What Is High Performance Liquid Chromatography (HPLC)?

High-performance liquid chromatography (HPLC; formerly referred to as high-pressure liquid chromatography), is a technique in analytical chemistry used to separate, identify, and quantify each component in a mixture. It relies on pumps to pass a pressurized liquid solvent containing the sample mixture through a column filled with a solid adsorbent material. Each component in the sample interacts slightly differently with the adsorbent material, causing different flow rates for the different components and leading to the separation of the components as they flow out the column.

What Is Ultra Performance Liquid Chromatography (UPLC Technology)?

In 2004, further advances in instrumentation and column technology were made to achieve very significant increases in resolution, speed, and sensitivity in liquid chromatography. Columns with smaller particles [1.7 micron] and instrumentation with specialized capabilities designed to deliver mobile phase at 15,000 psi [1,000 bar] were needed to achieve a new level of performance. A new system had to be holistically created to perform ultra-performance liquid chromatography, now known as UPLC technology. Basic research is being conducted today by scientists working with columns containing even smaller 1-microndiameter particles and instrumentation capable of performing at 100,000 psi [6,800 bar]. This provides a glimpse of what we may expect in the future.

What is High Performance Liquid Chromatography(HPLC)?

HPLC stands for High Performance Liquid Chromatography.
Its earlier name was High Pressure Liquid Chromatography because it involved use of liquid mobile phase requiring higher pressures than gases used in Gas Chromatography. The technique has found immense scope of applications in both academic and industrial laboratories requiring identification and quantification of mixtures of organic compounds.
Why not start with a short video?
Are you new to High Performance Liquid Chromatography? Simply spend about 2 to 3 min on the introductory video giving you an insight into what constitutes a HPLC system.  



Brief History and Definition :What Is HPLC (High Performance Liquid Chromatography)?

Liquid chromatography was defined in the early 1900s by the work of the Russian botanist, Mikhail S. Tswett. His pioneering studies focused on separating compounds [leaf pigments], extracted from plants using a solvent, in a column packed with particles. Tswett filled an open glass column with particles. Two specific materials that he found useful were powdered chalk [calcium carbonate] and alumina. He poured his sample [solvent extract of homogenized plant leaves] into the column and allowed it to pass into the particle bed. This was followed by pure solvent. As the sample passed down through the column by gravity, different colored bands could be seen separating because some components were moving faster than others. He related these separated, different-colored bands to the different compounds that were originally contained in the sample. He had created an analytical separation of these compounds based on the differing strength of each compound’s chemical attraction to the particles. The compounds that were more strongly attracted to the particles slowed down, while other compounds more strongly attracted to the solvent moved faster. This process can be described as follows: the compounds contained in the sample distribute, or partition differently between the moving solvent, called the mobile phase, and the particles, called the stationary phase. This causes each compound to move at a different speed, thus creating a separation of the compounds. Tswett coined the name chromatography [from the Greek words chroma, meaning color, and graph, meaning writing—literally, color writing] to describe his colorful experiment. [Curiously, the Russian name Tswett means color.] Today, liquid chromatography, in its various forms, has become one of the most powerful tools in analytical chemistry.

simple animation about High Perfermance Liquid Chromatography for beginners

simple animation about High Perfermance Liquid Chromatography for beginners https://www.youtube.com/embed/IUwRWn9pEdg

High performance liquid chromatography (HPLC)

High Performance Liquid Chromatography (HPLC) is a form of column chromatography that pumps a sample mixture or analyte in a solvent (known as the mobile phase) at high pressure through a column with chromatographic packing material (stationary phase). The sample is carried by a moving carrier gas stream of helium or nitrogen. HPLC has the ability to separate, and identify compounds that are present in any sample that can be dissolved in a liquid in trace concentrations as low as parts per trillion. Because of this versatility, HPLC is used in a variety of industrial and scientific applications, such as pharmaceutical, environmental, forensics, and chemicals.

Sample retention time will vary depending on the interaction between the stationary phase, the molecules being analyzed, and the solvent, or solvents used. As the sample passes through the column it interacts between the two phases at different rate, primarily due to different polarities in the analytes. Analytes that have the least amount of interaction with the stationary phase or the most amount of interaction with the mobile phase will exit the column faster.