HPLC is one of the most common separation techniques in many sciences. This paper first refers to the history of chromatography, its terms and relationships, and then to the careful selection of instrumental and laboratory parameters in order to perform an efficient separation. Parameters such as the choice of column type and the choice of moving phase, which depends on the polarity of the analyte, or the selection of a suitable detector, which depends on the structure and chemical properties of the sample, and finally the quantitative and qualitative analysis of the chromatogram. High performance liquid chromatography is a convenient way to separate, measure, and determine the type of material. This technique has many applications in various sciences in combination with other methods and advanced detectors such as mass spectrometers. Moving and stationary phase selection, detector selection, and moving phase current velocity are among the key factors in setting up a suitable HPLC method.