Sains Malaysiana 49(4)(2020): 765-774

http://dx.doi.org/10.17576/jsm-2020-4904-05

Characterization on Phenolic Acids and Antioxidant Activity of Microalgae Chlorella sp. using Subcritical Water Extraction

(Pencirian ke atas Asid Fenol dan Aktiviti Antioksidan Mikroalga Chlorella sp. menggunakan Pengekstrakan Air Subkritikal)

SITI MAISURAH ZAKARIA¹, SITI MAZLINA MUSTAPA KAMAL^1*, RAZIF HARUN², ROZITA OMAR² & SHAMSUL IZHAR SIAJAM²

¹Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

²Department of Chemical and Environmental Engineering, Faculty of Engineering, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

Received: 27 June 2019/Accepted: 30 December 2019

ABSTRACT

Chlorella sp. microalgae contain phenolic compounds with good functional properties. In this study, two extraction methods, soxhlet and subcritical water extraction (SWE) were applied and compared in terms of phenolic compounds recovery from Chlorella sp. microalgae and characterization of the phenolic acid components. Phenolic acid analysis demonstrated that the main components of the Chlorella sp. extracts were ferulic, caffeic and p-coumaric acids. The comparative study indicated that SWE gave higher extraction yield compared to conventional soxhlet method. High recoveries of phenolic acids were obtained at 175 °C with 3.20, 3.05 and 3.33 mg/100 g of ferulic, p-coumaric and caffeic acid, respectively, compared to soxhlet extraction using methanol with 2.10, 2.29 and 2.37 mg/100 g of ferulic, p-coumaric and caffeic acid, respectively. This proved that subcritical water treatment could effectively be used for the release of phenolic acids from Chlorella sp. using safe and green solvent. Analysis by Fourier transform infrared spectroscopy (FTIR) was also performed to observe the effect of subcritical water on the functional groups of the extracts. The current study demonstrated that SWE provided a better way of utilising Chlorella sp. as a source of phenolic acids and natural antioxidants.

Keywords: Extraction; microalgae; phenolic; subcritical; water

ABSTRAK

Mikroalga Chlorella sp. mengandungi sebatian fenol dengan ciri pelbagai fungsi yang bagus. Dalam kajian ini, dua kaedah pengekstrakan, soxhlet dan pengekstrakan air subkritikal (SWE) telah diaplikasikan dan dibandingkan daripada segi hasil sebatian fenol daripada mikroalgaChlorella sp. serta pencirian ke atas komponen asid fenol. Analisis asid fenolik menunjukkan bahawa komponen utama bagi ekstrakChlorella sp. adalah asid ferulik, kafeik dan ρ-kumarik. Kajian perbandingan menunjukkan SWE memberi hasil ekstrak yang lebih tinggi berbanding kaedah konvensional soxhlet. Hasil asid fenolik yang tinggi dicapai pada 175 °C dengan 3.20, 3.05 dan 3.33 mg/100 g masing-masing bagi asid ferulik, ρ-kumarik dan kafeik berbanding pengekstrakan soxhlet menggunakan metanol dengan 2.10, 2.29 dan 2.37 mg/100 g masing-masing bagi asid ferulik, ρ-kumarik dan kafeik. Ini membuktikan bahawa rawatan air subkritikal boleh digunakan secara berkesan bagi pengeluaran asid fenol daripada Chlorella sp. menggunakan pelarut yang selamat dan hijau. Analisis dengan spektroskopi transformasi Fourier inframerah (FTIR) juga dijalankan untuk melihat kesan air subkritikal ke atas kumpulan berfungsi bagi ekstrak. Kajian menunjukkan bahawa SWE adalah kaedah yang lebih baik bagi penggunaan Chlorella sp. sebagai sumber asid fenol dan antioksidan semula jadi.

Kata kunci: Air; fenol; mikroalga; pengekstrakan; subkritikal

REFERENCES

Blois, M.S. 1958. Antioxidant determinations by the use of a stable free radical. Nature 181(4617): 1199-1200.

Cao, W., Chen, W., Sun, S., Guo, P., Song, J. & Tian, C. 2007. Investigating the antioxidant mechanism of violacein by density functional theory method. Journal of Molecular Structure: Theochem 817(3): 1-4.

Cheng, Y., Xu, Q., Liu, J., Zhao, C., Xue, F. & Zhao, Y. 2014. Decomposition of five phenolic compounds in high temperature water. Journal of the Brazilian Chemical Society 25(11): 2102-2107.

Dey, S. & Rathod, V.K. 2013. Ultrasound assisted extraction of β-carotene from Spirulina platensis. Ultrasonics Sonochemistry 20(1): 271-276.

Goiris, K., Muylaert, K., Voorspoels, S., Noten, B., De Paepe, D., E Baart, G.J. & De Cooman, L. 2014. Detection of flavonoids in microalgae from different evolutionary lineages. Journal of Phycology 50(3): 483-492.

Hajimahmoodi, M., Faramarzi, M.A., Mohammadi, N., Soltani, N., Oveisi, M.R. & Nafissi-Varcheh, N. 2010. Evaluation of antioxidant properties and total phenolic contents of some strains of microalgae. Journal of Applied Phycology 22(1): 43-50.

Hartwig, V.G., Brumovsky, L.A., Fretes, R.M. & Boado, L.S. 2012. A novel procedure to measure the antioxidant capacity of yerba maté extracts. Food Science and Technology (Campinas) 32(1): 126-133.

Hasmida, M., Nur Syukriah, A., Liza, M. & Mohd Azizi, C. 2014. Effect of different extraction techniques on total phenolic content and antioxidant activity of Quercus infectoria galls. International Food Research Journal 21(3): 1075-1079.

He, L., Zhang, X., Xu, H., Xu, C., Yuan, F., Knez, Ž., Novak, Z. & Gao, Y. 2012. Subcritical water extraction of phenolic compounds from pomegranate (Punica granatum L.) seed residues and investigation into their antioxidant activities with HPLC–ABTS+ assay. Food and Bioproducts Processing 90(2): 215-223.

Herrero, M., Cifuentes, A. & Ibanez, E. 2006. Sub-and supercritical fluid extraction of functional ingredients from different natural sources: Plants, food-by-products, algae and microalgae: A review. Food Chemistry 98(1): 136-148.

Huie, C. 2002. A review of modern sample-preparation techniques for the extraction and analysis of medicinal plants. Analytical and Bioanalytical Chemistry 373(2): 23-30.

Ina, A. & Kamei, Y. 2006. Vitamin B 12, a chlorophyll-related analog to pheophytin a from marine brown algae, promotes neurite outgrowth and stimulates differentiation in PC12 cells. Cytotechnology 52(3): 181-187.

Joana Gil-Chávez, G., Villa, J.A., Fernando Ayala-Zavala, J., Basilio Heredia, J., Sepulveda, D., Yahia, E.M. & González-Aguilar, G.A. 2013. Technologies for extraction and production of bioactive compounds to be used as nutraceuticals and food ingredients: An overview. Comprehensive Reviews in Food Science and Food Safety 12(1): 5-23.

Kashif, A., Muhammad, S.H. & Joong-Ho, K. 2017. Green extraction methods for polyphenols from plant matrices and their byproducts: A review. Comprehensive Reviews in Food Science and Food Safety 16(2): 295-315.

Khan, M.I., Shin, J.H. & Kim, J.D. 2018. The promising future of microalgae: Current status, challenges, and optimization of a sustainable and renewable industry for biofuels, feed, and other products. Microbial Cell Factories 17(1): 36.

Klejdus, B., Kopecký, J., Benešová, L. & Vacek, J. 2009. Solid-phase/supercritical-fluid extraction for liquid chromatography of phenolic compounds in freshwater microalgae and selected cyanobacterial species. Journal of Chromatography A 1216(5): 763-771.

Knekt, P., Kumpulainen, J., Järvinen, R., Rissanen, H., Heliövaara, M., Reunanen, A., Hakulinen, T. & Aromaa, A. 2002. Flavonoid intake and risk of chronic diseases. The American Journal of Clinical Nutrition 76(3): 560-568.

Liang, S., Liu, X., Chen, F. & Chen, Z. 2004. Current microalgal health food R & D activities in China. In Asian Pacific Phycology in the 21st Century: Prospects and Challenges. Netherlands: Springer.

Liu, J., Lu, J.F., Kan, J., Tang, Y.Q. & Jin, C.H. 2013. Preparation, characterization and antioxidant activity of phenolic acids grafted carboxymethyl chitosan. International Journal of Biological Macromolecules 62(1): 85-93.

López, A., Rico, M., Rivero, A. & Suárez de Tangil, M. 2011. The effects of solvents on the phenolic contents and antioxidant activity of Stypocaulon scoparium algae extracts. Food Chemistry 125(3): 1104-1109.

Onofrejová, L., Vašíčková, J., Klejdus, B., Stratil, P., Mišurcová, L., Kráčmar, S., Kopecký, J. & Vacek, J. 2010. Bioactive phenols in algae: The application of pressurized-liquid and solid-phase extraction techniques. Journal of Pharmaceutical and Biomedical Analysis 51(2): 464-470.

Rice-Evans, C., Miller, N. & Paganga, G. 1997. Antioxidant properties of phenolic compounds. Trends in Plant Science 2(4): 152-159.

Rodríguez-Meizoso, I., Jaime, L., Santoyo, S., Señoráns, F., Cifuentes, A. & Ibáñez, E. 2010. Subcritical water extraction and characterization of bioactive compounds from Haematococcus pluvialis microalga. Journal of Pharmaceutical and Biomedical Analysis 51(2): 456-463.

Safafar, H., van Wagenen, J., Møller, P. & Jacobsen, C. 2015. Carotenoids, phenolic compounds and tocopherols contribute to the antioxidative properties of some microalgae species grown on industrial wastewater. Marine Drugs 13(12): 7069.

Scherer, R. & Godoy, H. 2014. Effects of extraction methods of phenolic compounds from Xanthium strumarium L. and their antioxidant activity. Revista Brasileira de Plantas Medicinais 16(1): 41-46.

Shian, T.E. & Abdullah, A. 2012. Antioxidant properties of three banana cultivars (Musa acuminata 'Berangan', 'Mas' and 'Raja') extracts. Sains Malaysiana 41(3): 319-324.

Uematsu, M. & Franck, E. 1980. Static dielectric constant of water and steam. Journal of Physical and Chemical Reference Data 9(4): 1291-1306.

Wan Mahmood, W.M.A., Lorwirachsutee, A., Theodoropoulos, C. & Gonzalez-Miquel, M. 2019. Polyol-based deep eutectic solvents for extraction of natural polyphenolic antioxidants from Chlorella vulgaris. ACS Sustainable Chemistry & Engineering 7(5): 5018-5026.

Wettasinghe, M. & Shahidi, F. 1999. Evening primrose meal:  A source of natural antioxidants and scavenger of hydrogen peroxide and oxygen-derived free radicals. Journal of Agricultural and Food Chemistry 47(5): 1801-1812.

Zakaria, S.M. & Kamal, S.M.M. 2015. Subcritical water extraction of bioactive compounds from plants and algae: Applications in pharmaceutical and food ingredients. Food Engineering Reviews 8(1): 23-34.

Zakaria, S.M., Kamal, S.M.M., Harun, M.R., Omar, R. & Siajam, S.I. 2017. Subcritical water technology for extraction of phenolic compounds from Chlorella sp. microalgae and assessment on its antioxidant activity. Molecules 22(7): 1105.

*Corresponding author; email: smazlina@upm.edu.my