Influence of Physicochemical Parameters and Perfusate Flow-Rate On the Distribution of Solutes in the Isolated-Perfused Rat Hindlimb Determined by the Impulse-Response Technique

The relationship between solute distribution, physicochemical properties, and tissue physiology was determined by the impulse-response (IR) technique and statistical moment analysis in the isolated perfused rat hindlimb. The concentration of bovine serum albumin (BSA; 2,4,7, and 7%, w/v), perfusate flow rate (4 and 8 mL/min), and solute physicochemical properties (lipophilicity, P; fraction unbound to protein, f(u); fraction ionized, f(i); and molecular weight MW) were varied to better understand the underlying determinants of solute distribution. An apparent low availability was found for a number of the solutes as a consequence of tissue sequestration. This low availability precludes the estimation of an apparent volume of distribution (V) for these solutes. The V of solute and tissue (skin, fat, and muscle) blood flow increased with perfusion flow rate (p< 0.01). The unbound distribution volume (V-u) of basic solutes was significantly linear with respect to P. Multiple linear regression analysis showed that the distribution volume of solute in tissue was significantly related to f(u) (p < 0.01), but not improved by including relationships to P, MW, and f(i). Data obtained with this IR technique yield results consistent with in vivo studies in terms of the importance of f(u) as a determinant of V. This work has shown that the estimations of solute V by the IR technique in a single-pass preparation are unreliable for solutes with a low availability due to apparent solute sequestration into tissue. The parameter V may also be affected by changes in the perfused limb physiology associated with the perfusion conditions used. The Ifs for lidocaine and diazepam vary with f(u) in accordance with deductions based on the results of steady-state studies