Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter September 23, 2020

Evaluation of Moroccan microalgae: Spirulina platensis as a potential source of natural antioxidants

  • Touria Ould Bellahcen EMAIL logo , Abderrahmane AAmiri , Ikram Touam , Fouzia Hmimid , Abdelaziz El Amrani , Abdelmjid Cherif and Mounia Cherki

Abstract

Background

Ethanolic, aqueous and lipidic extracts of Spirulina platensis were evaluated for their bioactive substances (polyphenols and fatty acids) and antioxidant activities using two different assays diphenyl-1-picrylhydrazyl (DPPH) and azino-bis (ethylbenzthiazoline-6- sulfonicacid (ABTS)).

Methods

The phenolic compounds amounts were determined by colorimetric assays and were analyzed by high-performance and liquid chromatography (HPLC) method. The identification of phenolic compounds in the extract was accomplished by comparison of their retention times with those of pure standards. The chemical composition of fatty acids in lipidic extract was determined by GC/FIDs and GC/MS method.

Results

The obtained results revealed that using DPPH, the ethanolic extract recorded the highest activity with (IC50=449 μg/mL ± 83). It was characterized with a high levels of phenolic and flavonoid content 0.33 ± 0.01 mg GAE/g dw and 0.21 ± 0.01 mg quercetin/g dw, respectively. The ethanolic extract showed the presence of caffeic acid, syringic acid, ferulic acid, p-coumaric acid, chlorogenic acid, kaempferol, quercetin and apigenin. Those entire phenolic compounds seem participle synergistically to the pronounced higher activity. To the best of our knowledge, the apigenin (4′, 5, 7,-trihydroxyflavone), a natural compound which is famously known by its potent antiradical activity, was identified for the first time in Moroccan S. platensis only in the ethanolic extract. A high positive correlation (r=0.895) between the antioxidant activity and the chemical composition of ethanolic extract was observed. For ABTS, lipidic extract showed the highest activity with (IC50=740 μg/mL ± 12). A total of 15 fatty acids compounds, amounting 89.73% of the extract were identified. γ-linolenic acid (GLA) (39.02%) and α-linolenic acid (ALA) (13.85%) were the major component and contribute greatly to the antioxidant activity observed. The lipidic extract has a high content of unsaturated fatty acids (70.95%) which are often exhibited very high antioxidant activity since they have two or more double bonds and chain lengths. The aqueous extract either with DPPH or ABTS methods, recorded low antioxidant activity. Its correlation was r=0.499, lower than found in the ethanolic extract.

Conclusions

These results showed that Spirulina platensis could be considered as a valuable source of fatty acids and phenolics with potent antioxidant activity. It may be employed in the manufacture of pharmaceutical drugs as an alternative source of natural antioxidants.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: On behalf all coauthors, I confirm that we not have conflict of interest with any reviewers or any financial and personal relationships with other people or organizations that could inappropriately influence our work.

References

[1] Blagojević D, Babić O, Rašeta M, Šibul F, Janjušević L, Simeunović J. Antioxidant activity and phenolic profile in filamentous cyanobacteria: the impact of nitrogen. J Appl Phycol 2018;30:2337–46. DOI:10.1007/s10811-018-1476-4.Search in Google Scholar

[2] Valko M, Rhodes CJ, Moncol J, Izakovic M, Mazur M. Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chem Biol Interact 2006;160:1–40. DOI:10.1016/j.cbi.2005.12.009.Search in Google Scholar PubMed

[3] Kancheva V, Kasaikina O. Bio-antioxidants. A chemical base of their antioxidant activity and beneficial effect on human health. Curr Med Chem 2013;20:4784–805. DOI:10.2174/09298673113209990161.Search in Google Scholar PubMed

[4] Goh SH, Yusoff FM, Loh SP. A comparison of the antioxidant Properties and total phenolic content in a Diatom, Chaetoceros sp and a Green Microalga, Nannochloropsis sp. J Agr Sci 2010;2:123–30. DOI:10.5539/jas.v2n3p123.Search in Google Scholar

[5] Goiris K, Muylaert K, Fraeye I, Foubert I, De Brabanter J, De Cooman L. Antioxidant potential of microalgae in relation to their phenolic and carotenoid content. J Appl Phycol 2012;24:1477–86. DOI:10.1007/s10811-012-9804-6.Search in Google Scholar

[6] Hajimahmoodi M, Faramarzi MA, Mohammadi N, Soltani N, Oveisi MR, Nafissi-Varcheh N. Evaluation of antioxidant properties and total phenolic contents of some strains of microalgae. J Appl Phycol 2010;22:43–50. DOI:10.1007/s10811-009-9424-y.Search in Google Scholar

[7] Kadota K, Nishiyama T, Shimiz K. A normalization strategy for comparing tag count data. Algorithms Mol Biol 2012;7:1–13. DOI:10.1021/jf901070g.Search in Google Scholar PubMed

[8] Sguera S. Spirulina platensis et ses constituants: intérêts nutritionnels et activités thérapeutiques. Doctoral dissertation, UHP-Université Henri Poincaré, 2008.Search in Google Scholar

[9] Pratt DE. Natural antioxidants from plant material. ACS Symp Ser 1992;507:54–71. DOI:10.1021/bk-1992-0507.ch005.Search in Google Scholar

[10] Ould Bellahcen T, Bouchabchoub A, Massoui M, El Yachioui M. Culture et production de Spirulina platensis dans les eaux usées domestiques. Larhyss J 2013;14:107–22.Search in Google Scholar

[11] Madkour Fadel F, Kamil AEW, Nasr HS. Production and nutritive value of Spirulina platensis in reduced cost media. Egypt J Aquat Res 2012;38:51–7.10.1016/j.ejar.2012.09.003Search in Google Scholar

[12] Pascaud M. The essential polyunsaturated fatty acids of Spirulina and our immune response. Bulletin De l’Institut Oceanographique 1993;12:49–57.Search in Google Scholar

[13] Gad AS, Khadrawy YA, El-Nekeety AA, Mohamed SR, Hassan NS, Abdel-Wahhab MA. Antioxidant activity and hepatoprotective effects of whey protein and Spirulina in rats. J Nutr 2011;27:582–9. DOI:10.1016/j.nut.2010.04.002.Search in Google Scholar PubMed

[14] Vonshak A, Taylor & Francis p. In Spirulina platensis (Arthrospira). Physiology, Cell-Biology and Biotechnology. CRC Press, 1997:131–58. Taylor & Francis Ltd, 1 Gunpowder Square, London EC4A 3DE.10.1201/9781482272970Search in Google Scholar

[15] Zarrouk C. Contribution a l’etude d’une Cyanophycee. Influence de Divers Facteurs Physiques et Chimiques sur la croissance et la photosynthese de Spirulina mixima. Thesis, University of Paris, France, 1966.Search in Google Scholar

[16] Babadzhanov AS, Abdusamatova N, Yusupova FM, Faizullaeva N, Mezhlumyan LG, Malikova MK. Chemical composition of Spirulina platensis cultivated in Uzbekistan. Chem Nat Compd 2004;40:276–9. DOI:10.1023/B:CONC.0000039141.98247.e8.Search in Google Scholar

[17] Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F. Colorimetric method for determination of sugars and related substances. Anal Chem 1956;28:350–6. DOI:10.1021/ac60111a017.Search in Google Scholar

[18] Bernal JL, Del Nozal MJ, Jiménez JJ. Use of a high-pressure Soxhlet extractor for the determination of organochlorine residues by gas chromatography. Chromatographia 1992;34:468–74. DOI:10.1007/BF02290238.Search in Google Scholar

[19] Ebeler SE. Analytical chemistry: Unlocking the secrets of wine flavor. Food Rev Inter 2001;17:45–64. DOI:10.1081/FRI-100000517.Search in Google Scholar

[20] Ellison TM. The iterative learning of phonological constraints. JACL 1991;20:1–8.Search in Google Scholar

[21] Bennett A, Bogorad L. Properties of subunits and aggregates of blue-green algal biliproteins. Biochem 1971;10:3625–34. DOI:10.1021/bi00795a022.Search in Google Scholar

[22] Silveira ST1, Burkert JF, Costa JA, Burkert CA. K S. Optimization of phycocyanin extraction from Spirulina platensis using factorial design. Bioresour Technol 2007;98:1629–34.10.1016/j.biortech.2006.05.050Search in Google Scholar

[23] Maadane A, Merghoub N, Ainane T, El Arroussi H, Benhima R, Amzazi S, et al. Antioxidant activity of some Moroccan marine microalgae: Pufa profiles, carotenoids and phenolic content. J Biotechno 2015;13–19. doi:10.1016/j.jbiotec.2015.06.400.Search in Google Scholar

[24] Dyer WJ, Bligh EG. A rapid method of total lipid extraction and purification. Can J Biochem Phys 1959;37:911–17.10.1139/y59-099Search in Google Scholar

[25] Brand-Williams MEC, Berset C. Respostas perceptivas E. LWT. J Food Sci Technol 1995;28:25–30. DOI:10.1016/S0023-6438(95)80008-5.Search in Google Scholar

[26] Baviskar JW, Khandelwal SR. Extraction, detection and identification of flavonoids from microalgae : an emerging secondary metabolite. IJCMAS 2015;2:110–17.Search in Google Scholar

[27] Engida AM, Kasim NS, Tsigie YA, Ismadji S, Huynh LH. Extraction, identification and quantitative HPLC analysis of flavonoids from sarang semut (Myrmecodia pendan). Ind Crops Prod 2013;41:392–6.10.1016/j.indcrop.2012.04.043Search in Google Scholar

[28] AFNOR - NF EN 14148. Foodstuffs - determination of vitamin k1 by HPLC, 2004.Search in Google Scholar

[29] Lopes-Lutz D, Dettmann J, Nimalaratne C, Schieber A. Characterization and quantification of polyphenols in Amazon grape (Pourouma cecropiifolia Martius). Molecules 2010;15:8543–52.10.3390/molecules15128543Search in Google Scholar

[30] Huber L, Waldbronn D. Validation of HPLC Methods BioPharm Validation of HPLC Methods, 1999; 1–8.Search in Google Scholar

[31] Kazusaki M, Ueda S, Takeuchi N, Ohgami Y. Validation of analytical procedures by high− performance liquid chromatography for pharmaceutical analysis. J Chromatogr 2012;33:265–73.10.15583/jpchrom.2012.005Search in Google Scholar

[32] Zheng G, Li C, Guo L, Ruo W, Wang S. Purification of extracted fatty acids from the microalgae Spirulina. J Am Oil Chem Soc 2012;89:561–6. DOI:10.1007/s11746-011-1956-.Search in Google Scholar

[33] Chen J, Lee Y. Optimization of the transesterification reaction of microalgal Monoraphidium sp. Renew Energ 2018;129:717–23.10.1016/j.renene.2017.06.012Search in Google Scholar

[34] Burits M, Bucar F. Antioxidant activity of Nigella sativa essential oil. Phytother Res 2000;14:323–8.10.1002/1099-1573(200008)14:5<323::AID-PTR621>3.0.CO;2-QSearch in Google Scholar

[35] Choi HS, Sun Song H, Ukeda H, Sawamura M. Radical-scavenging activities of citrus essential oils and their components: detection using 1,1-diphenyl-2-picrylhydrazyl. J Agric Food Chem 2000;48:4156–61.10.1021/jf000227dSearch in Google Scholar

[36] Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 1999;26:1231–7.10.1016/S0891-5849(98)00315-3Search in Google Scholar

[37] Jourdan JP. Cultivez votre spiruline. Edt Antenna Technologie 2006;5:146.Search in Google Scholar

[38] Falquet J, Hurni JP. The nutritional aspects of Spirulina. Antenna Foundation, 1997.Search in Google Scholar

[39] Hosseini E, Mohamadamini M, Akbari M, Nejati F, Mazdapour M, Ghasemian M, et al. The laboratory scale evaluation of multiple pH ranges on Spirulina platensis culture in the production of dry biomass, chlorophyll, phycocyanin, & carotenoids. Bull Env Pharmacol Life Sci 2015;4:13–18.Search in Google Scholar

[40] Ali HEA, Shanab SMM, Shalaby EAA, El Demerdash UMN, Abdullah MA. Evaluation of antioxidants, pigments and secondary metabolites contents in Spirulina platensis. Appl Mech Mater 2014;625:160–3.10.4028/www.scientific.net/AMM.625.160Search in Google Scholar

[41] Shanab SMM, Mostafa SSM, Shalaby EAA, Mahmoud GI. Aqueous extracts of microalgae exhibit antioxidant and anticancer activities. Asian Pac J Trop Biomed 2012;2:608–15. DOI:10.1016/S2221-1691(12)60106-3.Search in Google Scholar

[42] Paoletti C, Pushparaj B, Florenzano G, Capella P, Lercker G. Unsaponifiable matter of green and blue-green algal lipids as a factor of biochemical differentiation of their biomasses: I. Total unsaponifiable and hydrocarbon fraction. J Lipids 1976;11:258–65. DOI:10.1007/BF02544051.Search in Google Scholar

[43] Durand-Chastel H. Production and use of Spirulina in Mexico. Algae biomass: production and use. sponsored by the National Council for Research and Development, Israel and the Gesellschaft fur Strahlen-und Umweltforschung (GSF), Munich, Germany; editors, Gedaliah Shelef, Carl J. Soeder, 1980.Search in Google Scholar

[44] Santos TD, Bastos de Freitas BC, Moreira JB, Zanfonato K, Vieira Costa JA. Development of powdered food with the addition of Spirulina for food supplementation of the elderly population. Innov Food Sci Emerg Technol 2016;37:216–20.10.1016/j.ifset.2016.07.016Search in Google Scholar

[45] Čeksterytė V, Kurtinaitienė B, Venskutonis PR, Pukalskas A, Kazernavičiūtė R, Balžekas J. Evaluation of antioxidant activity and flavonoid composition in differently preserved Bee products. Czech J Food Sci 2016;34:133–42.10.17221/312/2015-CJFSSearch in Google Scholar

[46] Wang L, Pan B, Sheng J, Xu J, Hu Q. Antioxidant activity of Spirulina platensis extracts by supercritical carbon dioxide extraction. Food Chem 2007;105:36–41.10.1016/j.foodchem.2007.03.054Search in Google Scholar

[47] Shalaby Emad A, Shanab S. Comparison of DPPH and ABTS assays for determining antioxidant potential of water and methanol extracts of Spirulina platensis. Indian J Mar Sci 2013;42:556–64.Search in Google Scholar

[48] Formica JV, Regelson W. Review of the biology of quercetin and related bioflavonoids. Food Chem Toxicol 1995;33:1061–80.10.1016/0278-6915(95)00077-1Search in Google Scholar

[49] Riyaz AD, Pradeep K, Brahman N, Khurana Javed A, Wagay KSP. Evaluation of antioxidant activity of crocin, podophyllotoxin and kaempferol by chemical, biochemical and electrochemical assays. Arab J Chem 2017;10:1119–28.10.1016/j.arabjc.2013.02.004Search in Google Scholar

[50] Chen AY, Chen YC. A review of the dietary flavonoid, kaempferol on human health and cancer chemoprevention. Food Chem 2013;138:2099–107.10.1016/j.foodchem.2012.11.139Search in Google Scholar PubMed PubMed Central

[51] Eldahshan OA, Azab SS. Anti-inflammatory effect of apigenin-7-neohesperidoside (rhoifolin) in carrageenin-induced rat oedema model. J Appl Pharm Sci 2012;2:74.10.7324/JAPS.2012.2811Search in Google Scholar

[52] Liu R, Zhang T, Yang H, Lan X, Ying JDG. The flavonoid apigenin protects brain neurovascular coupling against amyloid-beta 25-35-induced toxicity in mice. Alzheimers Dis 2011;24:85–100.10.3233/JAD-2010-101593Search in Google Scholar PubMed

[53] Rithidech KN, Tungjai M, Reungpatthanaphong P, Honikel LSS. Attenuation of oxidative damage and inflammatory responses by apigenin given to mice after irradiation. Mutat Res 2012;749:29–38.10.1016/j.mrgentox.2012.08.001Search in Google Scholar PubMed

[54] Shoubaky GAE, Abdel-Daim MM, Mansour MH, Salem EA. Isolation and identification of a flavone apigenin from marine red alga Acanthophora spicifera with antinociceptive and anti-inflammatory activities. J Exp Neurosci 2016;10:21–9.10.4137/JEN.S25096Search in Google Scholar PubMed PubMed Central

[55] Ama Moor VJ, Pieme1 CA, Nya Biapa PC, Ngo Matip ME, Moukette Moukette B, Tankeu Nzufo F, et al. Chemical composition of Spirulina platensis of Nomayos-Yaounde (cameroon). Food Sci Tech 2016;17:524–52.Search in Google Scholar

[56] Gholivand MB, Rahimi-Nasrabadi M, Batooli H, Ebrahimabadi AH. Chemical composition and antioxidant activities of the essential oil and methanol extracts of Psammogeton canescens. Food Chem Toxicol 2010;48:24–8. DOI:10.1016/j.fct.2009.09.007.Search in Google Scholar PubMed

[57] Sawa T, Nakao M, Akaike T, Ono K, Maeda H. Alkylperoxyl radical-scavenging activity of various flavonoids and other phenolic compounds: Implications for the anti-tumor-promoter effect of vegetables. J Agric Food Chem 1999;47:397–402. DOI:10.1021/jf980765e.Search in Google Scholar PubMed

[58] Ravi M, De SL, Azharuddin S, Paul SF. The beneficial effects of Spirulina focusing on its immunomodulatory and antioxidant properties. Nutr Diet Suppl 2010;2:73–83. DOI:10.2147/NDS.S9838.Search in Google Scholar

[59] Romay C, Armesto J, Remirez D, González R, Ledon N, García I. Antioxidant and anti-inflammatory properties of C-phycocyanin from blue-green algae. Inflamm Res 1998;47:36–41. DOI:10.1007/s000110050256.Search in Google Scholar PubMed

[60] Furuno K, Akasako T, Sugihara N. The contribution of the pyrogallol moiety to the superoxide radical scavenging activity of flavonoids. Biol Pharm Bull 2002;25:19–23. DOI:10.1248/bpb.25.19.Search in Google Scholar PubMed

[61] Ambrozova JV, Misurcova L, Vicha R, Machu L, Samek D, Baron M$, et al. Influence of extractive solvents on lipid and fatty acids content of edible freshwater algal and seaweed products, the green microalga Chlorella kessleri and the cyanobacterium Spirulina platensis. Molecules 2014;19:2344–60.10.3390/molecules19022344Search in Google Scholar PubMed PubMed Central

[62] Ortega-Calvo JJ, Mazuelos C, Hermosin B, Saiz-Jimenez C. Chemical composition of Spirulina and eukaryotic algae food products marked in Spain. J Appl Phycol 1993;5:425–35.10.1007/BF02182735Search in Google Scholar

[63] Herrero M, VicenteM J, Cifuentes A, Ibáñez E. Characterization by high performance liquid chromatography/electrospray ionization quadrupole time of flight mass spectrometry of the lipid fraction of Spirulina platensis pressurized ethanol extract. Rapid Commun Mass Spectrom 2007;21:1729–38.10.1002/rcm.3017Search in Google Scholar PubMed

[64] Geneive Henry E, Momin RA, Nair MG, Dewitt DL. Antioxidant and cyclooxygenase activities of fatty acids found in food. J Agr Food Chem 2002;50:2231–4.10.1021/jf0114381Search in Google Scholar PubMed

[65] Cohen Z. Spirulina Platensis: Physiology, Cell Biology and Biotechnology, Vol. 175. London: Taylor and Francis, 1997:175–204.Search in Google Scholar

[66] Mahajan G, Kamat M. γ-Linolenic acid production from Spirulina platensis. Appl Microbiol Biotechnol 1995;43:466–9.10.1007/BF00218450Search in Google Scholar

[67] Sergeant S, Rahbar E, Chilton FH. Gamma-linolenic acid, Dihommo-gamma linolenic, Eicosanoids and inflammatory processes. Eur J Pharmacol 2016;785:77–86.10.1016/j.ejphar.2016.04.020Search in Google Scholar PubMed PubMed Central

[68] Nilsen DW, Aarsetoey H, Pønitz V, Brugger-Andersen T, Staines H, Harris WS, et al. The prognostic utility of dihomo-gamma-linolenic acid (DGLA) in patients with acute coronary heart disease. Int J Cardiol 2017;249:12–17.10.1016/j.ijcard.2017.09.202Search in Google Scholar PubMed

[69] Abd El-Baky H, El Baz FK, El-Baroty GS. Enhancement of antioxidant production in Spirulina platensis under oxidative stress. Acta Physiol Plant 2009;31:623–31. DOI:10.1007/s11738-009-0273-8.Search in Google Scholar

[70] Belay A, Ota Y, Miyakawa K, Shimamatsu H. Current knowledge on potential health benefits of Spirulina. J App Phycol 1993;235–41. doi:10.1007/BF00004024.Search in Google Scholar

[71] Cepoi L, Rudi L, Miscu V, Cojocari A, Chiriac T, Sadovnic D. Antioxidative activity of ethanol extracts from Spirulina platensis and Nostoc linckia measured by various methods. Fascicula Bio 2009;16:43–8.Search in Google Scholar

Received: 2019-05-11
Accepted: 2019-09-16
Published Online: 2020-09-23

© 2020 Walter de Gruyter GmbH, Berlin/Boston

Downloaded on 21.5.2024 from https://www.degruyter.com/document/doi/10.1515/jcim-2019-0036/html
Scroll to top button