High Performance Liquid Chromatography instruments have emerged as one of the most useful tools for industries such as life sciences, pharmaceuticals, petrochemicals, pesticides and lot more. HPLC devices and systems are considered to be a brilliant invention that has helped mankind get better products and services. Using HPLC systems, researchers and scientists can easily segregate molecules from mixtures which would otherwise have been impossible. HPLC systems help separate molecules in the most accurate manner than Gas chromatography devices. Let us understand about the main parts which comprises of a full-fledged HPLC system.
The main function of the HPLC Pump is to force a liquid (which is also known as the mobile phase) via a liquid chromatograph at a specific flow rate, expressed in milliliters per min (mL/min). The solvent is passed through the column using high pressure. Pumps are a vital part of HPLC systems as these help maintain flow rates and high pressure. The flow rate, in addition to how a molecule reacts with the beads in the column, determines how fast a particular chemical will migrate through the column. The migration time is used to identify individual chemicals.
The injector does the task of introducing the liquid sample into the flow stream of the mobile phase. It is mainly used to place a sample into the solvent stream flowing through the column. The Injector is specifically designed to place a liquid sample into the solvent stream with as little perturbation as possible.
Column is considered to be the most important part of the chromatograph which separates the sample components of interest using various physical and chemical parameters. It comprises of a metal housing in a tube shape which is packed with tiny beads that have an affinity for the chemicals that are being analyzed. The chemicals interact with the beads as soon as they flow through the column in a solvent. The minute molecular differences in similar compounds cause them to migrate through the column at different velocities, and separate them according to migration time through the column. The column also has a filter which lets the solvent to flow through but keeps the beads from leaving the column.
It does the work of detecting individual molecules that come out from the column. There are different varieties of detectors that are use for deciding when the chemical should exit the column (the main part of the HPLC system). For instance, the optical HPLC detectors use a beam of light to determine when a particular chemical passes out of the column. When the solvent exits the column, it gets passed through the detector’s light path. Next, there are Ultraviolet and visible light detectors which operate by recording the absorption of light caused by the chemical of interest. Now, the change in absorption caused by the chemical indicates the presence of the chemical. Other detectors that are available are fluorescence and electrochemical detectors.
This is a computer that helps controls all the modules of the HPLC instrument, takes the signal from the detector and uses it to determine the time of elution (retention time) of the sample components (qualitative analysis) and the amount of sample (quantitative analysis).
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