Über die Carotinoide von Hämatococcus pluvialis. III .(Carotinoide der Süßwasseralgen. IX. Teil)

Abstract

The pure culture of H. pluvialis used was Strain 60 from the Pflanzenphysiol. Inst., U. Prag. From Apr. 30 to June 11, 1943, >900 100- to 300-ml. flasks of nutrient soln. were inoculated with the culture. The nutrient soln. was that used previously (Hoppe-Seyler''s Zeitschr. Physiol. Chem. 252: 225. 1938). In Oct. the algae were harvested by pouring the contents of a number of flasks into a large sedimentation cylinder. After a few hrs., the clear supernatant soln. was decanted, the red algae mass mixed with sand and filtered over sand on Buchner funnels. The mixture was dried in vacuo, and after subtracting the weight of sand used, 118.5 g. of dry algae substance was obtained from 130.2 1. of nutrient soln. The substance was ground with acetone in a Block-Rosetti ball mill, the mixture filtered and the acetone extract concd. under N2 in vacuo. The residue was extracted with benzine (b.p. 70[degree]-80[degree]). This soln. was chro-matographed on a column of CaCO3 and fractions containing astaxanthin esters A, B and C obtained. To effect purification, the esters were repeatedly chromatographed on columns of CaCO3, powdered sugar and sucrose. The esters were saponified with saturated methyl alcoholic KOH at room temp., and astacin obtained from the 3 esters. Esters A and C contained fatty acids with the composition C16H30O2, while ester B contained the fatty acid C18H34O2. The absorption maxima for esters A, B and C in benzine, were, respectively: 485, 452 m[mu]; 478, 448 m[mu]; 485, 452 m[mu]. The extracts also contained the pigments aphanin, aphanicin and [beta]-carotene with their absorption maxima in benzine being, respectively: 494, 460 m[mu].; 495, 462 m[mu]; 485, 452 and 424 m[mu].