A Basic Guide To High Performance Liquid Chromatography

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In recent years Liquid Chromatography (LC) has assumed much importance for industries like pharmaceuticals, R&D, life sciences and food & beverages. The market for Liquid Chromatography is increasing rapidly at never seen before rates. Well, LC is a separation technique which involves injecting a small amount of liquid sample into a tube that is filled with porous particles. The individual components are then transported with the packed tube. hplc components have been widely used across the globe because of the many advantages these offer, the foremost being higher efficiency in terms of separating complex technical mixtures. In recent years, HPLC is also referred to as UPLC, RRLC, UFLC, and RSLC. Sometimes HPLC is also known as High Pressure Liquid Chromatography.

What is High Performance Liquid Chromatography?
HPLC is an analytical technique and is a vital part of Chromatography. The HPLC technique involves the usage of a liquid mobile phase for segregating the components of a chemical blend. The stationary ...
... phase can either be a liquid or a solid phase. The components initially are dissolved in a solvent. Next, these are then faced via a chromatographic column under a high pressure. Now, in the column, the blend is separated into different components.

Here it is important to remember that the amount of resolution is crucial which is dependent upon the extent of interaction between the solute components and the stationary phase. The stationary phase is termed as the immobile packing material in the column. The interaction of the solute with mobile and stationary phases can be manipulated through different choices of both solvents and stationary phases. Therefore, HPLC is a highly efficient technique which is not found in other chromatographic systems. It has the versatility of separating a wide range of chemical mixtures.

Major parts of HPLC Instruments
Pump – The main function of the 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).

Injector - The injector does the task of introducing the liquid sample into the flow stream of the mobile phase.

Column – Column is known as the main part of the chromatograph which separates the sample components of interest using various physical and chemical parameters.

Detector – It does the work of detecting individual molecules that come out from the column.

Data System – This is computer which 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).

Application of HPLC
HPLC instruments are mainly used for segregating non-volatile biological and chemical compounds. Non volatile compounds include –

Polymers (e.g. polystyrene, polyethylene)
Pharmaceuticals like aspirin, ibuprofen, or acetaminophen (Tylenol)
Salts like sodium chloride and potassium phosphate
Ginseng, plant extracts, and herbal medicines
Trinitrotoluene (TNT), enzymes
Egg white or blood protein
Asphalt or motor oil