Abstract
Seaweed polyphenols are potent antioxidants and have also been shown to have α-glucosidase inhibiting activity. In our continuous efforts to develop new marine-based nutraceuticals and functional food ingredients, we have investigated many algal species collected on the Atlantic coast of Canada. A simple method for estimating the total polyphenol content in seaweeds and their extracts was developed based on the classic Folin-Ciocalteau colorimetric reaction. By using the 96-well microplate and a microplate reader, this new method saves experimental time, significantly reduces the amount of sample required, handles large number of samples in one experiment, and also improves the repeatability of the results.
A number of algal samples collected on the seashore of Nova Scotia, Canada, were analyzed for their levels of polyphenol content using this microplate-based method. The antioxidant activity of these samples was also assessed by using DPPH (2, 2-diphenyl-1-picrylhydrazyl) radical scavenging assay. The results showed that there is a strong correlation between the total polyphenol content and the potency of antioxidant effect.
Similar content being viewed by others
References
Ahn MJ, Yoon KD, Min S-Y, Lee JS, Kim JH, Kim TG, Kim SH, Kim N-G, Huh H, Kim J (2004) Inhibition of HIV-1 reverse transcriptase and protease by phlorotannins from the brown alga Ecklonia cava. Biological and Pharmaceutical Bulletin 27: 544–547.
Chkhikvishvili ID, Ranazanov ZM (2000) Phenolic substances of brown algae and their antioxidant activity. Prikladnaya Biokhimiya I Mikrobiologiya 36: 336–338.
Folin O, Ciocalteu V (1927) On tyrosine and tryptophane determinations in proteins. Journal of Biological Chemistry 73: 627–650.
Folin O, Denis W (1912) On phosphotungstic-phosphomolybdic compounds as color regents. Journal of Biological Chemistry 12: 239–243.
Frei B (ed.) (1994) Natural Antioxidants in Human Health and Disease. Academic Press, San Diego.
Fukumoto LR, Mazza G (2000) Assessing antioxidant and prooxidant activities of phenolic compounds. Journal of Agricultural and Food Chemistry 48: 3597–3604.
Jimenez-Escrig A, Jimenez-Jimenez I, Pulido R, Saura-Calixto F (2001) Antioxidant activity of fresh and processed edible seaweeds. Journal of the Science of Food and Agriculture 81: 530–534.
Kang HS, Chung HY, Jung JH, Son BW, Choi JS (2003a) A new phlorotannin from the brown alga Ecklonia stolonifera. Chemical & Pharmaceutical Bulletin 51: 1012–1014.
Kang HS, Chung HY, Kim JY, Son BW, Jung HA, Choi JS (2004) Inhibitory phlorotannins from the edible brown alga Ecklonia stolonifera on total reactive oxygen species (ROS) generation. Archives of Pharmacal Research 27: 194–198.
Kang K, Park Y, Hwang HJ, Kim SH, Lee JG, Shin H-C (2003b) Antioxidative properties of brown algae polyphenolics and their perspectives as chemopreventive agents against vascular risk factors. Archives of Pharmacal Research 26: 286–293.
Kurihara H, Ayaki T, Takahashi Y, Sasaki S, Ota T, Obori T, Yoshikawa S (2002) α-Glucosidase inhibitors containing brown algae. JP (patent application), No. 2001045778, 6pp.
Lee SK, Mbwambo ZH, Chung HS, Luyengi L, Gamez EJC, Mehta RG, Kinghorn AD, Pezzuto JM (1998) Evaluation of the antioxidant potential of natural products. Combinatorial Chemistry and High Throughput Screening 1: 35–46.
Nagayama K, Iwamura Y, Shibata T, Hirayama I, Nakamura T (2002) Bactericidal activity of phlorotannins from the brown alga Ecklonia kurome. Journal of Antimicrobial Chemotherapy 50: 889–893.
Nagai T, Yukimoto T (2003) Preparation and functional properties of beverages made from sea algae. Food Chemistry 81: 327–332.
Nakamura T, Nagayama K, Uchida K, Tanaka R (1996) Antioxidant activity of phlorotannins isolated from the brown alga Eisenia bicyclis. Fisheries Science 62: 923–926.
Ragan MA, Jensen A (1977) Quantitative studies on brown algal phenols. I. Estimation of absolute polyphenol content of Ascophyllum nodosum (L.) Le Jol. and Fucus vesiculosus (L.). Journal of Marine Biology and Ecology 30: 209–221.
Ragan MA, Jensen A (1978) Quantitative studies on brown algal phenols. II. Seasonal variation in polyphenol content of Ascophyllum nodosum (L.) Le Jol. and Fucus vesiculosus (L.). Journal of Marine Biology and Ecology 34: 245–258.
Shahidi F, Naczk M (2004) Phenolics in Food and Nutraceuticals. CRC Press, Boca Raton.
Shibata T, Fujimoto K, Nagayama K, Yamaguchi K, Nakamura T (2002) Inhibitory activity of brown algal phlorotannins against hyaluronidase. International Journal of Food Science and Technology 37: 703–709.
Shibata T, Nagayama K, Tanaka R, Yamaguchi K, Nakamura T (2003) Inhibitory effects of brown algal phlorotannins on secretory phospholipase A2s, lipoxygenases and cyclooxygenases. Journal of Applied Phycology 15: 61–66.
Singleton VL, Orthofer R, Lamuela-Raventos RM (1999) Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. In Abelson JN, Simon M, Sies H (eds), Methods in Enzymology. Vol. 299, Oxidants and antioxidants, Part A. Academic Press, Orlando: 152–178.
Singleton VL, Rossi JA, Jr (1965) Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American Journal of Enology and Viticulture 16: 144–158.
Swain T, Hillis WE (1959) The phenolic constituents of Prunus domestica. I.-The quantitative analysis of phenolic constituents. Journal of the Science of Food and Agriculture 10: 63–68.
Van Alstyne KL (1995) Comparison of three methods for quantifying brown algal polyphenolic compounds. Journal of Chemical Ecology 21: 45–59.
Wei Y, Xu Z (2003) Studies on antioxidative activity of high molecular weight polyphenols from two kinds of brown algae. Zhongcaoyao 34: 3170–319.
Yan X, Fan X, Lou Q, Fang G, Chen Y, Song H (1998) Preparation and application of seaweed polyphenol antioxidant. Faming Zhuanli Shenqing Gongkai Shumingshu (Chinese Patent Application), No. 98110252.2, 4 pp.
Yan X, Li X, Fan X, Zhou C (1997) Studies on extraction procedure and antioxidative activity of phlorotannins from Sargassum kjellmanianum. Chinese Journal of Oceanology and Limnology 15: 42–45.
Yan X, Li X, Zhou C, Fan X (1996) Prevention of fish oil rancidity by phlorotannins from Sargassum kellmanianum. Journal of Applied Phycology 8: 201–203.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhang, Q., Zhang, J., Shen, J. et al. A Simple 96-Well Microplate Method for Estimation of Total Polyphenol Content in Seaweeds. J Appl Phycol 18, 445–450 (2006). https://doi.org/10.1007/s10811-006-9048-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10811-006-9048-4