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Overview of the HPLC industry

Among the various technologies developed for chromatography, devices dedicated for molecular separation, called columns and high-performance pumps for delivering solvent at a stable flow rate, are some of the key components of chromatographs. As related technologies became more sophisticated, the system, commonly referred to as high-performance liquid chromatography, became referred to as LC. Nowadays, ultra high-performance liquid chromatography (UHPLC), capable of high-speed analysis, has also become more wide-spread.

Only compounds dissolved in solvents can be analyzed with HPLC. HPLC separates compounds dissolved in a liquid sample, and allows qualitative and quantitative analysis of what components and how much of each component are contained in the sample.

High-performance liquid chromatography (HPLC) is a broad analytical chemistry technique used to separate compounds in a chemical mixture. These separations utilize the pressure-driven flow of a mobile phase through a column packed with a stationary phase. The specific intermolecular interactions between the molecules of a sample and the packing material define their time on-column. Hence, different constituents of a sample are eluted at different times. Thereby, the separation of the sample ingredients is achieved. HPLC columns are mostly normal phase, reverse phase, and ion exchange columns. In the pharmaceutical industry mostly, reverse columns are used for analysis. This is a very vast and deep topic when going into detail about each type of column specification. The purpose of HPLC analysis of any drug is to confirm the identity of a drug, provide quantitative results, and monitor the progress of a disease. HPLC offers a fast, automated, and extremely precise technique for recognizing certain chemical components in a sample. High-performance liquid chromatography offers a quick and precise quantitative analysis. Trace components are pre-concentrated. Chromatography is based on ligand exchange. For example, in a medical setting, HPLC can be used to determine the contents and concentrations of substances in biological materials. This could include drug analysis of urine or detection of vitamin levels in blood serum. The Rf value, or retention factor, is a ratio used to describe the distance a component in a mixture travels in relation to the distance the solvent travels. It is calculated by dividing the distance a component travels by the distance the solvent travels. HPLC is used to quantitatively determine plasma levels of drugs and their metabolites. This is of special importance during the development of new drugs and for monitoring therapy. The measurement of concentrations of certain biogenic substances helps in diagnosis. The Rf value of a compound is equal to the distance traveled by the compound divided by the distance traveled by the solvent front (both measured from the origin). The purpose of HPLC analysis of any drugs is to confirm the identity of a drug and provide quantitative results and also to monitor the progress of the therapy of a disease. The ultraviolet (UV) or visible (VIS) detector is the most common in HPLC because it provides good stability, is simple to operate, and has a high sensitivity. Fixed wavelength detectors, variable wavelength detectors, and diode array detectors are the three types of UV/VIS detectors. In HPLC chromatography, the column is where the separation of the sample takes place. This is where the sample is passed through the stationary phase with the mobile phase, resulting in the separation of the sample components. A detector, or radiation transducer, is any device that converts an amount of radiation into some other measurable phenomenon. Ultimately, most of these other measurable phenomena will be tied to an electrical signal. There are two main types of detectors – photon detectors and thermal detectors. Some measure photons, particles of light, such as the visible light from stars, or the invisible X-rays are used to examine broken bones. Other particle detectors identify protons, neutrons and electrons – the particles that make up atoms – or even entire atoms. Some detectors are designed to detect antimatter.

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