Equine plasma luteinizing hormone (LH) possesses both biological (in vitro bioassay, B) and immunological (radioimmunoassay, I) activities and the ratio of B:I varies with stage of the oestrous cycle. To estimate the contribution made by pituitary secretion and peripheral metabolism to changes in the B:I ratio, pituitary venous effluent and circulating plasma from 5 dioestrous and 2 oestrous mares were analyzed using both an in vitro bioassay and a radioimmunoassay. During dioestrus, LH was released in a pulsatile fashion with a frequency of 1.4 pulses/24 h and a pulse duration of 20-40 min (centrally) or 2-4 h (peripherally). Between pulses, further secretion of LH from the pituitary was undetectable. During spontaneous pulses, the B:I ratio increased as much as 2- to 3-fold and remained elevated for at least 1 h. A low dose of 10 ng/kg bodyweight (bwt) of the GnRH agonist Buserelin provoked similar changes, unless the exogenous stimulus was close to an endogenous LH discharge. A high dose (50 micrograms) of Buserelin reduced the B:I ratio significantly, in spite of a massive LH release. Samples taken from mares during oestrus showed constantly elevated B:I ratios, a consequence of much greater LH pulse frequency.