Decline in assayable progesterone in bovine plasma: effect of time, temperature, anticoagulant, and presence of blood cells.

Jugular vein blood samples collected from 5 Holstein-Friesian cows during the luteal phase of the estrous cycle were incubated at 4 C, 22 C, and 37 C for 0, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours. Blood samples were centrifugated after each incubation period, and plasma was frozen for later progesterone assay. Mean progesterone concentration in blood samples incubated at 4 C, 22 C, and 37 C decreased from 5.7 ng/ml at 0 hour to minimum concentrations of 3.5 ng/ml at 12 hours, 0.9 ng/ml at 12 hours, and 1.0 ng/ml at 8 hours, respectively. The first progesterone concentration which was significantly different from the initial value (0 hour) was at the 8-hour incubation period for blood samples incubated at 4 C and at the 1-hour incubation period for blood samples incubated at 22 C and 37 C. Progesterone concentration decreased similarly when blood samples were collected into heparinized tubes or EDTA tubes and incubated at 22 C for 0, 6, and 12 hours. Incubating harvested plasma or serum for 0 to 6 hours at 37 C had no effect on progesterone concentrations (7.9 and 8.2 ng/ml for plasma, and 6.9 and 6.6 ng/ml for serum.