The effect of irradiance over the range 5 to 70 μmol photon m-2 s-1 on production of antibiotically active metabolites was assessed for male and female gametophytes and tetrasporophytes of the red alga Sphyridia filamentosa (Wulfen) Harvey in Hooker in culture. Whole-algal extracts and ten recognizable TLC-separable zones were assayed against five human microorganisms pathogenic to humans. For all experimental irradiance conditions, the ten TLC zones displayed activity against four of the microorganisms. The maximum number of TLC zones with activity under any of the culture conditions was six each for male and female Spyridia at 70 μmol photon m-2 s-1. Small changes in irradiance resulted both in different activities against specific microorganisms and degree of activity. The fact that every TLC zone showed differing activities at different light conditions or when extracted from different life history stages strongly suggests the presence of multiple antibiotic principals in individual TLC zones.