Abstract
There are still limitations in the pigment extraction methods used in microalgae biomass, especially for laboratory scale. This work aimed to develop a simple method for simultaneous extraction of chlorophylls and carotenoids from the microalgae Spirulina sp. and Scenedesmus obliquus. The extraction was performed using a methanol/chloroform (2:1 v/v) mixture with different homogenization times (30, 60 and 120 min); after exhaustive extraction with chloroform, acetone, ethyl acetate, and ethanol were tested and compared with the reference maceration method (MM). Satisfactory recovery values, mostly between 101.1 and 124.2%, were obtained with 30 min of homogenization and extraction with ethanol after chloroform (PM30E). In general, the chlorophyll and carotenoid profiles obtained by the PM30E presented fewer isomers and possible degradation compounds. PM30E showed a reduction of about five times the volume of organic solvents and was four times faster than the MM. In addition, this method provided greater analytical frequency and security for the analyst, since PM30E is performed in a closed system of mechanical homogenization.
Similar content being viewed by others
References
Ambati RR, Gogisetty D, Aswathanarayana RG, Ravi S, Bikkina PN, Bo L, Yuepeng S (2019) Industrial potential of carotenoid pigments from microalgae: current trends and future prospects. Crit Rev Food Sci Nutr 59:1880–1902
Amorim-Carrilho KT, Cepeda A, Fente C, Regal P (2014) Review of methods for analysis of carotenoids. TrAC-Trend Anal Chem 56:49–73
Barsanti L, Gualtieri P (2018) Is exploitation of microalgae economically and energetically sustainable? Algal Res 31:107–115
Begum H, Yusoff FM, Banerjee S, Khatoon H, Shariff M (2016) Availability and utilization of pigments from microalgae. Crit Rev Food Sci Nutr 56:2209–2222
Biazotto KR, de Souza Mesquita LM, Neves BV, Braga ARC, Tangerina MMP, Vilegas W, Mercadante AZ, De Rosso VV (2019) Brazilian biodiversity fruits: discovering bioactive compounds from underexplored sources. J Agric Food Chem 67:1860–1876
Britton G, Liaaen-Jensen S, Pfander H (2012) Carotenoids: handbook. Birkhäuser, Basel
Camacho F, Macedo A, Malcata F (2019) Potential industrial applications and commercialization of microalgae in the functional food and feed industries: a short review. Mar Drugs 17:312–336
Casale M, Oliveri P, Casolino C, Sinelli N, Zunin P, Armanino C, Forina M, Lanteri S (2012) Characterization of PDO olive oil Chianti Classic by non-selective (UV–visible, NIR and MIR spectroscopy) and selective (fatty acid composition) analytical techniques. Anal Chim Acta 712:56–63
Chen K, Ríos JJ, Pérez-Gálvez A, Roca M (2017) Comprehensive chlorophyll composition in the main edible seaweeds. Food Chem 228:625–633
Craft NE, Soares JH (1992) Relative solubility, stability, and absorptivity of lutein and beta.-carotene in organic solvents. J Agric Food Chem 40:431–434
D’Alessandro EB, Antoniosi Filho NR (2016) Concepts and studies on lipid and pigments of microalgae: a review. Renew Sust Energ Rev 58:832–841
da Silva Ferreira V, Sant’Anna C (2017) Impact of culture conditions on the chlorophyll content of microalgae for biotechnological applications. World J Microbiol Biotechnol 33:20–27
Davies BH (1976) Carotenoids. In: Goodwin TW (ed) Chemistry and biochemistry of plant pigments. Academic Press, New York, pp 38–165
de Rosso VV, Mercadante AZ (2007) Identification and quantification of carotenoids, by HPLC-PDA-MS/MS, from Amazonian fruits. J Agric Food Chem 55:5062–5072
do Nascimento TC, Nass PP, Fernandes AS, Vieira KR, Wagner R, Jacob-Lopes E, Zepka LQ (2020) Exploratory data of the microalgae compounds for food purposes. Data Brief 29:105182
Egeland ES (2016) Carotenoids. In: Borowitzka MA, Beardall J, Raven JA (eds) The physiology of microalgae. Springer, Dordrecht, pp 507–563
Esquivel-Hernández DA, Ibarra-Garza IP, Rodríguez-Rodríguez J, Cuéllar-Bermúdez SP, Rostro-Alanis MDJ, Alemán-Nava GS, García-Pérez JS, Parra-Saldívar R (2017) Green extraction technologies for high-value metabolites from algae: a review. Biofuels Bioprod Biorefin 11:215–231
Faegri K, Kaland PE, Krzywinski K (1989) Textbook of pollen analysis. Wiley, Hoboken
Fernandes AS, Nogara GP, Menezes CR, Cichoski AJ, Mercadante AZ, Jacob-Lopes E, Zepka LQ (2017) Identification of chlorophyll molecules with peroxyl radical scavenger capacity in microalgae Phormidium autumnale using ultrasound-assisted extraction. Food Res Int 99:1036–1041
Galasso C, Gentile A, Orefice I, Ianora A, Bruno A, Noonan DM, Sansone C, Albini A, Brunet C (2019) Microalgal derivatives as potential nutraceutical and food supplements for human health: a focus on cancer prevention and interception. Nutrients 11:1226–1247
Gałuszka A, Migaszewski Z, Namieśnik J (2013) The 12 principles of green analytical chemistry and the significance mnemonic of green analytical practices. TrAC-Trend Anal Chem 50:78–84
Giuffrida D, Zoccali M, Giofrè SV, Dugo P, Mondello L (2017) Apocarotenoids determination in Capsicum chinense Jacq. cv. Habanero, by supercritical fluid chromatography-triple-quadrupole/mass spectrometry. Food Chem 231:316–323
Gong M, Bassi A (2016) Carotenoids from microalgae: a review of recent developments. Biotechnol Adv 34:1396–1412
Huang SC, Hung CF, Wu WB, Chen BH (2008) Determination of chlorophylls and their derivatives in Gynostemma pentaphyllum Makino by liquid chromatography–mass spectrometry. J Pharm Biomed Anal 48:105–112
Jacob-Lopes E, Maroneze MM, Deprá MC, Sartori RB, Dias RR, Zepka LQ (2019) Bioactive food compounds from microalgae: an innovative framework on industrial biorefineries. Curr Opin Food Sci 25:1–7
Kapoore RV, Butler TO, Pandhal J, Vaidyanathan S (2018) Microwave-assisted extraction for microalgae: from biofuels to biorefinery. Biology 7:18–42
Khalid N, Barrow CJ (2018) Critical review of encapsulation methods for stabilization and delivery of astaxanthin. J Food Bioact 1:104–115
Khan MI, Shin JH, Kim JD (2018) The promising future of microalgae: current status, challenges, and optimization of a sustainable and renewable industry for biofuels, feed, and other products. Microb Cell Factories 17:36–56
Klassen V, Blifernez-Klassen O, Hoekzema Y, Mussgnug JH, Kruse O (2015) A novel one-stage cultivation/fermentation strategy for improved biogas production with microalgal biomass. J Biotechnol 215:44–51
Lanfer-Marquez UM, Barros RM, Sinnecker P (2005) Antioxidant activity of chlorophylls and their derivatives. Food Res Int 38:885–891
Lim H, Tanaka A, Tanaka R, Ito H (2019) In vitro enzymatic activity assays implicate the existence of the chlorophyll cycle in chlorophyll b-containing cyanobacteria. Plant Cell Physiol 60:2672–2683
Loh CH, Inbaraj BS, Liu MH, Chen BH (2012) Determination of chlorophylls in Taraxacum formosanum by high-performance liquid chromatography–diode array detection–mass spectrometry and preparation by column chromatography. J Agric Food Chem 60:6108–6115
Louda JW, Li J, Liu L, Winfree MN, Baker EW (1998) Chlorophyll-a degradation during cellular senescence and death. Org Geochem 29:1233–1251
Mackenzie G, Boa AN, Diego-Taboada A, Atkin SL, Sathyapalan T (2015) Sporopollenin, the least known yet toughest natural biopolymer. Front Mater 2:66–70
Mandelli F, Miranda VS, Rodrigues E, Mercadante AZ (2012) Identification of carotenoids with high antioxidant capacity produced by extremophile microorganisms. World J Microbiol Biotechnol 28:1781–1790
Marcinkowska R, Namieśnik J, Tobiszewski M (2019) Green and equitable analytical chemistry. Curr Opin Green Sust 19:19–23
Maroneze MM, Zepka LQ, Lopes EJ, Pérez-Gálvez A, Roca M (2019) Chlorophyll oxidative metabolism during the phototrophic and heterotrophic growth of Scenedesmus obliquus. Antioxidants 8:600–614
Matos ÂP (2017) The impact of microalgae in food science and technology. J Am Oil Chem Soc 94:1333–1350
Mussagy CU, Winterburn J, Santos-Ebinuma VC, Pereira JFB (2019) Production and extraction of carotenoids produced by microorganisms. Appl Microbiol Biotechnol 103:1095–1114
Nabavi SM, Saeedi M, Nabavi SF, Silva AS (2020) Recent advances in natural products analysis. Elsevier, Amsterdam
Novoveská L, Ross ME, Stanley MS, Pradelles R, Wasiolek V, Sassi JF (2019) Microalgal carotenoids: a review of production, current markets, regulations, and future direction. Mar Drugs 17:640–660
Paliwal C, Ghosh T, George B, Pancha I, Maurya R, Chokshi K, Ghosh A, Mishra S (2016) Microalgal carotenoids: potential nutraceutical compounds with chemotaxonomic importance. Algal Res 15:24–31
Patias LD, Fernandes AS, Petry FC, Mercadante AZ, Jacob-Lopes E, Zepka LQ (2017) Carotenoid profile of three microalgae/cyanobacteria species with peroxyl radical scavenger capacity. Food Res Int 100:260–266
Porra RJ, Thompson WA, Kriedemann PE (1989) Determination of accurate coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochem Biophys Acta 975:384–394
Ramluckan K, Moodley KG, Bux F (2014) An evaluation of the efficacy of using selected solvents for the extraction of lipids from algal biomass by the Soxhlet extraction method. Fuel 116:103–108
Ribeiro JA, Almeida ES, Neto BA, Abdelnur PV, Monteiro S (2018) Identification of carotenoid isomers in crude and bleached palm oils by mass spectrometry. LWT Food Sci Technol 89:631–637
Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. Microbiology 111:1–61
Rodrigues DB, Menezes CR, Mercadante AZ, Jacob-Lopes E, Zepka LQ (2015) Bioactive pigments from microalgae Phormidium autumnale. Food Res Int 77:273–279
Rodriguez-Concepcion M, Avalos J, Bonet ML, Boronat A, Gomez-Gomez L, Hornero-Mendez D, Limon MC, Meléndez-Martínez AJ, Olmedilla-Alonso B, Palou A, Ribot J, Rodrigo MJ, Zacarias L, Zhu C (2018) A global perspective on carotenoids: metabolism, biotechnology, and benefits for nutrition and health. Prog Lipid Res 70:62–93
Saini RK, Keum YS (2018) Carotenoid extraction methods: a review of recent developments. Food Chem 240:90–103
Simon E, Schifferli A, Bucher TB, Olbrich D, Werner I, Vermeirssen EL (2019) Solid-phase extraction of estrogens and herbicides from environmental waters for bioassay analysis-effects of sample volume on recoveries. Anal Bioanal Chem 411:2057–2069
Vendruscolo RG, Facchi MMX, Maroneze MM, Fagundes MB, Cichoski AJ, Zepka LQ, Barin JS, Jacob–Lopes E, Wagner R (2018) Polar and non-polar intracellular compounds from microalgae: methods of simultaneous extraction, gas chromatography determination and comparative analysis. Food Res Int 109:204–212
Vendruscolo RG, Fagundes MB, Maroneze MM, do Nascimento TC, de Menezes CR, Barin JS, Zepka LQ, Jacob-Lopes E, Wagner R (2019) Scenedesmus obliquus metabolomics: effect of photoperiods and cell growth phases. Bioprocess Biosyst Eng 42:727–739
Wellman CH, Osterloff PL, Mohiuddin U (2003) Fragments of the earliest land plants. Nature 425:282–285
Zhang X, Yin W, Qi Y, Li X, Zhang W, He G (2017) Microencapsulation of astaxanthin in alginate using modified emulsion technology: preparation, characterization, and cytostatic activity. Can J Chem Eng 95:412–419
Zhang R, Parniakov O, Grimi N, Lebovka N, Marchal L, Vorobiev E (2019) Emerging techniques for cell disruption and extraction of valuable bio-molecules of microalgae Nannochloropsis sp. Bioprocess Biosyst Eng 42:173–186
Zhu Z, Wu Q, Di X, Li S, Barba FJ, Koubaa M, Roohinejad S, Xiong X, He J (2017) Multistage recovery process of seaweed pigments: Investigation of ultrasound assisted extraction and ultra-filtration performances. Food Bioprod Process 104:40–47
Funding
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) - Finance Code 001.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
ESM 1
(DOCX 24 kb)
Rights and permissions
About this article
Cite this article
Vendruscolo, R.G., Fernandes, A.S., Fagundes, M.B. et al. Development of a new method for simultaneous extraction of chlorophylls and carotenoids from microalgal biomass. J Appl Phycol 33, 1987–1997 (2021). https://doi.org/10.1007/s10811-021-02470-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10811-021-02470-8