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Article history:
Received 15 December 2010
Received in revised form 12 July 2011
Accepted 13 August 2011
Available online 26 August 2011
Keywords:
Lycopene
Tomato peel by-product
Tomato seed
Supercritical CO2 extraction
a b s t r a c t
This work discusses the extraction of lycopene from tomato peel by-product containing tomato seed
using supercritical carbon dioxide. The presence of tomato seed in the peel by-product improved the
yield of extracted lycopene. Extraction was carried out at temperatures of 7090 C, pressures of
2040 MPa, a particle size of 1.05 0.10 mm and ow rates of 24 mL/min of CO2 for 180 min extraction
time. Oil from tomato seed was extracted under similar operating conditions and analyzed using GCMS
and GCFID, while carotenoids extracted were analyzed by HPLC. The optimum operating condition to
extract lycopene, under which 56% of lycopene was extracted, was found to be 90 C, 40 MPa, and a ratio
of tomato peel to seed of 37/63. The presence of tomato seed oil helped to improve the recovery of lycopene from 18% to 56%. The concentration of lycopene in supercritical carbon dioxide as a function of density at various temperatures was determined.
2011 Elsevier Ltd. All rights reserved.
1. Introduction
The tomato is one of the most widely cultivated vegetable crops
and is used globally for human nutrition. Millions of tons of tomatoes are processed yearly to produce products such as ketchup and
sauce, resulting in large amounts of tomato peel, pulp, and seed as
industrial waste, an amount almost equivalent to 40% of the raw
material. Concentrations of lycopene in tomato vary from 30 to
200 mg/kg in fresh fruit and from 430 to 2950 mg/kg on a dry basis
(Vasapollo et al., 2004). Lycopene represents more than 85% of the
total carotenoids; tomato peel contains about ve times more lycopene than tomato pulp (Lazos and Kalathenos, 1988; Sharma and
Le Maguer, 1996).
Carotenoids are important compounds in the human diet as anticarcinogens, cardiovascular disease preventers, and immune system
regulators. Moreover, they can act as antioxidants and precursors of
vitamin A (Singh and Goyal, 2008). Due to their association with human health, carotenoids have also been used in medicines and cosmetics. One of the most important carotenoids is lycopene (C40H56)
an open chain hydrocarbon with 11 conjugated double bonds. It is a
Corresponding author. Address: Bioelectrics Research Center, Kumamoto University, Kurokami 2-39-1, Kumamoto shi, Kumamoto 860-8555, Japan. Tel.: +81 96
342 3664; fax: +81 96 342 3665.
E-mail addresses: sitimach@yahoo.com (S. Machmudah), mgoto@kumamoto-u.ac.jp
(M. Goto).
0260-8774/$ - see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jfoodeng.2011.08.012
291
Jasco, Japan); collection vials; and a wet gas meter (Sinagawa Co.,
Japan).
Initially, 4 g of material was placed in the extractor. Glass beads
were placed in the bottom and top of the extractor vessel to avoid
channeling during the extraction. Liquid CO2 from a cylinder tank
with a siphon was passed through a chiller kept at 5 C and compressed to the operating pressure by a high pressure pump.
Compressed CO2 was owed continuously into the extractor placed
in the heating bath and maintained at the operating pressure. The
exit uid from the extractor was expanded to the atmospheric
pressure by the back pressure regulator. The extract was collected
in vials every 5 min for the rst 30 min and then every 30 min for
180 min of extraction, weighed immediately after the collection,
and refrigerated at 20 C prior to analysis. To minimize decomposition and oxidation of extracted compounds, all samples were protected from light and ambient oxygen with aluminum foil.
The extractions were carried out at temperatures ranging from
70 to 90 C, pressures of 2040 MPa, CO2 ow rates of 24 mL/min,
and various ratios of peel, seeds and particle sizes. To investigate
the composition of tomato seed oil, oil from tomato seeds was extracted under identical operating conditions. Each experiment was
conducted in triplicate and cited as mean standard deviation.
2.3. Chemical analysis
Analysis for tomato seed oil was performed using a gas chromatography-ame ionization detector (GC-FID). Tomato oil consists of
polyunsaturated fatty acids (PUFAs) that must be analyzed by high
temperature gas chromatography. Before injection, the extracted
oil was converted to methyl esters according to the method of
Roy et al. (1996). Two ml of hexane was added to 50 mg of extracted oil. Then, 0.2 ml of 2 M KOH solution in methanol was
added to the sample and shaken at 100 C for 10 min. The sample
was centrifuged at 1500 rpm (309.5 G-force) for 15 min. Methyl esters dissolved in the hexane layer were separated by a funnel and
analyzed by GC-FID (Gas Chromatography GC-14A, Shimadzu, Japan) equipped with a DB-5 capillary column (15 m 0.250 mm
0.25 lm). The oven temperature was increased from 70 to
320 C at 5 C/min intervals. The injector and detector temperatures were controlled at 250 and 300 C, respectively. The injection
volume was 0.6 ll. All samples were injected in duplicate. The peak
area percentage of each PUFA in the extract at each extraction condition is cited as mean standard deviation.
Carotenoids in the extract were analyzed by high performance-liquid chromatography (HPLC) equipped with STR-ODS-II column
(5 lm; 4.6 mm 250 mm) (Nacalai Tesque, Inc., Japan) and UVvisible detector (UV-970, Jasco, Japan) as reported by Machmudah et al.
(2008a). A mixture of 2-propanol:acetonitrile:methanol:water in
52:39:5:4 v/v ratio was used as a mobile phase t a ow rate of
1.2 mL/min. The sample dissolved in the solvent mixture, consisting
of 1:2 chloroform:acetone, was injected in 20 ll units. The column
temperature was controlled at 30 C by a column heater (Sugai U620, Japan) and detection was measured at 450 nm. Detector signals
were recorded using BORWIN Chromatography software. Peaks of
lycopene and b-carotene were identied by comparing the retention
time in the extract with standard compounds. The content of lycopene and b-carotene in the extract were estimated by comparing
the peak areas with their respective standards. Calibration curves
of the standards were prepared by plotting standard concentrations
(0.254 mg/mL) versus their peak areas detected at 5.7 and 8.9 min
for lycopene and b-carotene, respectively. All samples were analyzed in duplicate. The recovery of lycopene and b-carotene in the
extract at each extraction condition is cited as mean standard
deviation.
Recovery of lycopene and b-carotene extracted was calculated
based on the maximum lycopene and b-carotene extracted with
100
292
80
60
40
20
0
0
30
90
120
150
180
Time (min)
60
Fig. 1. Effect of tomato seed oil as co-solvent on the recovery of lycopene and bcarotene at 90 C, 40 MPa, and 3 ml/min.
293
100
80
100
P = 20 MPa (-carotene)
P = 30 MPa (-carotene)
P = 40 MPa (-carotene)
P = 20 MPa (Lycopene)
P = 30 MPa (Lycopene)
P = 40 MPa (Lycopene)
60
40
20
80
60
40
20
T = 70 C (Lycopene)
o
T = 80 C (Lycopene)
o
T = 90 C (Lycopene)
30
60
90
120
150
180
30
60
Time (min)
100
P = 20 MPa
P = 30 MPa
P = 40 MPa
60
40
20
0
0
30
60
90
120
150
180
120
150
180
(a)
100
80
90
Time (min)
(a)
T = 70 C (-carotene)
o
T = 80 C (-carotene)
o
T = 90 C (-carotene)
120
150
180
T = 70 C
o
T = 80 C
o
T = 90 C
80
60
40
20
0
0
30
60
Time (min)
(b)
Fig. 2. Effect of pressure on the recovery of lycopene and b-carotene (a) and tomato
seed oil (b) at 90 C, particle size of 1.05 0.10 mm and 3 ml/min.
90
Time (min)
(b)
Fig. 3. Effect of temperature on the recovery of lycopene and b-carotene (a) and
tomato seed oil (b) at 40 MPa, particle size of 1.05 0.10 mm and 3 ml/min.
remove the extracts left in the tubing line and back pressure regulator; unfortunately, may also contaminate the product. Rinsing
product of carotenoids can, though, be used for analytical purposes
(Britton, 1985 and Meyers and Bligh, 1981). Food grade ethanol
solvent may be substituted for THF as the rinsing solvent because
of its non-toxic effect, which is ideal for food products (Kassama
et al., 2008). Moreover, ethanol as modier in supercritical CO2
extraction could improve the solubility of carotenoids that resulted
in 93% recovery of lycopene (Huang et al., 2008). In comparison
with previous work, extraction with tomato seed without rinsing
had a higher efciency than that of Topal et al. (2006), indicating
that even without the use of THF as a rinsing method, yield of
carotenoid can be improved by the addition of tomato seed oil in
the supercritical CO2 extraction. In addition, seed oil may also be
used to replace the toxic organic solvent used to remove carotenoids remaining in the extraction system.
3.5. Effects of tomato peel to seed ratio
100
Peel/Seed=25/75 (Lycopene)
Peel/Seed=37/63 (Lycopene)
Peel/Seed=50/50 (Lycopene)
Peel/Seed=25/75 (-carotene)
Peel/Seed=37/63 (-carotene)
Peel/Seed=50/50 (-carotene)
80
294
60
40
20
0
0
30
60
90
120
150
180
Time (min)
Fig. 5. Effect of tomato peel and seed ratio on the recovery of lycopene and bcarotene at 90 C, 40 MPa, particle size of 1.05 0.10 mm and 3 ml/min..
The effects of tomato peel to seed ratio on the lycopene and bcarotene recovery are shown in Fig. 5, which show recovery of
lycopene and b-carotene increased with increasing seed ratio from
50 to 63. Increasing the amount of tomato seed led to an increase
both in oil produced in the extractor as well as, in solvent power
(Brunner, 1994; Sovova et al., 2001; Vasapollo et al., 2004). As a result, recovery signicantly increased with an increased amount of
tomato seed. On the other hand, increasing the seed to peel ratio
from 63 to 75 signicantly decreased the recovery. This could be
due to higher extraction of seed oil which may have hindered
supercritical CO2 transport to penetrate into the solid matrix.
Moreover, the effect of vegetable oil as an entrainer is limited
due to its low solubility in CO2 (Sovova et al., 2001). Furthermore,
the recovery was signicantly decreased.
3.6. Effects of particle size
Raw material pretreatment is important in order to signicantly
increase the yield of lycopene extraction (Baysal et al., 2000;
Vasapollo et al., 2004; Ciurlia et al., 2009). For example, drying and
grinding processes inuence the physical properties and quality of
raw material. The effect of particle size (ungrounded,
1.59 0.12 mm and 1.05 0.01 mm) on the recovery of lycopene
and b-carotene is shown in Fig. 6. As seen, the recovery of both lycopene and b-carotene signicantly increased with decreasing particle
size. Decreasing the particle size decreases mass transfer resistance
ln C k ln q
a
T
80
100
100
60
40
20
Unground (-carotene)
1.59 mm (-carotene)
1.05 mm (-carotene)
Unground (Lycopene)
1.59 mm (Lycopene)
1.05 mm (Lycopene)
80
60
40
20
0
0
30
60
90
120
150
0
0
30
60
90
120
150
180
Time (min)
Fig. 4. Effect of rinsing method on the recovery of lycopene and b-carotene at 90 C,
40 MPa, particle size of 1.05 0.10 mm and 3 ml/min..
molecules associating with one molecule of solute to form the solvate complex, a and b are constants taking into account the total
heat of the reaction and the molecular weight of the species. Using
different temperature values, it is possible to obtain the corresponding density values and plotting ln (C) versus ln (q) at different
temperatures a straight line is obtained whose slope is equal to k.
Fig. 7 shows the concentration of lycopene in the supercritical
phase as a function of the solvent density at various temperatures.
Once the value of k is known, it is possible to calculate the constants
a and b and the following Chrastil equation can be obtained:
C q10:21 e
calc
n exp
1X
C C
AAD %
100
n i1 C exp
0.10
o
T = 70 C
o
T = 80 C
o
T = 90 C
T = 70 C (Chrastil)
o
T = 80 C (Chrastil)
o
T = 90 C (Chrastil)
C (g/L)
0.06
0.04
0.02
0.00
500
550
4. Conclusions
500057:34
T
0.08
295
600
650
700
750
800
850
900
(g/L)
CO2
296
Machmudah, S., Sulaswatty, A., Sasaki, M., Goto, M., Hirose, T., 2006b. Supercritical
CO2 extraction of nutmeg oil: Experiments and modeling. Journal of
Supercritical Fluids 39 (1), 3039.
Machmudah, S., Kawahito, Y., Sasaki, M., Goto, M., 2007. Supercritical CO2
extraction of rosehip seed oil: Fatty acids composition and process
optimization. Journal of Supercritical Fluids 41 (3), 421428.
Machmudah, S., Kawahito, Y., Sasaki, M., Goto, M., 2008a. Process optimization and
extraction rate analysis of carotenoids extraction from rosehip fruit using
supercritical CO2. Journal of Supercritical Fluids 44 (3), 308314.
Machmudah, S., Kondo, M., Sasaki, M., Goto, M., Munemasa, J., Yamagata, M., 2008b.
Pressure effect in supercritical CO2 extraction of plant seeds. Journal of
Supercritical Fluids 44 (3), 301307.
Meyers, S.P., Bligh, D., 1981. Characterization of astaxanthin pigment from heat
processed crawsh waste. Journal of Agricultural and Food Chemistry 29 (3),
505508.
NIST Chemistry WebBook, NIST Standard Reference Database Number 69, National
Institute of Standards and Technology. <http://webbook.nist.gov/chemistry>.
Roy, B.C., Goto, M., Hirose, T., 1996. Temperature and pressure effects on
supercritical CO2 extraction of tomato seed oil. International Journal of Food
Science and Technology 31 (2), 137141.
Rozzi, N.L., Singh, R.K., Vierling, R.A., Watkins, B.A., 2002. Supercritical uid
extraction of lycopene from tomato processing byproducts. Journal of
Agricultural and. Food Chemistry 50 (9), 26382643.
Sabio, E., Lozano, M., Montero de Espinosa, V., Mendes, R.L., Pereira, A.P., Palavra,
A.F., Coelho, J.A., 2003. Lycopene and b-carotene extraction from tomato
processing waste using supercritical CO2. Industrial and Engineering Chemistry
Research 42 (25), 66416646.
Saldana, M.D.A., Temelli, F., Guigard, S.E., Tomberli, B., Gray, C.G., 2010. Apparent
solubility of lycopene and b-carotene in supercritical CO2, CO2 + ethanol and
CO2 + canola oil using dynamic extraction of tomatoes. Journal of Food
Engineering 99 (1), 18.
Sharma, S.K., Le Maguer, M., 1996. Lycopene in tomatoes and tomato pulp fractions.
Italian Journal of Food Science 8 (2), 107113.
Shi, J., Yi, C., Xue, S.J., Jiang, Y., Ma, Y., Li, D., 2009a. Effects of modiers on the prole
of lycopene extracted from tomato skins by supercritical CO2. Journal of Food
Engineering 93 (4), 431436.
Shi, J., Khatri, M., Xue, S.J., Mittal, G.S., Ma, Y., Li, D., 2009b. Solubility of lycopene in
supercritical CO2 uid as affected by temperature and pressure. Separation and
Purication Technology 66 (2), 322328.
Singh, P., Goyal, G.K., 2008. Dietary lycopene: its properties and anticarcinogenic
effects. Comprehensive Reviews in Food Science and Food Safety 7 (3), 255270.
Sovova, H., Stateva, R.P., Galushko, A.A., 2001. Solubility of b-carotene in
supercritical CO2 and the effect of entrainers. Journal of Supercritical Fluids
21 (3), 195203.
Tonthubthimthong, P., Chuaprasert, S., Douglas, P., Luewisutthichat, W., 2001.
Supercritical CO2 extraction of nimbin from neem seeds- an experimental
study. Journal of Food Engineering 47 (4), 289293.
Topal, U., Sasaki, M., Goto, M., Hayakawa, K., 2006. Extraction of lycopene from
tomato skin with supercritical carbon dioxide: effect of operating conditions
and solubility analysis. Journal of Agricultural and Food Chemistry 54 (15),
56045610.
Vagi, E., Simandi, B., Vasarhelyine, K.P., Daood, H., Kery, A., Doleschall, F., Nagy, B.,
2007. Supercritical carbon dioxide extraction of carotenoids, tocopherols and
sitosterols from industrial tomato by-products. Journal of Supercritical Fluids
40 (2), 218226.
Vasapollo, G., Longo, L., Rescio, L., Ciurlia, L., 2004. Innovative supercritical CO2
extraction of lycopene from tomato in the presence of vegetable oil as cosolvent. The Journal of Supercritical Fluids 29 (12), 8796.
Wu, J., Hu, G., Wang, X., Li, D., Yu, H., Han, X., 2010. The reproductive toxicity of
organic compounds extracted from drinking water sources on Sprague dawley
rats: An in vitro study. Environmental Toxicology 25 (3), 284293.