Seaweed phytosterols are associated with potential health benefits, affording them and the seaweeds that produce them commercial interest. However, little is known about how their production is affected by the cultivation environment, limiting the efficiency with which these compounds can be exploited. Therefore, we performed a pilot study on the effect of nutrient availability and light stress on the sterol content of Saccharina latissima, a rapid growing brown alga of increasing interest in western mariculture. Individuals of S. latissima were subjected to a nutrient-replete and nutrient-depleted regime for 5 weeks, followed by the introduction of light-limited and light-saturated conditions in the sixth week; sampling occurred each week. No significant inter-treatment differences were found in the sterol content in week 1–5. However, significant intra-treatment differences were found in weeks 3–5 regardless of nutrient treatment, wherein the fucosterol, 24-methylenecholesterol, and squalene contents of both treatment groups were found to correlate inversely with photosynthetic performance. Factorial treatment of differential nutrient availability and light stress resulted in marked differences between the sterol content of all groups in week 6. Here, squalene and cycloartenol increased in concentration with increasing irradiance regardless of nutrient treatment. Concentrations of all other sterolic components increased with increasing irradiance and low nutrient conditions while decreasing or remaining unchanged with increasing irradiance and high nutrient conditions. Our data shows that within our cultivation conditions and time frame, the sterol content of S. latissima is unaffected by nutrient availability alone but changes with combined alterations in irradiance and nutrient availability.Graphical abstract