The present work examines the usefulness of the combination of multivariate analyses and further one-way analysis of variance in the search for links between causes and effects in heterologous stimulation of spawning in fish. Stimulations with both 20 and 50 μg.kg–1 doses of [D Trp6] luteinizing hormone-releasing hormone analogue (LH-RHa) were effective in inducing and synchronizing spawning of red drum, Sciaenops ocellatus. Oocyte development monitoring using ovarian biopsy allowed treatment injections at different stages of maturation. The influence at stimulation time of female oocyte development state upon further development was shown. An interesting potential of second stimulation, leading to high quality spawns, was established and factors of success were identified. The best conditions for stimulation success were simultaneously late stage of oocyte development characterized by the beginning of oil droplet coalescence, high ovarian homogeneity quoted by the proportion of most developed oocytes in the biopsy sample, and low dose of hormone (20 μg.kg–1) for both first and second stimulations.