Bioanalyses and pharmacokinetics of nafronyl in the dog

Improved specific and sensitive reverse-phase HPLC assays of nafronyl (I) and its acidic metabolite and hydrolysis product (II) in biological fluids were developed with sensitivities of 3-6 ng/mL using fluorometric detection with 225 nm excitation and 330 nm emission wavelengths. There were no significant differences in the stabilities and assays of I and II in plasma obtained using heparin, citrate phosphate dextrose solution, EDTA, citrate, or oxalate as anticoagulant. Inordinately high membrane binding did not permit the quantification of the high plasma protein binding of I by ultrafiltration; its instability precluded the use of equilibrium dialysis. Plasma protein binding of II by ultrafiltration was 76.4% and was not concentration dependent. The apparent red blood cell-plasma partition coefficients for I and II were 2.00 and 0.49, respectively, with almost all anticoagulants; the red blood cell-plasma water partition coefficient for II was 2.08 when corrected for plasma protein binding. Thus, both I and II had erythrocyte binding sites in addition to simple volume partitioning. Only heparin-treated blood gave anomalously low erythrocyte-plasma partition coefficients, indicating that heparin inhibited the partitioning of I and II into red blood cells from plasma water. The total body clearance of nafronyl (I) referenced to total plasma concentration [1295 +/- 65 (SEM) mL/min] was dose independent (35-70-mg range) and showed biphasic plasma half-lives (intravenous) of 12 and 100 min. Only 34% of the nafronyl appears as systemically circulating II in the plasma. Apparent volumes of distribution similarly referenced were 39.8 and 163 L for the central compartment and total body, respectively. Renal clearances referenced to total plasma concentration were 8.3 and 0.18 mL/m for I and II, respectively. The respective total urinary excretions of I, II, and the glucuronide of II (III) were 0.48, 0.021, and 0.32% of the administered intravenous doses. The respective total urinary excretions of I, II, and III for a bile-cannulated dog were 0.005, 0.16, and 0.40%. The total body clearance of intravenously administered II was 225 mL/min, with a renal clearance of 0.057 mL/min referenced to total plasma concentration. The respective total urinary excretions of II and III were 0.027 and 0.44% of the intravenous dose of II. Respective plasma half-lives of II (intravenous) were 2.5, 10.9, and 225 min. The apparent volume of distribution referenced to total plasma concentration was 2.2 L (9.1 L referenced to plasma water concentration). The apparent overall volume of distribution referenced to plasma concentration was 73 L.(ABSTRACT TRUNCATED AT 400 WORDS)