Monitoring the misuse of drugs and the abuse of substances and methods potentially or evidently improving athletic performance by analytical chemistry strategies is one of the main pillars of modern anti-doping efforts. Owing to the continuously growing knowledge in medicine, pharmacology, and (bio)chemistry, new chemical entities are frequently established and developed, various of which present a temptation for sportsmen and women due to assumed/attributed beneficial effects of such substances and preparations on, for example, endurance, strength, and regeneration. By means of new technologies, expanded existing test protocols, new insights into metabolism, distribution, and elimination of compounds prohibited by the World Anti-Doping Agency (WADA), analytical assays have been further improved in agreement with the content of the 2013 Prohibited List. In this annual banned-substance review, literature concerning human sports drug testing that was published between October 2012 and September 2013 is summarized and reviewed with particular emphasis on analytical approaches and their contribution to enhanced doping controls.