Black cohosh [Actaea racemosa L.; Cimicifuga racemosa (L.) Nutt.] is a perennial herb native to North America that is commonly used for the treatment of menopausal symptoms. The plant is almost exclusively harvested from the wild and is being threatened by overharvesting in some regions. As demand for this plant continues to increase, the potential for profitable cultivation of this species is becoming realistic. Little is known about the effect of various cultivation practices, soils, environments, and harvest times on the multitude of phytochemicals that occur in black cohosh. Furthermore, although the rhizome is the organ that is traditionally consumed, other tissues also contain various quantities of important phytochemicals, but this has not been well documented. The objectives of this study, therefore, were to ascertain any environmental effects on the production of two representative phytochemicals (23-epi-26-deoxyactein and cimiracemoside A) and to elucidate any season-long patterns or variations in the production of these compounds within five black cohosh tissues (leaf, rachis, rhizome, root, and inflorescence). All black cohosh tissues contained 23-epi-26-deoxyactein with substantially more, as a percentage of dry weight, detected in inflorescence (28,582 to 41,354 mg·kg−1) and leaf (8250 to 16,799 mg·kg−1) compared with rhizome (2688 to 4094 mg·kg−1), and all tissues experienced a linear season-long decrease in occurrence of this compound. Cimiracemoside A was not detected in leaf tissues. The highest levels were found in rhizome (677 to 1138 mg·kg−1) and root (598 to 1281 mg·kg−1), which likewise experienced a significant season-long decrease in this compound, whereas levels in the rachis (0 to 462 mg·kg−1) increased over time. In general, environmental factors did not affect production of either compound. Varying seasonal patterns in phytochemical production, combined with differences in phytochemical content among plant tissues, point to the potential for more targeted horticultural production of these and other medicinal compounds within black cohosh.