A COMPARATIVE STUDY ON THE ESTIMATION OF EXTRACTED PIGMENTS FROM SOME ALGAE USING DIFFERENT SOLVENTS

Document Type : Original Article

Authors

1 Fertilization Technology Department, National Research Centre, Cairo-Dokki, Egypt

2 Botany Department, Faculty of Science, Ain Shams University, Egypt

Abstract

Four different solvents were used to study their efficiency on pigments extraction from three different algal species of Chlorella vulgaris, Spirulina platensis and Nannochloropsis oculata. Dimethylsulfoxide, acetone, ethanol and methanol were used in the extraction process. The experiment was performed in both of fresh and oven dried algal samples. Chlorophylls a, b, total chlorophyll and total carotene concentrations were determined spectrophotometrically and by HPLC technique.  The spectrophotometric method revealed that, dimethylsulfoxide surpasses all other used solvents in all species and all pigments in fresh samples except for chlorophyll a of N. oculata, whereas acetone was more effective than the others in fresh and dry sample. For the dried sample of C. vulgaris, methanol gave the maximum extractability with chlorophyll a while chlorophyll b and total chlorophyll were found be maximum in dimethylsulfoxide solvent. In Spirulina, methanol represented the maximum chlorophyll a and total chlorophyll for the dried samples. Total carotene was found to be higher in ethanol solvent with both C. vulgaris and N. oculata but dimethylsulfoxide surpasses the other solvents with S. platensis. HPLC results emphasized that, acetone surpasses all other used solvent in fresh samples and methanol was ideal solvent in dry samples of C. vulgaris.  Dimethylsulfoxide represented the maximum chlorophyll a and total carotene with Spirulina in both samples. Methanol was the best solvent for extraction of all pigments with N. oculata except with β-carotene of dry sample, ethanol was the best solvent.

Keywords


Abalde, J., Betancourt, L., Torres, E., Cid, A. and Barwell, C. (1998). Purification and characterization of phycocyanin from the marine cyanobacterium Synechococcus sp. IO9201. Plant Science, 136:109-120.
Buchaca, T., Felip M. and Catalan, J. (2005). A comparison of HPLC pigment analyses and biovolume estimates of phytoplankton groups in an oligotrophic lake. J. Plankton Res.; 27:91-101.
Cartaxana, P., Mendesm C., Van Leeuwe, M. and Brotas, V. (2006). Comparative study on the microphytobenthic pigments of muddy and sandy intertidal pigments of the Tagus estuary. Est. Coast. Shelf Sci.;66:225-230.
Davies, H. (1976). Carotenoids. In Chemistry and Biochemistry of Plant Pigments. 2nd Eddition Goodwin. T. W. Academic Press, 2:38-165.
Devesa, R., Moldes, A., Díaz-Fierros, F. and Barral, M. (2007). Extraction study of algal of algal pigments in river bed sediments by applying factorial designs. Talanta,72:1546-1551.
El-Sayed, A. B. and El Fouly, M. M. (2005). Recovery of outdoor mass culture bleached Scendesmus sp. Pakistan journal of biological science, 8(3):470-474.
Gantt, E. (1981). Phycobilisomes. Ann. Rev. Plant Physiol.; 32:327-47.
Gerloff-Elias, A., Spijkerman, E. and Schubert, H. (2005). Light acclimation of Chlamydomonas acidophila accumulating in the hypolimnion of an acidic lake (pH 2.6). Freshwater Biol.; 50:1301-1314.
Grinham, A., Carruthers, T., Fisher, P., Udy, J. and Dinnison, W. (2007). Accurately measuring the abundance of benthic microalgae in spatially variable habitats. Limnol. Oceanogr. Methods,5:119-125.
Hasni, Z., Yaacob, J. S., Yusoff, A. I. M., Taha, R. M., Yahya, R., Bakrudeen, A. Ahmed, A. and Ramesh, K. (2011). Effect of different solvents extraction on recovery of pigments in Xylocarpus granatum, endangered medicinal plant Materials Research Innovations, 15(2):1-4.
 
 
Jeffrey, S. W., Mantoura, R. F. C. and Bjørnland, T. (1997). Data for the identification of 47 key phytoplankton pigments. – In: Jeffrey, S.W., Mantoura, R.F.C., Wright, S.W. (ed.): Phytoplankton pigments in oceanography: Guidelines to modern methods, 449-559.
Khalil, I. A. and F. R. Varananis. (1996). Carotenoid extraction and analysis by reversed phase HPLC system. Sarhad J. Agric.; 105(67): 15-21.
Koh, C., Khim, J., Araki, H., Yamanishi, H. and Koga, K. (2007). Within-day and seasonal patterns of microphytobenthos biomass determined by co-measuremen of sediment and water column chlorophylls in the intertidal mudflat of Nanaura Sea, Ariake Sea, Japan. Est.Coast. Shelf Sci.; 72:42-52.
Kumar, N. J. I.; Kumar, R. N., Bora, A.; Kaur Amb, M. and Chakraborthy, S. (2009). An Evaluation of the Pigment Composition of Eighteen Marine Macroalgae Collected from Okha Coast, Gulf of Kutch, India, Our Nature; 7:48-55.
Li, D. X., Guo, Y. X., Yun, H. Y., Zhang, M., Gong, X. Y. and Mu., F. (2005). Methods of chlorophyll determination from maize (in Chinese, with English abstract). Chin. Agri. Sci. Bull.; 21:153-155.
Lichtenthaler, H. K., Wellburn, A. R. (1983). Determination of total carotenoids and chlorophylls a and b of leaf in different solvents. Biol. Soc. Trans.; 11:591-592.
Jodłowska, S. and Latała, A. (2011). The comparison of spectrophotometric method and high performance liquid chromatography in photosynthetic pigments analysis. Online Journal of Biological Sciences, 11: 63–69.
Minkova, K. M., Tchernov, A. A., Tchorbadjieva, M. I., Fournadjieva, S. T., Antova, R. E. and Busheva, M. C. (2003). Purification of C-phycocyanin from Spirulina (Arthrospira) fusiformis. J. Biotechnol.; 102:55-59.
Murray, A. P., Gibbs, C. F., Shiroyama, A. R. and Flett, D. J. (1986). Determination of chlorophyll in marine waters: intercomparison of a rapid HPLC method with full HPLC, spectrophotometric and fluorometric methods. Mar. Chem.; 19:211-227
Nakamura, A. and Watanabe, T. (2001). Separation and determination of minor photosynthetic pigments by reversed-phase HPLC with minimal alteration of chlorophylls. Anal. Sci., 17: 503–508.
Pangestuti, R., and Kim, S-K. (2011). Biological activities and health benefit effects of natural pigments derived from marine algae, J. Func. Food., 3(4), 255-266.
Pompelli, M. F.; França,S. C.; Tigre, R. C.; de Oliveira, M. T.; Sacilot, M. and Pereira, E. C. (2013). Spectrophotometric determinations of chloroplastidic pigments in acetone, ethanol and dimethylsulphoxide. R. bras. Bioci., Porto Alegre, 11(1), 52-58.
Ritchie, R. J. (2006). Consistent sets of spectrophotometric chlorophyll equations for acetone, methanol and ethanol solvents. Photosynth. Res.; 89:27-41.
Ritchie, R. J. (2008).Universal chlorophyll equations for estimating chlorophylls a, b, c, and d and total chlorophylls in natural assemblages of photosynthetic organisms using acetone, methanol, or ethanol solvents. Photosynthetica, 46: 115–126.
Sarada, R., Pillai, M. G. and Ravishankar, G. A. (1999). Phycocyanin from Spirulina sp: influence of processing of biomass on phycocyanin yield, analysis of efficacy of extraction methods and stability studies on phycocyanin. Process Biochemistry, 34:795-801 
Sarkar, C. R. ; Das, L.; Bhagawati, B. and Goswami, B. Ch. (2012). A comparative study of carotenoid extraction from algae in different solvent systems Asian Journal of Plant Science and Research, 2 (4):546-549.
Sartory, D. P. and Grobbelaar, J. E. (1984). Extraction of chlorophyll a from freshwater phytoplankton for spectrophotometric analysis. Hydrobiol.; 114:177-187.
Shoaf, T. W. and Lium, B. W. (1976). Improved extraction of chlorophyll aand bfrom algae using dimethylsulphoxide. Limnol Oceanogr.; 21: 926-928.
Sigelman, H. W. and Kycia, J. H. (1978). Algal biliproteins. In: Hellebust JA, Craigie JS, editors. Handbook of Phycological Methods. Cam-bridge: Cambridge University Press, 72–81.
Simon D. and Helliwell, S. (1985). Extraction and quantification of chlorophyll a from freshwater green algae. Water Research, 32, 7: 2220-2223.
Stainer, R. Y., Kunisawa, R., Mandel, M. and Cohin-Bazire, G. (1971). Purification and properties of unicellular blue-green algae (order Chrococcales). Bacteriol. Rev.; 35:171-205.
 
Strain, H. H. and Svec. W. A. (1966). Extraction, separation, estimation and isolation of chlorophylls. In L. P. Vernon and G. R. Seeley [eds.], The chlorophylls. Academic. 21-66.
Sumanta, N.; Haque, I. C.; Nishika, J. and Suprakash, R. (2014). Spectrophotometric analysis of chlorophylls and carotenoids from commonly grown fern species by using various extracting solvents. Research Journal of Chemical Sciences, 4(9), 63-69.
 Vimala, T. and Poonghuzhali, T. V. (2015).  Estimation of pigments from seaweeds by using acetone and DMSO. International Journal of Science and Research, 4 (10), 1850-1854.
Wellburn, AR. (1994). The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. J. Plant Physiol.; 144:307-313.
Wright, S. W., Jeffrey, S. W. and Mantoura, F. R. C. (1997). Evaluation of methods and solvents for pigment extraction. In: Jeffrey, S.W., Mantoura, R.F.C., Wright, S.W. (eds.): Phytoplankton pigments in oceanography: Guidelines to Modern Methods, 261-282.