Comparison of three methods for quantifying brown algal polyphenolic compounds.
Three methods of quantifying total phenolic compounds in marine brown algae were compared for their ability to measure accurately known concentrations of phloroglucinol in the presence of a protein, bovine serum albumin (BSA). These methods were: (1) a Folin-Ciocalteu assay for compounds dissolved in 80% methanol, (2) a Folin-Ciocalteu assay for compounds dissolved in 75% methanol-25% trichloroacetic acid, and (3) an assay in which polyvinylpolypyrrolidone (PVPP) was used to remove phenolic compounds in order to quantify nonphenolic, Folin-Ciocalteu-reactive materials. The Folin-Ciocalteu assay used with compounds dissolved in 80% methanol was the method that produced the most consistent results. It was also the assay in which the presence of proteins interfered least with the measurements of phloroglucinol and the assay that was easiest to conduct. The presence of TCA decreased the pH of the extracting solution and increased the amounts of proteins in solution. These proteins later interfered with the Folin-Ciocalteu reaction. TCA effectively bound proteins when protein concentrations were low but not when protein concentrations were high. The amount of phloroglucinol removed by PVPP varied widely and was dependent on phloroglucinol concentration, making this an unreliable method for quantifying phenolic concentrations. The Folin-Ciocalteu assay will measure some nonphenolic substances when brown algae are extracted in 80% methanol; however, inFucus gardneri, nonphenolic substances were at most 5% of the material quantified by the assay.
Department of Biology, Kenyon College, 43022, Gambier, Ohio.
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van Alstyne