On the basis of the physiopathological mechanisms involved in the brain damage related to hypoxic-ischemic encephalopathy of the newborn, a number of metabolic parameters have been studied with the aim to provide an early and reliable marker of tissue injury for diagnostic and prognostic purpose. Some authors have even suggested that biochemical indicators may be more effective than the results of clinical, Apgar score, pH in cord blood, electroencephalographic and neuroimaging data. In the present review, we discuss the diagnostic and prognostic value of a series of biochemical parameters, with special reference to the diagnosis of neonatal hypoxic-ischemic encephalopathy. For methodological purpose the exposition is structured in the following sections: creatine kinase isoenzymes; lactate; lactate dehydrogenase, aspartate aminotransferase and hydroxibutirate dehydrogenase; excitatory amino acids; glial fibrillary acidic protein; cytokines; neuron-specific enolase; oxypurines; cyclic adenosine monophosphate; and others. The current role of the above mentioned biochemical parameters as predictors of brain damage and the future perspectives on this topic are discussed.