See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/366066755

Phytochemical screening and fourier transform of Sweet Orange peels The

phytochemical screening and fourier transform infrared spectrum analysis
(ftir) of Sweet Orange peels (Citru...

Article · June 2018

CITATIONS

5 authors, including:

George Sambo
Ahmadu Bello University
4 PUBLICATIONS 0 CITATIONS

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

INFLUENCE OF CHLORELLA VULGARIS ON THE GROWTH, BIOMASS AND YIELD PARAMETERS OF MAIZE View project

The herbicidal potential of sweet orange (Citrus sinensis L.) peel extracts on the germination and seedling growth of Senna obtusifolia l. (sicklepod) View project

All content following this page was uploaded by George Sambo on 07 December 2022.

The user has requested enhancement of the downloaded file.

Phytochemical screening and fourier transform of Sweet Orange peels

Journal of Tropical Biosciences

ISSN:0795-3089
Volume 13: (June, 2018)

The phytochemical screening and fourier transform infrared spectrum analysis (ftir) of
Sweet Orange peels (Citrus sinensis)

A.N. Adewale1, D. N. Iortsuun2, S. O. Alonge2, G. Y. Sambo2* and A. M. Chia2

2
Department of Botany, Ahmadu Bello University, Zaria,.
*Author for correspondence: georgeyabosambo@gmail.com

Abstract

This study was carried out to determine the presence of secondary metabolites and Fourier
Transform Infrared Spectrum Analysis (FTIR) were also carried out on sweet orange peels
extracts. Carbohydrate, Cardiac Glycosides, Alkaloids, Flavonoids, Saponins, Tannins,
Steroids and Triterpenes were all assessed using ethanol, petroleum ether, ethyl acetate,
chloroform, n-butanol and aqueous as solvents following standard procedures. FTIR was
carried out using the Shimadzu Spectrophotometer 40008. The phytochemical screening
showed the presence of seven secondary metabolites in orange peels extracts. They include:
carbohydrates, glycosides, saponins, steroids and triterpenes, flavonoids, tannins and alkaloids.
The ethanolic extract showed the presence of maximum number of compounds. FTIR analysis
indicates the presence of carbonyl, hydroxyl, amine, aliphatic and organic hydrocarbon
functional groups. This study demonstrated the biochemical composition of sweet orange peels
which could be further exploited for various uses.

Keywords: Tannins, FTIR, saponins, steroids, carbohydrates and glycosides

Introduction The phytochemical screening of methanol extract

Sweet orange (Citrus sinensis) fruits are mainly
used by juice processing industries but the peels
constitute to a large chunk of waste materials
(Manthey and Grohmann, 2001). In other to
address this problem there is increased attention
being given to the recycling and extraction of
useful products from waste materials.
Recently, efforts were made in the utilisation of
this in agriculture and medicine such as
commercial production of hesperidins (Kanaze et
al., 2009), and development of natural biocides
from Citrus waste as a new wood preservative
(Macías et al., 2005). Locally it has been found
useful in production of bathing powder,
mosquito repellant, bath oil, household cleaner,
scent and keeping of brown sugar soft (Valli,
2007).
© Journal of tropical Biosciences
of orange peel showed that it contains flavonoids,
phenol compounds and essential oils (Kanaze et
al., 2009). The presence of these phytochemicals
in the peel has made it useful as natural
nematicide (Abolusoro et al., 2010), insecticide
(Abolusoro, 2001), adsorbent for removal of
metals in industrial waste water (Rafique and
Nazir, 2013), and as main source of antioxidant
in food to prevent rancidity, fostering stability as
well as inhibiting lipid oxidation (Rehman, 2006,
Yassari and Yasari, 2013). Ezejiofor et al. (2011),
in one of their waste to wealth series, explored the
potential of sweet orange (Citrus sinensis) peels,
usually available in great abundance in parts of
Nigeria and other countries of the world, as a
major plant source of essential oils, a vital
resource, very useful in foods, cosmetics, and
pharmaceuticals, as well as other industrial and
domestic sectors of the economy. Researches are
Phytochemical screening and fourier transform of Sweet Orange peels
also on going on how to make plastic from orange
peels as a result of the presence of carbon
compound (Limonene) which is eco-friendly
(Valli, 2007). Suitable methods have to be
adopted to convert orange peels and pulp into
value-added products (Nand, 1998) which would
reduce environmental pollution. With this in
view, there is the need to know the phytochemical
composition of orange peels in Zaria.
Materials and methods
Collection of material and preparation of
extracts
About 1kg fresh peels of Citrus sinensis was
collected from orange sellers and put in a thimble
and placed into soxhlet apparatus and extracted
according to the method described by Fatope et
al. (1993), using 2L of 95 % ethanol at 60 °C
repeatedly for 12 cycles. The crude extract was
then partitioned successively into five fractions
using petroleum ether, chloroform, ethyl acetate,
n-butanol and water. The partitioning was
achieved by first dissolving the 30 g of ethanol
extract in 500 ml of water after which 500 ml
petroleum ether was added to the solution. The
mixture was shaken vigorously and poured into a
separatory funnel and then allowed to rest; two
layers were observed, the lower layer (organic)
was transferred into 500 ml conical flask. The
same procedure was performed subsequently for
chloroform, ethyl acetate and n-butanoll. All the
extracts collected were allowed to evaporate
under reduced pressure at 30°C and stored at 4°C
until when needed.
Phytochemical screening of orange peel
extracts
Test for carbohydrates: Test for the presence
of carbohydrates was carried out as described by
Trease and Evans (1989).
Test for Anthraquinones: Test for free
anthraquinones was carried out using Bontrager′s
test as described by Rahman et al. (2017).
© Journal of tropical Biosciences
Test for cardiac glycosides: This was carried out
using Kella-Killani test as described by Trease
and Evans (1989).
Test for alkaloids: This test was carried out
using Wagner’s test (Wagner, 1993).
Test for flavonoids: This was carried out using
Shinoda test (Yisa, 2009).
Test for steroid and triterpenes: This was
carried out as described by Trease and Evans
(1989).
Test for saponins: This was carried out using
Frothing test (Akinjogunla et al., 2010).
Test for tannins: This was carried out using
ferric chloride test as described by Ciulci (1994).
Fourier transform infrared (FTIR) spectrum
analysis
The FTIR analysis was conducted at the National
Research Institute for Chemical Technology,
Zaria, (NARICT) using Shimadzu
Spectrophotometer 40008. A drop of the extracts
each at a time was placed in the FTIR
compartment and scanned in accordance with
ASTM (America Standard Testing Method)
through infrared region for about 25 scans and
chromatograph was obtained.
Results
Phytochemical Screening of Orange Peels
Qualitative screening confirmed the presence of
seven secondary metabolites in the orange peels.
In all the extracts anthraquinone was absent while
carbohydrates and flavonoids were present in all.
Glycosides were present in all the extract and
fractions except in petroleum ether fractions.
Steroid was absent in n-butanollic fractions and
aqueous fractions, while triterpene, alkaloid and
saponin were present only in the ethanolic
extract, n-butanol and aqueous fractions. Tannins
was absent in petroleum ether and chloloform
fractions (Table 1).
Phytochemical screening and fourier transform of Sweet Orange peels

Table 1. The phytochemical composition of orange peel extracts

Solvents
Constituent Test method ETH N-BUT PE CH EA AQ
Carbohydrate Molisch + + + + + +
Anthraquinones Bontragers - - - - - -
Cardiac glycoside Kellankillani + + - + + +
Alkaloid (a) Mayer - - - - - -
Alkaloid (b) Wagner - - - - - -
Alkaloid (c) Dragendorf + + - - - +
Flavonoid Shinoda + + + + + +
Steroid Steroid + - + + + -
Triterpene Triterpene + + - - - +
Saponin Froth + + - - - +
Tannin Fecl3 + + - - + +
No. of constituents 8 7 3 4 5 7
Key: + = present; - = absent *ETH= ethanolic, N-BUT= n-butanollic, PE= petroleum ether,
CH=chloroform, EA=ethyl acetate, AQ=aqueous

Fourier Transform Infrared (FTIR) Analysis
The Fourier Transform Infrared (FTIR)
analysis within the range of 500-4000 cm-1
revealed the presence of various functional
groups in all the extracts. The broad bands at
3383.26cm-1 in ethanolic extract, 3387.11
cm-1 in petroleum ether, 3371.68cm-1 in
aqueous, 3400.62cm-1 in chloroform and
3566.5 cm-1 in n-butanol were assigned to O-
H stretching. This peak was totally absent
from ethyl acetate extract (Fig 4). The Peaks
at 2980.04 cm-1, 2928.04cm-1, 2924.18cm-1,
2854.74cm-1, 2926.11cm-1, 2854.74cm-1,
2929.74cm-1, 2854.74cm1 and 2928.04cm-1
in the extracts are assigned to C-H stretching
vibration (Fig 1 to 6). In petroleum ether,
chloroform and ethyl acetate extracts, two
peaks of C-H stretching were observed and
each is distinct.
The peaks at 1653.87cm-1, 1629.9cm-1 and
1626.05cm-1 in n-butanol, petroleum ether
and aqueous extracts (Fig 1, 5 and 6)
respectively indicate a C=C stretching
vibration, while the presence of aromatic
C=C were recognised at 1614.47cm-1 in
ethanol extract,1602.91cm-1 in petroleum
ether extract and 1600.97cm-1 in chloroform
extract.
The C=O stretching of carbonyl groups were
observed in all extracts at bands 1716.70cm-1
in ethanol extract, 1716.7cm-1 in n-butanol
extract, 1735.99cm-1 in petroleum ether
extract, 1716.7 cm-1 in ethyl acetate
extract,1722.49cm-1 in aqueous extract and
1732.13cm-1 in chloroform extract (Fig 1 to
6). This band was absent in all other extracts.
C-N stretching was found at frequencies
2362.88cm-1 and 2320.4cm-1 in ethanol
extract and aqueous extract (Fig 3)
respectively and absent in all other extracts.

© Journal of tropical Biosciences

 Phytochemical screening and fourier transform of Sweet Orange peels
Percentage transmittance

Wave number (cm-1)

Functional groups peaks (cm-1)

C=O C-H C=C O-H
1732.13 2854.74 1600.97 3400.62
2928.04

Figure 1: FTIR spectrum of chloroform fractions from ethanol extracts of orange peels

© Journal of tropical Biosciences

 Phytochemical screening and fourier transform of Sweet Orange peels

Percentage transmittance

Wave number (cm-1)

Functional groups peaks (cm-1)
C=O C-H C=C O-H
1722.49 2929.97 1626.05 3371.68

Figure 2: FTIR spectrum of aqueous fractions from ethanol extracts of orange peels

© Journal of tropical Biosciences

 Phytochemical screening and fourier transform of Sweet Orange peels
Percentage transmittance

Wave number (cm-1)

Functional Groups peaks (cm-1)

C=O C-H C=C O=H C=C C-N
1716.70 2928.04 1614.47 3383.26 2110.19 2362.88

Figure 3: FTIR spectrum of ethanol fractions from ethanol extracts of orange peels

© Journal of tropical Biosciences

Percentage transmittance Phytochemical screening and fourier transform of Sweet Orange peels

Wave number (cm-1)

Functional groups peaks (cm-1)

C=O C-H
1716.7 2926.11
2854.74

Figure 4: FTIR spectrum of ethyl acetate fractions from ethanol extracts of orange peels

© Journal of tropical Biosciences

Percentage transmittance Phytochemical screening and fourier transform of Sweet Orange peels

Wave number (cm-1)

Functional Groups peak (cm-1)

C=O C-H C=C O-H C-N
1716.7 2928.04 1653.05 3566.5 2320.4

Figure 5: FTIR spectrum of n-butanol fractions from ethanol extracts of orange peels

© Journal of tropical Biosciences

 Phytochemical screening and fourier transform of Sweet Orange peels
Percentage transmittance

Wave number (cm-1)

Functional Groups peaks (cm-1)
C=O C-H C=C O-H
1735.99 2924.18 1629.9 3387.11
2854.74 1602.9

Figure 6: Spectrum of petroleum ether fractions from ethanol extracts of orange peels

© Journal of tropical Biosciences

Phytochemical screening and fourier transform of Sweet Orange peels
Discussion
The qualitative screening confirmed the
presence of glycoside, saponins, steroids and
triterpenes, flavonoids and tannins. The
presence of these compounds in plants
support the use of such plant extracts as
bioherbicide (Macias et al., 1996; Azania et
al., 2003; Khanh et al., 2007). Ethanol, n-
butanol and aqueous extracts extracted more
of the phytochemicals which suggest that,
they are very good and efficient extractors.
The presence of saponins which serve as a
stimulant could stimulate the growth of plant,
while cardiac glycosides which is toxic could
discourage plant growth, flavonoids protects
plants from biotic and abiotic stresses and
serve as an antimicrobial agent and steroids
helps to alter the membrane fluidity (Vinken
et al., 2007). The Fourier Transform Infrared
(FTIR) analysis showed the presence of
aromatic alkene, carbonyl, hydroxyl,
aliphatic alkene, alkane, alkyne and amine
functional groups. The presence of amine
groups indicated the presence of alkaloid
which are derived from amino acids and
contain nitrogen (Zeige, 2004). Alkaloids are
found in ethanol, n-butanolic and aqueous
extracts. The presence of aromatic alkenes,
carbonyl and hydroxyl groups in petroleum
ether, ethanolic and chloroform extracts may
indicate the presence of phenolics. Phenolics
are chemically heterogeneous group of nearly
10,000 individual compounds. In keeping
with their chemical diversity, phenolics play
a variety of role in the plants such as defence
against herbivores and pathogens, others
function in mechanical support, in absorbing
harmful ultraviolet radiation or in reducing
the growth of nearby competing plants
(Vince and Zoltan, 2011). Phenolics have
also been found to reduce root initiation in
plants (Khanh et al., 2005).
Conclusion
Ethanol, n-butanol and aqueous extracts
extracted the highest number of
phytochemical constituents. Alkaloids and
anthraquinones were absent in all the extracts
used in extractions. Petroleum ether extract
gave the highest peak of C=O group (1735.99
© Journal of tropical Biosciences
cm-1), aqueous extract gave the highest peak
of C-H group (2929.97cm-1), n-butanol
extract gave the highest peak of C=C group
(1653.05 cm-1) and O-H group (3566.5 cm-1).
Reference
Abolusoro, S. A. (2001). Efficacy of some
plant materials for the control of
cowpea Bruchid Callosobruchus
maculatues (Fab). Journal of
Agricultural Technology, 9:1-6.
Abolusoro, S. A., Oyedunmade, E. A. and
Olabiyi, T .I. (2010). Evaluation of
sweet orange peel aqueous extract
(Citrus sinensis) as root–knot
nematode suppressant. Journal of
Tropical Agriculture, Food,
Environment and Extension,
9(3):170-175.
Akinjogunla, O. J., Yah, C. S., Eghafona, N.
O. and Ogbemudia, F. O. (2010).
Antibacterial activity of leaves
extracts of Nymphaea lotus
(Nymphaeaceae) on Methicillin-
resistant Staphylococcus
aureus (MRSA) and Vancomycin
resistant Staphylococcus aureus
(VRSA) isolated from clinical
samples. Annals of Biological
Research, 1(2):174-184.
Azania, A. A. P. M., Azina, C. A. M., Alves,
P. L. C. A., Palaniraj, R., Kadian, H.
S., Sati, S. C., Rawat, L. S., Dahiya,
D. S. and Narwal, S. S. (2003).
Allelopathic plants- sunflower
(Helianthus annus L.). Allelopathy
Journal, 11(1):1-20.
Ciulci, I. (1994). Methodolody for the
Analysis of Vegetable Drugs.
Chemical Industry Branch, Division
of Industrial Operations. Unido,
Romania, 24:26-67.
Ezejiofor, T. I. N., Eke, V., Okechukwu, R.
I., Nwoguikpe, R. N. and Duru, C. M.
(2011). Waste to wealth:Industrial
raw materials potential of peels of
Nigerian sweet orange (Citrus
sinensis). African Journal of
Biotechnology, 10(33):6257-6264.
Phytochemical screening and fourier transform of Sweet Orange peels
Fatope, M. O., Ibrahim, H. and Takeda, Y.
(1993). Screening of higher plants
reputed as pesticides using the brine
shrimp lethality assay. International
Journal of Pharmacognosy,
31(4):250-254.
Kanaze, F. I., Termentzi, A., Gabneli, C.,
Georgarakis, M. and Kokkalou, E.
(2009). The phyochemical analysis
and antioxidant activity assessment of
orange peel (Citrus sinensis)
cultivated in Greece-Crete indicates a
new commercial source of hesperidin.
Biomedical Chromatography,
23(3):239-249.
Khanh, T. D., Chung, M.I. and Xaun, T.D,
and Tavata, S. (2005). The
exploitation of crop allelopathy in
sustainable agricultural production.
Journal of Agronomy and Crop
Science, 191:172-184.
Khanh, T. D., Xuan, T. D and Chung, I. M.
(2007). Rice allelopathy and the
possibility for weed management.
Annual Applied Biology, 151: 325-
339.
Macias, F. A., Torres, A., Maya, C. C.,
Fernandez, B., Harper, J. D. I., An, M.
and Kent, J. H. (2005). Natural
biocides from Citrus waste as new
wood preservatives. In: Fourth World
Congress on Allelopathy, 21-26.
Macias, F. A., Torres, A., Molinllo, J. G.,
Varela, R. M. and Castellano, D.
(1996). Potential allelopathic
sesquiterpene lactones from
sunflower leaves. Phytochemistry,
43:1205-1215.
Manthey, A. and Grohmann, K. (2001).
Phenols in Citrus peel byproducts:
concentrations of
hydroxycinnamates and
polymethoxylated flavones in Citrus
peel molasses, Journal. of
Agricultural Food Chemistry. 49:32-
68..
© Journal of tropical Biosciences
Nand, K. (1998): Recent advances in the
treatment of liquid and solid wastes of
food processing industries for
biogas production and pollution
abatenront. Proc. 4th International.
Food Convention, Mysore, 35-37.
Rafique, U. and Nazir, S. (2013). Heavy
Metal Removal from Wastewater
using Environment Friendly Bio-
materials: A Cost Effective and Eco-
friendly Technique. International
Journal of Chemical and
Environmental Engineering, 4 (2):90-
99.
Rahman, G., Syed, U. J., Syed, F., Samiullah,
S. and Nusrat, J. (2017) “Preliminary
Phytochemical Screening,
Quantitative Analysis of Alkaloids,
and Antioxidant Activity of Crude
Plant Extracts from Ephedra
intermedia indigenous to
Balochistan,” The Scientific World
Journal, 7-11.
Rehman, Z. U. (2006). Citrus Peel Extract –
A Natural Source of Antioxidant.
Food Chemistry, 99: 450-454.
Trease, G. E. and Evans, W. C. (1989). A
Textbook of Pharmacognosy. 13th.
Edition. Bailliere Tindall Ltd.,
London, 21 pp.
Valli, G. P. (2007). 10 uses of orange peels.
Phytochemistry, 2:123-134.
Vince, O. and Zoltan, M. (2011). Plant
Physiology: Secondary metabolites.
Retrieved from
http://www.tankonyvtar.hu /
hu/tartalom/tamop425
/0010_1A_Book_angol_01_novenye
let tan /ch3s05.html.
Vinken, J. P., Heng, L., DeGroot, A. and
Gruppen, H. (2007). Phytochemical
and Biological Assessment of
Medicinal Important Plant
Ochradenus arabicus. Pakistan
Journal of Botany, 46(6):2027-2034.
 Phytochemical screening and fourier transform of Sweet Orange peels

Wagner, H. (1993). Pharmazeutische Yisa, J. (2009). "Phytochemical Analysis and

biologie, 5th edition. Gustav fisher Antimicrobial Activity Of Scoparia
Vwelag, Stuttgart, Germany. dulcis and Nymphaea lotus".
Yang, H., Yan, R., Chen, H., Lee, D. H. and Australian Journal of Basic and
Zheng, C. (2006). Characteristics of Applied Sciences, 3(4):3975–3979.
hemicellulose, cellulose and lignin Zeige, L. T. (2004). “Plant physiology: plant
pyrolysis. Energy Fuels, 1781-1788. defenses” p349-376.
Yassari, S. and Yasari, E. (2013) .Effects of
extracts of Thompson orange peels on
the stability of canola oil.
International Journal of Agriculture
and Crop Sciences, 5(4):410-420.

© Journal of tropical Biosciences

View publication stats