Feasibility and effect of midazolam administration on blood analytes and for sedation of Passeriformes being collected in a larger study of genetic biodiversity was assessed. Midazolam (5.6±2.7 mg/kg) was administered intranasally prior to sampling, euthanasia, and specimen preparation of 104 passerine birds. Each bird was assessed for sedation score and then multiple analytes were determined from jugular blood samples using the i-STAT^® point of care analyzer at "bird side". Most birds were acceptably sedated, sedation became more pronounced as midazolam dose increased, and only a single bird died. Electrolyte concentrations and venous blood gas analytes were affected by midazolam administration while blood pH, packed cell volume, hemoglobin, and calculated hematocrit were not. Intranasal midazolam gives adequate sedation and is safe for short-term use in free-living Passeriformes. Based on venous blood analyte data, sedation of Passeriformes prior to handling appears to reduce stress but also produces venous blood gas differences consistent with hypoventilation relative to birds which were not given midazolam. Further study is recommended to investigate midazolam's continued use in free-living avian species. Studies should include safety, reversal and recovery, effect upon additional endogenous analytes, and compatibility with studies of ecology and toxicology associated with pollution or other environmental degradation in Passeriformes.