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Column is the major component in all HPLC techniques. This is where the separation of the interested particles from a sample takes place. The mobile phase (solvent) is connected to one end of the column while waste is collected from the other end. Types of mobile phase used have a big effect on the retention. The main requirement for the mobile phase is that it has to dissolve the sample up to the concentration suitable for the detection. To keep the mobile phase moving through the column at steady flow with controlled pressure a pump is added in the flow path. Pump is designed to draw the solvent from the reservoir and deliver it to the column with required flow rate. An injector is added to introduce the sample in the flow path of the mobile phase prior to the column. It consists the liquid material of few microliters. It must not interfere or stop the mobile phase- maintaining the pressure. 

The sample mixture is added into the injector and it encounters the stationary phase on the column. Certain component in the mixture sample spends more time in the mobile phase meaning leaving the column early while other components stays longer in the column depending on the solvent used and the retention time between the components. This phase separates the sample mixture into separate components and as they go along the column they become more separated. A detector is introduced after the sample mixture passes through the column in order to detect the separated components. Detector is designed to be able to recognize some characteristics of sample components. The most commonly used detector is the UV (ultra-violet) detector, although different ranges of detectors are used in HPLC. Detector is also connected to a processor such as a computer to convert the results of the detector into the useable data. Peaks are recorded on the processor called chromatogram. Components spending longer time with mobile phase ends up having broader peak than the ones that spends less time. Different types of softwares can be used in the computer to record and analyze the data. Vacuum degassed is also included in HPLC system since all air needs to be removed from the mobile phase for optimum performance

High performance liquid chromatography is widely used in various industrial such as pharmaceutical, chemical and environmental. While it uses sample principles as thin layer chromatography or column chromatography. However, it is highly advanced form of column chromatography which analyte in a solvent (known as mobile phase). HPLC use high pressures of up to 400 atmospheres. It uses a very smaller particle size for the column packing material (known as stationary material) which provides a greater surface area for the interaction between stationary phase and mobile phases. HPLC is extremely sensitive and highly automated compares to column chromatography which uses detection methods. The sample is moved by moving carrier gas stream of nitrogen or helium. HPLC identify and sperate any compounds in a sample can be dissolved in a liquid. Separation of mixture depends on the different degree of retention of component in the column. The component in the column is determined by separation between the stationary phase and the liquid mobile phase. The sample retention time depends on the interaction between the stationary phase. Due to different polarities in the analytes, the sample interacts between two phases during the it moves through the column. The most amount of interaction with the mobile phase or the smallest amount of interaction with the stationary phase will leave the column faster. The detector identifies the separated compound bands when it elutes from the high-pressure column. The information which generate the chromatogram is delivered to computer from the detector. The mobile phase leaves the detector. It is wasted or collected. When the solvent is evaporated from the sample in Evaporative Light Scattering Detector (ELSD), nitrogen is used as a nebulization.