|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|
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, astaxanthin, zeaxanthin, violaxanthin, and fucoxanthin (Cha et al., 2008; Machmudah and Goto, 2013). Carotenoids produced by microalgae are commonly found in chloroplasts and bound to membranes and other macromolecules in the intracellular system. The cell wall and membrane surrounding the cell become barriers that limit the rate of mass transfer of the carotenoids during extraction, so carotenoid extraction requires disruption of cell walls, plasma membranes, and chloroplasts (Poojary et al., 2016). Carotenoids are easily damaged and degraded by exposure to light, heat, and oxygen. Therefore, extraction methods must be considered that reduce or eliminate these harmful effects (Mertz et al., 2010; Aflaki, 2012; Machmudah and Goto, 2013; Mäki-Arvela et al., 2014; Mulia et al., 2018; Saini and Keum, 2018).
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.
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