Solid-phase microextraction (SPME) is a simple, fast, and efficient sample preparation technique that eliminates the need for solvents and integrates sampling, isolation, and enrichment in one step. In SPME, the extraction is based on the partitioning of the analyte between the organic phase on the fused-silica fiber or nanomaterials coated fiber and the matrix. Fibers or capillary tubes coated with an appropriate stationary phase are key players in the process. By controlling the polarity and thickness of the coating on fibers or tubes, sampling time, and other extraction parameters, an analyst can achieve very reliable, accurate, and highly quantifiable robust results at subfemtogram level, for gaseous to macromolecular target solutes, with a shorter analysis time compared to conventional extraction methods. This solvent-free technique requires small samples, saves sample preparation time as well as solvent purchase and disposal costs. Moreover, it is environmental friendly and poses low health hazard risk to the laboratory associates. Today, SPME has been integrated to gas chromatography (GC), gas chromatography–mass spectrometry (GC–MS), high-performance liquid chromatography (HPLC or LC), LC–MS, or capillary electrophoresis (CE) for analysis of complex mixtures of samples that include environmental, chemical, biochemical, biological, pharmaceuticals, clinical, biomedical, natural products, food abstracts, plant extracts, and organic compounds in media containing nanoparticles.