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HPLC And Its Variations



HPLC is the process used to identify components of complex mixtures. This process was launched in the year 1980 through many software and types of columns used in complex analysis. Application of HPLC is in many areas like an industrial application – environmental, food, pharmaceutical, R&D. The main instruments in an HPLC are solvent reservoir, pump, injector, column temperature control over, and lastly the integrator. Some of the leading manufacturers and makers of HPLC include Agilent, Waters, and HP. The best part about HPLC is that it can work with several detectors and software.

Types of HPLC used:

Ultraviolet: UV detectors are the most affordable and common type of HPLC detector used in the field. This detector has the ability to respond to substances and components that absorb light. Their main application is in the biomedical and pharmaceutical industry to separate the recognize the active components of a complex mixture. The Ultraviolet detectors are very efficient as they can test substances which are low in chromophores.

Fluorescence: This is the most sensitive HPLC detector mainly used in the LC or liquid chromatography. These detectors only sense the substances which emit light. They are most commonly used in tracing analysis in the field of environmental science. But they are very sensitive so the response is linear over any limited concentrated range.
Mass Spectrometry: This detector is the best amongst the lot. The mass spectrometry together with HPLC is called HPLC-MS. These detectors are the most powerful ones especially relevant for use in pharmaceutical labs and research and development. They have the ability to analyze and provide molecular identity to a varied range of components.
Refractive Index Detection: The RI or refractive index applies a monochromator and is supposedly the least sensitive LC detector. They are very efficient in detecting components that are non-ionic and don’t absorb UV light and fluoresce. Components that are commonly detected with refractive index detection include alcohol, sugar, polymers, and fatty acids.

A laboratory process used in the application of separating mixtures by absorbing the individual components is called Chromatography. Many forms of chromatography exist including paper chromatography, column, and thin-layer chromatography. HPLC or high-performance liquid chromatography is popular of column chromatography. Refurbished HPLC  is commonly used for recognizing an unknown mixture through its organic chemical components. They are also used for removing impurities from any single component substance.