Columns plays a very important role for the use of High Performance Liquid Chromatography(HPLC). There are different types of HPLC Columns Used in Pharmaceutical Analysis. According to the difference of stationary phase and mobile phase, we can be divided them into below 4 different types that commonly used.
Normal-Phase HPLC Columns
Normal-Phase HPLC Columns is chromatography columns in which there is more polar stationary phase than the mobile phase. This type of HPLC columns are used at early stage of HPLC development. Silica gel without any functional group was used as stationary phase. Normal-phase are mainly used for analysis of neutral and ionic pharmaceutics. For example, it is used for determination of enantiomer in tofacitinib cirate.
Reversed-phase HPLC columns
Reversed-phase HPLC columns is chromatography columns that contain more non-polar stationary phase.Silica gel with functional group are used as stationary phase. C18 is the most popular used non-polar reversed-phase column. Reversed-phase HPLC columns are more frequently used than normal phase nowadays, and are widely used in pharmaceutical analysis. It is suitable for the separation and analysis of substances containing non-polar groups, such as proteins, peptides, amino acids, nucleic acids, steroids, lipids, fatty acids, sugars, and alkaloids.
Hydrophilic Interaction Chromatography Columns
Hydrophilic Interaction Chromatography Column(HILIC) uses a polar stationary phase in conjunction with a mobile phase containing at least 2.5% water combined with a higher proportion of a less polar solvent. HILIC is popularly used for the separation and analysis of carbohydrates. For example, HILIC can be used for determination of saponins in herbal medicines.
Ion exchange HPLC columns
Ion exchange HPLC columns contains stationary phase with negative or positive charge, and mobile phase as inorganic salt solution. Ion exchange HPLC columns are used for the analysis of substance based on differences in the net charge of the molecules. It can be used for purification and anslysis of protein, oligonucleotides, peptides,as well as other charged molecules. For example, Ion exchange HPLC column can be used for evaluation of glycosylated hemoglobin.