A New Protocol Using Acidification for Preserving DMSP in Macroalgae and Comparison with Existing Protocols.

Abstract

Dimethylsulfoniopropionate (DMSP) plays many important physiological and ecological roles in macroalgae. The most common method to measure DMSP is by gas chromatography analysis of the dimethylsulfide (DMS) produced after NaOH hydrolysis (pH>12). Storage of DMS, however, is not recommended for more than a week. We investigated if acidification can be a suitable method to preserve DMSP in macroalgal samples over three months of storage, compared to widely used protocols such as drying and freezing at -20°C. The DMSP content of green (Ulva sp. and Ulva compressa), red (Chondrus crispus), and brown (Bifurcaria bifurcata) macroalgae were analyzed: 24h after NaOH addition (control values); and after acidification (0.2mol · L HCl-1 ) for 24h of fresh material, followed by NaOH addition for 24h. These values were compared to measurements after 3-month storage of samples that had been either dried in a heater (60°C for a night, and storage at room temperature), or frozen at -20°C, or kept in 0.2mol · L HCl-1 . There was no significant difference between DMSP measurements on freshly collected material and after acidification of the samples, whether 24h later or after 3months of storage. This was in contrast to 3-month storage protocols involving overnight drying at 60°C (75-98% DMSP loss), and to a lesser degree freezing at -20°C (37-80% DMSP loss). We thus advise to acidify macroalgal samples for preservation over long periods of time rather than drying or freezing, when assaying DMSP content.