*Article* **The Pharmacokinetics of Medetomidine Administered Subcutaneously during Isoflurane Anaesthesia in Sprague-Dawley Rats**

### **Leila T. Kint 1, Bhedita J. Seewoo 2,3,4, Timothy H. Hyndman 5, Michael W. Clarke 6, Scott H. Edwards 7, Jennifer Rodger 3,4, Kirk W. Feindel 2,8 and Gabrielle C. Musk 1,9,\***



**\*** Correspondence: gabrielle.musk@uwa.edu.au

Received: 28 May 2020; Accepted: 17 June 2020; Published: 18 June 2020

**Simple Summary:** Rodents, including rats, are used as animal models for research investigating neurological diseases in humans. To enable this research the animals are anaesthetized to facilitate imaging of the brain, but the anaesthetic drugs impact the results of the research. To minimize the variation between studies anaesthetic protocols should be similar. A common anaesthetic regime is the combination of two drugs (medetomidine and isoflurane); however, there is much variation in the doses of these drugs and the way in which they are administered. To provide some evidence to facilitate the standardization of anaesthetic protocols this study was performed to elucidate the details of what the body does to these drugs when they are administered in a certain way. Three groups of rats were studied to determine the desired dose of medetomidine when isoflurane is used at a low dose (approximately 0.5%). The results of the study are an evidence-based suggestion for medetomidine and isoflurane anaesthesia during functional magnetic resonance imaging (fMRI) studies.

**Abstract:** Anaesthetic protocols involving the combined use of a sedative agent, medetomidine, and an anaesthetic agent, isoflurane, are increasingly being used in functional magnetic resonance imaging (fMRI) studies of the rodent brain. Despite the popularity of this combination, a standardised protocol for the combined use of medetomidine and isoflurane has not been established, resulting in inconsistencies in the reported use of these drugs. This study investigated the pharmacokinetic detail required to standardise the use of medetomidine and isoflurane in rat brain fMRI studies. Using mass spectrometry, serum concentrations of medetomidine were determined in Sprague-Dawley rats during medetomidine and isoflurane anaesthesia. The serum concentration of medetomidine for administration with 0.5% (vapouriser setting) isoflurane was found to be 14.4 ng/mL (±3.0 ng/mL). The data suggests that a steady state serum concentration of medetomidine when administered with 0.5% (vapouriser setting) isoflurane can be achieved with an initial subcutaneous (SC) dose of 0.12 mg/kg of medetomidine followed by a 0.08 mg/kg/h SC infusion of medetomidine. Consideration of these results for future studies will facilitate standardisation of medetomidine and isoflurane anaesthetic protocols during fMRI data acquisition.

**Keywords:** functional MRI; rat anaesthesia; refinement
