Madagascar's radiated tortoises (Astrochelys radiata) are critically endangered, threatened by illegal collection, and confiscated in alarming numbers in recent years. Robust population- and technique-specific hematology and biochemistry reference intervals are valuable yet heretofore missing tools for triage, rehabilitation, and reintroduction of confiscated radiated tortoises. We determined reference intervals in 120 previously confiscated, clinically healthy subadult radiated tortoises living under human care within their native habitat at the Tortoise Conservation Center (TCC). Specific analytes measured were manual packed cell volume, total solids, white blood cell (WBC) count and differentials, and biochemistry analytes using a point of care system. To evaluate the effects of different commonly used techniques on these analytes, we compared results between two venipuncture sites (subcarapacial sinus and brachial vein) and three different WBC quantification methods (Natt and Herrick, Leukopet, and slide estimate). Reference intervals were narrower for most analytes, and sodium and potassium were qualitatively higher in the TCC population compared to previously published values from radiated tortoises housed in North American institutions. Creatine kinase, aspartate aminotransferase, glucose and inorganic phosphorus were all significantly greater in brachial samples than in subcarapacial samples. There was poor agreement and evidence of constant and/or proportional bias between all WBC quantification methods. Differences based on time of sample collection were incidentally found in some analytes. These results highlight the need for considering technique, demographic, and environmental factors in creating and applying reference intervals, and contribute foundational knowledge for improving care of radiated tortoises throughout the confiscation-to-release pathway.