Important avonoids and limonin in selected Thai

citrus residues
Vaniya Chinapongtitiwat, Saranya Jongaroontaprangsee, Naphaporn Chiewchan
*
,
Sakamon Devahastin
Department of Food Engineering, Faculty of Engineering, King Mongkuts University of Technology Thonburi, 126 Pracha u-tid Road, Tungkru,
Bangkok 10140, Thailand
A R T I C L E I N F O
Article history:
Received 2 November 2012
Received in revised form
14 March 2013
Accepted 14 March 2013
Available online 6 April 2013
Keywords:
Citrus fruits
Dietary bre
Flavanones
Limonin
Polymethoxyavones
A B S T R A C T
The distributions of important avonoids and limonin in selected Thai citrus residues were
investigated in this study. The residues of interest were peels of pomelo (Citrus grandis (L.)
Osbeck cv. Kao Yai and cv. Kao Nampheung), residues after juice extraction of tangerine
(Citrus reticulata Blanco cv. Sainamphueng and cv. Bangmod) and peels and residues after
juice extraction of kafr lime (Citrus hystrix DC). Flavonoids were characterized and quan-
tied in terms of avanones, i.e., naringin and hesperidin, and polymethoxyavones
(PMFs), i.e., nobiletin, sinensetin and tangeretin. Naringin was a dominant avanone in
polmelo peels. Three PMFs were found in all samples with varying contents. Comparing
the samples, residues from tangerine cv. Bangmod exhibited the highest value of each
PMFs. Limonin was present in small amounts in pomelo peels and residues after juice
extraction of tangerine and kafr lime. All samples were good sources of dietary bre, with
a total dietary bre content of more than 60 g/100 g on a dry weight basis with high propor-
tion of soluble dietary bre.
2013 Elsevier Ltd. All rights reserved.
1. Introduction
Citrus fruits are consumed either in their fresh or processed
form due to their pleasant avour, refreshing juice and health
benets. Many phytochemicals have been identied in citrus
fruits; these include vitamin C, carotenoids, avonoids and
limonoids (Igual, Garc a-Mart nez, Camacho, & Mart nez-
Navarrete, 2013). Citrus avonoids have gained much interest
due to their chemoprotective effects. Citrus avonoids exhibit
antioxidant, antimicrobial, anticarcinogenic, antiviral, anti-
allergic and anti-inammatory activities (Benavente-Garc a,
Castillo, Mar n, Ortun o, & Del Ro, 1997; Ram & Singh, 2006).
They also inhibit human platelet aggregation (Benavente-
Garc a et al., 1997; Kanadaswami et al. 2005). Moreover, citrus
fruits also contain high amounts of dietary bre (DF) with well
balanced proportion of soluble dietary bre (SDF) and insolu-
ble dietary bre (IDF) (Baker, 1994).
There are many classes of avonoids with avanones
being the abundant group in citrus fruits (Peterson et al.,
2006; Ram & Singh, 2006). The most prevalent avanones in
tissues and peels of citrus fruits are naringin and hesperidin
(Gattuso, Barreca, Gargiulli, Leuzzi, & Caristi, 2007; Stuetz,
Prapamontol, Hongsibsong, & Biesalski, 2010). Naringin
exhibits many health benets, including an ability to prevent
cancer by suppression of carcinogenesis and inducing cell
apoptosis (Meiyanto, Hermawan, & Anindyajati, 2012).
Hesperidin has also been reported to reduce the proliferation
of many cancer cells (Nazari, Ghorbani, Hekmat-Doost,
1756-4646/$ - see front matter 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jff.2013.03.012
* Corresponding author. Tel.: +66 2 470 9243; fax: +66 2 470 9240.
E-mail address: naphaporn.rat@kmutt.ac.th (N. Chiewchan).
J O U R N A L O F F U N C T I O N A L F O O D S 5 ( 2 0 1 3 ) 1 1 5 1 1 1 5 8
Avai l abl e at www. sci encedi r ect . com
j our nal homepage: www. el sevi er . com/ l ocat e/ j f f
Jeddi-Tehrani, & Zand, 2011; Park, Kim, Ha, & Chung, 2008)
and also possesses anti-inamatory effect (Benavente-Garc a
et al., 1997).
Flavones are also found in citrus fruits but at lower con-
centrations than avanones. Flavones are generally found in
the oil glands of citus avedo (Chen, Montanari, & Widmer,
1997). Polymethoxyavones (PMFs), avones bearing two or
more methoxy groups on their basic benzo-c-pyrone (15-car-
bon, C
6
C
3
C
6
) skeleton with a carbonyl group at the C
4
posi-
tion (Li et al., 2009), are of particular interest due to their
broad biological activities, including anticancer, anti-athero-
genic and anti-inammatory activities (Du & Chen, 2010;
Lee et al., 2013; Li, Lo, & Yo, 2006; Ram & Singh, 2006). PMFs
that have typically been identied in citrus fruits include
nobiletin, sinensetin and tangeritin (Robards, Li, Antolovich,
& Boyd, 1997).
In addition to being a rich source of avonoids, citrus fruits
also contain signicant amounts of limonoids (Manners,
2007), which are a group of chemically related triterpene
derivatives found in citrus fruits and exist in many forms
(Hasegawa, 2000). Limonin is a major limonoid and is the pri-
mary cause of bitterness of citrus fruits (Hasegawa, Dillberger,
& Choi, 1984); the amount of limonin varies greatly depending
on the variety and part of the fruit (Ohta & Hasegawa, 1995).
Although limonoids are a major cause of bitterness in citrus
fruits, these compounds have been reported to possess
substantial anticancer and antiviral activities (Lam, Li, &
Hasegawa, 1989; Roy & Sara, 2006).
Many researchers have shown that citrus by-products are
a good source of DF (Jongaroontaprangsee et al., 2007;
Ubando-Rivera, Navarro-Ocan a, & Valdivia-Lo pez, 2005) as
well as phytochemicals (Pichaiyongvongdee & Haruenkit,
2009a, 2009b; Sun et al., 2010) and there is a potential to trans-
form citrus by-products into functional DF powder possessing
antioxidant and anticarcinogenic activities (Kuljarachanan,
Devahastin, & Chiewchan, 2009). Thailand has a wide variety
of citrus fruits that the bioactive compounds as well as DF
content in processing residues have never been fully charac-
terized. This work was therefore aimed at studying the
proles of important avonoids and limonin in selected Thai
citrus residues. The dietary bre content in the residues was
also determined.
2. Materials and methods
2.1. Sample preparation
Fresh citrus fruits, i.e., pomeloes (Citrus grandis (L.) Osbeck cv.
Kao Nampheung and C. grandis (L.) Osbeck cv. Kao Yai), tan-
gerines (Citrus reticulata Blanco cv. Bangmod and C. reticulata
Blanco cv. Sainamphueng) and kafr lime (Citrus hystrix DC)
with fresh appearance, free of rotting and bruising or any
other signs of deterioration were purchased from Pracha u-
tid 61 Market in Bangkok, Thailand. The sizes of pomeloes,
tangerines and kafr limes used in the experiments were
170200, 3234 and 2530 mm, respectively. For pomeloes,
only their peels (albedo and avedo) with an average thick-
ness of 2025 mm were used. After purchasing the samples
were kept at 4 C until the time of experiment, which was
on the same day of the purchase.
Prior to experiment, fruit samples were washed with tap
water and gently rubbed by a sponge. The washed fruits were
air dried at ambient temperature (28 C). Peels of pomeloes
and kafr limes were taken off from the fruits. The peels were
cut into small pieces before being chopped by a chopper (War-
ing, Torrington, CT, USA). For the preparation of tangerine and
kafr lime residues, fruits were cut into half and squeezed by
a hand-pressed juice extractor. The residues were cut into
small pieces and subsequently chopped by the chopper. The
particle size of the samples after chopping was approximately
13 mm. The prepared citrus residues were dried in a freeze
dryer until the moisture content was less than 10% on a dry
basis. The dried samples were vacuum-packed in aluminum
foil packets and kept at 18 C until further determination
of avonoids, limonin and DF contents. All analyses were per-
formed within one week.
2.2. Determination of avonoids contents
The avonoids extraction method was as suggested by Sene-
virathne, Joen, Ha and Kim (2009) with slight modications.
One g of a sample was rst mixed with 50 mL of methanol
and placed in an incubator shaker (New Brunswick Scien-
tic, Edison, NJ, USA) at 120 rpm at ambient temperature
(28 C) for one day; the content was then ltered through
Whatman No. 1 lter paper with the aid of a vacuum pump
(Gast, Benton Harbor, MI, USA). The extract solution was dis-
solved in 50 mL of methanol (AR grade) before further
analysis.
The avonoid constituents were determined using high
performance liquid chromatography (HPLC) following the
method suggested by Nogata et al. (2006) with some modica-
tions. In brief, the sample extract was ltered through a 0.45-
lm nylon lter. Ten microlitres of the ltrate was injected into
a Symmetry C
18
5 lm (3.9 150 mm) HPLC column (Waters,
Milford, MA, USA). The HPLC system consisted of a pump
and controller (Waters, Milford, MA, USA) and photodiode ar-
ray detector (Waters, Milford, MA, USA). The mobile phase
was 0.01 M phosphoric acid (H
3
PO
4
) and methanol (MeOH,
HPLC grade). The gradient program was as follows:
(1) 055 min, 7055% (v/v) H
3
PO
4
and 3045% (v/v) MeOH;
(2) 5595 min, 550% (v/v) H
3
PO
4
and 45100% (v/v) MeOH;
(3) 95100 min, 100% (v/v) MeOH; the ow rate was set at
0.6 mL/min. A UV spectrophotometer detector at a wave-
length of 285 nm was used for detecting avonoids. The
column temperature was maintained at 40 C.
The concentration of each avonoid was calculated from
an integrated chromatographic peak area of the sample and
the corresponding standard; standard avonoids consisted
of hesperidin, naringin, sinensetin, nobiletin and tangeretin.
2.3. Determination of limonin content
The limonin content was determined using HPLC following
the method of Sun, Chen, Chen, and Chen (2005) with slight
modications as proposed by Kuljarachanan et al. (2009).
One gram of sample was mixed with 60 mL of dichlorometh-
ane and placed in the incubator shaker at 120 rpm at ambient
temperature (28 C) for 4 h; the content was then ltrered
through Whatman No. 1 lter paper with the aid of a vacuum
1152 J O U R N A L O F F U N C T I O N A L F O O D S 5 ( 2 0 1 3 ) 1 1 5 1 1 1 5 8
pump. The extract solution was evaporated to close to com-
plete dryness with a rotary evaporator (Resona Technics, Gos-
sau, Switzerland) at 30 C for 20 min. The residues were
dissolved in 10 mL of acetonitrile (HPLC grade) and ltered
through a 0.45-lm syringe lter. Ten microlitres of the ltered
solution were then injected into a liquid chromatography
column.
Symmetry C
18
5 lm (4.6 150 mm) HPLC column (Waters,
Milford, MA, USA) was used for limonin analysis. The HPLC
system consisted of a pump and controller (Waters, Milford,
MA, USA) and a tunable absorbance detector (Waters, Milford,
MA, USA). A mixture of methanol, acetonitrile and water
(1:37:62, v/v/v) was used as the mobile phase and its ow rate
was set at 1 mL/min. A UV spectrophotometer detector at a
wavelength of 210 nm was used for detecting limonin. The
mobile phase was degassed using an ultrasonic generator.
2.4. Determination of dietary bre contents
Total dietary bre (TDF), soluble dietary bre (SDF) and insol-
uble dietary bre (IDF) contents of a sample were determined
according to AOAC (2000) Method 991.43. In brief, a dried pow-
dery sample was rst gelatinized with heat stable a-amylase
(SigmaAldrich, Steinheim, Germany). After gelatinization
the sample was digested with protease (SigmaAldrich,
Steinheim, Germany) and amyloglucosidase (SigmaAldrich,
Steinheim, Germany) to remove proteins and starch present
in the sample. Subsequently, IDF was ltered and the residue
was washed with warm distilled water. The ltrate and wash
water were combined and added with 4 volumes of 95% (v/v)
methanol at 60 C to precipitate SDF. The precipitate (residue)
was then ltered and weighed after drying at 105 C in a hot
air oven (Memmert, Schwabach, Germany). Both IDF and
SDF residues were corrected for proteins (Kjeldahl procedure),
ash (incinerating the sample at 525 C) and blank for the nal
calculation of SDF and IDF contents. A blank sample was
evaluated along with the tested sample to measure any
contribution of the reagents on the residues.
2.5. Statistical analysis
All experiments were performed in duplicate and mean val-
ues (on dry basis) with standard deviations are reported.
The experimental data were analyzed using an analysis of
variance (ANOVA). Differences between mean values were
established using the Fishers test at 95% condence. SPSS

software (version 17, SPSS Inc., Chicago, IL, USA) was used
to perform all the statistical calculations.
3. Results and discussion
3.1. Flavonoids contents in Thai citrus residues
Fig. 1 shows a typical chromatogram of important avonoids
in selected citrus residues. The quantitative determination of
the important avonoids on a dry basis is given in Tables 1
and 2. The results showed that the type and contents of avo-
noids varied widely depending on the citrus species. It is
Sinensetin
Nobiletin
Tangeretin
Naringin
(a)
(b)
Hesperidin
Fig. 1 Typical chromatograms of important avonoids in (a) pomelo peels (Citrus grandis (L.) Osbeck cv. Kao Yai) and (b)
residues after juice extraction of tangerine (Citrus recticulata Blanco cv. Bangmod).
J O U R N A L O F F U N C T I O N A L F O O D S 5 ( 2 0 1 3 ) 1 1 5 1 1 1 5 8 1153
noted that naringin and hesperidin were the targeted ava-
nones while sinensetin, nobiletin and tangeretin were the tar-
geted polymethoxyavones in this study.
Table 1 lists the naringin and hesperidin contents of the
selected citrus residues. Naringin was noted to be a major a-
vonoid in the peels of both varieties of pomeloes. Naringin
has also been reported to be a predominant avanone in peels
and edible portions of many varieties of C. grandis (Nogata
et al., 2006; Wang, Chuang, & Ku, 2007; Zhang, Duan, Zang,
Huang, & Liu, 2011). Although high concentrations of naringin
were noted in the peels of both varieties of pomeloes, the
amounts were still lower than those detected in grapefruit
(C. paradisi) peels (McIntosh & Mansell, 1997; Xu, Ye, Chen, &
Liu, 2007). Naringin contents reported here were also lower
than those reported previously for the same cultivars of
pomeloes. Pichaiyongvongdee and Haruenkit (2009a) deter-
mined the naringin contents in different parts of Thai pomelo
cultivars, including Kao Yai and Kao Nampheung. Naringin in
avedo and albedo were separately determined; the sum of
the naringin contents in avedo and albedo of pomeloes cv.
Kao Yai and Kao Nampheung were 16,929 and 25,071 ppm,
respectively. Cultural practice as well as cultivation area
might contribute to the variation of naringin in pomelo peels
of the same cultivar. Chaiwong and Theppakorn (2010) re-
ported that esh of C. grandis cv. Thong Dee grown in different
parts of Thailand possessed naringin content in the range of
254719 ppm.
On the other hand, only minute content of naringin was
observed in the residues after juice extraction of tangerine
cv. Sainamphueng, while no naringin was detected in the res-
idues after juice extraction of tangerine cv. Bangmod. The ab-
sence of naringin in many tangerine cultivars has also been
reported (Kawaii, Tomono, Katase, Ogawa, & Yano, 1999;
Nogata et al., 2006; Peterson et al., 2006).
In terms of hesperidin, this avanone was not observed in
the selected pomelo peels. Previous works have indeed shown
that pomeloes contained small amounts of hesperidin
(Pichaiyongvongdee & Haruenkit, 2009b; Wang, Chuang, &
Ku, 2008; Wang et al., 2007), while some possessed no
hesperidin at all (Zhang et al., 2011). Pichaiyongvongdee and
Haruenkit (2009) also reported that many Thai cultivar of
pomeloes contained no hesperidin.
In contrast, hesperidin was note to be a major avanone in
the residues after juice extraction of tangerine. Manthey and
Grohmann (1996) also reported that hesperidin is a main av-
anoid in orange peels. The hesperidin contents in the resi-
dues after juice extraction of tangerines reported here were
in similar order to those previously reported in orange peels
(19,17027,781 ppm) (Manthey & Grohmann, 1996) and much
higher than those reported in sour orange (C. Aurantium) peels
(Bocco, Cuvelier, Richard, & Berset, 1998) and Satsuma man-
darin (C. Unshiu Marc.) peels (Ma et al., 2008). Hesperidin con-
tent in the residues of tangerine cv. Bangmod was higher than
that of tangerine cv. Sainampheung. Tangerine cv. Sai-
nampheung nevertheless contained higher amount of hes-
peridin than hand-pressed juice and peeled fruit of the
same cultivar (Stuetz et al., 2010). Residues after juice extrac-
tion are indeed known to be a rich source of hesperidin than
the other fruit parts and its juice (Sun et al., 2010). Hesperidin
was also much more dominant than naringin in kafr lime
residues. Berhow, Fong, and Hasegawa (1996) indeed reported
that hesperidin is abundant in the leaves of C. Hystrix, while
naringin was not detected.
Since it has been reported that sinensetin, nobiletin and
tangeretin are PMFs commonly found in a wide variety of cit-
rus fruits (Manthey & Grohmann, 2001; Robards et al., 1997),
these PMFs were determined in this study. Table 2 lists the
amounts of PMFs found in various residues. All residues con-
tained three PMFs at different contents. The two tangerine
residues possessed more signicant amounts of the three
PMFs comparing to the pomelo peels and kafr lime residues.
The results were similar to those reported by Kawaii et al.
Table 1 Amount of avanones (ppm) in Thai citrus residues.
Scientic name Cultivar Sample type Naringin content Hesperidin content
Citrus grandis (L.) Osbeck Kao Yai Peels 10.884 608
a
ND
Citrus grandis (L.) Osbeck Kao Namphueng Peels 11.875 955
a
ND
Citrus reticulata Blanco Sainamphueng Residues after juice extraction 176 1
b
17.680 97
a
Citrus reticulata Blanco Bangmod Residues after juice extraction ND 23.327 3856
a
Citrus hystrix DC Kafr lime Peels 149 43
b
2.210 852
b
Citrus hystrix DC Kafr lime Residues after juice extraction 90 24
b
5.326 152
b
Same letters in the same column indicate that values are not signicantly different (p > 0.05).
Table 2 Amount of polymethoxyavones (ppm) in Thai citrus residues.
Scientic name Cultivar Sample type Sinensetin content Nobiletin content Tangeretin
content
Citrus grandis (L.) Osbeck Kao Yai Peels 29 19
b
12 9
c
14 2
c
Citrus grandis (L.) Osbeck Kao Namphueng Peels 17 7
b
11 3
c
3 2
Citrus reticulata Blanco Sainamphueng Residues after juice extraction 201 40
a
702 55
b
498 20
b
Citrus reticulata Blanco Bangmod Residues after juice extraction 208 5
a
1.566 72
a
1.361 276
a
Citrus hystrix DC Kafr lime Peels 28 17
b
83 12
c
7 3
c
Citrus hystrix DC Kafr lime Residues after juice extraction 46 32
b
220 95
bc
7 1
c
Same letters in the same column indicate that values are not signicantly different (p > 0.05).
1154 J O U R N A L O F F U N C T I O N A L F O O D S 5 ( 2 0 1 3 ) 1 1 5 1 1 1 5 8
(1999) who noted that Hirado Buntan (C. grandis) contained
only small amounts of nobiletin (1 ppm on a dry weight basis)
and tangeritin (7 ppm on a dry weight basis), while no sinese-
tin was detected in the edible parts (juice sac and segment
epidermis) of the fruit. Wang et al. (2007) also reported that
the edible portions of the two varieties of C. grandis, i.e., Wen-
dun and Peiyou, contained sinensetin of only 2543 ppm on a
dry weight basis.
The above results illustrate that the residues obtained
from the two cultivars of Thai tangerine are good sources of
PMFs. Nobiletin and tangeretin contents noted in these two
samples were indeed much higher than those reported for
the edible parts (juice sac and segment epidermis) of Ponkan
(C. recticulata) and Ota Ponkan (C. recticulata) (Kawaii et al.,
1999). Sinensetin contents in the residues of both tangerine
cultivars were in the same order as those reported in the peels
of Ponkan (C. recticulata) (Wang et al., 2008).
Similar PMFs, i.e., sinensetin, nobiletin and tangeretin,
were observed previously in hand-pressed juice and peeled
fruits of tangerine cv. Sainampheung (Stuetz et al., 2010) but
the contents reported were much lower than those reported
here. The differences may arise from the different fruit parts
used in the experiments. Naturally, avonoids distribute
throughout the fruit with varying amounts; peels, however,
accumulate the highest contents of avonoids (Sun et al.,
2010). Residues after juice extraction of tangerine included
all parts of the fruit except juice and seeds. This made the res-
idues become the richer source of PMFs.
To the best of our knowledge, the proles of PMFs in tan-
gerine cv. Bangmod have not previously been reported. It is
interesting that their PMFs contents were signicantly higher,
especially nobiletin and tangeretin, than those of tangerine
cv. Sainampheung.
3.2. Limonin contents of Thai citrus residues
Fig. 2 exemplies a typical chromatogram of limonin in citrus
samples; the shown chromatogragphs belongs to the residues
after juice extraction of kafr lime. Limonin contents in all
investigated citrus residues are given in Table 3. In general,
limonin is present in all citrus fruit tissues and the amount
varies widely depending on the variety, stage of fruit growth
and fruit part (McIntosh & Mansell, 1997; Sun et al., 2005).
Only small amounts of limonin were noted in peels of both
pomelo cultivars and the contents were similar to that found
in peels of grapefruit (C. Paradisi), which was in the range of
17.7107 ppm on a dry weight basis (McIntosh & Mansell,
1997). However, the limonin content noted in the present
work was much lower than that in Thai pomelo peels
(350535 ppm on a dry weight basis) as reported by
Pichaiyongvongdee and Haruenkit (2009). The limonin
contents in the residues after juice extraction of tangerine
Limonin
Fig. 2 Typical chromatogram of limonin in residues after
juice extraction of kafr lime (Citrus hystrix DC).
Table 4 Amount of dietary bre (g/100 g) in Thai citrus residues.
Scientic name Cultivar Sample type IDF content SDF content TDF content IDF:SDF (w/w)
Itrus grandis (L.) Osbeck Kao Yai Peels 52.13 4.95
ab
30.55 0.76
bc
82.69 2.83
a
1.71:1
Citrus grandis (L.) Osbeck Kao Namphueng Peels 48.51 0.30
bc
33.95 1.03
a
82.46 0.73
a
1.43:1
Citrus reticulata Blanco Sainamphueng Residues after juice extraction 36.38 0.09
d
31.82 0.05
ab
68.20 0.02
c
1.14:1
Citrus reticulata Blanco Bangmod Residues after juice extraction 36.97 1.32
d
27.76 1.86
c
64.73 0.54
c
1.33:1
Citrus hystrix DC Kafr lime Peels 54.01 1.48
a
28.13 0.52
c
82.14 0.96
a
1.92:1
Citrus hystrix DC Kafr lime Residues after juice extraction 46.93 2.67
c
28.10 1.63
c
74.94 4.31
bc
1.67:1
Same letters in the same column indicate that values are not signicantly different (p > 0.05).
Table 3 Limonin content (ppm) of Thai citrus residues.
Scientic name Cultivar Sample type Limonin content
Citrus grandis (L.) Osbeck Kao Yai Peels 86 10
b
Citrus grandis (L.) Osbeck Kao Namphueng Peels 41 5
ab
Citrus reticulata Blanco Sainamphueng Residues after juice extraction 90 34
b
Citrus reticulata Blanco Bangmod Residues after juice extraction 85 12
b
Citrus hystrix DC Kafr lime Peels 194 34
c
Citrus hystrix DC Kafr lime Residues after juice extraction 14 2
a
Same letters in the same column indicate that values are not signicantly different (p > 0.05).
J O U R N A L O F F U N C T I O N A L F O O D S 5 ( 2 0 1 3 ) 1 1 5 1 1 1 5 8 1155
cv. Sainamphueng and cv. Bangmod were not so high. The
limonin contents reported in this work were similar to those
reported by Jungsakulrujirek and Noomhorm (1998) for peels
(albedo and avedo) of Thai tangerine fruits (C. reticulata Blan-
co). Kafr lime peels, on the other hand, possessed much
higher content of limonin.
3.3. Dietary bre contents of Thai citrus residues
Dietary bre (DF) contents and compositions of Thai citrus res-
idues are listed in Table 4. The results showed that all samples
were good sources of dietary bre, containing total dietary -
bre of higher than 60 g/100 g on a dry weight basis with high
proportion of soluble dietary bre which were even higher
than many other agricultural by-products such as residues
after juice extraction of peach (Grigelmo-Miguel & Mart n-
Belloso, 1999), carrot pomace (Nawirska & Kwasniewska,
2005) and carrot peels (Chantaro, Devahastin, & Chiewchan,
2008). Pomelo peels of both cultivars and kafr lime peels con-
tained similar DF contents of approximately 82 g/100 g on a
dry weight basis. The TDF content in peels of pomelo cv. Kao
Yai was similar to that in albedo of the same cultivar (78
80 g/100 g on a dry weight basis), while the SDF content was
lower than that reported by Naowakul, Wirjantoro, and
Phianmongkhol (2013). The TDF contents of the residues after
juice extraction of both varieties of tangerine were approxi-
mately 6568 g/100 g on a dry weight basis, which were higher
than the TDF content of peels of Thai Tangerine (C. reticulata
Blanco) (Attavanich & Anprung, 2003).
It is interesting to note that the SDF contents of all sam-
ples were in a similar order, while there was a wide variation
in the TDF contents. This was then resulted in the variation in
the IDF content and IDF:SDF ratio. Residues after juice extrac-
tion of tangerine possessed a better balance of IDF content
and SDF content than the other tested samples. Fibre source
suitable for use as a food ingredient should have an IDF:SDF
ratio close to 12.3 (Grigelmo-Miguel & Mart n-Belloso, 1999).
Comparing between peels and residues after juice extrac-
tion of kafr lime, it was observed that the peels contained
higher amount of IDF than the residues after juice extraction;
the SDF contents were nevertheless similar. In general, IDF is
a structure of natural cell walls, which are the main composi-
tion of fruit; IDF makes up about 2/3 of the bre in most foods.
This is why the peels of citrus residues provided the higher
amount of IDF.
4. Conclusion
Important avonoids and limonin in selected Thai citrus res-
idues were investigated. The results showed that types and
amounts of the phytochemicals of interest varied widely
depending on citrus variety as well as fruit part used for the
analysis. In terms of avanones, pomelo peels was found to
be a rich source of naringin, while tangerine residues after
juice extraction were abundant in hesperidin. Three PMFs,
namely, sinensetin, nobiletin and tangeretin, were detected
in all samples with varying amounts; residues after juice
extraction of tangerine were again rich sources of PMFs. Only
small amount of limonin were detected in pomelo peels,
tangerine and kafr lime residues. All samples also contained
well-balanced proportion of IDF and SDF in the range of 1.14
1.92.
Due to the high contents of benecial phytochemicals and
DF, the investigated by-products may be used as raw materi-
als to produce DF associated with various bioactive com-
pounds. Study of the effects of pretreatment as well as
drying methods and conditions on the retention of the bioac-
tive compounds contained in the DF powder is suggested for
further study. The functional properties of the DF powder
should also be investigated.
Acknowledgements
The authors express their sincere appreciation to the Thai-
land Research Fund (TRF) and King Mongkuts University of
Technology Thonburi for supporting the study nancially.
Author Jongaroontaprangsee thanks the TRF, through its Roy-
al Golden Jubilee (RGJ) Scholarship Program, and the Commis-
sion on Higher Education for supporting her doctoral study.
R E F E R E N C E S
AOAC. (2000). Ofcial methods of analysis. Washington DC:
Association of Ofcial Analytical Chemists.
Attavanich, N., & Anprung, P. (2003). Dietary bre powder from
Citrus reticulata Blanco peel and its application. Food, 33, 4555.
Baker, R. A. (1994). Potential dietary benets of citrus pectin and
bre. Food Technology, 48, 133139.
Benavente-Garca, O., Castillo, J., Marn, F. R., Ortun o, A., & Del R o,
J. A. (1997). Uses and properties of citrus avonoids. Journal of
Agricultural and Food Chemistry, 45, 45054515.
Berhow, M. A., Fong, C. H., & Hasegawa, S. (1996). Limonoid and
avonoid composition in varieties of papeda and
papedocitrus. Biochemical Systematics and Ecology, 24, 237242.
Bocco, A., Cuvelier, M.-E., Richard, H., & Berset, C. (1998).
Antioxidant activity and phenolic composition of citrus peel
and seed extracts. Journal of Agricultural and Food Chemistry, 46,
21232129.
Chaiwong, S., & Theppakorn, T. (2010). Bioactive compounds and
antioxidant capacity of pink pomolo (Citrus grandis (L.) Osbeck)
cv. Thong Dee) in Thailand. ISSAAS Journal, 16, 1016.
Chantaro, P., Devahastin, S., & Chiewchan, N. (2008). Production of
antioxidant high dietary bre powder from carrot peels. LWT
Food Science and Technology, 41(2008), 19871997.
Chen, J., Montanari, A. M., & Widmer, W. W. (1997). Two new
polymethoxylated avones, a class of compounds with
potential anticancer activity, isolated from cold pressed Dancy
tangerine peel oil solids. Journal of Agricultural and Food
Chemistry, 45, 364368.
Du, Q., & Chen, H. (2010). The methoxyavones in Citrus reticulata
Blanco cv. Ponkan and their antiproliferative activity against
cancer cells. Food Chemistry, 119, 567572.
Gattuso, G., Barreca, D., Gargiulli, C., Leuzzi, U., & Caristi, C. (2007).
Flavonoid composition of citrus juices. Molecules, 12,
16411673.
Grigelmo-Miguel, N., & Martn-Belloso, O. (1999). Comparison of
dietary bre from by-products of processing fruits and greens
and from cereals. LWT Food Science and Technology, 32,
503508.
Hasegawa, S. (2000). Biochemistry of limonoids in citrus. In M. A.
Berhow, S. Hasegawa, & G. D. Manners (Eds.), Citrus limonoids:
1156 J O U R N A L O F F U N C T I O N A L F O O D S 5 ( 2 0 1 3 ) 1 1 5 1 1 1 5 8
Functional chemicals in agriculture and food. Washington DC:
American Chemical Society.
Hasegawa, S., Dillberger, A. M., & Choi, G. Y. (1984). Metabolism of
limonoids: Conversion of nominlin to obacunone in
Corynebacterium fascians. Journal of Agricultural and Food
Chemistry, 32, 457459.
Igual, M., Garca-Martnez, E., Camacho, M. M., & Martnez-
Navarrete, N. (2013). Jam processing and storage effects on b-
carotene and avonoids content in grapefruit. Journal of
Functional Foods. http://dx.doi.org/10/1016/j.jiff:2013.01.019.
Jongaroontaprangsee, S., Tritrong, W., Chokanaporn, W.,
Methacanon, P., Devahastin, S., & Chiewchan, N. (2007). Effects
of drying temperature and particle size on hydration
properties of dietary bre powder from lime and cabbage by-
products. International Journal of Food Properties, 10, 887897.
Jungsakulrujirek, S., & Noomhorm, A. (1998). Effect of harvesting
time and fruit size on titratable acidity, soluble solid and
distribution of limonin in Thai tangerine juice. International
Journal of Food Science and Technology, 33, 367374.
Kanadaswami, C., Lee, L.-T., Lee, P.-P., Hwang, J.-J., Ke, F.-C.,
Huang, Y.-T., et al. (2005). The antitumor activities of
avonoids. In vivo, 19, 895910.
Kawaii, S., Tomono, Y., Katase, E., Ogawa, K., & Yano, N. (1999).
Quantitation of avonoid constituents in citrus fruit. Journal of
Agricultural and Food Chemistry, 47, 35653571.
Kuljarachanan, T., Devahastin, S., & Chiewchan, N. (2009).
Evolution of antioxidant compounds in lime residues during
drying. Food Chemistry, 113, 944949.
Lam, L. K. T., Li, Y., & Hasegawa, S. (1989). Effects of citrus
limonoids on glutathione S-transferase activity in mice.
Journal of Agricultural and Food Chemistry, 37, 878880.
Lee, A. C.-L., Hsiao, W.-C., Wrigh, D. E., Chong, S. Y., Leow, S. K.,
Ho, C.-T., et al. (2013). Induction of GADD45a expression
contributes to the anti-proliferative effects of
polymethoxyavones on colorectal cancer cells. Journal of
Functional Foods. http://dx.doi.org/10.1016/j.jff.2013.01.003.
Li, S., Lo, C.-Y., & Yo, C.-T. (2006). Hydroxylated
polymethoxyavones and methylated avonoids in sweet
orange (Citrus sinensis) peel. Journal of Agricultural and Food
Chemistry, 54, 41764185.
Li, S., Pan, M.-H., Lo, C.-Y., Tan, D., Wang, Y., Shahidi, F., et al.
(2009). Chemistry and health effects of polymethoxyavones
and hydroxylated polymethoxyavones. Journal of Functional
Foods, 1, 212.
Ma, Y.-Q., Ye, X.-Q., Fang, Z.-X., Chen, J.-C., Xu, G.-H., & Liu, D.-H.
(2008). Phenolic compounds and antioxidant activity of
extracts from ultrasonic treatment of Satsuma mandarin
(Citrus unshiu Marc.) peels. Journal of Agricultural and Food
Chemistry, 56, 56825690.
Manners, G. D. (2007). Citrus limonoids: Analysis, bioavailability
and biomedical prospects. Journal of Agricultural and Food
Chemistry, 55, 82858294.
Manthey, J. A., & Grohmann, K. (1996). Concentrations of
hesperidin and other orange peel avonoids in citrus
processing by-products. Journal of Agricultural and Food
Chemistry, 44, 811814.
Manthey, J. A., & Grohmann, K. (2001). Phenols in citrus peels by-
products. Concentration of hydroxycinnamates and
polymethoxylated avones in citrus peel molasses. Journal of
Agricultural and Food Chemistry, 49, 32683273.
McIntosh, C. A., & Mansell, R. L. (1997). Three-dimensional
distribution of limonin, limonoate A-ring monolactone and
naringin in the fruit tissues of three varieties of Citrus paradise.
Journal of Agricultural and Food Chemistry, 45, 28762883.
Meiyanto, E., Hermawan, A., & Anindyajati (2012). Natural
products for cancer-targeted therapy: Citrus avonoinds as
potent chemoprotective agents. Asian Pacic Journal of Cancer
Prevention, 13, 427436.
Naowakul, B., Wirjantoro, T. I., & Phianmongkhol, A. (2013).
Effects of speed and time of wet milling on properties of
dietary bre from pomelos albedo. Food and Applied Bioscience
Journal, 1, 3448.
Nawirska, A., & Kwas niewska, M. (2005). Dietary bre fractions
from fruit and vegetable processing waste. Food Chemistry, 91,
221225.
Nazari, M., Ghorbani, A., Hekmat-Doost, A., Jeddi-Tehrani, M., &
Zand, H. (2011). Inactivation of nuclear factor-jB by citrus
avanone hesperidin contributes to apoptosis and chemo-
sensitizing effect in Romos cells. European Journal of
Phamacology, 650, 526533.
Nogata, Y., Sakamoto, K., Shiratsuchi, H., Ishii, T., Yano, M., &
Ohta, H. (2006). Flavonoid compostion of fruit tissues of
citrus species. Bioscience, Biotechnology and Biochemistry, 70,
178192.
Ohta, H., & Hasegawa, S. (1995). Limonoids in pummelos [Citrus
grandis (L.) Osbeck]. Journal of Food Science, 60, 12841285.
Park, H. J., Kim, M.-J., Ha, E., & Chung, J.-H. (2008). Apoptosis effect
of hesperidin through caspase 3 activation in human colon
cancer cells, SNU-C4. Phytomedicine, 15, 147151.
Peterson, J. J., Dwyer, J. T., Beecher, G. R., Bhagwat, S. A., Gebhardt,
S. E., Haytowitz, D. B., et al. (2006). Flavanones in oranges,
tangerines (mandarins), tangors and tangelos: A compilation
and review of the data from the analytical literature. Journal of
Food Composition and Analysis, 19, S66S73.
Pichaiyongvongdee, S., & Haruenkit, R. (2009a). Comparative
studies of limonin and naringin distribution in different
part of pummelo (Citrus grandis (L.) Osbeck) cultivars
grown in Thailand. Kasetsart Journal (Natural Science), 43,
2836.
Pichaiyongvongdee, S., & Haruenkit, R. (2009b). Investigation of
limonoids, avanones, total polyphenol content and
antioxidant activity in seven Thai pummel cultivars. Kasetsart
Journal (Natural Science), 43, 458466.
Ram, L., & Singh, S. (2006). Medicinal importance of citrus
products and by-products a review. Agricultural Reviews, 27,
170180.
Robards, K., Li, X., Antolovich, M., & Boyd, S. (1997).
Characterisation of citrus by chromatographic analysis of
avonoids. Journal of the Science of Food and Agriculture, 75,
87101.
Roy, A., & Sara, S. (2006). Limonoids: Overview of signicant
bioactive triterpenes distributed in plants kingdom. Biological
and Pharmaceutical Bulletin, 29, 191201.
Seneviranthne, M., Jeon, Y.-J., Ha, J.-H., & Kim, S.-H. (2009).
Effective drying of citrus by-products by high speed drying: A
novel drying technique and their antioxidant activity. Journal of
Food Engineering, 92, 157163.
Stuetz, W., Prapamontol, T., Hongsibsong, S., & Biesalski, H.-K.
(2010). Polymethoxylated avones, avanone glycosides,
carotenoids and antioxidants in different cultivation types of
tangerines (Citrus reticulata Blanco cv. Sainampueng) from
northern Thailand. Journal of Agricultural and Food Chemistry, 58,
60696074.
Sun, C., Chen, K., Chen, K., & Chen, Q. (2005). Contents and
antioxidant capacity of limonin and nomilin in different
tissues of citrus fruit of four cultivars during fruit growth and
maturation. Food Chemistry, 95, 599605.
Sun, Y., Wang, J., Gu, S., Liu, Z., Zhang, Y., & Zhang, X. (2010).
Simultaneous determination of avonoids in different parts of
Citrus reticulata Chachi fruit by high performance liquid
chromatography-photodiode array detection. Molecules, 15,
53785388.
Ubando-Rivera, J., Navarro-Ocan a, A., & Valdivia-Lo pez, M. A.
(2005). Mexican lime peel: Comparative study on contents of
dietary bre and associated antioxidant activity. Food
Chemistry, 89, 5761.
J O U R N A L O F F U N C T I O N A L F O O D S 5 ( 2 0 1 3 ) 1 1 5 1 1 1 5 8 1157
Wang, Y.-C., Chuang, Y. C., & Ku, Y. H. (2007). Quantication of
bioactive compounds in citrus fruits cultivated in Taiwan. Food
Chemistry, 102, 11631171.
Wang, Y.-C., Chuang, Y. C., & Ku, Y. H. (2008). The avonoid,
carotenoid and pectin content in peels of citrus cultivated in
Taiwan. Food Chemistry, 106, 277284.
Xu, G., Ye, X., Chen, J., & Liu, D. (2007). Effect of heat treatment on
the phenolic compounds and antioxidant capacity of citrus
peel extract. Journal of Agricultural and Food Chemistry, 55,
330335.
Zhang, M., Duan, C., Zang, Y., Huang, Z., & Liu, G. (2011). The
avonoid composition of avedo and juice from the pomelo
cultivar (Citrus grandis (L.) Osbeck) and the grapefruit cultivar
(Citrus paradisi) from China. Food Chemistry, 129, 15301536.
1158 J O U R N A L O F F U N C T I O N A L F O O D S 5 ( 2 0 1 3 ) 1 1 5 1 1 1 5 8