Plasma biochemical profiles were studied in 112 mature (3 to 5-year-old) healthy cattle comprised of 61 Thai indigenous and 51 Simmental x Brahman crossbred male and cyclic female cattle at Nongkwang (Central Thailand) Livestock Research and Breeding Center, Thailand. Data were analysed for the effect of breed and sex. The results showed that the plasma glucose and gamma-glutamyl transferase (GGT) in the two breeds were significantly (P < 0.05) different. Furthermore, the urea, creatinine, albumin, total protein, aspartate amino transferase (AST), alanine amino transferase (ALT) and alkaline phosphatase (ALP) levels in Thai indigenous were significantly (P < 0.01) higher than in crossbred cattle. However, creatine kinase did not significantly differ in crossbred and indigenous animals. A sex difference was found in glucose level with male Thai indigenous having significantly higher levels (P < 0.05) than the other three groups. Plasma urea concentration in male crossbred cattle was lower than in the other groups (P < 0.05). Female crossbred cattle had significantly (P < 0.05) lower plasma creatinine levels than the other animals. Furthermore, levels of albumin in male and total protein in female crossbred were the lowest (P < 0.05) among the groups. The AST, ALT, ALP and GGT levels were significantly (P < 0.05) different between male and female. Female crossbred cattle had the lowest (P < 0.05) AST and GGT levels, whereas lowest (P < 0.05) ALT and ALP concentration was determined in male individuals of these breeds.