Investigation of globulin fractions by serum protein electrophoresis (SPE) is the first step towards evaluation of the proteome in the southern white rhinoceros (Ceratotherium simum simum). Furthermore, identification of changes in globulins in animals with poaching and other injuries can guide discovery of potentially useful biomarkers of inflammation. The aim of this study was to develop reference intervals for agarose gel SPE in healthy white rhinoceros and to compare these serum protein electrophoresis results to those from animals with tissue trauma. Reference intervals for total serum protein and agarose gel electrophoretic albumin and globulin fractions were generated using serum samples from 49 healthy free-ranging adult white rhinoceros. A standardised gating system together with identification of specific proteins by mass spectrometry aided in fraction identification. Six globulin fractions were identified: α1a, α1b, α2, β1, β2 and γ. Reference intervals were generated for total serum protein (76-111 g/L), albumin (10-27 g/L) and globulin fractions (α1a: 1.6-3.2 g/L; α1b: 1.7-3.6 g/L; α2: 16.1-26.6 g/L; β1: 6.6-18.2 g/L; β2: 11.8-30.4 g/L; γ: 10.4-23.1 g/L; albumin: globulin ratio: 0.12-0.39). Results were compared to those from 30 animals with various degrees and chronicities of tissue trauma. Wounded animals had lower concentrations of total serum protein, albumin, total globulin, α and β1 globulins, lower percentages of α2 and β1 globulins, and higher percentages of β2 and γ globulins. These protein changes are similar to those seen in human patients with wounds rather than classic acute phase or chronic inflammatory responses.