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

net/publication/272494799

Extraction of Orange Oil by Improved Steam Distillation and its

Characterization Studies

Article · February 2015

CITATIONS READS

11 26,754

2 authors, including:

Mercy Nisha Pauline

Government College of Technology
13 PUBLICATIONS   51 CITATIONS   

SEE PROFILE

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

Biodiesel View project

All content following this page was uploaded by Mercy Nisha Pauline on 20 February 2015.

The user has requested enhancement of the downloaded file.

 International Journal of Engineering Technology, Management and Applied Sciences

www.ijetmas.com February 2015, Volume 3 Issue 2, ISSN 2349-4476

Extraction of Orange Oil by Improved Steam Distillation

and its Characterization Studies
Mercy Nisha Pauline. J*, Nithyalakshmi. B, Aadhithiya Lakshmi. R
Department of Industrial Biotechnology, Government College of Technology,
Coimbatore, Tamil Nadu
Abstract
The orange peel which is considered as a waste can be used for the extraction of essential oil
which has many applications ranging from food flavoring agent to cosmetics. Orange oil can be
extracted by various conventional methods like steam distillation, solvent extraction. Novel methods
like super critical CO2 extraction, turbo distillation also has been employed. The conventional
methods though simple are robust and the yield percentage is less whereas the novel methods are not
cost effective. This paper focuses on improved steam distillation, where the orange peels are
preheated before subjecting to distillation. The preheating enhances the oil yield and the water
distillate can be used as such for further applications. This extraction procedure can also be used for
the extraction of aromatic oils from other sources such as leaves, flowers, stem, wood as well.
Key Words: Improved Steam distillation, Conventional steam distillation, orange peels,
Limonene
Introduction
Essential oils contain highly volatile substances that can be isolated by a physical method or
process from plants of a single botanical species. The oils normally bear the name of the plant
species from which they were derived. Few essential oils have been identified to possess a very
good anti microbial effect. Immune system needs support and these essential oils can give the
required endorsement. [D. Pandey et al.]. Orange is an excellent source of Vitamin C. Besides,
oranges constitute Vitamins (B1, B2, B3, B5, and B6), flavonoids, terpenes, potassium and calcium.
They are also a very good source of dietary fiber [Milind et al., 2012]. Orange peel oil is the major
oil produced worldwide and is used extensively in the food industry, primarily as a flavoring agent. It
possesses a light, sweet, fresh top note with fruity and aldehydic character. Many household and
personal-care products employ orange oil owing to its pleasing character, ability to blend with other
aroma components, low cost and availability.
Citrus peel and/or essence oils are being commonly employed as a top note component in
some perfumes and colognes [Glen et al, 2007]. Orange fruit contains 1.5% essential oil. D-limonene
(1-methyl-4-(1-methylethanyl cyclohexane) is a monoterpene with a lemon-like odor and is a major
constituent in several citrus oils. Limonene is a naturally occurring chemical which is used in many
food products, soaps and perfumes for its lemon-like flavor and odor. Limonene also is a registered
active ingredient in 15 pesticide products used as insecticides, insect repellents, and dog and cat
repellents.

1 Mercy Nisha Pauline. J, Nithyalakshmi. B, Aadhithiya Lakshmi. R

 International Journal of Engineering Technology, Management and Applied Sciences

www.ijetmas.com February 2015, Volume 3 Issue 2, ISSN 2349-4476

Orange oil can be extracted from the peel by conventional methods like simple distillation,
solvent extraction, maceration, cold pressing and modern techniques like effleurage, Super Critical
CO2 extraction and Turbo Distillation Extraction. These methods of extraction has its own pros and
cons. The modern methods are costly and laborious. Maceration is not desirable as it changes the
composition of oil. Solvent extraction leaves traces of solvent in the extract which causes allergies
and affect immune system. The residues of solvent is also undesirable for either organoleptic and/or
health reasons. Steam distillation, is also a type of distillation or a separation process for temperature
sensitive materials like oils, resins, hydrocarbons, etc., which are insoluble in water and may
decompose at their boiling point. The main advantages of steam distillation is the properties of the
oils obtained by this method are not altered. It is also a economical and a faster process. The oil is
not decomposed, as steam reduces the boiling point of it. The objective of this paper is the extraction
of orange oil by both simple conventional and improved steam distillation. The yield of orange oil
obtained by both methods was compared and the presence of orange oil and limonene was confirmed
using pH test, Thin layer chromatography, FTIR and Gas chromatography.
Materials and Methods
Two different species of oranges, viz. Citrus sinensis and Citrus reticulate were taken for our
study.
Orange rinds were peeled off carefully with the help of a sharp razor blade. The samples were
checked to ensure that none of the white flesh under the rind was included in the sample because
white flesh contains little or no limonene. Each rind sample was cut into smaller pieces and 100g
mass of the sample was taken. The sample was initially rinsed with distilled water
Conventional Steam Distillation
Steam distillation was carried out similar to the extraction of various essential oils including
that from spices like cinnamon, star anise [Pavia et al., 2005], aromatic plants like granium
[Rabesiaka JR et al., 2012].
Improved Steam Distillation (ISD)
The water washed orange peels were pre-heated in a vessel for about 30 minutes at a
temperature of 50°C. Fresh orange peels are taken and preheated at a temperature below 50 οC for
about 30 minutes prior to steam distillation procedure. A little amount of distilled water that is
enough to immerse the orange peels is added. The pre-heated sample was then loaded into the round
bottomed flask and two-third of it was filled with the distilled water including the water that retained
after preheating the cut orange peels. The condenser water was turned on and heated up to 100°C for
two hours. Water loss occurred due to continuous heating. If the water level got too low, the sugar
present in oranges would caramelize and burn. To avoid this two-third of the round bottomed flask
was filled with water.The distillation was then run, for about 120 minutes, at varied temperature
ranges (10°C, 20°C and 30°C) of the heating mantle.The distillate was collected and the presence of
the orange oil in the condensate was confirmed by the cloudy appearance on the top of the distillate.
Extraction Of Orange Oil Using Chloroform
The Orange Oil was separated from water by chloroform extraction. The distillate was poured
into a 250ml separating funnel and 2 g of sodium chloride was added in it because sodium chloride
helps to minimize emulsions during the extractions by making the organic layer less soluble in the

2 Mercy Nisha Pauline. J, Nithyalakshmi. B, Aadhithiya Lakshmi. R

 International Journal of Engineering Technology, Management and Applied Sciences

www.ijetmas.com February 2015, Volume 3 Issue 2, ISSN 2349-4476

aqueous layer. The separating funnel was stoppered, shaken and vented for four or five times. 15 ml
chloroform was used to rinse the apparatus and it was allowed to drain into the separating funnel.
The orange oil was extracted from the upper organic layer by inverting the separating funnel and
shaking it back and forth gently. The layers were allowed to separate before draining the lower
aqueous layer. The topmost layer was collected and the extraction was repeated by using another 10
ml of chloroform. The chloroform extracts were combined in a single beaker and kept in a waterbath
to vaporize the chloroform present in the extract. After the chloroform had evaporated completely,
orange oil remained and it was collected and stored for further analysis [Mukhtar.H, 2012]. The yield
of the essential oil obtained from the orange peels is calculated using the formula,
%yield = Wt. of oil * 100 (g/g)
Wt. of orange peels taken
Wt. of oil = Wt. of beaker with oil - Wt. of empty beaker (g)
Characterization Studies
The qualitative and quantitative analysis was done to identify the constituents in the oil and the
percentage of components present in the oil respectively. [H.J. Williams et al., 1993].
Iodine Test
This test is performed to check the presence of unsaturated bonds in a molecule. A few drops
of the orange oil were taken in a test tube. A few crystals of iodine were added and shaken well.
pH Test
The pH of any liquid sample is generally measured using pH meter. A few ml of the orange
oil was taken for pH analysis
Thin Layer Chromatography (TLC)
TLC is used to identify the compound in a mixture using the Rf value. Initially, the mobile
phase was prepared by dissolving the iodine in Hexane and the mixture was taken in a beaker,
covered with a watch glass, without the TLC slide. This was meant for saturation, and allowed for
about 15 minutes. Readymade TLC plates were used. A drop of Orange Oil was loaded at a point
0.5cm above the bottom of the slide, using a capillary tube. The slide, along with the sample, was
now placed in the beaker containing the solvent, in a perpendicular fashion. The solvent was allowed
to travel upto 80% of the slide. The slide was then taken out and iodine solution was sprayed on it. A
dark brown coloured line appeared on it after spraying, which indicated the presence of limonene at
that point and the retention factor (Rf) was calculated using the formula,
Rf =Distance travelled by the compound / Distance travelled by the solvent
Fourier Transform Infrared Spectroscopy (FTIR)
FTIR is used to determine the presence d-limonene in the extracted oil [NIST, 2011].
Gas Chromatography (GC)
GC is used to measure the desired component both quantitatively and qualitatively.Nitrogen gas
at approximately 60.9 kPa constant pressure was used as a carrier gas [Villen, 1998]. RTX-5 column
and capillary type injector port were used.
Results and Discussion
Extraction of Orange oil
The orange peels were cut into small size and were pre heated at 50°C for 30 mins. The
heated sample was subjected to improved steam distillation. Conventional steam distillation was also

3 Mercy Nisha Pauline. J, Nithyalakshmi. B, Aadhithiya Lakshmi. R

 International Journal of Engineering Technology, Management and Applied Sciences

www.ijetmas.com February 2015, Volume 3 Issue 2, ISSN 2349-4476

carried out. Though the distillation procedure allowed only the separation of volatile compounds
(essential oils), which, to a greater or lesser extent, are transformed under the influence of the
elevated temperature, the absence of light and air during extraction reduce the risk of degradation
reactions [Zoran P. Zeković et al., 2009].
It has been observed that yield of the orange oil was higher in improved steam distillation for
both the orange species than that of the CSD of the orange peels. ISD, produced orange oil of almost
two times more than CSD in C. sinensis. ISD in C. reticulate also showed that the orange oil yield
was five times more than the oil extracted by CSD. It was also inferred that more oil was extracted
from C. reticulate to C. sinensis. The % yield of orange oil obtained by conventional and improved
steam distillation are presented in table 1.
The increase in orange oil yield (%) by ISD may be higher due to the reason that the orange
peels were pre-heated which softened the cell walls of the orange peels which are made up of
cellulose, hemicelluloses and pectin. When the preheated peels were further subjected to normal
steam distillation procedure, it further softened and opened the cell walls of the orange peels to
liberate all the components present in it in the form of orange oil. Similar phenomenon was observed
when the tea leaves were heated for 15min 100°C during the extraction of caffeine from tea leaves. It
was found that during heating, the cell walls of the tea leaves were opened to liberate more caffeine
from them.
The distillate was colourless and cloudy in nature. It was soft and had a very pleasant,
aromatic and lemon taste.

Table 1: % yield of the orange oil samples extracted by CSD and ISD

Type of Distillation C. sinensis (%) C. reticulate (%)

Improved Steam Distillation 4.237 5.865

Conventional Steam Distillation 2.475 0.98

Essential oils are mixtures of organic compounds that are dominated by the terpenes and the
terpenoids which are the oxygen containing derivatives of the terpenes [Villen at al., 1998]. The oil
turned to slight orange colour after few hours of solvent extraction.
Steam distillation method can be one of the promising techniques for the extraction of
essential Oil from plants as this process will preserve the original qualities of the plant. Steam
distillation for temperature sensitive materials like oils, resins, hydrocarbons, etc. which are insoluble
in water and may decompose at their boiling point can be prevented by using Steam Distillation
method at low pressure. Sahoo et al., [2010] observed the increase in volume of essential oil with
increase in temperature by keeping time of heating constant. They also found that the volume of
essential oil increased with increase in the time of heating keeping the temperature constant.

4 Mercy Nisha Pauline. J, Nithyalakshmi. B, Aadhithiya Lakshmi. R

 International Journal of Engineering Technology, Management and Applied Sciences

www.ijetmas.com February 2015, Volume 3 Issue 2, ISSN 2349-4476

Characterization of Orange oil

The orange oil so obtained from the peels of both the orange species by ISD was analyzed for
the presence of limonene as it is the major component of orange oil. About 97% of orange oil is
limonene [Rodriguez, 2011]. Hence, this compound was chosen to characterize the orange oil. For
analyses, the orange oil extracted was subjected to iodine test, pH test, Thin Layer Chromatography
(TLC), Fourier-Transform Infrared (FTIR) spectroscopy and Gas Chromatography (GC).

Iodine Test
Iodine test was the characterization test for the presence of any unsaturated compound in a
test sample. On addition of iodine, the colorless orange oil sample extracted by ISD turned into
brown color. The presence of limonene and other aromatic compounds in the sample was confirmed.
The brown color is due to the reaction of iodine with limonene and other unsaturated compounds like
α-pinene, β-pinene, myrcene, limonene and linalool.
pH Test
The oil samples after distillation of the pre-heated peels were collected and subjected for pH
analysis using a pH meter. The pH value of C. sinensis and C. reticulate was found to be 4.67 and
4.98 respectively, which are very much acidic in nature [7].

Thin Layer Chromatography

TLC was performed to check the retention factor (Rf) of the aromatic compound, limonene.
The TLC plates were developed by applying a few drops of iodine solution. The brown colored
markings were observed immediately. The color of the spot disappeared on exposure to air. Rf values
of the orange oil samples C. sinensis and C. reticulate were found to be 0.76 and 0.757 respectively.
The retention factor (Rf value) of the orange oil was found to be 0.75 which was found to be
in accordance with that of limonene [1].

FTIR Spectroscopy
For further confirmation of the presence of Limonene in the extract, the sample oil from C.
sinensis was taken for FTIR analysis. Since the sample was in the liquid form, NaCl plates were
used. The graph was smoothened using the software Origin Pro 8.0 to avoid the noise and to pin
point the peak corresponding to limonene.
The X-axis and Y-axis represented the wavenumber (cm-1) and absorbance respectively. Limonene,
in general, showed an infrared spectrum in the following range of wavenumber, 2850cm-1 to
3100cm-1 [NIST,2011]. From the graph, it was observed that the C-H stretching spectra of the
Limonene was found at 2900cm-1 (Approx.).

5 Mercy Nisha Pauline. J, Nithyalakshmi. B, Aadhithiya Lakshmi. R

 International Journal of Engineering Technology, Management and Applied Sciences

www.ijetmas.com February 2015, Volume 3 Issue 2, ISSN 2349-4476

Fig.1. Smoothened FTIR graph by Origin Pro 8.0

Gas Chromatography
This type of analysis, giving both qualitative and quantitative information, was used for
quality control of essential oils. A typical separation obtained by this method for the analysis of
orange oil sample from C. sinensis and C. reticulate is shown in Fig.2 and Fig.3. The chromatogram
shows a detailed picture of the main compounds and of minor constituents. From the database [David
et al., 2002], the Rf value of limonene was found to be at 35th minute. The chromatogram obtained
showed the peaks at 35th minute.

Fig 5.5.1. Chromatogram for orange oil Fig.5.5.2. Chromatogram of orange oil

sample from peels of C. sinensis samples obtained from peels of C.reticulate

6 Mercy Nisha Pauline. J, Nithyalakshmi. B, Aadhithiya Lakshmi. R

 International Journal of Engineering Technology, Management and Applied Sciences

www.ijetmas.com February 2015, Volume 3 Issue 2, ISSN 2349-4476

Thus orange oil was extracted using both conventional and improved steam distillation. It
was found that improved steam distillation produced more orange oil than the conventional method.
Limonene was qualitatively characterized. This can be considered as a green method of extraction as
it is less energy intensive process. The distillate containing water and orange oil can be used as such
for further applications and this method can also be used for the extraction of essential oils from
other fruits and flowers. This study can to be further optimized for the temperature and time.
Acknowledgements:
The authors wish to thank the Department of Industrial Biotechnology of Bannariamman
Institute of Technology, Sathyamangalam, Tamil Nadu, India and Chemistry Department of
Bharathiar University, Coimbatore, Tamil Nadu, India for enabling them to use the GC and FTIR
facilities.

References
1. Chemistry Practical Work: Part III: Useful Information & Data
2. David, F., Scalan, F., Sandra, P. and Szelewski, M. (2002) Analysis of Essential Oil Compounds
Using Retention Time Locked Methods and Retention Time Databases. Food and Flavours, Agilent
Technologies.
3. Glen O. Brechbill. (2007) Classifying Aroma Chemicals. Fragrance books Inc., New Jersey, First
Edition: pp. 31-44.
4. Milind, P., and Dev, C. (2012) Orange: Range of Benefits. IRJP, 3 (7): pp. 59-63.
5. Mukhtar, H., Shabbiri, M.K., Nadeemi, R., Farooq and Mumtazi, W. (2012) Physicochemical
analysis and Determination of various chemicals constituents of essential oils in Rosa cent foli,
University of Agriculture, Faisalbad, Pakistan.
6. No authors listed. D-Limonene, 2011, National Institute of Standards and Technology, Available
online at http://webbook.nist.gov/cgi/cbook.cgi?ID=C5989275&Mask=80.
7. No authors listed. D-Limonene, Biochem Corp. of Florida., Inc 2000, USA. Available online at
http://www.biochemcorp.com/dlimonene2.htm.
8. Pandey D., Rao Virendra, P.S., Extraction of essential oils from Eucalyptus leaves (B.Tech
Project, National Institute of Technology, Rourkela)
9. Rodriguez, A., Andres, V.S., Cervera, M., Redondo, A., Alquezar, B., Shimada, T., Gadea, J.,
Rodrigo, M.J., Zacarias, L., Palou, L., Lopez, M.M., Castanera, P. and Pena, L., 2011, “Terpene
Down-Regulation in Orange Reveals the Role of Fruit Aromas in Mediating Interactions with Insect
Herbivores and Pathogens”, American Society of Plant Biologists, 156 (2): pp. 793-802.

7 Mercy Nisha Pauline. J, Nithyalakshmi. B, Aadhithiya Lakshmi. R

 International Journal of Engineering Technology, Management and Applied Sciences

www.ijetmas.com February 2015, Volume 3 Issue 2, ISSN 2349-4476

10. Pavia, D., L.; Lampman, G., M.; Kriz, G., S.; Engel, R., G. Introduction to Organic Laboratory
Techniques: A Small Scale Approach; Thomson Brooks/Coles, 2005.
10. Prof. A. Sahoo, K. Satish Kumar. (2010) Extraction of essential oil using steam distillation (E-
thesis submitted for the partial fulfillment for the degree of Bachelor of Technology, National
Institute of Rourkela.
11. A. Sahoo et al., (2010), Extraction of essential oil using Steam Distillation, Thesis Submitted to
National Institute oif Technology, Rourkela
12. H. J. Williams, Ahmed, Mahmoud, A. I. Scott, J. H. Reibenspies, and T. J. Mabry. 1993. New
sesquiterpene a-methylene lactones from the Egyptian plant Jasonia candicans. J. Nat. Prod.
56:1276–1280.
13. Villen, J., Blanch, G.P., del Castillo, M.L.R. and Herriaz, M., 1998, “Rapid and Simultaneous
analysis of Free Sterols, Tocopherols, and Squalene in Edible Oils by Coupled Reversed-Phase
Liquid Chromatograpgy-Gas Chromatography”, J. Agric. Food Chem., 46 (4): pp. 1419-1422.
14. Zoran P. Zeković, Žika D. Lepojević, Ibrahim O. Mujić., 2009, Laurel Extracts Obtained by
Steam Distillation, Supercritical Fluid and Solvent Extraction, Journal of Natural Products Vol.
2(2009):104-109

8 Mercy Nisha Pauline. J, Nithyalakshmi. B, Aadhithiya Lakshmi. R

View publication stats