Adipocytes from white adipose tissue secrete cytokines and other bioactive proteins which are collectively termed adipokines. Adiposity has been linked with increased breast cancer risk as adipokines secreted by adipocytes significantly affect epithelial cells from which breast cancer arises. Measurement of extracellular adipokine concentrations that would be involved in signaling through mammary tissue is therefore of importance. In this work, microdialysis sampling was used to collect adipokines from the interstitial space of the mammary fat pad of female rats under isoflurane anesthesia. The adipokines CCL2 (MCP-1), leptin and IL-6 were quantified from dialysate samples and compared to total tissue concentrations surrounding the implanted probes. After three hours of microdialysis sampling at 1 μL min(-1), the respective median values for these adipokines in dialysate samples were approximately 175 pg mL(-1) (CCL2), 150 pg mL(-1) (IL-6) and 750 pg mL(-1) (leptin). Adipokine protein levels from dialysates were an order of magnitude lower than levels obtained directly from mammary tissue. However, the adipokine concentrations between excised tissue surrounding the microdialysis sampling probes and control tissue without implants did not differ. This work demonstrates the utility of microdialysis sampling to quantify mammary gland adipokine levels, with relevance to understanding mammary physiology.