REACTION OF 4-NITROBENZALDEHYDE AND 3-PENTANONE IN

ALKALINE SOLVENT

OOI AH GUAN

LOA WOEI JYH

LOW QIAU WEI

MARK TIHDNOE JOAL

SERENE SUNG SUYIN

SOONG MEE LAI

Report Submitted to Fulfill the Partial Requirements

for the Diploma in Pharmacy

UNIVERSITY KUALA LUMPUR

ROYAL COLLEGE OF MEDICINE PERAK

JAN 2010

i
 DECLARATION

I declare that this report is our original work and all references have been cited

adequately as required by the University.

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ii
 APPROVAL PAGE

We have examined this report and verify that it meets the program and University’

requirements for the Diploma in Pharmacy

Date: Signature: ………………………….

Supervisor’s Name: …………………

Official Stamp

Date: Signature: ………………………….

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iii
 ACKNOWLEDGEMENT

First of all, we would like to express our deep and sincere gratitude to our

supervisor, Mr. Maywan Hariono and our co-supervisor, Mrs Suryati Bt Syafri for

valuable guidance and advise. Their wide knowledge and their logical way of thinking

have been of great value for us.

Besides this, we would like to take this opportunity to thank our University

Kuala Lumpur Royal College of Medicine Perak to provide us a good environment to

complete our research under UniKL Short Term Research Grant(STRG)2009-2010

Project. It give us an opportunity to participate and learn about how to carry out a

research.

We will like to deeply grateful to our elective group leader always lead us in this

research. In addition, we would like to thanks our group members that always give co-

operation along this research. So that our research is done successfully.

We also would like to thank you to who involve direct and indirect in our

research.

Finally, an honorable mention goes to our families for their supports on us in

completing this research. Without help of the particular that mentioned above, we would

not succeed and will face many difficulties while doing this research.

iv
 ABSTRACT

Objective: A study have been conducted, aimed to differ the product after reaction
between 2,4-bis-(4-nitrobenzylidene)-3-pentanone from 4-nitrobenzaldehyde and 3-
pentanone using NaOH as a catalyst.
Method: By using aldol reaction, 4-nitrobenzaldehyde able to react with 3-pentanone
using NaOH as a catalyst. Once the crude product obtained, we isolate it using filtration
method.
Finding: The product, 2,4-bis-(4-nitrobenzylidene)-3-pentanone is differ from 4-
nitrobenzaldehyde as prove by TLC test, UV spectrophotometry, and physical properties
analysis.
Conclusion: It has been known that a disease infection is the most of pathogenesis
which cause an anatomy-physiological disorder. It needs more highly potency of
antibacterial agent to enhance its activity and more selective spectrum. Once this
hypothesis is confirmed, it will be a trigger to numerous other equivalent studies such us
its toxicity, pharmacokinetic, pharmacodynamic, etc. that would explore the prospect of
the product to be the new antibacterial agent.

v
 ABSTRAK

Objektif: Sebuah kajian dijalankan untuk membezakan 2,4-bis-(4-nitrobenzylidene)-3-

pentanone sebelum dan selepas tindak balas oleh 4-nitrobenzaldehyde dengan 3-
pentanone menggunakan NaOH sebagai pemangkin.
Kaedah: Dengan menggunakan tindak balas aldol, 4-nitrobenzaldehyde boleh bertindak
balas dengan 3-pentanone menggunakan NaOH sebagai pemangkin.
Dapatan: Produk (2,4-bis-(4-nitrobenzylidene)-3-pentanone) itu berbeza dengan 4-
nitrobenzaldehyde. Ini dibuktikan oleh ujian kromatografi lapisan nipis, UV
spectrophotometri, dan analisis ciri-ciri fizikal.
Kesimpulan: Jangkitan penyakit adalah paling bahaya, ia boleh menyebabkan
perubahan anatomi fisiologi. Oleh itu, ia perlu antibakteria yang lebih kuat untuk
meningkatkan activity dan spectrum yang lebih tepat. Bila hipotesis ini disahkan, ia
menjadi penjana permulaan kajian yang lain seperti tahap tosik, farmakokinetik,
farmakodinamik, dan lain-lain, yang akan mengembara dalam prospek produk anti
bakteria yang baru.

TABLE OF CONTENTS

vi
 Page

TITLE PAGE i

DECLARATION ii

APPROVAL iii

ACKNOWLEDGEMENTS iv

ABSTRACT v

ABSTRAK vi

TABLE OF CONTENTS vii

LIST OF TABLES x

LISTS OF FIGURES xi

LIST OF ABBEVATION xii

CHAPTER 1 INTRODUCTION

1.1 Background of the study 1

1.2 Problem statement 2

1.3 Research objective 2

1.4 Hypothesis 2

1.5 Significance 3

CHAPTER 2 Literature Review

2.1 Curcumin 4

2.2 Curcumin Analogues 5

2.3 Derivatives of benzylidene ketone 6

2.4 Aldehyde and Ketone

2.4.1 Aldehydes 7

2.4.2 Ketones 7

2.4.3 Chemistry of the carbonyl group 8

2.4.4 Naming the aldehyde and ketones 8

2.5 Aldol Condensation (Condensation of aldehydes and ketones) 8

2.6 Dehydration of aldol product: Synthesis of enones 9

2.7 Thin Layer Chromatgraphy 10

2.8 Ultraviolet-visible spectroscopy

vii
 2.8.1 Absorption by Organic Compounds 11

2.8.2 Qualitative Applications of Ultraviolet-visible spectroscopy 12

CHAPTER 3 Methodology

3.1 Material 13

3.2 Equipment 13

3.3 Method

3.3.1 The reaction (adopted from Rowland, 2004) 13

3.3.2 Isolation and Purification 14

3.3.3 Yields calculation 15

3.3.4 Analysis of physicochemical properties, purity and molecules

structure of product

3.3.4.1 Physical Appearance test 15

3.3.4.2 Solubility test 15

3.3.4.3 Thin layer Chromatography test 15

3.3.4.4 Maximum wave length test 16

CHAPTER 4 Results

4.1 Yields calculation 17

4.2 Physical appearance 18

4.3 Solubility test 18

4.4 Thin layer chromatography 19

4.5 Maximum wave length test 21

CHAPTER 5 Discussion

5.1 Synthesis and isolation 23

5.2 Physical appearance 24

5.3 Solubility test 24

5.4 Thin layer chromatography 24

5.5 Maximum wave length test 28

viii
CHAPTER 6 Conclusion and Recommendations

6.1 Summary of main findings 29

6.2 Implications of the study 29

6.3 Recommendations for future research 29

REFERENCES 31

APPENDICES 32

LIST OF TABLES

ix
 Page

1.1 The solubility result for sample product 18

1.2 The solubility result for 4-nitrobenzaldehyde 18
1.3 Indonesian Pharmacopeia 3rd Edition 19
1.4 Differenced between product, 4-nitobenzaldehyde and 3-pentanone 25

LIST OF FIGURE

Page

1.1 The reaction of 2,4-bis-(4-nitrobenzylidene)-3-pentanone 2

x
1.2 Chemical structure of curcumin 4

1.3 The structure of demethoxycurcumin and bisdemethoxycurcumin 5

1.4 The structure of Pentagamavunone, Hexagamavunone and Gamavutone 6

1.5 General structure and the examples of aldehyde and ketone 7

1.6 The reaction of aldol condensation 9

1.7 The dehydration of aldol product 10

1.8 The TLC plate A 19

1.9 The TLC plate B 20

2.0 The TLC plate C 21

2.1 Wavelength of 4-nitro-benzyladehyde 22

2.2 Wavelength of product (2-4-bis-4 nitrobenzylidene)- 3 pentanone 23

2.3 TLC plate (Plate A, plate B, plate C) under short wave Ultraviolet light 26

2.4 TLC plate (Plate A, plate B, plate C) under long wave Ultraviolet light 27

LIST OF ABBREVATION

TLC Thin Layer Chromatography

UV-vis spectrophotometer Ultra violet visible spectrophotometer

xi
 CHAPTER 1

INTRODUCTION

1.1 Background of the study

Curcumin is one of isolates from Curcuma sp plant (Aggarwal et al. 2005). It is

well-documented that curcumin is the major constituent of turmeric powder extracted

from the rhizome of Curcuma longa, possesses a broad spectrum of biological activity.

The most pronounced are: antioxidant, anti-inflammatory, anticancer, chemopreventive,

antibacterial, antifungal, antiparasitic, antiviral and antihistaminic activities (Al-Wabli,

2006).

Modification of curcumin molecule structure has been studied refers to its

instability toward light, pH, temperature and its poor pharmacokinetic profiles. It was

intended to enhance the stability and absorption of curcumin when administered orally.

Sardijman et al. (1997) have synthesized many curcumin analogues, one of them was

2,5-bis(4’-hydroxy-3’-methoxybenzylidene)cyclopentanone (pentagamavunone-0) which

were derivate from benzylidene. That compound has been examined as antioxidant

(Sardjiman, et al. 1997), anti-inflammatory (Masuda et al. 1993) and antitumor (Youssef

and El-Sherbeny, 2005).

In this study, it will be synthesized one of benzylidene ketone derivatives which

has nitro group at para position and alkyl modification at ketone chain. That compound

is predicted having biological activity. It will be a challenge to discover a new compound

which has a better biological activity than the previous drug.

1
1.2 Problem Statement

Is there any difference before (starting material) and after (product) reaction of 4-

nitrobenzaldehyde and 3-pentanone.

1.3 Research Objective

This study aimed to differ the product which is predicted as 2,4-bis-(4-

nitrobenzylidene)-3-pentanone from 4-nitrobenzaldehyde and 3-pentanone as starting

material using NaOH as a catalyst. The reaction is presented at Figure-1.

O

C O
H
NaOH
2 + H 3C C CH3
C C
O 2N H2 H2

4-nitrobenzaldehyde 3-pentanone

O
H H
C C C
C C
+ 2 H 2O
CH3 CH3 NO2
O2 N

2, 4-bis-(4-nitrobenzylidene)-3-pentanone

Figure 1.1 The reaction of 2,4-bis-(4-nitrobenzylidene)-3-pentanone

1.4 Hypothesis

Yes, there is a different before (starting material) and after (product) reaction of

4-nitrobenzaldehyde and 3-pentanone.

2
1.5 Significance

By synthesizing new compound, it will be a challenge to discover a new drug

which has a better biological activity than the previous one.

3
 CHAPTER 2

LITERATURE REVIEW

2.1 Curcumin

Curcuma longa or turmeric is a tropical plant native to southern and southeastern

tropical Asia. The parts used are the rhizomes. The most active component in turmeric is

curcumin, which may make up 2 to 5% of the total spice in turmeric. Curcumin is a

diferuloylmethane present in extracts of the plant. Curcuminoids are responsible for the

yellow color of turmeric and curry powder. They are isolated from turmeric by ethanol

extraction. The chemical structure of curcumin can be seen at Figure 2.

O O

H3CO OCH3

HO OH

Figure 1.2 Chemical structure of curcumin

Turmeric contains a wide variety of phytochemicals including curcumin,

demethoxycurcumin, bisdemethoxycurcumin, zingiberene, curcumenol, curcumol,

eugenol, tetrahydrocurcumin, triethylcurcumin, turmerin, turmerones and turmeronols

(Aggarwal et al. 2005).

The invention was initiated by the fact that curcumin was widely used for

medication as antiinflammation, antibacteria, antioxidant, anti-hepatotoxic,

hypercholesterolemia, anti-cancer, and antioxidant (Sardjiman et al. 1997).

4
2.2 Curcumin analogues

As indicated above turmeric contains two different analogues of curcumin i.e.,

demethoxycurcumin and bisdemethoxycurcumin (Figure 3). Whether both of two

analogues exhibit equal activity is not clear. While in most systems, curcumin was found

to be most potent but in some systems, bisdemethoxycurcumin were found to exhibit

higher activity than curcumin and demethoxycurcumin (Aggarwal et al. 2005).

O O

OCH3

HO Demethoxycurcumin OH

O O

HO Bisdemethoxycurcumin OH

Figure 1.3 The structure of demethoxycurcumin and bisdemethoxycurcumin

5
2.3 Derivatives of benzylidene ketone

Modification of the center part of curcumin using electron withdrawing as well

as electron donating group gave some novel compounds as derivatives of benzylidene

cyclohexanone, benzylidene cyclopentanone, and benzylidene acetone. The products

were proposed under the following patent names: Hexagamavunone, Pentagamavunone

and Gamavutone. They were patented by Sarjiman, et al. (1998). Figure 4 present the

chemical structure of the patent products.

O

H3CO OCH3

HO OH
Pentagamavunone
O

H3CO OCH3

HO OH

Hexagamavunone
O

H3CO OCH3

HO Gamavutone OH

Figure 1.4 The structure of Pentagamavunone, Hexagamavunone and Gamavutone

6
2.4 Aldehyde and Ketone

2.4.1 Aldehydes

Aldehydes are organic compound that contain terminal carbonyl group. The

carbonyl compound contain carbon which bond to hydrogen and double bond to oxygen

(O=CH). Any chemical contain this structure is known as aldehydes or formyl or

methanoyl group. The word aldehydes origin from alcohol dehydrogenated.

(www.wikipedia.org)

O
O C
H
C
R H
O2N
Aldehyde 4-nitrobenzaldehyde

O
H3C C CH3
C C
C H2 H2
R R

Ketone 3-pentanone

Figure 1.5 General structure and the examples of aldehyde and ketone

2.4.2 Ketones

Ketones is similar to aldehydes, which also contain carbonyl group. The carbon

bond to another two carbons and a double bond to oxygen (R-C=O-R). If the R group is

hydrogen, the compound is aldehydes. (www.wikipedia.org). Figure 5 presents general

structures and the example of aldehyde and ketone.

The carbon atom adjacent to a carbonyl group is called the α-carbon. Hydrogens

attached to this carbon are called α-hydrogens. In the presence of an acid catalyst the

7
ketone is subjected to so-called keto-enol tautomerism. The reaction with a strong base

gives the corresponding enolate. A diketone is a compound containing two ketone

groups. (McMurry and Simanek. 2007)

2.4.3 Chemistry of the carbonyl group

The carbon-oxygen double bond are polarised because of the high

electronegativity of oxygen relative to carbon. The carbonyl carbon which is positively

polarised, it is electrophilic and react with nucleophilic. The carbonyl oxygen is

negatively polarised and nucleophilic. (McMurry and Simanek. 2007)

2.4.4 Naming the aldehydes and ketones

Aldehydes are named by replacing the e in the alkane group of the parent chain

with al. The parent chain must contain –CHO group and the carbon in –CHO must

number as carbon 1.

Ketones are named by replacing the e in the alkane group of the parent chain

with one. The parent chain is the longest chain that contain ketone group (RCOR).

Numbering of the carbon begin with the carbon nearest to ketone group. (McMurry and

Simanek. 2007)

2.5 Aldol condensation ( Condensation of aldehydes and ketones)

When acetaldehydes is dissolved in alcohol solvent and treated with a base, such

as sodium hydroxide, a rapid reversible condensation reaction occur. The product is a β-

hydroxy product known commonly aldol (aldehydes + alcohol). Called aldol reaction,

base catalyse condensation is a general reaction of all aldehydes and ketone with α-

8
hydrogen atom. If the aldehydes or ketone does not have any -hydrogen atom, aldol

condensation cannot occur. The condensation product is generally favour for reaction of

monosubstitute acetaldehydes (RCH2CHO), but the starting material is favour for

reaction disubstitute aldehydes (R2CHCHO) and for ketones. (McMurry and Simanek.

2007)

O OH H O
NaOCH2CH3

2 H3C C H CH3CH2OH H3C CH CH C H

 

Aldol
Acetaldehyde
(a hydroxy aldehyde)

Figure 1.6 The reaction of aldol condensation

2.6 Dehydration of aldol product: synthesis of enones

The β-hydroxy ketones and β-hydroxy aldehydes form in aldol reaction are easier

dehydrated to yield conjugated (α, β unsaturated) enones (ene + one). In fact, its lost of

water that give the aldol condensation it name, since condense out the reaction. Most

alcohol are resistant to dehydration by diluted acid or base, but OH group that are two

carbon away from a carbonyl group are special. Under basic condition, an α-hydrogen is

extract and the resulted enolate ion expels the nearby OH- leaving group. Under acidic

condition, the OH group of enol protonated and H2O then expelled as the leaving group.

The reaction condition needed for aldol dehydration are often only a bit more vigorous

(slightly higher temperature for instant) then the condition needed aldol condensation

itself. As a result, conjugated enones are often obtain directly from aldol reaction without

ever isolating the intermediate β-hydroxy carbonyl compound. (McMurry and Simanek.

2007)

9
 O
OH O

C C
C -H2O H
C H3C
H C H
H3 C C H H
H2

Figure 1.7 The dehydration of aldol product

2.7 TLC- Thin Layer Chromatography

Thin-layer chromatography (TLC) is an important technique in organic

chemistry. TLC can be used to assess the course of a reaction, to assess the purity of a

sample and also to identify unknown compounds by comparison with standards.

Thin Layer Chromatography (TLC) is a chromatography that used to separate

mixtures of substances into their components. Thin layer chromatography is performed

on a sheet of glass, plastic or aluminium foil. This is coated with a thin layer of

adsorbent material usually made of silica gel, aluminium oxide or cellulose. All forms of

chromatography work on the same principle.

Stationary phase is the part of chromatography system through which a solvent

flows where distribution of solutes between the phases occurs. The stationary phase may

be a solid or a liquid that is immobilized or adsorbed on a solid. Mobile phase is the part

of the chromatography system which carries the solutes through the stationary phase.

The mobile phases are either liquids or gases.

2.8 Ultraviolet- visible spectroscopy

10
2.8.1 Absorption by Organic Compounds

Absorption of radiation by organic molecules in the wavelength region between

180-780 nm results from interactions between photons and electrons. The wavelength at

which an organic molecules absorbs depend on how tightly is electrons are bound. The

shared electrons in carbon/carbon or carbon/hydrogen angle bond are held that excitation

requires energies to wavelengths in vacuum ultraviolet region below 180nm. Single-

bond spectra have not been widely exploited for analytical purposes because of the

experimental difficulties working in this region.

Electrons involved in double and triple bonds of organic molecules are not

strongly held and it more easily excited by radiation. Species with unsaturated bond

generally exhibit useful absorption peaks. Unsaturated organic functional group that

absorbs in the ultraviolet or visible regions are known as chromophores.

Conjugation between two or more chromophores tends to cause shifts in peak

maxima to longer wavelengths. Finally, the vibrational effects will affects the absorption

peaks in the ultraviolet and visible region.

Saturated organic compounds containing such as heteroatoms as oxygen,

nitrogen, sulfur or halogens have nonbonding electrons that can be excited by radiation

in the 170-250nm range. Alcohols and ethers are common solvents, so their absorption in

this region prevents measuring absorption of analytes dissolved in these compounds at

wavelengths shorter than 180-200nm. (Skoog, et al. 2004)

2.8.2 Qualitative Applications of Ultraviolet/Visible Spectroscopy

11
 Spectrophotometric measurements with ultraviolet radiation are useful for

detecting chromophoric groups. Because large parts of even the most complex organic

molecules are transparent to radiation longer than 180nm, the appearance of one or more

peaks in the region from 200 to 400 nm is clear indication of the presence of unsaturated

groups of the atoms such as sulfur or halogens. Can get an idea to the identity of the

absorbing groups by comparing the spectrum of an analyte with those of simple

molecules containing various chromophoric groups. Usually, ultraviolet spectra do not

have sufficient fine structure to permit an analyte to be identified unambiguously.

Ultraviolet qualitative data must be supplemented with other physical or chemical

evidence such as infrared, nuclear magnetic resonance and mass spectra as well as

solubility and melting and boiling point information. (Skoog, et al. 2004)

12
 CHAPTER 3

METHODOLOGY

3.1 Material

4-nitrobenzaldehyde, 3-pentanone, sodium hydroxide, ethanol 95%, distilled water,

ether, chloroform, ethyl acetate, acetone, ethanol, water, silica gel F254, methanol,

universal pH paper, phenolphthalein. All the material used are analysis grade (C) which

was obtained from SIGMA ALDRICK.

3.2 Equipment

Analytical balance, glass beaker, conical flask, measuring cylinder, magnetic stirrer,

oven (Memmert), water bath, filter paper, funnel, capillary tube, bijor bottle, Ointment

jar 250gm, TLC chamber, UV254 lamp (UVP 55 handheld UV lamp), UV

spectrophotometer (Perkin Elmer- Lambda 25)

3.3 Method

3.3.1 The Reaction (adopted from Rowland, 2004)

10ml of Sodium Hydroxide 10% was mixed with 4ml of Ethanol, known as

Mixture 1. Then, 2ml of Pentanone was mixed with 1gm of 4-Nitrobenzaldehyde, known

as Mixture 2. Half of the mixture 2 was added into Mixture 1, these 2 mixtures were

stirred for about 15 minutes. The remaining of mixture 2 (another half) was added into

the stirred mixture. The complete mixture was stirred for about 1 ½ hours. The result of

the

13
mixture was observed and found out that the colour of mixture has changed into dark

brownish colour.

3.3.2 Isolation and purification

Before isolation test, the oven was heated at 70°c. The complete mixture was

filtered in a funnel with a filter paper.

Few drops of sulphuric acid were dropped into the filtrate until the lump of the

oily phase appeared/ seen to grow up the crystal it was scratched intensively. 9ml of

filtrate was taken out and added with 9ml of ethyl acetate. These 2 mixtures were mixed

in a separating funnel. Mixture was shaking vigorously and allowed it to form 2 layers.

The top layer was collected. Then, procedure repeated few times by adding with 9ml of

ethyl acetate into separating funnel until a clearer top layer appears.

Product was put into the oven to evaporated ethyl acetate at 70°c. After taken out

from oven (product), it formed crystals / solid form on the filter paper.

Crystals were rinsed with distilled water and the pH solution of the product was

tested with an indicator. If the indicator appeared pink colour, this indicated as an

alkaline condition. It needed to rinse for few times until neutral. Indicator must appear in

colourless.

Crystals were rinsed with ethanol. Put the filter paper that contained the product

(crystals) into an oven again for drying. After that, product was taken out to weight.

14
3.3.3 Yields calculation
The obtained product
Yields = x 100%
The theoretical product

3.3.4 Analysis of physicochemical properties, purity and molecule structure of

product

3.3.4.1 Physical Appearance Test

The product is described about physical appearance such as shape, color and odor.

3.3.4.2 Solubility Test

6 bijor bottles are prepared, weight and labeled for the appropriate solvent to be

mixed. A sufficient sample (product) was put into each bottle and weighted. Put each

different solution (drop by drop) into sample until the sample dissolved. Solution that

used was recorded. Method was repeated for 4-nitrobenzaldehyde.

3.3.4.3 Thin Layer Chromatography (TLC)

Prepared solvent in a ointment jar 250gm that contain chloroform, CHCl3 (2) :

Methanol (8). Jar was covered and waited for 10 minutes. While waiting, 4-

nitrobenzaldehyde (standard), curcumin analogue and sample (product), was dissolved

with ethanol. Put a drop from each (standard,curcumin analogue and sample product) on

the TLC plate and labeled down. TLC plate was placed into the jar by making it stand.

Mobile phase/ spot were observed and it rose and reached to the top (1cm before the

top). TLC plate was allowed to dry. The spot was detected under the UV lamp. (The spot

of the standard should be higher than the spot of sample).

15
3.3.4.4 Maximum wave length test

The sufficient amount of product is dissolved in ethyl acetate and then the

maximum wave length is measured by UV-Visible spectrophotometer.

16
 CHAPTER 4

RESULTS

4.1 Yields calculation

2 (4-nitrobenzaldehyde) + 3-pentanone product + 2 H2O

Initial: 2(6.617 x 10-3) = 0.0132 0.0189 0 0

Reaction: 0.0132 0.0132 0.0132 0.0132

Balance: 0 5.7 x 10-3 0.0132 0.0132

3-pentanone 4-nitrobenzaldehyde
mass mass
ρ =volume mol =
JMR
mass
0.813 = 2ml mol = 1gm
151.12
mass = 1.626gm mol = 6.617 x 10-3
mass
mol = JMR
1.626gm
mol = 86.13

mol = 0.0189

Theoretically = 0.0132 mol of product

mass
mol =
JMR
mass
0.0132 =
352.338
mass = 4.65 gm

17
 The obtained product
Yields = The theoretical product x 100%

0.061gm
= 4.65gm x 100%

= 1.3%

4.2 Physical appearance

The product is in dark brown crystalline odourless powder.

4.3 Solubility test

Table 1.1 The solubility result for sample product

Solvent Weight of Volume of ratio Term in solubility
compound solvents
Water 0.002g >5ml 1:2500 Very insoluble
Ethanol 0.001g >5ml 1:5000 Very insoluble
Acetone 0.003g 0.3ml 1:100 Slightly soluble

Ethyl acetate 0.002g 0.4ml 1:200 Insoluble

Chloroform 0.003g >5ml 1:1667 Very insoluble
n-hexane 0.002g >5ml 1:2500 Very insoluble

Table 1.2 The solubility result for 4-nitrobenzaldehyde

Solvent Weight of Volume of ratio Term in solubility

compound solvents
Water 0.004g >5ml 1:1250 Very insoluble
Ethanol 0.005g 0.2ml 1:40 Slightly soluble
Acetone 0.005g 0.2ml 1:40 Slightly soluble

Ethyl acetate 0.01g 0.2ml 1:20 Soluble

Chloroform 0.005g 0.2ml 1:40 Slightly soluble
n-hexane 0.007g >5ml 1:714 Insoluble

18
 Table 1.3 Indonesian Pharmacopeia 3rd Edition

Terms of solubility A ml of solvent needed to dissolve 1gm of substance

Very easily soluble 1: (<1)
Easily soluble 1: (1-10)
Soluble 1: (10-30)
Slightly soluble 1: (30-100)
Insoluble 1: (100-1,000)
Very insoluble 1: (1,000-10,000)
Practically insoluble 1: (>10,000)

4.4 Thin layer chromatography

This is the TLC call chromatogram.

Plate A - Mobile Phase: 2 part chloroform: 8 part methanol

Rf = 0.88

Rf = 0.80

A B
Figure 1.8 The TLC plate A.
Inside the plate is 4–nitrobenzaldehyde (A) and the product (B)

Plate A - Mobile Phase: 2 part chloroform: 8 part methanol

Rf = Distance travel by the substance

Distance travel by the solvent

Rf of 4-nitrobenzaldehyde = 4.6 cm
5.2 cm
= 0.887

Rf of product = 4.2 cm
19
 5.2 cm
= 0.808

Plate B - Mobile Phase: 5 part chloroform: 5 part methanol

Rf = 0.96

Rf = 0.90

A B
Figure 1.9 The TLC plate B.
Inside the plate is 4–nitrobenzaldehyde (A) and the product (B)

Rf = Distance travel by the substance

Distance travel by the solvent

Rf of 4-nitrobenzaldehyde = 5.2 cm
5.4 cm
= 0.963

Rf of product = 4.9 cm
5.4 cm
= 0.907

20
Plate C - Mobile Phase: 8 part chloroform: 2 part methanol
Rf = 0.98

Rf = 0.92

A B
Figure 2.0 The TLC plate C.
Inside the plate is 4–nitrobenzaldehyde (A) and the product (B)

Rf = Distance travel by the substance

Distance travel by the solvent

Rf of 4-nitrobenzaldehyde = 5.1 cm
5.5 cm
= 0.927

Rf of product = 5.4 cm
5.5 cm
= 0.982

Indicate that the structure of the product is different with 4-nitrobenzaldehyde as

indicated in Plate A.

21
4.5 Maximum wave length test

Figure 2.1 The wavelength of 4-nitrobenzaldehyde

22
Figure 2.2 The wavelength of product (2,4-bis-(4-nitrobenzylidene)-3 pentanone)
CHAPTER 5
23
 DISCUSSION

5.1 Reaction and Isolation

The reaction of 4-Nitrobenzaldehyde and 3-pentanone using Sodium Hydroxide

(NaOH) as a catalyst and ethanol as a solvent product reddish brown mixture. It is

because aldol condensation occur during this reaction. The obtained product still in

soluble so it was acidified by using acid sulphuric and helped by scratching technique.

Once the crystals grow, it is ready to isolate.

During isolation, the filtrate was mixed with ethyl acetate. Mixture was shake

and allowed to form 2 layers. The filtrate itself contains products, water and ethanol.

Thus, the upper layer is less polar and the bottom layer is more polar. Since the product

is predicted as a non-polar solvent, it was expected that product will be removed from

NaOH-ethanol to ethyl acetate. Once it is extracted, it will be easily purified.

First purification, it was used water to clean up any by products or contaminants

which soluble in it while neutralizing the product from acid pH. Second purification, it

was used ethanol to clean up a bit of 4-nitrobenzaldehyde that might be remained in the

product. Then, the product was dried to obtain the pure crystals.

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5.2 Physical appearance

Physical Product 4-nitrobenzaldehyade 3-pentanon

appearance
Shape Crystalline powder Crystal-like edges Colourless liquid
Colour Dark brown White or pale yellow Colourless
Odour Odourless Characteristic odour Characteristic odour
Table 1.4 Differences between Product and 4-nitrobenzaldehyde and 3-pentanone

From the data (Table 2.1) it gave the first positive result that the difference physical

appearance between the product, 4-nitrobenzaldehye and 3-pentanone describe the

successful of reaction

5.3 Solubility Test

The product is slightly soluble in acetone because it contains ketone group. The

product and acetone has a similarity in their structure, it will be easier to dissolve in the

acetone compare to others solvent used in this test. So it can enhance the positive result

of occur reaction. Refer to the Table 1.1 and Table 1.2, it provide the second information

that the product is different with 4-nitrobenzaldehyde as a starting material especially the

solubility in alcohol.

5.4 Thin Layer Chromatography

Thin layer chromatography is a technique to identify unknown compounds by

comparison with standards. We compare our product, 2, 4-bis-(4-nitrobenzylidene)-3-

pentanone with standard 4-nitrobenzaldehyde. The stationary phase is silica gel on

aluminium plate. The measurement of the TLC plate is 7cm height and 3cm wide. We

use two types of solvents (mobile phase) chloroform (non polar solvent) and methanol

(polar solvent) in different proportion. We use three proportion, two part chloroform and

25
8 part methanol, 5 part chloroform and 5 part methanol, and 8 part chloroform and 2 part

methanol. The TLC plate label as plate A, plate B, plate C accordingly. The different

proportions provide different polarity of the mobile phase (solvent).

Figure 2.4 From left Plate A, Plate B, Plate C under short wave ultraviolet light.
The dark spot is the 4-nitrobenzaldehyde.

26
Figure 2.5 From bottom; Plate A, Plate B, Plate C under long wave ultraviolet light.
The spot that florescence is the product.

For the plate A the height travel by the mobile phase is 5.2cm. The height travel

by the product is 4.2cm while the standard, 4-nitrobenzaldehyde is 4.6cm. The

Retardation factor (Rf) for 4-nitrobenzaldehyde is 0.887 while Rf for the product is

0.808. This indicate that the structure of the product is different with 4-

nitrobenzaldehyde.

27
 For the plate B The height travel by the mobile phase is 5.4cm. The height travel

by the product is 4.9cm while the standard, 4-nitrobenzaldehyde is 5.2cm. The

Retardation factor (Rf) for 4-nitrobenzaldehyde is 0.963 while Rf for the product is

0.907 This indicate that the structure of the product is different with 4-nitrobenzaldehyde

as indicated in Plate A.

For the plate C The height travel by the mobile phase is 5.5cm. The height travel

by the product is 5.4cm while the standard, 4-nitrobenzaldehyde is 5.1cm. The

Retardation factor (Rf) for 4-nitrobenzaldehyde is 0.927 while Rf for the product is

0.982 This indicate that the structure of the product is different with 4-nitrobenzaldehyde

as indicated in Plate A and Plate B.

The TLC plate is polar so the compound that have higher Rf will be less polar

compound. But the Rf of both compound do not have significant differences so we can

conclude that both the compound is similar in term of polarity.

The 4-nitrobenzaldehyde exhibit dark spot on all plate under short wave ultra

violet light. This due to the structure of the conjugated double bond in 4-

nitrobenzaldehyde absorb the short wave ultra violet light. 4-nitrobenzaldehyde has one

benzene ring and less conjugated double bond present in the compound. Thus the light

that 4-nitrobenzaldehyde absorb is short wave as it required more energy from the ultra

violet light to promote the electron to jump from pi bonding to pi anti-bonding. That why

4-nitrobenzaldehyde is colourless.

The product or 2,4-bis-(4-nitrobenzylidene)-3-pentanone exhibit florescence

under long wave ultra violet light illumination. The more double bond (pi bonding) or

benzene ring ( also known as delocalisation) there is, the smaller the gap between the

28
highest energy pi bonding orbital and the lowest energy pi anti-bonding orbital. Thus it

takes less energy to promote the electron from pi bonding orbital to pi anti-bonding

orbital. Thus, the energy required to absorb the ultraviolet light is low. Less energy

means a lower frequency of light gets absorbed - and that's equivalent to a longer

wavelength. This cause the spot to florescence under the long wave ultraviolet light.

This data enforce the positive result as expected as hypothesis stated.

5.5 Maximum wave length test

The sample was tested using UV-Vis Spectrophotometer. From the solubility test,

we found out that the product is quite soluble in ethyl acetate. Therefore, it was using

ethyl acetate as a solvent. By using the UV-Vis spectrophotometer, it can measure the

maximum wavelength of the product.

The result shows that the product have maximum wavelength 293.96nm and the

4-nitrobenzaldehyde have maximum wavelength 264.28nm. It show that the product is

different with the 4-nitrobenzaldehyde. Therefore, the structure of 4-nitrobenzaldehyde

was changed. From the result, between the 200nm and 250nm, it show unstable result

which caused by impurity of the product.

This shift of the product wavelength is due to the prolongation of conjugated double

bond as seen in figure 2.2.

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 CHAPTER 6

CONCLUSION AND RECOMMENDATIONS

6.1 Conclusion

Refer to the result of the physical that there are different physical and chemical

properties before and after reaction of 4-nitrobenzaldehyde and 3-pentanone due to the

predicted product of 2,4-bis-(4-nitrobenzylidene)-3-pentanone.

6.2 Implications of the study

Once is hypothesis is confirmed, it will be a trigger to numerous other equivalent studies

such us its toxicity, pharmacokinetic, pharmacodynamic, etc. that would explore the

prospect of the product to be the new antibacterial agent. The synthesis of compound is

relative efficient in time and cost, therefore it can be predicted that candidate will be

relative low in price. It will be benefit both for pharmaceutical producer and a patient as

a consumer, respectively.

6.3 Recommendations for future research

As the yield was low, still it is suggested to the next study to optimize the method by

using less reactive base catalyst such as pyridine to minimize the abundancy if the

product.

It also recommended that the next research should test for its biological, antimicrobial,

antioxidant, anti inflammatory and other activities.

30
 REFERENCES

Aggarwal, B. B., A. Kumar, M. S. Aggarwal, and S. Shishodia. Curcumin derived from

turmeric (Curcuma longa): A spice for all seasons. Phytopharmaceuticals in
Cancer Chemoprevention. 2005. CRC Press LLC

Al- Wabli, R. I. Synthesis of curcumin analogues biconjugates as potential antitumor

agents in isolated human cells. Thesis. 2006. College of Pharmacy. King Saud
Saudi. p. 10

Mcmurry J. and E. Simanek Fundamental of Organic Chemistry Sixth Edition 2007

Thomson Higher Education. Pp 357-360

Masuda, T., J. Isobe, A. Jitoe, and N. Nakatani. Antioxidative curcuminoids from

rhizomes of Curcuma xanthorrhiza Roxb. Phytochemistry. 1992. 31 (10). pp:
3645-3647

Rowland, A. T. A Multistep synthesis sequence. An aldol condensation, a Michael

addition, and Ethylene Ketal Formation.2004. CER Modular Program in
Chemistry. Thomson learning. pp: 1-5

Sardjiman, M. S. Reksohadiprodjo, and H. Timmerman. Derivatives of benzylidene

cyclohexanone, benzylidene cyclopentanone, and benzylidene acetone and their
synthesis. 1997. United States Patent Application

Skoog. D. A., D. M. West, J. Holler and S. R. Crouch. Fundamental of analytical

chemistry,2004. 8th Ed., International Student Edition. Thomson learning.
Pp 784-787, pp 811-817, pp 973-974 and 980-981

Youessef, K.M, M. El- Sherbeny, F. S. El-Shafie, H.A Farag, O. A. Al-Deeb.

Synthesis of curcumin analogues as potential antioxidant, cancer
chemopreventive agents. Arch Pharm (wePincheim). 2004. Vol. 337. Issue I. pp.
42-54

http://en.wikipedia.org/wiki/Aldehydes, accessed on 14.07.2009

http://en.wikipedia.org/wiki/Ketones, accessed on 14.07.2009

http://en.wikipedia.org/wiki/Thin_layer_chromatography, accessed on 29.03.2010

http://en.wikipedia.org/wiki/Solvent, accessed on 30.03.2010

http://www.chemguide.co.uk/analysis/uvvisible/theory.html, accessed on 30.03.2010

31
http://www.chemyq.com/En/xz/xz11/101813qypdq.htm, accessed on 10.04.10

http://www.inchem.org/documents/icsc/icsc/eics0874.htm, accessed on 10.04.10

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 APPENDICES

A. The apparatus and materials use

2,4-bis-(4-nitrobenzylidene)-3-pentanone 4-nitrobenzaldehyde and 3-pentanon

A dark brown crystalline odourless powder white or pale yellow crystal-like edges

B. Differences between two chemical physically

33
C. The reaction between 4-nitrobenzaldehyde and 3 pentanone

D. Isolation of our product

34
 FILTRATE

OILY PRODUCT (COMPLETE MIXTURE)

E. The isolation of product

F. Separating Funnel

35

G. Thin Layer Chromatography Plate