An in situ single-pass perfusion model for assessing absorption across the intestinal mucosa of the brushtail possum

Abstract

AIM: To develop an in situ animal model for assessing absorption of molecules across the intestinal mucosa of possums.

METHODS: A surgical preparation was used to perfuse known concentrations of reference compounds (fluorescein and luteinising hormone-releasing hormone; LHRH) through measured sections of selected regions (jejunum, caecum, proximal colon) of the intestinal tract of 19 possums, over a 2-h period. Plasma concentrations of the compounds, which were perfused either with or without co-administration of a permeation enhancer (sodium deoxycholic acid; SDA), were determined in the perfusion effluent, peripheral and in some instances in the pre-hepatic circulation by spectrofluorometry (fluorescein) or radio-immunoassay (LHRH). Pharmacokinetic parameters of both compounds in the possum were determined over a period of up to 4 h in a further 30 animals (fluorescein, n=6; LHRH n=24), from their plasma profiles following intravenous (I/V) administration of a bolus dose.

RESULTS: In animals perfused with 25 mg/ml fluorescein (Perfusion Experiment (PE) 1), the mean plasma concentration was 2.8 (SE 0.12) µg/ml in post-hepatic blood samples. When possums were perfused with 2.5 mg/ml fluorescein and 7 µg/ml LHRH (PE 2), mean plasma concentrations were 0.3 (SE 0.01) and 7.8 (SE 1.64) µg/ml fluorescein and 0.1 (SE 0.02) and 6.3 (SE 0.45) ng/ml LHRH, in the absence and presence of permeation enhancer, respectively. There was a poor correlation between pre-hepatic and post-hepatic concentrations.

CONCLUSIONS: The single-pass in situ perfusion technique provided a useful model for investigating basic information on the absorption of biocontrol agents across the intestinal tract of possums, but had limitations that must be recognised.

KEY WORDS:

• Brushtail possum
• intestinal mucosa
• fluorescein
• LHRH
• absorption
• permeation enhancer

Acknowledgements

This study was funded by the MAFPolicy division of the New Zealand Ministry of Agriculture and Forestry. We thank Tim Manley (AgResearch Invermay) for assistance with the radioim-munoassay of plasma concentrations of LHRH.