Eastern collared lizards (Crotaphytus collaris) are growing in popularity as exotic pets in the United States and worldwide. However, clinical biochemistry reference data to support the interpretation of health and disease for this species are lacking. This study evaluated 87 apparently healthy eastern collared lizards (C. collaris). Blood samples were collected from the ventral coccygeal vein and assayed via an Abaxis VetScan VS2 analyzer and avian/reptile profile plus rotor. Although lizard weight and snout–vent length (SVL) were normally distributed, most biochemical analytes, except albumin and total protein, were not. Many analytes were affected, albeit slightly, by lizard age, SVL, body condition score, gravidity and/or recent oviposition, sex, health status, and color and locality. However, except for calcium, phosphorous, and albumin for gravid or immediately postoviposition females, biochemical values remained within the generated reference interval. Limitations of this study included that the VetScan VS2 avian/reptile profile plus rotor was unable to successfully provide values for bile acids for most of these apparently healthy lizards. When compared to biochemistry analyte values of lizards from the suborder Iguania from the western hemisphere, eastern collared lizard biochemistries were similar for some analytes, but a relatively increased plasma glucose and uric acid occurs in this species, which could affect the diagnosis of clinical disease or other health abnormalities.