High Performance Liquid Chromatography is an analytical instrument that Separates, identifies and quantifies elements in a sample. It is a widely used system in analytical chemistry and biochemistry fields. Basically, the system carries the sample using a solvent or mixture of solvents into the static phase, where separation of compounds happens. A sensor captures the separated compounds and signals are sent to the integrator to create a graphic visual.
HPLC Contains the components below:
- Mobile Stage – This is the solvent or generally a mixture of solvents Used to transfer the samples throughout the entire system. The solvents need to be miscible in the mix; else the immiscible solvents will lead to pressure build-up from the HPLC system. The ratios of each solvent component in the mobile phase has an effect on the separation of compounds in addition to analysis length.
- Pump or solvent delivery unit – This element is to deliver the mobile phase and samples throughout the system at a constant flow rate or pressure. Usually, for analytical purposes, HPLC pump is set to operate in constant flow rate.
- Injector Port or auto sampler – Analytical samples are introduced through this component. Samples introduced through injector interface need to be manually injected using a proper HPLC syringes. Auto sampler enables an analyst to load all of the samples to the HPLC system and the system will automatically pick the proper sample to inject at present problems.
- Stationary phase – Also called column. This part of the system is Really the core of separation. It is constructed of tightly packed material in a stainless steel column. Because of the compactness of the packed material, higher pressure are expected to pump or send solvents through the system, hence HPLC occasionally are expression as High what is chromatography. Since the samples flow through the column, the chemicals in the sample will interact concurrently with the static and mobile phase in a distinct manner to yield distinct elution time of each chemical. The aim of every analysis is to separate the summit of interest from other existing compounds.
- Detector – This unit finds the split compounds in the sample. There are various detectors utilizing different manner of detection such as ultra-violet, fluorescence, mass spectroscopy and refractive index.
- Integrator – Integrator turns the signs conveyed from the sensor into visual output called chromatograms. Nowadays integrators come in the shape of computer systems rather than the traditional ones that use paper charts.