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Int. Journal of Applied Sciences and Engineering Research, Vol. 3, No.

2, 2014
Copyright 2014 by Authors Licensee IJASER Under Creative Commons License 3.0
Research article

www.ijaser.com
editorial@ijaser.com
ISSN 2277 8442

Quantitative and Qualitative analysis, and comparative study of


essential oil extracted from Nigerian orange, lemon and lime peels
1

Njoku V.I., 2Evbuomwan B.O.


Department of Chemical Engineering, college of Graduate Studies, University of Port Harcourt, Choba,
Nigeria.
DOI: 10.6088/ijaser.030200024
Abstract: Essential oils are a product obtained from vegetable raw materials and are complex mixtures
whose composition may include volatile terpenic compounds. In this study, the relationship between time
and amount of essential oil extracted from the citrus peels using steam distillation was shown. Orange peel
was shown to contain more amount of essential oil (20ml at 180minutes) than lemon (7ml at 180minutes)
and lime (5ml at 180minutes).Using GC-MS analysis, 11, 12 and 24 chemical constituents were identified
in orange, lemon and lime essential oil respectively. The 3 main components and their percentage
compositions are D-Limonene (54.151%), Pulegone (11.652%) and L-Carvone (2-cyclohexen-1-one)
(5.457%) for orange peel essential oil, D-limonene (64%), 2-cyclohexen-1-ol (6.21%) and -Pinene
(3.822%) for lemon peel essential oil, and -Pinene (23.124%), D-limonene (17.07%) and -Pinene
(10.399%) for lime essential oil. D-limonene was the most prevalent chemical constituent in orange and
lemon essential oil, but -Pinene was the most prevalent in lime peel essential oil. Although lemon
contained more D-limonene per unit volume of essential oil, application of these percentages on the
volume of essential oil obtained from 370g of the peels at 180minutes showed that orange peel can deliver
more D-limonene per unit mass of available peel. Hence, orange peel essential oil is a more economical
source of D-limonene in Nigeria.
Keywords: Quantitative, analysis, qualitative, comparative, extracted, essential oil, orange, lemon, lime.

1. Introduction
Essential oil from citrus is a large type of natural flavours and frangrances, which is popularly used in food
industries, daily chemical product and health care field (Sheng-min, et al., 2012). The citrus species are a
potential source of variable oil which might be utilized for edible and other industrial application (Maria C.
et al, 2012). Essential oils are broadly used as pharmaceutical components, in nutritious supplements and
for cosmetic industry and aromatherapy (Maria, et al., 2012). Guenther E., 1955, also stated that the oil is
also employed in perfumes, toilet waters, eaux de cologne, and in cosmetics to which it impacts a
refreshing top note.
Essential oils are a product obtained from vegetable raw materials (Berger, 2007) , and are complex mixtures
whose composition may include volatile terpenic compounds, which have the formula (C5H8)n , where the
compounds are monoterpenes if n=2, sesquiterpenes when n=3, diterpenes when n=4 etc. (Smith, et al.,
2001). These are secondary metabolites in plants (Mazen, 2002) and responsible for the characteristic aroma
on the fruit.

*Corresponding author (e-mail:)


Received on February, 2014; Accepted on March 2014; Published on April 2014

279

Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

Lemon essential oil are complex mixtures of chemical compounds like limonene, Y-terpinene, citral,
linalool and -caryophyllene among others, which can be represented by three main classes, namely
terpenes, oxygenates, and sesquiterpenes (Benvenuti, et al., 2001 and Van Straten et al., 1983). The most
significant flavour compound is citral, while linalool possesses highly distinctive organoleptic
characteristics. In addition, limonene, myrcene, octanol, and Y-terpene among others contribute with high
aroma flavour of lemon oil (Benvenuti, et al., 1983).
The composition of orange oil varies for several reasons. Region and seasonal changes as well as the
method used for extraction lead to these variation. Gamarra, et al., (2002) also stated that the quality of
essential oil depends on different factors; among them are the chemotype and biotype of the plant, the
climatic conditions as well as the extractive process. Several compounds have been identified in orange oil
with gas chromatograph-mass spectrometry. Most of the substances are part of the terpene group
(limonene, -pinene, sabinene, -pinene, myrcene and -3-carene) with limonene being the dominant one.
Long chain aliphatic hydrocarbon alcohols and aldehydes like octanol, decanal, and octanal are second
important group of substances (Verzera, et al., 2004).
Citrus fruits have a rough, robust and bright (green to yellow) coloured skin. They are usually 4 to 30cm
long and 4 to 20cm in diameter, with a leathery surrounding rind or skin known as epicarp (or flavedo) that
cover the fruits and protect it from damages. Citrus fruits are notable for their fragrance, partly due to
flavanoids and limonoids contained in the rind (Manthey, 2004). The endocarp is rich in soluble sugar and
contains significant amounts of vitamin C, pectin, fibres, different organic acids and potassium salt which
give the fruits its characteristic citrus flavour (Ezejiofor, et al., 2011). Citrus juice also contains a high
quality of organic acids (citric, malic, acetic and formic acids).
In Nigeria and other parts of the world, citrus sinensis (sweet orange) are cheaply available, thus serves as
a major source of vitamins in diets. Orange fruit and its juice have several beneficial, nutritive and health
properties (Okwu and Emenike, 2006). They are rich in vitamins especially ascorbic and folic acids. Over
the last decades, many other virtues and medicinal benefits of orange fruits have been discovered besides
their anti-scurvy property (Rapisararda, 1999). There is convincing epidemiological evidence that the
consumption of orange fruit is beneficial to health and contributes to the prevention of degenerative
process, particularly lowering incidence of degenerative process, particularly cardio and cerebro- vascular
diseases (Rapisararda, 1999). The protection that orange fruit provides against these diseases has been
attributed to the various antioxidant phytonutrients contained in citrus species (Okwu and Emenike, 2006;
Rapisararda, 1999).
Ezejiofor, et al., 2011 reports that of the 36.0 million metric tonnes world production, Nigeria produces 0.3
million metric tonnes and has the potential to produce more orange wastes ( orange peels) in high
production.

2. Materials and methods


Oranges used in this work were bought from Rumuokoro market in Portharcourt, Rivers state, Nigeria.
They were washed with de-ionized water, peeled and cut into small pieces. The orange peel was then
weighed using RADWAG WAGI electronic top loading balance ( AS 220/C/2) to obtain exactly 370grams.
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Njoku V.I., Evbuomwan B.O.
Int. Journal of Applied Sciences and Engineering Research, Vol. 3, No. 2, 2014

Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

The orange peels were then pureed using MARLEX ELECTROLINE (Excella) blender.
The pureed orange peel was then carefully transferred into a 1000ml flat bottom flask. 650ml of de-ionized
water was measured using a 1000ml measuring cylinder and then added to the 1000ml flat bottom flask
(distilling flask). Boiling chips was then added to the distilling flask to ensure that boiling occurs calmly
without bumping. The experiment was then set-up as shown in Figure 1 below;

Figure 1: laboratory set-up for the steam distillation of citrus peel


Heating of the distilling flask was then slowly commenced using STUART HOT PLATE/ MAGNETIC
STIRER. The heat source was adjusted so that the distilling rate is approximately 20 drops per minutes. As
the mixture boils and distils, de-ionized water was added via the separation funnel in the set-up just to keep
the water level at the operation level preventing the water level from going too low which can cause the
sugar in the puree to caramelize and burn and also keeping the heat at a low steady level. The experiment
was conducted for the following times 60, 100, 140 and 180 minutes, in each case, the distillate was
collected and transferred into a 250ml separating funnel. 20ml of diethyl ether was added to the distillate in
the separating funnel to extract the oil. It was ensured that there was no flame source or hot plate on
during the use of diethyl ether because diethyl ether is very, very flammable. The water layer in the
separating funnel was then drained off. The diethyl ether layer was collected and small quantity of
anhydrous magnesium sulphate was added to dry off the water content. The oil was then kept in a fume
cupboard for 12 hours to allow all the diethyl ether vapourize. The oil extracted was then measured and
recorded. The procedure was repeated using lemon peel and lime peel, and in each case, the volume of oil
extracted was collected, measured and recorded.

3. Experiments
3.1 Determination of the solubility of citrus peel essential oil in water
Approximately 6 drops of water was added to the test tube containing 3 drops of orange peel essential
oil. The test tube was stirred thoroughly with a glass stirring rod. Two separate phases was observed. The pH
of the water was measured to determine if the essential oil is partially soluble in water and whether it has
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Int. Journal of Applied Sciences and Engineering Research, Vol. 3, No. 2, 2014

Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

changed the pH of the water. It was observed that the pH paper did not change colour. Thus, the orange peel
essential oil is a water insoluble compound. The above experiment was carried out on lemon and lime
essential oil, and same result was obtained.

3.2. Determination of the boiling point of citrus peel essential oil


5ml of the essential oil was placed in a small test tube. A capillary, sealed at one end is placed open-end down
into the essential oil. The test tube is firmly attached to a thermometer with a rubber band such that the
thermometer bulb should be even with the test tubes bottom, and this entire assembly immersed in an oil
bath (half filled 100ml beaker). As the temperature is slowly increased, a rapid evolution of bubbles from the
end of the tube begins. Heating was continued for about 5-10 seconds to be sure that all of the air has been
expelled from the capillary, and the vapours of the essential oil become equal to the atmospheric pressure. As
the temperature decreases, the bubbles slowed down and the essential oil starts rising into the capillary. At
the point when the bubble stops, the thermometer was read and recorded. The above process was repeated 2
more times, and the temperature reading in each case was recorded.The above experiment was carried out on
orange, lemon and lime essential oil at atmospheric pressure.

3.3. Determination of specific gravity of orange peel essential oil


Density bottle was used for determining the density of the oil. A clean and dry bottle of 25ml capacity was
weighed (W0) and then filled with the oil, stopper inserted and reweighed to give (W1). The oil was
substituted with water after washing and drying the bottle and weighed to give (W2).The above experiment
was done using orange, lemon, lime essential oil

3.4. Determination of refractive index of citrus peel essential oil


The Abb's refractometer was used for the determination of refractive index. It gives values up to the 4th
decimal place. The refractive index is denoted by nD25 where n is the refractive index at 25C taken with
sodium light (D - line). First, the refractometer was standardized with distilled water which has refractive
index of nD29.5= 1.3315. Then it was cleaned with acetone and dried with cotton. After this, a drop of
orange peel essential oil was placed between the prisms of refractometer. The telescope was rotated to
bring the border line of total refraction to the junction of cross-wire in the telescope. The refractive index
was recorded at room temperature.
The above experiment was repeated for lemon and lime peel essential oil.

3.5. Determination of peroxide value of orange, lime and lemon essential oil
30ml of acetic acid chloroform solution was measured into a flask containing 2g of the oil sample. A
0.5ml saturated solution of potassium iodide was then added, followed closely by the addition of 30ml of
distilled water. The flask content was then titrated against 0.1M sodium thiosulphate (Na2S2O3) until the
yellow colour almost disappeared. 0.5ml starch indicator was added and the titration continued until the
end-point (where the blue-black colour just disappeared). A blank titration was also performed.
Where S and B represent sample and blank titrations respectively.
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Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

The peroxide value is thus calculated;


PEROXIDE VALUE =
Where S= Sample titration.
B= Blank titration.

3.6. Determination of saponification value of citrus peel essential oil


5.3716g of orange peel essential oil were weighed into a conical flask separately. 25ml of 0.1N ethanolic
potassium hydroxide was added to the conical flask, and content constantly stirred for 1 hour followed by
reflux. Phenolpthalein indicator was then added to the conical flask and titrated with 0.5M HCl till the
solution changes to colourless. The same procedure was repeated for the blank. The above experiment was
repeated using 5.3014g of lemon and 5.3006g of lime essential oil.

3.7. Determination of acid value of citrus peel essential oil


25ml of diethyl ether and 25ml of ethanol was mixed in a 250ml beaker. The resulting mixture was added
to 5.0858g orange peel essential oil in a 250ml conical flask, and few drops of phenolphthalein were added
to the mixture. The mixture was then titrated with 0.1M KOH to the end point with consistent shaking for
which a dark pink colour was observed and the volume of 0.1M KOH (Vo) was noted.
The above experiment was repeated using 5.0808g lemon and 5.0826g lime essential oil

4. Results and discussion


The time and quantiy of essential oil extracted from the citrus peels are presented in Tables 1, 2 and 3, and
Figure 2, 3, 4, 5 for orange, lemon and lime respectively.
From Table 1, 2 and 3 for orange, lemon and lime respectively, it was observed that the volume of essential
oil extracted from the peels increases with time. It was also noticed that more essential oil (20ml) at 180
minutes of heating was extracted from 370g of the orange peels than from equal mass of lemon and lime
peel at the same time of heating, while lime had the least quantity of essential oil extracted (5 ml). These
observations are in agreement with the findings of Kamal, et al.(2011) who reported that among the citrus
species tested, C. Sinensis exhibited the maximum oil yield (0.24-1.07%) followed by C. reticulata
(0.30-0.50%) and C.paradisii (0.20-0.40%).
Gamarra, et al. (2002) stated that the quality of essential oil depends on different factors; among them are
the chemotype and biotype of the plant, the climatic conditions as well as the extractive process. Soumaya,
et al.(2012) also reports that essential oil yield varied during ripening to reach maximum values during the
middle stage of maturity( stage 2) for mandarin and orange, while the highest lemon yield was found at the
beginning of fruit maturation and decreased after that. Also Des Gachons, et al. (2005) stated that water
supply during ripening was reported to influence considerably the essential oil content with an
enhancement of yield under moderate water shortage conditions. All these will possibly be responsible for
the high yield of orange essential oil over that of lemon and lime.

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Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

Table 1: Variation of volumes of essential oil extracted from orange peel.


S/N

Plant
material

Weight
(gm.)

Volume of
distilled
water(ml)

Time of
heating(min)

Temp.( c)

Volume of
essential
oil (ml)

ORANGE
PEEL

370

650

60

100

14

370

650

100

100

16

370

650

140

100

18

370

650

180

100

20

Table 2: Variation of volumes of essential oil extracted from lime peel.


S/N

Plant
material

Weight
(gm.)

Volume of
distilled
water(ml)

Time of
heating(min)

Temp.(0c)

Volume of
essential
oil (ml)

LIME
PEEL

370

650

60

100

370

650

100

100

370

650

140

100

4.5

370

650

180

100

Table 3: Variation of volumes of essential oil extracted from lemon peel.


S/N

Plant
material

Weight
(gm.)

Volume of
distilled
water(ml)

Time of
heating(min)

Temp.(0c)

Volume of
essential
oil (ml)

LEMON
PEEL

370

650

60

100

370

650

100

100

4.5

370

650

140

100

370

650

180

100

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Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

Figure 2: Graph of Time of heating (min) Vs Volume of Essential oil for orange peel.

Figure 3: Graph of Time of heating (min) Vs Volume of Essential oil for lemon peel.

Figure 4: Graph of Time of heating (min) Vs Volume of Essential oil for lime peel.

Figure 5: Multiple bar chart showing Volume of Essential oil Extracted from citrus
peel at different times.
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Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

The GC-MS analysis of the citrus peel essential oil was done using (5975 Series MSD with an Agilent
7890A) shown in Figure 4.5. 0.5ml of each of the oil sample was diluted to 5ml using 4.5ml of methylene
chloride (diluents). Helium was used as the carries gas because of it inert nature. The result of the GC-MS
analysis of the orange, lemon and lime peels essential oil are shown in Table 4, Table 5 and Table 6
respectively.

Figure 6: 5975 Series MSD with an Agilent 7890A


The result showed that the orange peel, lemon peel and lime peel essential oil contains a total of 11, 12 and
24 components respectively. Table 4, 5, and 6 shows the total components and their percentage
composition for orange, lemon and lime respectively. The GC-MS Chromatogram for orange, lemon and
lime are shown in Figure 7, 8 and 9 respectively.

Figure 7: Gas chromatographic plot for orange peel essential oil


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Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

Figure 8: Gas chromatographic plot for lemon peel essential oil

Figure 9: Gas chromatographic plot for orange peel essential oil


Table 4: Components and percentage composition in orange peel essential oil
S/N

COMPONENT

PERCENT
COMPOSITION

1R--Pinene

4.452

Cyclohexene

1.912

D-Limonene

54.151

(Z)-3-Carene

3.707

1,3,8-p-Menthatriene

2.808

Pulegone

11.652
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Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

1-Methylcyclohexa-1,3-diene

4.348

1,3-Cycloheptadiene

3.982

2-Cyclohexen-1-ol

5.231

10

1,4-Cyclohexadiene

2.299

11

2-Cyclohexen-1-one
(L-Carvone)

5.457

Table 5: Components and percentage composition in lemon peel essential oil


S/N

COMPONENT

PERCENTAGE
COMPOSITION

1R--Pinene

3.197

-Phellandrene

3.523

-Pinene

3.822

D-Limonene

64.000

1,4-Cyclohexadiene

1.957

Cis-p-Mentha-2,8-dien-1-ol

3.150

1,3,8-p-Menthatriene

2.909

1,4-Cyclohexadiene

2.968

3-Cyclohexen-1-ol

2.461

10

2-Cyclohexen-1-ol

6.210

11

1,4-Cyclohexadiene

2.539

12

2-Cyclohexen-1-one (L-Carvone)

3.265

Table 6: Components and percentage composition in lime peel essential oil


S/N

COMPONENT

PERCENTAGE
COMPOSITION

-Phellandrene

1.331

1R--Pinene

10.399

Ocimene

0.733

-Pinene

23.124

D-Limonene

17.070

1,4-Cyclohexadiene

5.516

Bicyclo[4.1.0]hept-2-ene

1.108

3-Carene

2.030

Bicyclo[3.1.0]heptan-3-ol

1.320

10

3-Cyclohexen-1-ol

4.955

11

3-Cyclohexene-1-methanol

3.611

12

E,Z-4-Ethylidenecyclohexene

1.149

13

2-Cyclohexen-1-ol

1.116

14

1,3,8-p-Menthatriene

0.834

15

2-Cyclohexen-1-one

0.810

16

2,6-Octadienal

1.320
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Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

17

1,3-Cyclohexadiene

1.006

18

1S--Pinene

0.793

19

1H-Cycloprop[e]azulene

1.237

20

Caryophyllene

4.800

21

Trans--Bergamotene

4.670

22

Bicyclo[3.1.1]hept-2-ene

2.475

23

Cyclohexene

6.924

24

Caryophyllene oxide

1.668

From Table 4, the 3 highest occurring components in orange peel essential oil are D-Limonene (54.151%),
Pulegone (11.652%) and L-Carvone (2-cyclohexen-1-one) (5.457%). Table 5 showed that the 3 highest
occurring components in lemon peel essential oil are D-limonene (64%), 2-cyclohexen-1-ol (6.21%) and
-Pinene (3.822%), while Table 6 showed that the 3 highest occurring components in lime peel essential
oil are -Pinene (23.124%), D-limonene (17.07%) and -Pinene (10.399%). The result of lemon peel
essential oil for D-limonene and -Pinene is in agreement with Soumaya, et al.(2012) findings that
D-Limonene (37.63-69.71%) , -Pinene (0.63-31.49%), -Terpene(0.04-9.96%), and p-Cymene
(0.23-9.84%) were the highest ones in lemon. However percentage composition of sweet orange (Citrus
sinensis) did not agree with that of Soumaya, et al., (2012), which had a percentage of (81.52-86.43%).
This discrepancy could possibly be as a result of Gamarra, et al. (2002) report that the quality of essential
oil depends on different factors; among them are the chemotype and biotype of the plant, the climatic
conditions as well as the extractive process. Figueiredo, et al. (2008) and Schmidt (2010) also stated that it
needs to be kept in mind that the chemical composition may already vary in the starting material, being
influenced by plant health, growth stage, habitat including climatic, edaphic factors, as well as harvest
time.
Also, Widmark and Blohm (1957) stated that oxygen consumption upon storage of different monoterpenes
has been recorded. El. Nikeety and others (1998); Turek and others (2012) went further to state that
changes in composition as well as physicochemical properties of essential oils were generally more
pronounced in half-filled containers than when only little or no headspace were present.
Asnaashari, et al. (2010), stated that the GC-MS analysis of the essential oils of lime ( C. aurantifolia) was
also performed and approximately 22 main components, with Limonene (28.27%) being the principal one,
were identified and quantified, as against 24 components identified in this work with -pinene ( 23.124%)
being the principal one, and Limonene (17.07%) being the next.
However, from Table 4, 5 and 6, it can be deduced that lemon peel essential oil has the highest percentage
of D-Limonene (64%) when comparing same quantity of the three (3) oil samples. But, from Table 1, 2 and
3, it can also be seen that orange peel produced the highest volume of essential oil when comparing same
mass of the 3 citrus peels and same time of heating.
Therefore, taking the highest time of heating (180 minutes) in Table 1, 2 and 3, and the percentage
composition of D-Limonene in the citrus peels in Table 4, 5 and 6, orange peel produced 20ml of essential
oil, while lemon produced 7ml and lime 5ml. Working out the quantity of D-Limonene in the volume of
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Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

essential oil obtained from the citrus peels at 180 minutes, using the percentage of D-Limonene in the
citrus peels as shown in tables 4, 5 and 6, lime peel essential oil contains 0.85ml D-Limonene, lemon
4.48ml D-Limonene, while orange peel essential oil contains 10.83 ml D-Limonene.

5. Conclusion/ Recommendation
The results obtained showed that lemon peel contains more amount of D-limonene per unit volume of
essential oil, while orange peel contains more amount of essential oil than lime and lemon per unit mass of
peel. However, by obtaining percentage of D-limonene in volume of essential oil obtainable from unit
mass of peel, it was concluded that orange peel is a more viable and economical source of essential oil
which is very rich in D-limonene needed for the production of insecticide. In handling the insecticide
prepared in this work, it is recommended that caution should be applied as no laboratory analysis has been
conducted to ascertain its side effect on man.
The shelf life of citrus peel essential oil have been established from literature to be greatly affected by its
storage conditions such as wrong storage containers, storage temperature and head space in storage
containers. Thus, citrus essential oil should be stored in amber bottles, and below room temperature, with
the avoidance of headspace in storage containers.
Further research to evaluate the effect of fertilizer application on;
a) The physicochemical properties of orange peel essential oil.
b)The chemical constituents in orange peel essential oil.
c) Volume of essential oil in orange peel.
d)Pectin content in orange peel.

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Quantitative and Qualitative analysis, and comparative study of essential oil extracted from Nigerian orange, lemon and lime
peels

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