Headspace microextraction has already been established as the method of choice for analyzing volatiles blended in complex matrices, such as environmental, food and biological samples. The modern trend of analytical chemistry for 'going small' has led to the successful development of various sorbing materials and microextraction techniques. As it is anticipated, microextraction is usually combined with powerful separation and optical techniques permitting enhanced recoveries of analytes, selectivity and sensitivity. In addition, derivatization reactions are often employed for improved detectability of several classes of compounds. Volatile compounds of biological significance are key substances due to the fact that they may constitute a characteristic of the status of the organism. A closer look at the biological applications of the headspace microextraction techniques (solid-phase and single drop microextraction) is the primary aim of this review. The variability of biological samples and analytes are considered primarily, while derivatization and optimization strategies are also discussed.