Different wavelengths of sunlight either drive or inhibit macroalgal production. Ultraviolet radiation (UVR) effectively disrupts photosynthesis, but since UVR is rapidly absorbed in coastal waters, macroalgal photoinhibition and tolerance to UVR depend on the depth of attachment and acclimation state of the individual. The inhibition response to UVR is quantified with a biological weighting function (BWF), a spectrum of empirically derived weights that link irradiance at a specific wavelength to overall biological effect. We determined BWFs for shallow (0 m, mean low water [MLW]) and deep (10 m) Laminaria hyperborea (Gunnerus) Foslie collected off the island of Finnøy, Norway. For each replicate sporophyte, we concurrently measured both O2 evolution and 13C uptake in 48 different light treatments, which varied in UV spectral composition and irradiance. The relative shape of the kelp BWF was most similar to that of a land plant, and the absolute spectral weightings and sensitivity were typically less than phytoplankton, particularly in the ultraviolet radiation A (UVA) region. Differences in BWFs between O2 and 13C photosynthesis and between shallow (high light) and deep (low light) kelp were also most significant in the UVA. Because of its greater contribution to total incident irradiance, UVA was more important to daily loss of production in kelp than ultraviolet radiation B (UVB). Photosynthetic quotient (PQ) also decreased with increased UVR stress, and the magnitude of PQ decline was greater in deepwater kelp. Significantly, BWFs assist in the comparison of biological responses to experimental light sources versus in situ sunlight and are critical to quantifying kelp production in a changing irradiance environment.