The term neurosteroid was coined by Baulieu and colleagues in Paris towards the end of the last century to describe steroids which are synthesised in the central or peripheral nervous system [E.E. Baulieu, Psychoneuroendocrinology 23 (1998) 963-87]. This definition was restricted to side-chain "shortened" steroids with 21 carbon atoms or less, and excluded sterols and their carboxylic acids with an intact side-chain. By analogy, we now use the term neurosterol to describe C(27) sterols synthesised in the nervous system. In this review we discuss the biological importance of neurosterols, and how they are extracted, isolated, and analysed by GC-MS and LC-MS/MS, from brain and relevant body fluids. We present applications of methodology employed for analysis of specific sterols and comment on the relative merits of the methods employed. Finally, the importance of future in-depth "sterolomic" investigations of brain is highlighted.