The working group “Analyses in Biological Materials” of the German Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area (MAK Commission) describes the current status of headspace-gas chromatography with respect to its potential applications in human biomonitoring. Particular focus is given to the review and discussion of newly developed methods for headspace sample collection as well as analyte enrichment. The article gives an overview on internationally published headspace methods for the matrices urine, blood, serum and plasma, existing assessment values for headspace parameters, background exposure levels in the non-occupationally exposed general population as well as half-lives of the most prominent hazardous substances measurable by headspace methods. In addition, critical requirements for and possible pitfalls of the preanalytical phase and of the calibration of headspace analyses are also discussed. The review shows that headspace methods have been continuously improved in recent decades and thus continue to make an important contribution to human biomonitoring of occupational and environmental exposure to volatile hazardous compounds.