The use of alcohol, despite it being a psychoactive substance, is widely accepted. This acceptance and the ease of access may lead to alcohol abuse and dependence. Misuse of alcohol is one of the leading causes of preventable illness, injury or death, has social consequences, and can be linked to various types of criminal activities. Upon monitoring alcohol consumption, the question is not necessarily solely whether or not someone is or was abstinent, but may also imply what kind of drinking behavior or patterns are displayed. To reveal the answer, a portfolio of different biomarkers for alcohol (ab)use has been established. A distinction is made between indirect and direct markers. Indirect biomarkers are proteins, enzymes, or cells of which the level or activity undergoes typical changes as a result of (excessive) alcohol intake [e.g., carbohydrate‐deficient transferrin (CDT)], while direct biomarkers are ethanol itself and molecules which are directly related to the metabolism of alcohol [e.g., ethyl glucuronide (EtG), ethyl sulphate (EtS), fatty acid ethyl esters (FAEE), and phosphatidylethanol (PEth)]. These direct biomarkers have the potential to differentiate between drinking behaviors because they allow a higher sensitivity and specificity, in particular for the detection of abstinence or any drinking, and because they can be measured in different matrices, with different detection windows. These assets open the possibilities for testing strategies based on a combination of different markers that can be used both in retrospective and prospective assessments of alcohol intake. This article is categorized under: Toxicology > Alcohol