Summary. This series of papers describes methods for measuring solute partition coefficients between membrane suspensions and water, and applies these methods to the system dimyristoyl lecithin:water. Knowledge of partition coefficients is relevant to analyzing permeability coefficients, probing membrane structure, and evaluating permeation models. The theoretical section of this first paper derives equations to correct measured partition coefficients for the effects of trapped water in the precipitate, tritium exchange with lipids or membranes, and nonsolvent water adjacent to membranes. The experimental section presents methods for measuring partition coefficients in liposomes of dimyristoyl lecithin, including techniques for minimizing effects of counter drift and quenching on double-label radioactive counting. These methods offer advantages in measuring membrane:water partition coefficients that are not high.A partition coefficient (K) is the ratio, at equilibrium, of a solute's concentrations in two immiscible phases between which solute can migrate. In the present series of papers we describe methods for measuring nonelectrolyte partition coefficients between membrane suspensions and water, and we apply these methods to aqueous suspensions of dimyristoyl lecithin liposomes. The methods should also be appficable to determining partition coefficients between biological membranes and water.These measurements are relevant to at least three general problems in membrane biology. First, the solute permeability coefficient (P), as commonly measured in biological membranes, is a black-box quantity which actually depends upon the solute's membrane:water K, the solute's diffusion coefficient within the membrane D, the membrane thickness Xo, and the resistances r' and r" of the two membrane:water interfaces to the * These papers are dedicated to the memory of Aharon Katchalsky, admired teacher and friend.