This section provides an overview for preparative hplcs as well as their applications and principles. Also, please take a look at the list of 10 preparative hplc manufacturers and their company rankings.
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A preparative HPLC is an instrument for collecting components separated by high-performance liquid chromatography (HPLC).
It can be used to purify and improve the purity of the main components and to collect trace amounts of impurities in a sample. Unlike other purification methods such as column purification or recrystallization, preparative HPLC can selectively collect each component separated on HPLC.
Since the principle of preparative HPLC is the same as that of normal HPLC analysis, it is possible to separate and purify each component by various characteristics of the compound, such as chemical structure, molecular weight, and steric structure, if an appropriate column is selected.
Moreover, by changing the column size and instrument configuration, preparative HPLC can change the amount of the target compound obtained from milligrams to kilograms.
Preparative HPLCs are characterized by the fact that each peak separated on HPLC can be obtained separately.
In the fields of organic chemistry and biochemistry, samples obtained by synthesis contain many trace components such as impurities and by-products in addition to the main components. However, it is difficult to selectively extract only one component by general purification methods such as column purification and recrystallization.
With preparative HPLCs, each peak separated by HPLC can be collected as a separate fraction, allowing selective extraction of the main component and impurities with high purity.
For example, the materials industry uses preparative HPLCs to evaluate the purity of materials and physical properties.
In the biological field, preparative HPLCs are used to purify proteins using size-exclusion chromatography (SEC) columns.
In the field of polymers, preparative HPLCs are used for purification of polymers using gel permeation chromatography columns (GPC), molecular weight fractionation, and characterization of individual components, sometimes utilizing the high resolution of HPLC to extract target compounds from natural product samples containing a variety of compounds.
The separation mechanism of preparative HPLCs are similar to that of conventional HPLC analysis.
The HPLC column is packed with a material called stationary phase, which has functional groups such as alkyl groups and hydroxyl groups modified on its surface. When a sample solution is injected into the column, each component of the sample is adsorbed to the column surface by the interaction between the sample and the stationary phase.
Since the magnitude of this interaction and adsorption depends on the chemical structure of each component, each component in the sample is separated as it is repeatedly adsorbed and desorbed in the column. Components that interact weakly with the stationary phase elute early from the column, while components that interact strongly elute late.
The type of interaction depends on the stationary phase of the column. For example, in the case of a stationary phase with a modified alkyl group, such as a C18 column, the separation is affected by the magnitude of the hydrophobic interaction between the sample and the stationary phase. Therefore, the more hydrophobic or carbon-rich a compound is, the slower it will elute.
In preparative HPLCs, a device called a fraction collector can be connected to the back of the HPLC system.
The fractions are divided by retention time, and can be collected in detail according to the elution behavior of the peaks.
The recycling preparative HPLCs system is equipped with a switching valve behind the column, allowing eluates that have passed through the column once to be passed through the same column again.
Generally, the longer the column, the better the HPLC resolution. Therefore, by passing the eluent through the column multiple times, it is possible to separate and recover components that could not be separated in a single column. However, due to the configuration of the device with the switching valve, it is necessary to prepare a dedicated device separate from the normal HPLC device.
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