Stability and partitioning of closantel and rafoxanide in ruminal fluid of sheep

Abstract

The stability and the partitioning of closantel and rafoxanide in ruminal fluid (RF) was examined in vitro. Stability was evaluated in two studies in a ruminal fluid-artificial saliva (RF-AS) mixture containing either drug. Drug concentrations were measured in samples collected sequentially from four batches of RF-AS fortified with either closantel or rafoxanide in one study and in four separately incubated aliquots of a RF-AS mixture of each drug in the second study at the start and at various intervals during a 24 h incubation period. The viability of the in vitro RF-AS incubation model was validated by the presence of digoxin degradation (T1/2 of 39.1±13 h) and by the absence of significant time related differences (P>0.5) in volume of gas produced, pH and methylene blue reduction time of the RF-AS drug mixture. Partitioning of closantel and rafoxanide was determined by measuring the relative drug concentration of the fluid and particulate phases in RF fortified with either drug at different concentrations. Closantel and rafoxanide were shown to be stable in a RF-AS mixture and were not subjected to any significant biodegradation. An initial marked reduction in drug concentration measured in the RF-AS mixture during the first 2 h of incubation was attributed to the attachment of both drugs onto particulate matter. This was subsequently confirmed in the partitioning study. More than 80% of closantel and rafoxanide was shown to be associated with the particulate phase of RF.