Abstract. The aim of this study was to investigate whether the interstitial colloid osmotic pressure (COP i ) of peritoneum is of importance for peritoneal fluid reabsorption during peritoneal dialysis (PD). For testing this hypothesis, a method to isolate interstitial fluid from the peritoneal membrane and to measure the interstitial COP i in the normal peritoneum and directly after the initiation of PD needed to be developed and validated. Eighteen female rats were anesthetized subcutaneously in the neck region with fentanyl-midazolam (1:1). Nylon wicks were implanted postmortem by means of a plastic catheter in the tissue just underneath the peritoneal surface. The characteristics of this fluid were compared with that isolated from wicks that were implanted in intermuscular spaces in hindlimb muscle and back subcutis. All wicks were removed after 20 min and centrifuged. The wick fluid was collected and analyzed in a colloid osmometer constructed for submicroliter samples, and interstitial fluid COP was compared with that of plasma. PD was initiated by injecting 20 ml of 3.86% glucose-containing PD fluid (Dianeal) into the peritoneal cavity, over a dwell time of 4 h. Control rats received no PD. The ratio of COP i to that of plasma (COP p ) during control was 0.65 Ϯ 0.05 in peritoneum, 0.53 Ϯ 0.04 in muscle, and 0.59 Ϯ 0.05 in skin. After a PD dwell, the ratio was 0.29 Ϯ 0.03 in peritoneum, 0.54 Ϯ 0.08 in muscle, and 0.41 Ϯ 0.06 in skin. Thus, the COP ratio in peritoneum fell by 55% (P ϭ 0.014) and in skin by 30% (P ϭ 0.03), whereas the COP ratio in muscle was unchanged. The results suggest that acute PD results in a marked fall of the COP i in the peritoneal membrane, shifting the Starling equilibrium toward an absorptive state. The effect was restricted to the peritoneal membrane and, to some extent, the skin. It is speculated that the increase in peritoneal hydrostatic pressure after PD causes an increase in interstitial tissue volume, with primarily a dilution of interstitial colloids.