The newest achievements in the application of adsorption and reversed‐phase high‐performance liquid chromatography (HPLC) used for the separation and quantitative determination of various pesticides is discussed in detail. A short overview of the basic principles of the technique is given, and the main theoretical equations employed for the characterization of retention processes and the quality of separation are presented. Instrumentation and troubleshooting are outlined to the extent to give a working knowledge for the solution of practical separation problems for the analysts. The sample preparation strategies such as liquid–liquid, supercritical fluid, solid‐phase and solid‐phase microextraction suitable for the enrichment, and prepurification of pesticides in complicated matrices are surveyed. The techniques employed for the measurement of pesticides classes such as organochlorine, organophosphorous, carbamate, urea, triazine, phenoxyalkyl acid, and pyrethroid pesticides are compiled and critically evaluated. The newest and the most adequate methods are described in detail, whereas the previous HPLC procedures are shortly referred.