Characteristics of High-Performance Liquid Chromatography
High-performance liquid chromatography is based on classic liquid chromatography, with liquid as the mobile phase, and a high-pressure infusion system is used to pump a single solvent with different polarities or a mixed solvent in different proportions, buffer, and other mobile phases into the particles containing particles. The chromatographic column with the ultra-thin high-efficiency stationary phase, after the components in the column, are separated, enters the detector for detection, to realize the analysis of the sample.
The separation principle of High-performance liquid chromatography is: when the components dissolved in the mobile phase (mobile phase) pass through the stationary phase, the size and strength of the action (adsorption, distribution, exclusion, affinity) of the stationary phase (station phase) are different. The retention time in the stationary phase is different, so they flow out of the stationary phase successively. Also known as HPLC chromatography.
As an important branch of HPLC chromatography, high-performance liquid chromatography (HPLC) has become an important application of separation and analysis technology in the fields of chemistry, medicine, industry, agriculture, commodity inspection, and law inspection. Hawach supplies PFP HPLC Columns, SCX HPLC Columns, Polar-RP HPLC Columns, and other HPLC-Columns for your use. Now let’s take a look at the characteristics of HPLC.
Composition
Chromatographic column: Separate the substances.
Sampler: The samples are introduced into the chromatographic system.
High-pressure infusion pump: Drive the mobile phase and sample through the chromatographic separation column and detection system.
Detector: Changes in the concentration of the column effluent are converted into optical or electrical signals.
HPLC Column Characteristics
1. High sensitivity;
2. Small sample size;
3. Fast analysis speed;
4. High separation efficiency;
5. Wide application range;
6. Columns can be used repeatedly.
Silica Gel Characteristics of HPLC Column
Better reproducibility
Provides a better peak shape
Available in different particle sizes
Better strength of filler particles
Decreased activity of free silicon hydroxy groups
Types of High-Performance Liquid Chromatography
1. Ion chromatography;
2. Affinity chromatography;
3. Partition chromatography;
4. Adsorption chromatography;
5. Size exclusion chromatography;
6. Bonded-phase chromatography.
Classification of HPLC Column
The frequently used HPLC columns are packed with silica, which is divided into normal-phase columns and reversed-phase columns. Other packing columns are also available in polymer fillers and inorganic fillers.
1. Normal phase
The stationary phase for normal phase HPLC chromatography is typically silica gel and other bonded phase fillers with polar functional groups. As the silicon hydroxy group (SiOH) or other polar groups on the surface of the silica gel are relatively polar, the order of separation is based on the polarity of each component in the sample, that is, the component with weak polarity is first washed out.
2. Reverse phase
The packing for the reverse phase HPLC column is usually based on silica gel with a bonded phase bonded to a relatively weakly polar functional group. The mobile phase used in the reverse phase HPLC column is more polar, which is usually a mixture of water, buffer and methanol, acetonitrile, and the like.
The order in which the sample exits the column is that the more polar component is first washed out, while the less polar component is more retained on the column. Commonly used reverse packing materials are C18 (ODS), C8 (MOS), C4 (Butyl), C6H5 (Phenyl), and the like.
Fraction Collector
Data acquisition and processing system: display the signals.
The emergence of high-performance liquid chromatography is only more than 30 years, but this separation analysis technology is developing rapidly and widely.
Click here to learn about Bonded-Phase Chromatography
