Sphingolipids are a complex family of compounds that perform structural and regulatory functions for eukaryotes and some prokaryotes and viruses. They share a common structural feature, a so-called sphingoid base backbone that is biosynthesized by condensation of serine and a long-chain fatty acyl-coenzyme A (typically palmitoyl-CoA) and, after several additional reactions, produces ceramides (N-acyl-sphingosines). Ceramides are incorporated into more complex phosphosphingolipids (in mammals, to ceramide 1-phosphates, sphingomyelins, and ceramide phosphoethanolamines), glycosphingolipids with a single to dozens of complex carbohydrates (comprised of mainly glucose, galactose, N-acetylglucosamine, N-acetylgalactosamine, fucose, sialic acid, and glucuronic acid, for mammals), derivatives of some of these glycosphingolipids (such as sulfation to form sulfatides), and protein adducts. Atypical sphingolipids have been recently discovered where alanine or glycine is utilized in the initial step of sphingoid base biosynthesis, resulting in 1-deoxy-sphingolipids. Several diseases result from disruption of de novo sphingolipid biosynthesis by hereditary defects or environmental factors, and sphingolipid biosynthesis is abnormal in many diseases (including cancer), which is being explored in greater depth with the availability of new sphingolipidomic tools to understand better how they contribute to the disease etiology and/or might be potential targets for nutritional and pharmaceutical intervention.