Effect of anticoagulants and sampling time on results of progesterone determination in canine blood samples

The concentration of plasma progesterone was measured by ELISA, in serum and samples prepared with three different anticoagulant agents - namely ethylenediaminetetraacetic acid (EDTA), heparine and sodium fluoride oxalate potassium(NaFK). Forty clinically healthy bitches were selected based on the signs of pro-oestrus or oestrus. Values of progesterone concentration were significantly higher in serum than in EDTA-plasma (p < 0.0005); heparin-plasma (p < 0.05) and NaFK-plasma (p < 0.005). During pro-oestrus and oestrus until the time of ovulation, progesterone exhibited a conspicuous and statistically verified diurnal pattern (p < 0.05), its serum concentration being higher during 6.00-7.00 p.m. than 8.00-9.00 a.m. By the time of ovulation tendency of higher p.m. progesterone level reverses and from this point on the a.m. progesterone concentration is higher. The results of these experiments indicate that the concentration of canine progesterone assayed with ELISA may be affected by the time of collection and the method of preservation used.