HPLC Column Configurator

HPLC Columns for every application

Find the right column from over 50,000 HPLC columns from more than 10 different manufacturers. We also offer our own brand of Altmann Analytik HPLC columns as a high-quality and cost-effective alternative. Compare prices and the application possibilities of the various manufacturers. If you are unable to find a suitable HPLC column, please feel free to contact us.

NP columns for Normal Phase Chromatography

Prior to the development of the Reversed Phase (RP), Normal Phase Chromatography was the most common separation mode. We offer Normal Phase (NP) HPLC columns by all well-known manufacturers here.

In Normal Phase Chromatography, the mobile phase is non-polar and the stationary phase is polar (common: silica gel). This technique relies on the interaction of analytes with polar functional groups on the surface of the stationary phase. This interaction is strongest when using non-polar solvents as the mobile phase. The least polar compounds elute first and the most polar compounds elute last.

Normal Phase Chromatography is a very effective separation method because a wide range of solvents can be used to fine-tune the selectivity of a separation. However, it has become less popular with many chromatographers because of the complexity involved. Under certain conditions, long equilibration times or reproducibility problems may occur. This is due mainly to the sensitivity of the technique to the presence of low concentrations of polar contaminants in the mobile phase. Controlling these problems effectively produces better chromatograms than reverse phase methods as the commonly used solvents have a lower viscosity.

HPLC columns for Reversed Phase (RP) Chromatography

In Reversed Phase Chromatography, the polarity of the phases are "reversed". The mobile phase is polar and the stationary phase is non-polar. As it is one of the most popular separation modes in chromatography, all related products of all well-known manufacturers are available here.

The non-polar side chains in Reversed Phase Chromatography columns are bound either to a polymer or to a structure made of silica gel, which leads to them being hydrophobic. The longer the chain, the more non-polar the phases are. Polar analytes are eluted from the column first, followed by the non-polar analytes. Reversed phase is more widely practised than normal phased as they can be used universally for polar and non-polar analytes. In addition, this method is very sensitive and flexible as small changes in the composition of the mobile phase (e.g. salts, pH, organic solvents) or the temperature can completely change the separation properties of the system. RP-HPLC is used especially in numerous applications of UV spectroscopy (LC-UV).

Chiral columns for Chiral/Enantiomeric chromatography

Unlike normal HPLC columns, chiral or enantiomeric HPLC can also be used to separate and determine chiral compounds. This requires special chiral stationary phases that have fixed chiral functionalities. In the Analytics Shop, we have over 1,400 different chiral columns from different manufacturers, including the high-quality columns from Chiral Technologies as well as lower-priced alternatives of the same quality from YMC and our own Altmann Analytik brand.

Special columns such as chiral or enantiomeric HPLC columns are required to separate chiral compounds. With suitable interaction, these enable the separation of the enantiomers which have almost identical physical and chemical properties. Enantiomers are molecules with a chiral center. Diastereomers are molecules with two or more chiral centers, differ in their chemical properties and can be separated using chromatography. The separation of enantiomers is based on the formation of diastereomers and their interaction with other chiral molecules like chiral stationary phases (e.g. silica gels) in chromatographic separations. A conventional solvent is used as the eluent.

UHPLC and UPLC^® columns

With Ultra High Performance Liquid Chromatography (UHPLC), dramatic increases in resolution, speed and sensitivity of HPLC can be achieved by using short and thin columns. The particles of the filling material have diameters of less than 2 µm, which results in an improved separation performance compared to standard HPLC. The increased total surface area of the filling material gives the analyte a higher adsorption ability. Analysis times are reduced due to the shorter columns and less solvent is required.

Columns for Hydrophilic Interaction Chromatography (HILIC)

Hydrophilic interaction chromatography (HILIC) is a popular alternative to normal phase and reverse phase chromatography. Similar to NP, buffer systems are used as stationary phases in HILIC mode. These are aqueous buffer systems with organic solvents, e.g. B. acetonitrile. In HILIC mode, water is the strongest eluant. HILIC columns are suitable for the separation of polar compounds such as carbohydrates, polar metabolites and hydrophilic compounds regardless of charge and molecular size. An extensive range of HILIC columns from established quality manufacturer YMC is available in our shop.

Columns for Supercritical Fluid Chromatography (SFC)

Significant advantages of Supercritical Liquid Chromatography (SFC) columns compared to HPLC columns:

• Inexpensive: reduced solvent comsumption
• Environmentally friendly
• Time-saving: quick separation with high resolution
• Flexible: chiral and achiral phases

Supercritical fluids show low viscosities and higher diffusivities when used as a mobile phase, resulting in narrower peaks due to rapid diffusion and faster elution and less pressure drop through the column. Supercritical fluids combine the benefits of liquids and gases, hence enabling the SFC technique to combine the best aspects of HPLC and gas chromatography (GC). In most cases, a supercritical fluid such as carbon dioxide is used as the mobile phase. The low viscosity of supercritical carbon dioxide enables analytical separations that are 3-5 times faster than those for normal phase HPLC. The speed of SFC separations, the preservation of organic solvents and more concentrated product fractions make SFC a preferred chromatographic technique for separating and purifying chemical mixtures.

SFC is an environmentally friendly separation technique that makes use of CO2-based mobile phases. The use of high-performance preparative columns (internal diameter of 10 - 50 mm) with a large number of particle sizes from 3 - 20 μm leads to the quick separation and recovery of cleaned components.

Nano HPLC columns

Nano HPLC uses columns with very small inner diameters. Columns with internal diameters of 75 µm, 100 µm or 150 µm are commonly used. Due to the reduction in the internal diameter, injection and flow rates must be reduced which is particularly advantageous when only small or diluted sample quantities are available. The smaller nano HPLC columns lead to an increased sensitivity with lower solvent consumption. Nano columns are able to maintain a high concentration of the injected sample and to direct approx. 40-50 % of the sample to the detector. Nano HPLC has a high separation efficiency compared to the traditional HPLC technique.

Columns for Gel Permeation Chromatography (GPC) & Size Exclusion Chromatography (SEC)

Gel permeation chromatography (GPC) is particularly suitable for the separation of non-polar molecules and can be used for a wide range of solvents, from non-polar organics to aqueous applications. GPC/SEC columns are packed with very small porous beads. In GPC/SEC, the molecules are separated according to size (size exclusion chromatography, SEC). The smaller molecules can enter the pores more easily and therefore spend more time in these pores, eluting last. Conversely, larger molecules spend little if any time in the pores and are eluted quickly. The GPC columns are filled with a microporous packing material and gel, hence the name gel permeation.

Ion Exchange Chromatography (IEC)

Ion exchange chromatography (IEC) separates molecules based on the respective charged groups on the surface of a protein. Molecules undergo electrostatic interactions wirh opposite charges on the stationary phase matrix.

The pH value of the buffer, the buffer concentration, packing of the columns and the salt gradient, in which the salt ions compete with the desired proteins to bind to charged groups on the surface of the medium, are important aspects affecting the results of IEC. At any pH, a protein has a net positive or negative charge due to the state of charge of the amino acids. If this protein is positively charged, it will bind to negatively charged materials and vice versa. The separation is based essentially on the charge and size of the ions.

Resins such as polystyrene, cellulose and crosslinked polyacrylamide or polydextran gels are used as mediums for ion exchange. The two types of ion chromatography are anion-exchange and cation-exchange. Cation-exchange chromatography is used when the molecule of interest is positively charged and anion-exchange chromatography is when the stationary phase is positively charged. Ion exchange chromatography is usually performed at pH values in which the protein has a net charge opposite to that of the stationary phase matrix.

Columns with polar bound phases

Polar-bound phases are based on silica gels which have chains attached onto the silica structure with functional groups. Chromatography makes use of the difference in polarity to separate a mixture. Separation takes place through different mechanisms and often as a result of the combination of several processes like adsorption, distribution, ion exchange, molecular size exclusion etc.

Permitted changes according to USP (United States Pharmacopeia)

The U.S. Pharmacopeia Convention is a scientific non-profit company that sets the standards for ingredients, concentration, quality and purity of medicines, food ingredients and food supplements that are manufactured, distributed and consumed all over the world. According to USP regulations, the following deviations may occur:

USP Chapter

1. Column Length: ± 70%
2. Column Inner Diameter: can be changed if the flow rate is kept constant.
3. Particle Size: - 50%
4. Flow Rate: ± 50%
5. Ratio of components in the mobile phase: ± 30% or ± 10%
6. pH value of the mobile phase: ± 0,2
7. Salt concentration in the buffer: ± 10%
8. Column Temperature: ± 10°C
9. Wave Length of the UV-Vis detector: ± 3 nm
10. Injection Volume: can be reduced until precision and detection limits are reached.

Permitted changes according to EP (European Pharmacopeia)

European Pharmacopeia Ph. Eur., Chapter 2.2.46

The European Pharmacopeia is a published collection of monographs describing the individual and general quality standards of ingredients, dosages and analytical methods of medicine. The aim is to set common quality standards across Europe to control the quality of medicines and other chemical products. According to the EP regulation, the following deviations can occur:

Isocratic elution

1. Column Length: ± 70%
2. Column Inner Diameter: ± 25%
3. Particle Size: - 50%
4. Flow Rate: ± 50%
5. Ratio to components in the mobile phase: ± 30% or ± 2% absolute
6. pH value of the mobile phase: ±0,2
7. Salt concentration in the buffer: ± 10%
8. Column Temperature: ± 10°C
9. Wave Lenght of the detector: ± 3 nm
10. Injection Volume: can be reduced until precision and detection limits are reached.

Gradient elution

1. Column Length: ± 70%
2. Column Inner Diameter: ± 25%
3. Particle Size: no changes permitted
4. Flow Rate: changes acceptable if column size is changed
5. Ratio to components in the mobile phase: small changes in the composition of the mobile phase and the (density) grade are acceptable if the system configuration still meets the requirements.
6. Dwell Volume: gradient instants can be used to compensate for differences in dwell volume between different systems.
7. pH value of the mobile phase: no changes permitted
8. Salt concentration in the buffer: no changes permitted
9. Column Temperature: ± 5°C
10. Wave Lenght of the detector: no deviations permitted
11. Injection Volume: can be reduced until precision and detection limits are reached.