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ISSN: 2320-5407 Int. J. Adv. Res. 12(01), 1261-1267
1261
Journal Homepage: -www.journalijar.com
ArticleDOI:10.21474/IJAR01/18244
DOI URL: http://dx.doi.org/10.21474/IJAR01/18244
RESEARCH ARTICLE
USING FOURIER TRANSFER INFRARED SPECTROSCOPY TO ANALYSIS EFFECTS OF CADMIUM
TREATMENT ON RICE (ORYZA SATIVA L.) SEEDLINGS
Prity Basak1, Ankita Ghosh2, Dipshikha Biswas3, Soumi Ghosh4 and Rohit Kumar Ghosh5
1. Guru Nanak Institute of Pharmaceutical Science and Technology, Kolkata-700114, West Bengal, India.
2. Guru Nanak Institute of Pharmaceutical Science and Technology, Kolkata-700114, West Bengal, India.
3. Guru Nanak Institute of Pharmaceutical Science and Technology, Kolkata-700114, West Bengal, India.
4. University of Calcutta, Kolkata- 700073, West Bengal, India.
5. Guru Nanak Institute of Pharmaceutical Science and Technology, Kolkata-700114, West Bengal, India.
……………………………………………………………………………………………………....
Manuscript Info Abstract
……………………. ………………………………………………………………
Manuscript History
Received: 30 November 2023
Final Accepted: 31 December 2023
Published: January 2024
Keywords:-
Oryza SativaL., FTIR Spectroscopy,
Functional Groups, DMSO
The present study is aimed to analyse the DMSO (Dimethyl sulfoxide)
extract of shoots and roots of Oryza sativaL. (var IET 4786) through
FTIR spectroscopy method. These studies contain different
characteristic of the peak values with various functional compounds in
the extracts. This analysis revealed the presence of different types of
biomolecules in plant extracts. The FTIR analysis of DMSO leaf
extracts of Oryza sativaL. confirmed the presence of alcohols, phenols,
amides, alkanes, carboxylic acids, aldehydes, ketones, alkenes,
aromatics, esters, alkyl halides and aliphatic amines compounds; each
of the functional group showed major peaks. This method was
performed on a spectrophotometer system which was used to detect the
characteristics peak values and their functional groups. In this present
study, the FTIR spectrum profile for the plant Oryza sativa L. is
generated andcan be used in the industry.
Copy Right, IJAR, 2024,. All rights reserved.
……………………………………………………………………………………………………....
Introduction:-
Cadmium (Cd) is non-essential heavy metal and also it is hazardous for living organism (Bhattacharya, 2015;
Aiqing, 2021; Shanmugaraj, 2019). The common symptoms of the cadmium toxicity are chlorosis, growth inhibition
and necrosis. In order to the functional groups present in these chemical constituents of plant are usually identified
by FTIR. It is used to elucidate the structure of isolated compounds.The presence of phenols, alkanes, alcohol, alkyl
halides, carboxylic acid and aromatic compounds in DMSO extracts of Oryza sativaL. was also studied by FTIR
spectroscopy method. Identify the chemical nature of the constituents that present in the rice plants. And the
properties are related to certain compounds such as flavonoids, alkaloids and other things.Thus, FTIR spectroscopy
method has become one of the avenues for the identification of compounds. Hence, an attempt is made in the present
study to analyse the functional groups of Phyto active compounds present in the shoot and root extracts (in solvent
such as, DMSO). This also helps to know the influence of solvents on functional group present in the plant
(Raghavendran, et al., 2011)
Corresponding Author:- Rohit Kumar Ghosh
Address:- Guru Nanak Institute of Pharmaceutical Science and Technology, Kolkata-
700114, West Bengal, India.
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Materials and Methods:-
Seed collection and treatment
The rice seedlings Oryza sativaL. (var. IR 4786) was obtained from the Bidhan ChandraKrishi Vishwavidyalaya
(BCKV), which is also known as the Bidhan Chandra AgriculturalUniversity and is located in Mohanpur,West
Bengal, India. After being surface
sterilizedwithasolutioncontaining1%sodiumhypochlorite,thisvarietywassoakedindistilledwaterat 4oC for 24 hours to
absorb water. The 15 seeds were then transferred to three petriplates and surrounded by three filter papers. The
seedlings are then given Cadmium chloride(CdCl2) treatments (0, 100, 500 µM). For 10 days, the experiment was
carried out in thelaboratory under room temperature conditions. By irrigation with distilled water twiceper week, the
Cadmium (Cd) treatment was administered. According to [4], three sets weremaintained throughout the entire
experiment. Every day, the nutrient solutions were changed[2].After sevendays,theshootsandrootsof three rice
plantsampleswere usedfor FTIRexperiments.
FTIR Spectroscopic Analysis:
Fourier Transform Infrared Spectrophotometer (FTIR) is most significant tool for identifying the types of functional
groups that present in chemical compounds. The plant material was dried in hot air oven. The dried plants shoot and
root material was ground to powder and stored in air tight container for further use.The dried powdered shoot and
root (1 g) of Oryza sativaL. was ground in mortal and pestle in order to obtain fine powder and mixed with DMSO
(95% v/v) for 24 hours. After 24 hours the plant extract was loaded in FTIR spectroscope ((Shimadzu, IR Affinity 1,
Japan) and Scan range from 400 to 4000 cm- 1 with a resolution of cm- 1(Nair, Sar, Arora, & Mahaptra, 2019)
Result:-
The FTIR spectrum of shoot and root extracts of the data on the peak values and the functional groups of Oryza
sativaL. presented in table 1 to 4
In control, the intense band occurring at 2920 cm-1, 2854 cm-1, 2226 cm-1, 1746 cm-1, 1672 cm-1, 1437 cm-1, 1316
cm-1, 1014 cm-1, 704.6 cm-1 and 610.5 cm-1 corresponding to C-H/ O-H/ -C#C-/ C=O/ C=C/ C-F/ C-O/ =C-H/ C-Br
stretch and bend are indicating the presence of Alkanes, Carboxylic acids, Alkynes, Amides, Alkenes, Aromatics,
Alkyl halides and Esters compounds respectively in shoots and 2919 cm-1,1672 cm-1,1315 cm-1, 1014 cm-1, 704.3
cm-1 and 617.3 cm-1 corresponding to O-H/ C=O/ C-C/ C-O/ =C-H/ C-H stretch and bend are indicating the presence
of Carboxylic acids, Ketones, Aromatics, Esters, Alkenes, Alkynes compounds respectivelyin roots IET 4786
variety of Oryza sativaL. (Rice).
In (100 µM) the low concentration of cadmium chloride, the intense band occurring at 2919 cm-1,1672 cm-1,1437
cm-1, 1316 cm-1, 1013 cm-1, 705.1 cm-1 and 609 cm-1 corresponding to C-H/ C=O/ C-C/ C-O/ C-F stretch, "oop", and
bend are indicating the presence of Alkanes, Ketones, Aromatics, Carboxylic acids, Alkyl halides, Alkynes
compounds respectivelyin shoots and 2919 cm-1,1672 cm-1,1437 cm-1, 1316 cm-1, 1013 cm-1, 705.4 cm-1 and 609.7
cm-1 corresponding to C-H/ C=O/ C-C/ C-N/ C-F, N-H wag and stretch and bend are indicating the presence of
Alkanes, Ketones, Aromatics, Carboxylic acids, Alkyl halides, Amines, Alkynes compoundsrespectively in roots the
IET 4786 variety of rice(Oryza sativaL.).
In (500 µM) the high concentration of cadmium chloride, the intense band occurring at 2919 cm-1,1676 cm-1,1437
cm-1, 1316 cm-1, 1014 cm-1, 704.8 cm-1 and 565.2 cm-1 corresponding to C-H/ C=O/ C-C/ C-N/ C-O/ C-Br stretch
and "oop" are indicating the presence of Alkanes, Amides, Aromatics, Carboxylic acids, Esters, Alkyl halides
compounds respectivelyin shoots and 2920 cm-1,1672 cm-1,1437 cm-1, 1316 cm-1, 1014 cm-1, 704.6 cm-1 and 610.5
cm-1 corresponding to C-H/ C=O/ C-C/ C-O/ C-F stretch, "oop", and bend are indicating the presence of Alkanes,
Ketones, Aromatics, Carboxylic acids, Alkyl halides, Alkynes compoundsrespectively in roots the IET 4786 variety
of Oryza sativaL. (Rice).
Table 1:- FTIR spectra without CdCl2 concentrations on 10 days old(var. IET 4786) shoot of Oryza sativa L.
riceseedlings.
Peak
number
Wave number cm-
1[Test sample]
Wave number cm-1
[Reference article]
Functional groups
Phyto compounds
1
2920
3000–2850
C–H stretch
Alkanes
2
2854
3300-2500
O–H stretch
Carboxylicacids
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3
2226
2260-2100
–C#C– stretch
Alkynes
4
1746
1750 -1740
C=O stretch
Amides
5
1672
1680-1640
C=C stretch
Alkenes
6
1437
1500-1400
C–C stretch (in-ring)
Aromatics
7
1316
1350 - 1000
C-F stretch
Alkyl halides
8
1014
1300-1000
C–O stretch
Esters
9
704.6
1000-650
=C–H bend
Alkenes
10
610.5
690-515
C–Br stretch
Alkyl halides
Table 2:- FTIR spectra without CdCl2 concentrations on 10 days old (var IET 4786) root of Oryza sativa L.
riceseedlings.
Peak
number
Wave number cm-1
[Test sample]
Wave number cm-1
[Reference article]
Functional groups
Phyto
compounds
1
2919
3300-2500
O-H stretch
Carboxylic
acids
2
1672
1715
C=O stretch, aliphatic
ketones
ketones
3
1315
1500-1400
C-C stretch (in-ring)
Aromatics
4
1014
1200-1000
C-O stretch
Esters
5
704.3
1000-650
=C–H bend
Alkenes
6
617.3
700-610
C–H bend
Alkynes
Table 3:- FTIR spectra low CdCl2 concentration (100 µM) on 10 days old (var IET 4786) shootof Oryza sativa L.
riceseedlings.
Peak
number
Wave number cm-1
[Test sample]
Wave number cm-1
[Reference article]
Functional groups
Phyto
compounds
1
2919
3000–2850
C–H stretch
Alkanes
2
1672
1685-1666
C=O stretch
Ketones
3
1437
1500-1400
C–C stretch
Aromatics
4
1316
1320-1210
C–O stretch
Carboxylic acids
5
1013
1350 - 1000
C-F stretch
Alkyl halides
6
705.1
900-675
C–H "oop"
Aromatics
7
609
700-610
C–H bend
Alkynes
Table 4:- FTIR spectra low CdCl2 concentration (100 µM) on 10 days old (var IET 4786) root of Oryza sativa L.
riceseedlings.
Peak
number
Wave number cm-1
[Test sample]
Wave number cm-1
[Reference article]
Functional groups
Phyto
compounds
1
2919
3000–2850
C–H stretch
Alkanes
2
1672
1685-1666
C=O stretch
Ketones
3
1437
1500-1400
C–C stretch
Aromatics
4
1316
1335-1250
C–N stretch (aromatic
amines)
Carboxylic acids
5
1013
1350 - 1000
C-F stretch
Alkyl halides
6
705.4
910-665
N–H wag (primary and
secondary amines only)
Amines
7
609.7
700-610
C–H bend
Alkynes
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Wavenumbers [cm-1 ]
3400 3200 3000 2800 2600 2400 2200 2000 1800 1600 1400 1200 1000 800 600 400 200
Transmittance [%]
130
120
110
100
90
80
70
60
50
40
30
20
10
2919 98.61
2317 100.1
1966 100.3
1672 101.3
1437 86.64
1315 91.76
1014 31.68
951.6 55.41
704.3 90.07
617.3 98.15
Sample 026 By PEService Date Wednesday, March 22 2023
Table 5:- FTIR spectra high CdCl2 concentration (500 µM) on 10 days old (var IET 4786) shoot of Oryza sativa L.
riceseedlings.
Peak
number
Wave number cm-1
[Test sample]
Wave number cm-1
[Reference article]
Functional groups
Phyto
compounds
1
2919
3000–2850
C–H stretch
Alkanes
2
1676
1685-1666
C=O stretch (free)
Amides
3
1437
1500-1400
C–C stretch
Aromatics
4
1316
1335-1250
C–N stretch (aromatic
amines)
Carboxylic acids
5
1014
1300-1000
C-O stretch
Esters
6
704.8
900-675
C–H "oop"
Aromatics
7
565.2
700-610
C–Br stretch
Alkyl halides
Table 6:- FTIR spectra high CdCl2 concentration (500 µM) on 10 days old (var IET 4786) root of Oryza sativa L.
riceseedlings.
Peak
number
Wave number cm-1
[Test sample]
Wave number cm-1
[Reference article]
Functional groups
Phyto compounds
1
2920
3000–2850
C–H stretch
Alkanes
2
1672
1680-1640
C=C stretch
Alkenes
2
1437
1500-1400
C–C stretch (in ring)
Aromatics
4
1316
1350 - 1000
C-F stretch
Alkyl halides
5
1014
1300-1000
C–O stretch
Esters
6
704.6
1000-650
=C–H bend
Alkenes
7
610.5
690-515
C–Br stretch
Alkyl halides
Figure 1:- FTIR analysis without CdCl2 concentrations of shoot and root on (var IET 4786) of 10 days old rice
seedlings Oryza sativa L.
Figure 2:- FTIR analysis low CdCl2 concentration (100 µM) on 10 days old (var IET 4786) shoot and root of Oryza
sativa L. riceseedlings
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Figure 3:- FTIR analysis high CdCl2 concentration (500 µM) on 10 days old (var IET 4786) shoot and root of Oryza
sativa L. riceseedlings.
Discussion:-
In this study we identify the functional group that present in the shoot and root in the variety (IET 4786) of Oryza
sativa L. with the help of FTIR analysis.In fig 1 to 3 it helps to identify the chemical components, interprets the
structure of chemical structure and helps to understand the importance of functional group.
The spectra of simple alkanes are characterized by absorptions due to C–H stretching and bending (the C–C
stretching and bending bands are either too weak or of too low a frequency to be detected in IR spectroscopy). In
simple alkanes, which have very few bands, each band in the spectrum can be assigned. However, each organic
compound has its own unique absorption pattern in this region and thus an IR spectrum can be used to identify a
compound by matching it with a sample of a known compound( MEENAMBAL, et al., 2012).
Carboxylic acids show a strong, wide band for the O–H stretch. Unlike the O–H stretch band observed in alcohols,
the carboxylic acid O–H stretch appears as a very broad band in the region 3300-2500 cm-1, cantered at about 3000
cm-1. This is in the same region as the C–H stretching bands of both alkyl and aromatic groups. Thus, a carboxylic
acid shows a somewhat "messy" absorption pattern in the region 3300-2500 cm-1, with the broad O–H band
superimposed on the sharp C–H stretching bands. The reason that the O–H stretch band of carboxylic acids is so
broad is because carboxylic acids usually exist as hydrogen-bonded dimers. The carbonyl stretch C=O of a
carboxylic acid appears as an intense band from 1760-1690 cm-1. The exact position of this broad band depends on
whether the carboxylic acid is saturated or unsaturated, dimerized, or has internal hydrogen bonding. (Sharma, et al.,
2011). The C–O stretch appears in the region 1320-1210 cm-1, and the O–H bend is in the region 1440-1395 cm-1
and 950-910 cm-1, although the 1440-1395 band may not be distinguishable from C–H bending bands in the same
region.
Alkynes are compounds that have a carbon-carbon triple bond (–C#C–). The –C#C– stretch appears as a weak band
from 2260-2100 cm-1. This can be an important diagnostic tool because very few organic compounds show an
absorption in this region. A terminal alkyne (but not an interchain alkyne) will show a C–H stretch as a strong,
narrow band in the range 3330-3270 cm-1. (Often this band is indistinguishable from bands resulting from other
functional groups on the same molecule which absorb in this region, such as the O-H stretch.). A terminal alkyne
will show a C–H bending vibration in the region 700-610 cm-1(Ncube , et al., 2008).
Amides functional group combines the features of amines and ketones because it has both the N-H bond and the
C=O bond. Therefore, amides show a very strong, somewhat broad band at the left end of the spectrum, in the range
between 3100 and 3500 cm-1 for the N-H stretch. At the same time, they also show the stake-shaped band in the
middle of the spectrum around 1710 cm-1 for the C=O stretch(Gaurav , et al., 2010). As with amines, primary
amides show two spikes, whereas secondary amides show only one spike.
Alkenes are compounds that have a carbon-carbon double bond, –C=C–. The stretching vibration of the C=C bond
usually gives rise to a moderate band in the region 1680-1640 cm-1. Stretching vibrations of the –C=C–H bond is of
higher frequency (higher wavenumber) than those of the –C–C–H bond in alkanes. Absorption peaks above 3000
cm-1 are frequently diagnostic of unsaturation. This is a very useful tool for interpreting IR spectra. Only alkenes and
aromatics show a C-H stretch slightly higher than 3000 cm-1. Compounds that do not have a C=C bond show C-H
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stretches only below 3000 cm-1. The strongest bands in the spectra of alkenes are those attributed to the carbon-
hydrogen bending vibrations of the =C–H group. These bands are in the region 1000-650 cm-1(THENMOZHI, et al.,
2011).
The =C–H stretch in aromatics is observed at 3100-3000 cm-1. This is at slightly higher frequency than is the –C–H
stretch in alkanes. This is a very useful tool for interpreting IR spectra: Only alkenes and aromatics show a C–H
stretch slightly higher than 3000 cm-1. Compounds that do not have a C=C bond show C–H stretches only below
3000 cm-1. Aromatic hydrocarbons show absorptions in the regions 1600-1585 cm-1 and 1500-1400 cm-1 due to
carbon-carbon stretching vibrations in the aromatic ring. Bands in the region 1250-1000 cm-1 are due to C–H in-
plane bending, although these bands are too weak to be observed in most aromatic compounds. Besides the C–H
stretch above 3000 cm-1, two other regions of the infrared spectra of aromatics distinguish aromatics from organic
compounds that do not have an aromatic ring(DEEPASHREE, et al., 2012). It is useful to remember that aromatics
in general show a lot more bands than compounds that do not contain an aromatic ring.
Alkyl halides are compounds that have a C–X bond, where X is a halogen: bromine, chlorine, fluorene, or iodine. In
general, C–X vibration frequencies appear in the region 850-515 cm-1, sometimes out of the range of typical IR
instrumentation. C–Cl stretches appear from 850–550 cm-1, while C–Br stretches appear at slightly lower
wavenumbers from 690-515 cm-1. In terminal alkyl halides, the C–H wag of the –CH2X group is seen from 1300-
1150 cm-1. Complicating the spectra is a profusion of absorptions throughout the region 1250-770 cm-1, especially in
the smaller alkyl halides(JANAKIRAMAN, et al., 2011) All of these bands are in the fingerprint region.
The carbonyl stretch C=O of aliphatic esters appears from 1750-1735 cm-1; that of alfa,beta-unsaturated esters
appears from 1730-1715 cm-1.The C–Ostretches appear as two or more bands in the region1300-1000cm-
1(MUTHANNA, et al., 2009).
The carbonyl stretching vibration band C=O of saturated aliphatic ketones appears at 1715 cm-1. Conjugation of the
carbonyl group with carbon-carbon double bonds or phenyl groups, as in alpha, beta-unsaturated aldehydes and
benzaldehyde, shifts this band to lower wavenumbers, 1685-1666 cm-1.
The most characteristic band in amines is due to the N-H bond stretch, and it appears as a weak to medium. This
band is positioned at the left end of the spectrum, in the range of about 3250 - 3400 cm-1. Primary amines have two
N-H bonds;therefore, they typically show two spikes that make this band resemble a molar tooth. Secondary amines
have only one N-H bond, which makes them show only one spike, resembling a canine
tooth(MURUGANANTHAM, et al., 2009) Finally, tertiary amines have no N-H bonds, and therefore this band is
absent from the IR spectrum altogether. The spectrum below shows a secondary amine.
Secondary amines (R2NH) show only a single weak band in the 3300-3000 cm-1 region, since they have only one N–
H bond. Tertiary amines (R3N) do not show any band in this region since they do not have an N–H bond. The N–H
bending vibration of primary amines is observed in the region 1650-1580 cm-1. Usually, secondary amines do not
show a band in this region and tertiary amines never show a band in this region. Another band attributed to amines is
observed in the region 910-665 cm-1. This strong, broad band is due to N–H wag and observed only for primary and
secondary amines.
Conclusion:-
In the current study from the results, after extraction the functional groups have been noticed in the sample. Using
FTIR spectrum we can confirm the functional groups are present in the given extract and indicate the presence of
phytoconstituents like carbohydrates, flavonoids, carotenoids, amino acids, amides, phosphates, lipids and phenols.
We know OH group present in the plant extract that has inhibitory activity against different microorganisms.Many
researchers applied the FTIR spectrophotometer as a tool for distinguishing the associated plants.Additionally, it has
accurate and sensitive technique for detecting the changing of biological molecular compounds.
Acknowledgment:-
I would like to extend my gratitude to Professor (Dr) Abhijit Sengupta (Director) and Professor (Dr) Lopamudra
Dutta (Principal) of Guru Nanak Institute of Pharmaceutical Sciences and Technology, Kolkata for allowing us the
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opportunity to participate in this research endeavor. I express my thanks to Dr. Bhaskar Choudhury, my mentor, for
his insightful counsel and inspiration during the production of the paper.
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