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A Simple 96-Well Microplate Method for Estimation of Total Polyphenol Content in Seaweeds

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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.

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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.

    Article  PubMed  CAS  Google Scholar 

  • Chkhikvishvili ID, Ranazanov ZM (2000) Phenolic substances of brown algae and their antioxidant activity. Prikladnaya Biokhimiya I Mikrobiologiya 36: 336–338.

    CAS  Google Scholar 

  • Folin O, Ciocalteu V (1927) On tyrosine and tryptophane determinations in proteins. Journal of Biological Chemistry 73: 627–650.

    CAS  Google Scholar 

  • Folin O, Denis W (1912) On phosphotungstic-phosphomolybdic compounds as color regents. Journal of Biological Chemistry 12: 239–243.

    CAS  Google Scholar 

  • Frei B (ed.) (1994) Natural Antioxidants in Human Health and Disease. Academic Press, San Diego.

    Google Scholar 

  • Fukumoto LR, Mazza G (2000) Assessing antioxidant and prooxidant activities of phenolic compounds. Journal of Agricultural and Food Chemistry 48: 3597–3604.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Nagai T, Yukimoto T (2003) Preparation and functional properties of beverages made from sea algae. Food Chemistry 81: 327–332.

    Article  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • Shahidi F, Naczk M (2004) Phenolics in Food and Nutraceuticals. CRC Press, Boca Raton.

    Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Chapter  Google Scholar 

  • Singleton VL, Rossi JA, Jr (1965) Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American Journal of Enology and Viticulture 16: 144–158.

    CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • Van Alstyne KL (1995) Comparison of three methods for quantifying brown algal polyphenolic compounds. Journal of Chemical Ecology 21: 45–59.

    Article  Google Scholar 

  • Wei Y, Xu Z (2003) Studies on antioxidative activity of high molecular weight polyphenols from two kinds of brown algae. Zhongcaoyao 34: 3170–319.

    Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

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Correspondence to Junzeng Zhang.

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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

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  • DOI: https://doi.org/10.1007/s10811-006-9048-4

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