameter Disc

Paper (mm)

E. coli

6.6

6.0

S. aureus

13.3

6.0

Cluster C = N in the Schiff base can be antibacterial

where the nitrogen atom has free electrons. The

presence of an imine group which has a cationic

charge that is able to bind the food source of the

bacterium thus inhibits food nutrition into bacterial

cells (Darmanto et al., 2010). Antibacterial strength is

classified into 3, which is strong if it produces a

diameter inhibition zone of more than 8 mm,

moderate activity if it produces a 7-8 mm inhibition

zone diameter, and weak activity if it has a diameter

inhibition diameter of less than 7 mm, thus that the

Schiff base is formed has strong antibacterial

strength. The wider the inhibition zone produced

shows the stronger Schiff's base ability in inhibiting

bacterial growth. The antibacterial test data showed

that the Schiff base had good antimicrobial activity

against the Staphylococcus aureus bacteria which is a

common bacterium outside the body. For Escherichia

coli bacteria, the Schiff base is not very good.

Escherichia coli bacteria are commonly found in the

body.

4 CONCLUSIONS

The condensation reaction between 6.6 g of

cinnamaldehyde and 5.9 g of glutamic acid produced

a Schiff base of 9.47 g obtained for a total yield of

49.39%. Supported by FT-IR spectroscopic data,

namely by the appearance of a vibration peak in the

area of the wave number 1627 cm

-1

which indicates

the group C = N (Imina). The UV-Vis data showed a

change in the wavelength of 238 nm for

cinnamaldehyde to 321 nm for the Schiff base. The

results of the Schiff alkaline antibacterial activity test

showed strong results against the Staphylococcus

aureus bacteria and were weak for Escherichia coli

bacterial.

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Synthesis 2-(3-Phenylallylidenamino) Pentanedioic Acid by Condensation of Cinnamaldehyde with Glutamic Acid and the Activity Test as

Antibact

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