Pyrrhoxanthin was reisolated on the mg scale from a natural bloom of Ceratium spp. and fully characterized spectroscopically. All-trans-(3S,5R,6R,3'S,5'R)-peridinin acetate was converted into all-trans pyrrhoxanthin acetate. Semisynthetic all-trans pyrrhoxanthin acetate could not be separated from all-trans pyrrhoxanthin acetate ex natural pyrrhoxanthin, and exhibited the same Cotton effect as natural pyrrhoxanthin. This result, supporting the (3R,3'S,5'R,6'R)-configuration for pyrrhoxanthin, is consistent with H-1 NMR evidence for the epoxidic end group. The dehydration reaction mimics the metabolic conversion. 9-cis Pyrrhoxanthin acetate and anhydroperidinin acetate, obtained as by-products in the POCl3-treatment, were characterized by spectrometry.