Columns are the main component in HPLC because the column is responsible for the separation of the sample c through the column with the mobile phase and separates in its components when it comes out from the column.
Different types of HPLC column are used in analysis of different pharmaceutical compounds according to their nature and column separatio capacity.
Generally, silica gel is filled in the high-performance liquid chromatography columns because of its particle size and of components and silica gel is also an inert material that does not react with mobile phases. Therefore silica colu compounds of different chemical natures. The material filled in the HPLC columns is known as a stationary phase. There are different types of chromatography columns on the basis of their composition and method of separation.
So…What is hplc column !
Normal Phase Columns // Reverse Phase Columns // Ion Exchange Columns // Size Exclusion Columns
Normal Phase HPLC Columns: This type of columns has more polar stationary phase than the mobile phase. The packing material of the colum mobile phase and this condition is fulfilled by the silica that is polar material. But water is more polar than the silica, methylene chloride, hexane and chloroform or a mixture of these with diethyl ether is used as mobile phase. Separation of the sample components occurs on the basis of the polarity of the sample components. Sample c interact more with polar stationary phase resulting in separation from the less polar component that interacts with columns are widely used in the pharmaceutical analysis. The chromatography column packing in which normal phas Normal Phase Chromatography.
- Reverse Phase HPLC Columns: In reverse phase columns as its name states, it is reverse of the normal phase columns. It has a non-polar or less more polar mobile phase. Bonded hydrocarbons like C8 and C18 and other non-polar hydrocarbons are used as s columns while aqueous organic solution like water-methanol or water-acetonitrile mixture is used as mobile phase. Separation of sample components in reverse phase columns also occurs on the basis on the polarity of the sample opposite of the normal phase HPLC columns, therefore, this type of chromatography is known as Reverse Phase C
Ion Exchange HPLC Columns: The compounds those can easily ionize are analyzed using these columns. Stationary phase in these columns negative or positive charge while mobile phase is a polar liquid as the salt solution in water. Separation of mole attractive ionic force between molecules and the charged stationary phase. Due to the exchange of ions d components, it is known as Ion Exchange Chromatography.
HPLC columns have a different length varying from 30 mm to 250 mm and their particle size or porosity from 3 analysis of sample, therefore, these are considered important during the HPLC analytical method development. Co the nature of the compound to be analyzed and the mobile phase. Column performance should also be evaluated runs or as required.