Pitfalls and Related Improvements of In Vivo Gas Uptake Pharmacokinetic Experimental Systems

Abstract

Gas uptake chamber studies have been widely used to study inhalation pharmacokinetics (PKs) in rodents, often for the ultimate purpose of developing physiologically-based pharmacokinetic (PBPK) models that can be used to describe human PKs and to support risk assessment for the chemical. In the course of our studies of gasoline PKs, we revisited several important issues heretofore not thoroughly addressed. Here, we report several refinements which will significantly improve future studies with this type of system, relating to the understanding of loss rates, the importance of carbon dioxide removal, and sampling of blood and chamber air at the same time. Losses of chemicals in gas uptake systems consist of leakage, adsorption to system components, and adsorption to the hair and skin (fur) of experimental animals. The loss rates were experimentally determined for a series of chemicals and mixtures including n-hexane, benzene, toluene, ethylbenzene, o-xylene, gasoline, and other gasoline components. The rate of loss to the animals' fur was similar to loss rates to system components and involved absorption to both hair and skin. Most of the absorption to fur was reversible when the chamber concentration was low enough. The amount of chemical that desorbed from the animal after an experiment was significant when compared to the amount of chemical in the chamber at the end of a gas uptake experiment, indicating that the rate of decline in concentrations can be influenced by a decrease in the fur absorption rate or desorption of chemicals. A modified gas uptake system design is described in which a steel ring improved the connections to an autosampler and allowed insertion of probes to monitor gases, such as carbon dioxide (CO₂), in the chamber. When CO₂ absorbent efficiency was inadequate, CO₂ concentrations rose to levels that significantly affected the animals' ventilation rate. Using a real-time CO₂ probe, an absorbent system was developed that adequately controlled CO₂ levels in the chamber. Attention to details of absorptive loss and CO₂ scrubbing can improve the reliability of kinetic constants inferred from closed chamber studies. We then describe a method for extending gas uptake experiments by simultaneously collecting blood to be analyzed for chemicals and/or metabolites.