A methylene blue assay for the quantification of fucoidan – A specific application for seaweed extracts

Fucoidan is a fucose-rich sulfated polysaccharide found mainly in brown seaweeds. Fucoidan has recently gained much attention due to its bioactive and medicinal properties, hence its high potential to be utilized in the pharmaceutical industry. Therefore, an accurate, reliable, and simple method for the specific determination of fucoidan would be a useful tool when looking to recover this valuable polysaccharide. This research focused on developing an assay utilizing the thiazine dye, methylene blue, for the quantification of fucoidan, with a specific application for fucoidan found in algal extracts. Fucoidan could be specifically determined at pH 1, to maximum concentrations of 4 g L^-1 in solution without the interference of alginate and other seaweed extract contaminants. At pH 7 and 11, quantification of fucoidan led to interference from alginate and gallic acid. A conversion factor of 1.8 allowed the quantification of fucoidan from Ecklonia maxima extracts using a commercial standard from Fucus vesiculosis. An Ecklonia maxima extract was found to contain 5.47 ± 0.061 g L^-1 of fucoidan. Measuring the fucoidan and L-fucose concentration in crude seaweed extract through the methylene blue assay and modified Dische and Shettles J Biol Chem 175:595–603, (1948) method, respectively, confirmed the accuracy and specificity of the method. The limit of quantification of the methylene blue assay at pH 1 was 0.62 g L^-1 of fucoidan.