Published at : 27 Nov 2020
Volume : IJtech
Vol 11, No 5 (2020)
DOI : https://doi.org/10.14716/ijtech.v11i5.4331
Dianursanti | Bioprocess Engineering Study Program, Department of Chemical Engineering, Faculty of Engineering, Universitas Indonesia, Kampus UI Depok, Depok 16424, Indonesia |
Aisyah Siregar | Bioprocess Engineering Study Program, Department of Chemical Engineering, Faculty of Engineering, Universitas Indonesia, Kampus UI Depok, Depok 16424, Indonesia |
Yoshiaki Maeda | Division of Biotechnology and Life Sciences, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan |
Tomoko Yoshino | Division of Biotechnology and Life Sciences, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan |
Tsuyoshi Tanaka | Division of Biotechnology and Life Sciences, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan |
Research
around using algae as a natural source of carotenoids has been intense in the
21st century, given the wide applications of carotenoids in the pharmaceutical,
health, and food industries. This study aimed to get the highest yield of
carotenoids from Chlorella vulgaris by ultrasound extraction. This study
evaluated two parameters: the extraction solvent (ethanol, acetone, and diethyl
ether were tested) and the solid-to-solvent ratio (1:30, 1:50, and 1:100 were
tested). The carotenoid extracted from C. vulgaris was lutein, and its
compounds were identified by UV-Vis spectroscopy. The highest carotenoid yield
was achieved using ethanol at 1.146±0.082 mg/g and a solid-to-solvent ratio of
1:100 (g/mL). This research shows the use of a specific extraction solvent
along with a solid-to-solvent ratio is significant to determine carotenoids
yield desired. Further study of other parameters (e.g., temperature and
ultrasound intensity) is necessary for the optimum extraction condition
Carotenoids; Chlorella vulgaris; Lutein; Ultrasound extraction; Solvent extraction
Carotenoids are pigments synthesized in plants and
microorganisms to give them a yellow orange to red color. The carotenoids
function alongside chlorophyll to absorb light energy in photosynthesis,
maintain the structure and function of photosynthesis, and anticipate excess
energy (Saini & Keum, 2018). Carotenoids
have antioxidant functions caused by long polyene chains, which have 35-40
carbon atoms (Chandi and Gill, 2011).
Because of their antioxidant properties, carotenoids are used in the
pharmaceutical and health fields to reduce the risk of certain cancers and
cardiovascular diseases, stimulate the immune response, and hamper cataract,
and atherosclerotic development (Alves-Rodriguez and
Shao, 2004). Moreover, carotenoids from Chlorophyta are important
natural food colorants and additives as they have high nutritional values and
are source of proteins, carbohydrates, lipids, and vitamins (Christaki et al., 2015).
Research
around using utilization as a natural source of carotenoids has been very
intensive in the 21st century. Microalgae, especially Chlorella,
are alternative and sustainable sources of various bioactive natural
carotenoids, including ?-carotene, lutein,
Chlorella vulgaris
is used as a source of carotenoid compounds because it has the highest total
content compared to the other Chlorophyta microalgae (Chandi and Gill, 2011). The most-used techniques
to extract carotenoid compounds from C. vulgaris in recent years have
been maceration, sonication, Soxhlet, and pressurized liquid extraction (PLE) (Machmudah and Goto, 2013; Mäki-Arvela et al., 2014; Saini
and Keum, 2018). Ultrasound extraction has been reported to be an
economical method for carotenoid extraction because it requires less energy and
solvent, which damaging carotenoid content because of light, heat, or oxygen (Cha et al., 2010). Increasing the extraction
yield from ultrasound treatment comes from cavitation, which facilitates the
disruption of the cell wall by the ultrasound waves (Mäki-Arvela
et al., 2014). Moreover, the carotenoid yield from C. vulgaris by
ultrasound extraction was higher than from maceration, Soxhlet extraction, and
PLE (Cha et al., 2010).
The objective of this study was to increase the yield of carotenoids by testing two
parameters and to provide a new set of data on the optimum conditions for extracting carotenoids from C. vulgaris. To achieve those results,
three solvents with various polarities (ethanol, acetone, and diethyl ether)
and three solid-to-solvent ratios (1:30, 1:50, and 1:100) were tested. Using a
solvent to extract carotenoids depends on whether carotenoids are polar or
non-polar (Cha et al., 2010; Machmudah and Goto,
2013; Mäki-Arvela et al., 2014; Mulia et al., 2015; Othman et al., 2017).
A previous study also showed that the increase of solid-to-solvent ratio in the
extraction on carotenoids from pumpkin resulted in a significant increase in
carotenoid content (Shahidan et al., 2017).
Thus, the present study is expected to achieve increased yields of carotenoids
by varying the parameters being tested. It is also expected to provide a new
set of data on the optimum conditions for carotenoid extraction from C.
vulgaris. The extract of carotenoids was found to be optimized at a 1:100
solid-to-solvent ratio by using ethanol.
The
results of the present study show that different solvents and different
solid-to-solvent ratios vary the carotenoid yield from C. vulgaris by
ultrasound extraction. It shows that ethanol
is a better extraction solvent than diethyl ether or acetone. The highest carotenoid yield was achieved
using ethanol as the extraction solvent and a solid-to-solvent ratio of 1:100
(g/mL). For further study, we recommend optimizing this extraction method by
testing extraction under other conditions, such as differences in temperature,
duration, ultrasound intensity, and ultrasound frequency.
The
authors would like to thank Tanaka, Arakaki & Yoshino Laboratory,
Biomolecular Engineering & Marine Biotechnology, Department of
Biotechnology and Life Sciences, Tokyo University of Agriculture and
Technology, for their research facilities and JASSO (Japan Student Service
Organization) and PUTI for providing financial aid for this research.
Aflaki, N., 2012. Optimization of
Carotenoid Extraction in Peel and Flesh of Cantaloupe (Cucumis melo L.), with
Ethanol Solvent. Master’s Thesis, Graduate Program, Laval University,
Quebec, Canada
Alves-Rodrigues, A., Shao, A., 2004. The Science
Behind Lutein. Toxicology Letters, Volume 150, pp. 57–83
Araya, B., Gouveia, L.,
Nobre, B., Reis, A., Chamy, R., Poirrier, P., 2014. Evaluation of the
Simultaneous Production of Lutein and Lipids using a Vertical Alveolar Panel
Bioreactor for Three Chlorella Species. Algal Research, Volume 6(B),
pp. 218–222
Bera, D., Das, S., 2013. Mathematical Model
Study on Solvent Extraction of Carotene from Carrot. International Journal
of Research in Engineering and Technology, Volume 2(09), pp. 343–349
Calo, P., Velazquez, J.B.,
Sieiro, C., Blanco, P., Longo, E., Villa, T.G., 1995. Analysis of astaxanthin and
other carotenoids from several Phaffia rhodozyma mutants. Journal of
Agricultural and Food Chemistry, Volume 43(5), pp. 1396–1399.
Cha, K.H., Koo, S.Y., Lee, D.U., 2008. Antiproliferative Effects of
Carotenoids Extracted from Chlorella ellipsoidea and Chlorella
vulgaris on Human Colon Cancer Cells. Journal of Agricultural and Food
Chemistry, Volume 56(22), pp. 10521–10526
Cha, K.H., Lee, H.J., Koo, S.Y., Song,
D.G., Lee, D.U., Pan, C.H., 2010. Optimization of Pressurized Liquid Extraction
of Carotenoids and Chlorophylls from Chlorella vulgaris. Journal of
Agricultural and Food Chemistry, Volume 58(2), pp. 793–797
Chandi, G.K., Gill, B.S., 2011.
Production and Characterization of Microbial Carotenoids as an Alternative to
Synthetic Colors: A Review. International Journal of Food Properties,
Volume 14(3), pp. 503–513
Christaki, E., Bonos, E., Florou-Paneri, P.,
2015. Innovative Microalgae Pigments as Functional Ingredients in Nutrition. In:
Handbook of Marine Microalgae. Christaki et al., Academic Press, Elsevier,
London, pp. 233–243
Danesi, E.D.G., Rangel-Yagui, C.O., Sato, S., de
Carvalho, J.C.M., 2011. Growth and Content of Spirulina platensis Biomass Chlorophyll
Cultivated at Different Values of Light Intensity and Temperature using
Different Nitrogen Sources. Brazilian Journal of Microbiology, Volume
42(1), pp. 362–373
de
Carvalho, L.M.J., Gomes, P.B., de Oliviera Godoy, R.L., Pacheco, S., do Monte,
P.H.F., de Carvalho, J.L.V., Nutti, M.R., Neves, A.C.L., Vieira, A.C.R.A.,
Ramos, S.R.R., 2012. Total Carotenoid Content, A-Carotene and ß-Carotene, of
Landrace Pumpkins (Cucurbita moschata Duch): A Preliminary Study. Food
Research International, Volume 47(2), pp. 337–340
Deenu, A., Naruenartwongsakul, S., Kim,
S.M., 2013. Optimization and Economic Evaluation of Ultrasound Extraction of
Lutein from Chlorella vulgaris. Biotechnology and Bioprocess
Engineering, Volume 18, pp. 1151–1162
Dianursanti., Santoso, A., Delaamira, M., 2016. Utilization of Chlorella
vulgaris to Fixate a High Concentration of Carbon Dioxide in a
Compost-based Medium. International Journal of Technology, Volume 7(1), pp.
168–175
Herrero,
M., Jaime, L., Martin-Alvarez, P.J., Cifuentes, A., Ibanez, E., 2006.
Optimization of the Extraction of Antioxidants from Dunaliella salina
Microalga by Pressurized Liquids. Journal of Agricultural and Food Chemistry, Volume 54(15), pp.
5597–5603
Lebovka, N., Vorobiev, E., Chemat, F., 2011. Enhancing
Extraction Processes in the Food Industry. 1st Edition. New
York, USA: CRC Press
Machmudah, S., Goto, M., 2013. Methods
for Extraction and Analysis of Carotenoids. In: Natural Products. Springer-Verlag Heidelberg,
Berlin, Germany pp. 3367–3411
Mäki-Arvela, P., Hachemi, I., Murzin,
D.Y., 2014. Comparative Study of the Extraction Methods for Recovery of
Carotenoids from Algae: Extraction Kinetics and Effect of Different Extraction
Parameters. Journal of Chemical Technology & Biotechnology, Volume
89(11), pp. 1607–1626
McClure, D.D.,
Nightingale, J.K., Luiz, A., Black, S., Zhu, J., Kavanagh, J.M.,
2019. Pilot-Scale Production of Lutein using Chlorella Vulgaris. Algal
Research, Volume 44, pp. 1–12
Mertz, C., Brat. P., Caris-Veyrat, C.,
Gunata, Z., 2010. Characterization and Thermal Lability of Carotenoids and
Vitamin C of Tamarin Fruit (Solanum betaceum Cav.). Food Chemistry, Volume
119, pp. 653–659
Mulia, K.,
Adam, D., Zahrina, I., Krisanti, E.A., 2018. Green Extraction of Palmitic Acid
from Palm Oil using Betaine-based Natural Deep Eutectic Solvents. International Journal of Technology, Volume 9(2), pp. 335–344
Mulia, K., Krisanti, E., Maulana, T.,
Dianursanti., 2015. Selective Polarity-guided Extraction and Purification of
Acetogenins in Annona muricata L. Leaves. International Journal of
Technology, Volume 6(7), pp. 1221–1227
Nguyen, T.D.P., Nguyen, D.H., Lim, J.W., Chang, C-K., Leong, H.Y., Tran,
T.N.T., Vu, T.B.H., Nguyen, T.T.C, Show, P.L., 2019. Investigation of the
Relationship between Bacteria Growth and Lipid Production Cultivating of
Microalgae Chlorella vulgaris in Seafood Wastewater. Energies,
Volume 12(12), pp. 1–12
Othman, R., Noh, N., Nurrulhidayah,
A.F., Anis Hamizah, H., Jamaludin, M.A., 2017. Determination of Natural Carotenoids Pigments from Fresh Water Green
Algae as Potential Halal Food Colorants. International Food Research
Journal, Volume 24 (Suppl), pp. S468–S471
Poojary, M.M., Barba, F.J.,
Aliakbarian, B., Donsi, F., Pataro, G., Dias D.A., Juliano, P., 2016.
Innovative Alternative Technologies to Extract Carotenoids from Microalgae and
Seaweeds. Marine Drugs, Volume 14(11), pp. 1–34
Reichardt, C., 2003. Solvents and
Solvent Effects in Organic Chemistry. 4th Edition. Germany: Wiley-VCH
Publishers
Rivera,
S., Canela, R., 2012. Influence of Sample Processing on the Analysis of
Carotenoid in Maize. Molecules, Volume 17, pp. 11255–11268
Rodriguez-Amaya, D.B., Kimura, M., 2004. HarvestPlus
Handbook for Carotenoid Analysis. 1st Edition. Washington DC,
USA: HarvestPlus
Saini, R.K., Keum, Y.S., 2018. Carotenoid extraction methods: A review
of recent developments. Food Chemistry,
Volume 240, pp. 90–103.
Shahidan, N., Koy, C.N., Rashidi, O., Ho,
L.H., Azrina, I., Nurul Zaizuliana, R.A., N.Z., Zarinah, Z., 2017. The Effect
of Time, Temperature and Solid-to-solvent Ratio on Pumpkin Carotenoids
Extracted using Food Grade Solvents. Sains Malaysiana, Volume 46(02),
pp. 231–237
Sharma, A.K., Sahoo, P.K., Singhal, S., Patel, A.,
2016. Impact
of Various Media and Organic Carbon Sources on Biofuel Production Potential
from Chlorella spp. Biotech, Volume 6(2),
pp. 1–12
Shi,
J., Mazza, G., Maguer, M.L., 2002. Functional Foods: Biochemical and
Processing Aspects. 1st Edition. New York, USA: CRC Press
Tan,
P.W., Tan, C.P., Ho, C.W., 2011. Antioxidant Properties: Effects of
Solid-to-solvent Ratio on Antioxidant Compounds and Capacities of pegaga (Centella
asiatica). International Food Research Journal, Volume 18(2), pp.
557–562
Warkoyo,
W., Saati, E.A., 2011. The Solvent Effectiveness on Extraction Process of
Seaweed Pigment. Makara Journal of Technology, Volume 15(1), pp.
5–8