Christ loc 1 m alpha 2 4

Crude fucoidan was extracted as described previously (13 (link)). Each extraction was carried out using sporophylls from separate plants harvested in September, 2011. Four replicate samples of dried algal biomass (15 g) was treated at room temperature for 24 h with a MeOH–CHCl₃–water mixture (4:2:1) to remove lipids, protein, and colored pigments. The treated algal biomass was filtered through a Whatman’s filter paper (90 mm GF/D), then washed with acetone and dried overnight at room temperature. Treated algae (10 g) were mechanically stirred with 2% aqueous CaCl₂ (100 mL) at 85°C for 5 h. The extract was centrifuged at 18500 g (Eppendorf Centrifuge 5810R V3.1, Eppendorf AG, Hamburg, Germany) and the supernatant was collected. A hexadecyltrimethylammonium bromide (cetavlon) solution (10%, 50 mL) was added to the extract and was left to precipitate at 4°C overnight. The precipitate was centrifuged at 18500 g, washed with water and mechanically stirred with 60 mL ethanolic NaI solution (20%) for 72 h. The precipitate was removed by centrifugation, washed with ethanol and lyophilized (Christ LOC 1-M Alpha 2-4, Martin Christ, Osterode am Harz, Germany) to give crude fucoidan (23 (link)).

Undaria pinnatifida was harvested from Port Underwood, New Zealand in September 2011 from two selected mussel farms (farms 106 and 327). The two seaweed samples were independently prepared for extraction. The sporophyll and blade were separated into labeled bags and lyophilized (Christ LOC 1-M Alpha 2-4, Martin Christ, Osterode am Harz, Germany) in bulk within 48 h of frozen storage. The dried sample was milled using a coffee grinder (Breville CG2B), sieved (<0.6 mm) and then stored in PET containers at room temperature before analyses (22 (link)). All chemicals and reagents were of analytical grade.