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Characterisation of musk lime (Citrus

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DOI: 10.1002/jsfa.3134

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Journal of the Science of Food and Agriculture J Sci Food Agric 88:676683 (2008)

Characterisation of musk lime (Citrus

microcarpa) seed oil
Yanty NA Manaf,1 Azizah Osman,1 Oi M Lai,2 Kamariah Long3 and
Hasanah M Ghazali1
1 Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
2 Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang,
Selangor, Malaysia
3 Malaysian Agriculture Research and Development Institute (MARDI), PO Box 12301, 50774 Kuala Lumpur, Malaysia

Abstract

BACKGROUND: The seeds of musk lime (Citrus microcarpa) represent a substantial waste product of small-scale
citrus-processing factories, as they constitute about 100.0 3.2 g kg1 of the whole fruit and contain a considerable
amount of crude fat (338.0 11.3 g kg1 ). Thus the aim of the present study was to determine the physicochemical
properties of this fat with a view to potential applications.

RESULTS: The iodine and saponification values and unsaponifiable matter and free fatty acid contents of the
freshly extracted oil were 118.0 g I2 per 100 g oil, 192.6 mg KOH g1 oil, 22 mg g1 oil and 18 mg oleic acid g1 oil
respectively. The oil had a Lovibond colour index of 33.1 Y + 1.1 B. Its fatty acid profile indicated that 73.6% of
the fatty acids present were unsaturated. Linoleic (L, 31.8%), oleic (O, 29.6%) and palmitic (P, 21.4%) acids were
the predominant fatty acids, existing mainly as the triacylglycerols POL (18.9%), PLL (13.7%) and OLL (11.9%).
The melting and cooling points of the oil were 10.7 and 45.2 C respectively. Electronic nose qualitative analysis
of the oil showed the presence of volatile (aroma) compounds, although the concentrations of the more volatile
compounds were lower than those present in the seeds.

CONCLUSION: Musk lime seeds are a rich source of oil, which is unusual in having linoleic, oleic and palmitic
acids dominating the fatty acid composition. This property should make the oil both relatively stable to thermal
oxidation owing to the combined presence of oleic and palmitic acids (61.0%) and highly nutritive owing to its high
concentration of unsaturated fatty acids (73.6%).
2007 Society of Chemical Industry

Keywords: musk lime; Citrus microcarpa; seed oil; physicochemical properties; characterisation

INTRODUCTION The fruit contains over 400 constituents, including

In Malaysia, musk lime (Citrus microcarpa, also
Citrofortunella microcarpa) is known as kasturi lime;
alternative common names are calamondin orange,
Chinese or China orange, Panama orange, golden
lime and scarlet lime.1 The fruit is also known
as kalamondin, kalamunding, kalamansi, calamansi,
limonsito or agridulce in the Philippines,2 jeruk
kasturi in Indonesia3 and ma-nao-wan in Thailand.1
Musk lime is native to tropical Asia and is widely grown
in India and throughout southern Asia and South East
Asia. It is commonly grown in gardens in Malaysia and
has become the most important citrus juice source in
the Philippines. The fruits are round, small (diameter
24 cm) and have a smooth and shiny skin. They are
green when unripe and turn to yellowish orange when
ripe. The pulp is orange in colour, has many seeds
embedded in it, is juicy and has a very sour taste.
carbohydrates, organic acids, amino acids, minerals
and small quantities of flavanoids, carotenoids,
volatiles and lipids, and its consumption is linked
to potential health-related benefits.4
Like its relatives such as mandarin orange, pomelo
and sweet orange, musk lime is rich in phosphorus
(0.79 g kg1 ), calcium (0.13 g kg1 ), iron (0.002 g
kg1 ) and vitamin C or ascorbic acid (0.42 g kg1 ).5
The ascorbic acid content in the peel has been found
to exceed that in the extracted juice.4,6 Nisperos-
Carriedo et al.2 reported that the ascorbic acid content
in musk lime juice was 0.45 g L1 . Ascorbic acid in
citrus juice is also known as a natural antioxidant
and is potentially safe for use in the food industry.7
Citrus seeds and peels have also been found to
be good sources of natural antioxidants.8 10 Lime
volatile extract has been reported to have insecticidal

Correspondence to: Hasanah M Ghazali, Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 Serdang,
Selangor, Malaysia
E-mail: hasanah@putra.upm.edu.my
Contract/grant sponsor: Malaysia IRPA Programme
(Received 23 July 2007; revised version received 21 September 2007; accepted 27 September 2007)
Published online 13 December 2007; DOI: 10.1002/jsfa.3134

2007 Society of Chemical Industry. J Sci Food Agric 00225142/2007/$30.00


properties.11 Essential oils obtained from citrus fruits
and fruit peels are used in perfumes, cosmetics,
soaps, detergents and as flavourings in food products,
confectionery and beverages.12 They are also used
as components of pharmaceuticals, antiseptics and
aromatherapy products.
Musk lime juice is primarily valued for mak-
ing acidic beverages. In the Philippines a home-
grown company established in 1995, Lozada
Juices, produces a commercial concentrate and
juice of musk lime. In Malaysia, several compa-
nies have commercialised musk lime juice, such
as Nestle (Malaysia) Bhd., Universal Nutribev-
erage Sdn. Bhd. and Greenlove Food Indus-
tries Sdn. Bhd. (http://www.asiaep.com/malaysian/
foodbeverageindustry.php).
In Malaysia, musk lime seeds constitute a waste
product of citrus-processing factories. Accordingly,
the present study was carried out to determine the
proximate composition of musk lime seeds and the
physicochemical properties of the oil derived from the
seeds so as to gauge potential applications of the oil.
MATERIALS AND METHODS
Materials
The musk lime fruits used in the experiment were
obtained from a local market in Serdang, Selangor,
Malaysia. The proportions of seed, skin and juice in
the fruits were determined as 100.0 3.2, 452.0 1.9
and 448.0 2.5 g kg1 respectively. The seeds were
removed, cleaned under running tap water and dried
overnight in an oven at 60 C. The dried seeds were
kept in a sealed bottle under cool dry storage. For
analysis, whole seeds were finely ground in a Waring
32BL80 blender (Dynamic Corporation of America,
New Hartford, CT, USA) before use. The chemicals
used for oil extraction and analysis were of general and
analytical grades. Five batches of seeds (from fruits
obtained at different times) were prepared, pooled
and used in the study.
Proximate analysis
Moisture, crude protein (micro-Kjeldahl), crude
fibre and oil (Soxhlet) contents of the seeds were
determined using the methods described by Pearson,13
ash content was determined using the method of
Pomeranz and Meloan14 and total carbohydrate
was calculated by difference. The solvent used in
Soxhlet extraction was petroleum ether (4060 C).
All determinations were done in triplicate.
Oil extraction
Dried and ground musk lime seeds with shells (150 g)
were placed in a cellulose paper cone and the oil was
extracted with petroleum ether (4060 C) in a 5 L
Soxhlet extractor for 8 h.15 The oil was recovered using
an Eyela N-1 rotary evaporator (Tokyo Rakakikal Co.,
Ltd, Tokyo, Japan). The extracted fat was placed in an
oven at 60 C for 1 h and then transferred into a capped
J Sci Food Agric 88:676683 (2008)
DOI: 10.1002/jsfa
Characterisation of musk lime (Citrus microcarpa) seed oil
reagent bottle and stored at 20 C until needed for
analysis. Prior to analysis the oil was removed from
frozen storage, left standing at room temperature for
1 h and then warmed at 60 C until completely melted.
Determination of iodine and saponification
values and unsaponifiable matter and free fatty
acid contents
The iodine and saponification values and unsaponifi-
able matter and free fatty acid contents of the oil were
determined by AOAC15 standard analytical methods.
Determination of fatty acid composition
The fatty acid composition of the oil was determined
using a Shimadzu GC-14A gas chromatograph
(Shimadzu Corp., Kyoto, Japan) fitted with a flame
ionisation detector (FID) and a BPX70 polar capillary
column (SGE International Pty, Ltd, Victoria,
Melbourne, Australia) at a column pressure of 10 psi,
after conversion of the oil into fatty acid methyl esters
(FAMEs) according to the method of Cocks and Van
Rede.16 The column temperature was increased from
90 C initially to 110 C, held for 1 min, then increased
to 220 C at 8 C min1 and held for 1 min.17 The
injector and detector temperatures were maintained at
240 C. Individual peaks of FAMEs were identified
by comparing their retention times with those of
standards.
Determination of triacylglycerol profile
The triacylglycerol (TAG) profile of the oil was
obtained by non-aqueous reverse phase high-
performance liquid chromatography (HPLC) using a
Shimadzu LC-10AD liquid chromatograph equipped
with a Shimadzu SLC-10A system controller (Shi-
madzu Corp.), an auto-injector, a 0.05 mL sample
loop and a commercially packed RP-18 column
(250 mm 4 mm, particle size 5 m; Merck, Darm-
stadt, Germany).18 The analysis was done without first
removing the free fatty acids from the sample. Where
possible, the TAG peaks were identified based on the
retention times of TAG standards (LLL, OLL, OOL,
POL, PLL, OOO, POO, PPO and SOO, where L,
O, P and S denote linoleic, oleic, palmitic and stearic
acids respectively; Sigma-Aldrich, Inc., St Louis, MO,
USA) and the results of Ghazali et al.18 Peak areas
produced by the data integrator were used to quantify
the components based on relative percentages.
Determination of thermal behaviour
A Perkin-Elmer Diamond differential scanning
calorimeter (Perkin-Elmer Corp., Shelton, CT, USA)
was used to determine the thermal properties of the oil
according to the procedure described by Abdulkarim
et al.17 The oil sample was first heated to 60 C, then
cooled to 60 C at 5 C min1 and held isothermally
for 2 min. It was subsequently heated to 60 C at 5 C
min1 , held isothermally for 2 min and finally cooled
to 60 C at 5 C min1 . The heating and cooling
677
YNA Manaf et al.
thermograms were recorded and the peak, onset and
offset temperatures were noted. The melting point,
determined as the temperature where the sample was
completely melted, was established from the heating
programme.
Determination of solid fat content
The solid fat content (SFC) of the oil was determined
using a Minispec mq20 wide-line pulse nuclear
magnetic resonance (NMR) spectrometer (Brucker,
Karlsruhe, Germany) via a direct measurement
method advocated by the Palm Oil Research Institute
of Malaysia (currently known as the Malaysian Palm
Oil Board).19 Sample temperature treatment was
done using a water bath. The frozen sample was
melted at 60 C for 30 min and then poured into six
separate SFC tubes up to one-third of the tube height.
The tubes were transferred into a water bath (JEIO
TECH VTRC-620 Desktop Refrigerated Circulator;
Jeio Tech Co., Ltd., Daejeon-city, Korea) set at 0 C
and allowed to remain there for 1 h. Then five of
the tubes were transferred individually into separate
water baths set at 5, 10, 15, 17 and 20 C. The tubes
were left to stand in the water baths for 30 min and
were subsequently inserted one after another into the
NMR spectrometer. The signals were recorded and
integrated by computer to obtain the SFC (mg g1 oil).
Colour determination
The liquid oil sample was placed in a 1 inch (2.54 cm)
cell and its colour was determined using a Lovibond
Model E tintometer (Lovibond, Salisbury, UK) at
30 C by achieving the best possible match with the
standard colour slides of blue (B) and yellow (Y )
indices.
Aroma (volatile compound) profiling
The aroma (volatile compound) profiles of musk lime
seeds and seed oil were analysed qualitatively using
an Ultrafast GC zNose 7100 analyser (Electronic
Sensor Technology Co., Newbury Park, CA, USA).
The analysis was done by placing clean seeds and
unrefined seed oil in different universal bottles up
to half capacity. Then the bottles were covered with
two layers of parafilm and the samples were heated
in an oven at 60 C for 15 min. This procedure was
done to allow the emission of aroma compounds.
The vapour sample was introduced into the electronic
Table 1. Proximate composition (g kg1 ) of Citrus microcarpa (musk lime) and other citrus seeds
C. microcarpa Hybrid
Component (musk lime) (tangelo)20
Moisture 55.0 2.5
Oil 338.0 11.3 393.0 2.5
Protein 116.0 7.6
Fibre 264.0 2.4
Ash 34.0 8.7
Carbohydrate 193.0 5.6
678
nose via an inlet. The column (DB-5) temperature
was programmed from 40 to 200 C and the surface
acoustic wave (SAW) quartz microbalance detector
temperature was set at 60 C. The helium gas
flow rate was 3.5 106 m3 min1 . The chemical
analysis of flavour was accomplished within 20 s by
a very fast separation of chemically sampled vapours.
Profiles obtained are presented as zNose (Ultrafast
GC) chromatograms and polar plots (Vaporprint ,
Electronic Sensor Technology Co., Newbury Park,
CA, USA).
Statistical analysis
Where relevant, values are presented as the mean
standard deviation of triplicate determinations.
Statistical analysis was carried out by Students t test
using SPSS software (Version 11.0 software, SPSS
Inc., Chicago, IL, USA) and by analysis of variance
(ANOVA, with Duncans multiple range test) using
SAS software Version 8e (SAS Institute, Cary, NC,
USA). Significance was defined at P < 0.05.
RESULTS AND DISCUSSION
Proximate analysis of seeds
Analysis of the musk lime seed showed that it con-
tained 55.0 2.5 g kg1 moisture, 338.0 11.3 g kg1
oil, 116.0 7.6 g kg1 protein, 264.0 2.4 g kg1
crude fibre, 34.0 8.7 g kg1 ash and 193.0
5.6 g kg1 carbohydrate (by difference) (Table 1). Aje-
wole and Adeyeye20 reported that citrus seeds in
general have high oil contents, ranging from 243 g
kg1 in tangerine to 411 g kg1 in lime. It has also
been reported that lemon grown in Tunisia contains
seeds with 789 g kg1 oil, while blood orange, sweet
orange, bergamot and bitter orange seeds contained
340, 518, 361 and 261 g kg1 oil respectively.12 The
high oil content and high proportion of seeds to fruit
make the musk lime seed a suitable candidate as an
additional vegetable oil source.
Iodine and saponification values and
unsaponifiable matter and free fatty acid
contents
The iodine and saponification values and unsaponifi-
able matter and free fatty acid contents of musk lime
and other citrus seed oils are shown in Table 2. Ajew-
ole and Adeyeye20 reported that all citrus seed oils have
C. paradisi C. aurantium C. reticulata C. aurantifolia
(grapefruit)20 (sour orange)20 (tangerine)20 (lime)20
299.0 2.9 324.0 2.9 243.0 2.4 411.0 2.2
J Sci Food Agric 88:676683 (2008)
DOI: 10.1002/jsfa

Table 2. Iodine and saponification values and unsaponifiable matter and free fatty acid (FFA) contents of Citrus microcarpa (musk lime) and other
citrus seed oils
Iodine value
Seed oil source (g I2 per 100 g)
C. microcarpa (musk lime) 118.1 0.72
C. aurantium (sour orange)20 109.0 0.49
C. reticulata (tangerine)20 108.0 0.50
C. aurantifolia (lime)20 100.0 1.31
Hybrid (tangelo)20 114.0 0.78
C. sinensis (sweet orange)20 102.0 0.65
C. paradisi (grapefruit)20 101.0 0.41
high iodine and saponification values. The iodine value
of musk lime seed oil was 118.1 0.72 g I2 per 100 g,
which is slightly higher than the range (100114 g
I2 per 100 g) reported for some other lime oils.20
The iodine value indicates that musk lime seed oil is
a highly unsaturated oil containing fatty acids such
as oleic and linoleic acids. The results suggest that,
besides food use, the oil may be suitable for the formu-
lation of resin, paint and polish and even for biodiesel
production.21,22
The saponification value of musk lime seed oil
(192.6 0.05 mg KOH g1 ) is comparable to the
saponification values of raspberry, grape and safflower
seed oils (191.0, 192.9 and 191.6 mg KOH g1
respectively).23 The unsaponifiable matter content of
musk lime seed oil was 22 mg g1 , while the free fatty
acid content was 18 mg oleic acid g1 . The amounts
of these materials are expected to decrease following
refining.
Fatty acid composition
The fatty acid compositions of musk lime and other
citrus seed oils are shown in Table 3. It can be
clearly seen that citrus oils are generally highly
unsaturated, with degrees of unsaturation ranging
from 60.3 to 86.2%. Musk lime seed oil contained
73.6% unsaturated fatty acids, with linoleic (31.8%)
and oleic (29.6%) acids predominant. Palmitic acid
was the most abundant (21.4%) saturated fatty acid.
Ajewole and Adeyeye20 reported that the citrus seed
oils they studied contained palmitic acid as the
predominant saturated fatty acid (ranging between
12.1% in tangelo and 28.0% in tangerine), while oleic
(ranging from 26.1 to 45.3%) and linoleic (ranging
from 29.0 to 37.8%) acids were the major unsaturated
fatty acids. Sadani et al.12 also reported that palmitic,
oleic and linoleic acids were the three main fatty acids
in several Tunisian citrus seed oils, while Kalayasiri
et al.22 found that palmitic acid was the major fatty
acid in C. aurantium (orange) (39.5%) and C. maxima
(pomelo) (37.1%).
The presence of large proportions of palmitic, oleic
and linoleic acids makes musk lime oil and a number
of other citrus oils rather unique in comparison with
other known natural oils. Table 3 shows the ratios
of these fatty acids in different types of citrus oils,
with musk lime oil having a ratio of 0.7:1.0:1.1. This
J Sci Food Agric 88:676683 (2008)
DOI: 10.1002/jsfa
Characterisation of musk lime (Citrus microcarpa) seed oil
Saponification value Unsaponifiable FFA
(mg KOH g1 ) matter (mg g1 ) (g oleic acid g1 )
192.6 0.05 22 23 18 4.5
186.0 0.64
188.0 0.59
196.0 0.65
193.0 0.52
186.0 0.85
192.0 0.67
property would make the oil excellent in terms of
oxidative stability (51.0% palmitic plus oleic acids)
and nutritive value (73.6% unsaturated fatty acids).
Similar palmitic/oleic/linoleic acid ratios in the other
citrus seed oils in Table 3 make these oils also worthy
of further investigation.
Musk lime seed oil was also found to contain
trace quantities of lauric and myristic acids (Table 3).
Because it contained a low proportion of saturated
fatty acids, the oil remained fluid at room temperature.
It had a yellowish green colour, with a Lovibond colour
index of 33.1 Y + 1.1 B.
Triacylglycerol composition
Table 4 shows the distribution of TAGs determined
by HPLC, while Fig. 1a shows a typical TAG profile of
musk lime oil. The oil contained POL (18.9%) as the
most prominent TAG, followed by PLL (13.7%) and
OLL (11.9%). Unknown TAGs constituted 22.5% of
the total TAG content.
The unique fatty acid composition of musk lime
seed oil is reflected in its TAG profile, which differs
from the TAG profiles of other oils, including palm
olein,18 sunflower oil,24 corn oil (Fig. 1b), olive oil
(Fig. 1c), Moringa oleifera seed oil17 and animal fats
such as tallow and chicken fat.25
Thermal behaviour
In differential scanning calorimetry (DSC) analysis the
ends of the melting and cooling ranges are determined
by the position of the offset of the last peak. Each
individual fatty acid has its own melting point and,
since fats and oils are essentially mixtures of various
fatty acids as TAGs, they do not have sharp melting
points. Musk lime seed oil has a relatively low melting
point (10.7 C) and crystallisation temperature point
(4.2 C). This is consistent with the fact that the oil
has a high degree of unsaturation and a high proportion
of linoleic and oleic acids. The DSC melting and
cooling thermograms are shown in Fig. 2.
Solid fat content
The SFC of musk lime seed oil is shown in Fig. 3. At
0 C the SFC was 1.6 mg g1 . This value is consistent
with the fact that the oil has a high proportion of
unsaturated fatty acids. The SFC was reduced to zero
when the oil temperature was raised to 15 C, with the
oil remaining liquid at that temperature and beyond.
679
 680
YNA Manaf et al.

Table 3. Fatty acid compositions (%) of Citrus microcarpa (musk lime) and other citrus seed oils

Citrus Hybrid Citrus Citrus Citrus Citrus Blood

Fatty acid microcarpa (tangelo)20 aurantium20 reticulata20 aurantifolia20 sinensis20 Lemon12 orange12 Bergamot12

Caprylic/octanoic acid (C8:0) 0.04 0.01 0.60 0.2

Capric/decanoic acid (C10:0) 0.05 0.005 0.20 0.1
Lauric/dodecanoic acid (C12:0) Trace 0.04 0.01 0.07 0.01
Myristic/tetradecanoic acid (C14:0) Trace 0.10 0.05 0.80 0.3
Myristoleic/tetradece-noic acid (C14:1) 0.10 0.01 0.20 0.1
Palmitic/hexadecanoic acid (C16:0) 21.4 12.1 0.68 24.8 0.94 27.5 0.74 24.6 0.65 25.2 0.45 21.40 1.9 33.20 1.4 34.10 2.5
Palmitoleic/hexadece-noic acid (C16:1) 3.7
Margoric/heptadeca-noic acid (C17:0) 0.1
Stearic/octadecanoic acid (C18:0) 4.9 1.7 0.22 3.0 0.59 1.2 0.42 8.6 0.60 4.2 0.54 2.30 1.1 3.40 0.6 3.60 1.2
Oleic/octadecenoic acid (C18:1) 29.6 45.3 0.52 27.2 1.14 29.0 0.82 33.9 0.60 26.1 0.45 36.60 2.8 28.50 0.8 27.10 2.2
Linoleic/octadeca-dienoic acid (C18:2) 31.8 36.4 0.50 37.6 0.79 29.0 0.87 30.0 0.90 37.8 1.11 31.40 3.3 26.20 1.1 32.50 3.3
Linolenic/octadeca-trienoic acid (C18:3) 7.8 4.5 0.40 7.4 0.94 13.2 0.42 3.4 0.39 6.7 0.51 6.90 0.8 5.40 0.1 2.20 1.1
Arachidic/eicosanoic acid (C20:0) 0.7 0.30 0.01
Eicosenoic acid (C20:1) 0.03 0.01
Unsaturated fatty acids 73.6 86.2 72.2 71.2 67.3 70.6 75.0 60.3 61.8
Saturated fatty acids 26.4 13.8 27.8 28.8 32.7 29.4 23.9% 38.6 37.7
C16:0/C18:1/C18:2 0.7:1.0:1.1 0.3:1.0:0.8 0.9:1.0:1.4 0.9:1.0:1.0 0.7:1.0:0.9 1.0:1.0:1.4 0.6:1.0:0.9 1.2:1.0:0.9 1.3:1.0:1.2

DOI: 10.1002/jsfa
J Sci Food Agric 88:676683 (2008)
 Characterisation of musk lime (Citrus microcarpa) seed oil

Table 4. Triacylglycerol (TAG) composition of Citrus microcarpa

(musk lime) seed oil

TAG componenta Content (relative %)

LLL 6.8
OLL 11.9
PLL 13.7
OOL 8.7
POL 18.9
OOO 2.8
POO 8.9
PPO 5.5
SOO 0.3
Others 22.5
a L, linoleic acid; O, oleic acid; P, palmitic acid; S, stearic acid.
Figure 2. DSC cooling (A) and heating (B) thermograms of musk lime
seed oil.

Figure 3. Solid fat content (SFC) of musk lime seed oil.

zNose is essentially an instrument which comprises

Figure 1. HPLC chromatograms of (a) musk lime seed oil, (b) corn oil
and (c) olive oil.
Aroma (volatile compound) profile
The aroma of fats and oils is one of the most important
criteria influencing quality and sensory characteristics
associated with foods. An electronic nose such as the
an array of electronic chemical sensors with partial
specificity and an appropriate pattern recognition
system capable of recognising simple or complex
odours.26 zNose analysis offers a rapid and simple
way of estimating similarities and differences between
samples without actually determining the identities
of the compounds involved. Gan et al.27 reported
on the application of an electronic nose for the
characterisation of the aroma/odour characteristics of
several vegetable oils.
In the present study the qualitative analysis of
the aroma profiles of musk lime seeds and seed oil
(unrefined) was also conducted using an electronic
nose, and results obtained are shown as both
zNose (Ultrafast GC) chromatograms and polar plots
(Vaporprint ) (Fig. 4). It can be clearly seen that,
compared with the seeds, the oil contained lower
concentrations of volatiles that were eluted from the
separation column in the first 10 s, whereas there was
a concentration effect of those that were eluted later.
The solvent extraction process used to obtain the seed
oil was likely to have caused some losses in the more
volatile compounds, especially during solvent removal.
A potential application of the unrefined oil is to use it
as a flavoured oil that can be drizzled onto foods such
as French fries, salads and finished entrees or onto
meats during grilling.28

J Sci Food Agric 88:676683 (2008) 681

DOI: 10.1002/jsfa
YNA Manaf et al.
(a)
(c)
Figure 4. (a, c) Electronic nose chromatograms and (b, d) polar plots (Vaporprint ) of (a, b) musk lime seeds and (c, d) musk lime oil.
To date, no studies have been reported on the aroma
compounds of musk lime. However, studies on the
peel oils of citrus fruits such as C. aurantifolia (lime)29
and C. junos (yuzu)30 have shown that limonene is
often a major aroma compound. In the case of yuzu
tea the most intense aroma-active compounds were
linalool (floral/lemon) and decanal (orange/waxy).31
Moufida and Marzouk32 reported that limonene, -
pinene, -pinene and p-cymene were present in blood
orange, sweet orange, lemon, bergamot and bitter
orange juices, with limonene being the most abundant
compound. The proportion of limonene has been
found to vary from 50% in lemon aroma to 93%
in bitter orange aroma and about 97% in grapefruit
aroma.33
CONCLUSION
Some citrus seeds, such as musk lime seeds, are
beverage industrial waste products which can be
collected in large quantities and their potential uses
explored. Analysis showed that musk lime seeds
are rich in oil (34%) with a high proportion of
unsaturated fatty acids (74%) due mainly to the
presence of oleic (30%) and linoleic (32%) acids.
It also contains a considerable amount of palmitic
acid (21%). The combination of fatty acids should
682
(b)
(d)
make the oil both relatively stable to thermal oxidation
owing to the combined presence of oleic and palmitic
acids and highly nutritive owing to the presence of
high levels of unsaturated fatty acids. Further research
is necessary to obtain information on the nutrient
content, antioxidant and other biological activities,
inherent stability and safety of the oil, as this should
provide evidence as to the suitability of musk lime seed
oil as a valuable agricultural commodity.
ACKNOWLEDGEMENT
The authors wish to acknowledge the grant awarded
to HM Ghazali under the Malaysia IRPA Programme,
which made this research possible.
REFERENCES
1 Allen BM, Musk lime, in Common Malaysian Fruits. Malaysian
Nature Handbook, ed. by. Tweedie MWF, Allen BM. Long-
man (Malaysia), Kuala Lumpur, pp. 3839 (1975).
2 Nisperos-Carriedo MO, Baldwin EA, Moshonas MG and Shaw
PE, Determination of volatile flavor components, sugars and
ascorbic, dehydroascorbic and other acids in calamondin
(Citrus mitis Blanco). J Agric Food Chem 40:24642466
(1992).
3 Sarwono B, Jeruk dan Kerabatnya. PT Penebar Swadaya,
Jakarta, pp. 368 (1986).
J Sci Food Agric 88:676683 (2008)
DOI: 10.1002/jsfa

4 Davies FS and Albrigo LG, Fruit quality, harvesting and
postharvest technology, in Citrus. Crop Production Science
in Horticulture, ed. by Davies FS. CAB International,
Wallingford, pp. 203224 (1994).
5 Tee ES, Noor MI, Azudin MS and Idris K, Nutrient Composition
of Malaysian Foods (4th edn). Institute for Medical Research,
Kuala Lumpur, pp. 7071 (1997).
6 Kale PN and Adsule PG, Citrus, in Handbook of Fruit
Science And Technology: Production, Composition, Storage and
Processing, ed. by. Salunkhe DK, Kadam SS. Marcel Dekker,
New York, NY, pp. 3965 (1995).
7 Ranganna S, Gobindarajan VS and Ramana KVR, Citrus
fruits varieties, chemistry, technology and quality evalua-
tion. Part II. CRC Crit Rev Food Sci Nutr 18:313386 (1983).
8 Moure A, Cruz JM, France D, Dominguez JM, Seneiro J,
Dominguez H, et al, Natural antioxidants from residual
sources. Food Chem 72:145171 (2001).
9 Bocco A, Cuvelier ME, Richard H and Berset C, Anti-oxidant
activity and phenolic composition of citrus peel and seed
extracts. J Agric Food Chem 46:21232129 (1998).
10 Rehman ZU, Citrus peel extract a natural source of antioxi-
dant. Food Chem 99:450454 (2006).
11 Ezeonu FC, Chidume GI and Udedi SC, Insecticidal properties
of volatile extracts of orange peels. Bioresource Technol
76:273274 (2001).
12 Sadani M, Dhifi W and Marzouk B, Lipid evaluation of some
Tunisian citrus seeds. J Food Lipids 11:242250 (2004).
13 Pearson D, The Chemical Analysis of Foods. Longman, London,
pp. 626 (1976).
14 Pomeranz Y and Meloan C, Food Analysis. Theory and Practice
(3rd edn). Chapman and Hall, New York, NY, p. 78 (1994).
15 AOAC, Official Methods of Analysis (14th edn). Association of
Official Analytical Chemists, Washington, DC (1984).
16 Cocks LV and Van Rede C, Laboratory Handbook for Oils and
Fats Analyses. Academic Press, London, pp. 113117 (1966).
17 Abdulkarim SM, Long K, Lai OM, Muhammad SKS and
Ghazali HM, Some physico-chemical properties of Moringa
oleifera seed oil extracted using solvent and aqueous enzymatic
methods. Food Chem 93:253263 (2005).
18 Ghazali HM, Hamidah S and Che Man YB, Enzymatic trans-
esterification of palm olein with nonspecific and 1,3 specific
lipases. J Am Oil Chem Soc 72:633639 (1995).
19 PORIM, PORIM Test Methods. Palm Oil Research Institute of
Malaysia/Ministry of Primary Industries, Bangi (1995).
20 Ajewole K and Adeyeye A, Characterisation of Nigerian citrus
seed oils. Food Chem 47:7778 (1993).
J Sci Food Agric 88:676683 (2008)
DOI: 10.1002/jsfa
View publication stats
Characterisation of musk lime (Citrus microcarpa) seed oil
21 Sonntag AV, Sources, Utilization and Classification of Oils and
Fats, in Baileys Industrial Oil and Fat Products (4th edn), ed.
by Swern D. Wiley, New York, NY, p. 284285 (1979).
22 Kalayasiri P, Jeyasoke N and Krisnangkura K, Survey of seed
oils for use as diesel fuels. J Am Oil Chem Soc 73:471474
(1996).
23 Oomah BD, Ladet S, Godfrey DV, Liang J and Girard B,
Characteristics of raspberry (Rubus idaeus L.) seed oil. Food
Chem 69:187193 (2000).
24 Lim LP, Ghazali HM, Lai OM and Chu BS, Comparison of
transesterified palm stearin/sunflower oil blends catalysed
by Pseudomonas and Mucor javanicus lipases. J Food Lipids
8:103114 (2001).
25 Marikkar JMN, Ghazali HM, Che Man YB, Peris TSG and
Lai OM, Distinguishing lard from other animal fats in admix-
tures of some vegetable oils using liquid chromatographic data
coupled with multivariate data analysis. Food Chem 91:514
(2005).
26 Gardner JW and Bartlett PN, A brief history of electronic noses.
Sens Actuat B 18/19:211220 (1994).
27 Gan HL, Che Man YB, Tan CP, NorAini I and Nazimah SAH,
Characterisation of vegetable oils by surface acoustic wave
sensing electronic nose. Food Chem 89:507518 (2005).
28 Sloan E, Top ten global food trends. Food Technol 59:2032
(2005).
29 Ramesh Yadav A, Chauhan AS, Rekha MN, Rao LJM and
Ramteke RS, Flavour quality of dehydrated lime [Citrus
aurantifolia (Christm.) Swingle]. Food Chem 85:5962 (2004).
30 Song HS, Sawamura M, Ito T and Ukeda H, Chemical com-
positions of volatile part of yuzu (Citrus junos Tanaka)
peel cold-pressed oil from Japan and Korea. Flav Fragr J
14:383389 (1999).
31 Kim TH, Lee SM, Kim SY, Kim KH, Oh S and Lee HJ,
Aroma dilution method using GC injector split ratio for
volatile compounds extracted by headspace solid phase
microextraction. Food Chem 83:151158 (2003).
32 Moufida S and Marzouk B, Biochemical characterization of
blood orange, sweet orange, lemon, bergamot and bitter
orange. Phytochemistry 62:12831289 (2003).
33 Dugo G, Verzera A, Stagno H, dAlcontres IS, Cotroneo A and
Ficarra R, On the genuineness of citrus essential oils. Part
XLI. Italian bitter orange essential oil: composition and
detection of contamination and additions of oils and terpenes
of sweet orange and of lemon. Flav Fragr J 8:2533 (1993).
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