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URL: https://dx.doi.org/DOI:10.17504/protocols.io.j6ncrde

Authors: Magdalena Gutowska, Sebastian Sudek, Alexandra Worden, Brateen Shome, Tadhg Begley

Group: Worden Lab

Summary: We use the described methods for determining the internal cellular quota of thiamin and its different phosphorylation states in marine algae, in the below case for the haptophyte Emiliania huxleyi. The chemical assay used – the thiochrome assay - was developed in the 1930s but has been modified over the decades to include instrumentation with improved detection (Backstrom et al. 1995, Reddick et al. 2001). The assay is based on cyclization/oxidation of thiamin to thiochrome under basic conditions in the presence of an oxidizing agent such as potassium ferricyanide. Thiochrome is intensely fluorescent and easy to detect and quantify with high sensitivity in an HPLC assay. This method has previously been used to determine cellular quotas of thiamin and its different phosphorylation states in algae from brackish waters (Pinto et al. 2002, Sylvander et al. 2013).

Affiliations: MBARI, MBARI, MBARI, TAMU, TAMU

Version: 1

Publication Date: 2017

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