HYPERCARB liquid chromatography column

Introduction to HYPERCARB Liquid Chromatography Column

Porous water permeable graphite (P) is a fixed phase structure in which carbon atoms are tightly bonded in a planar hexagonal structure, forming large molecules of polycyclic aromatic compounds. Compared with traditional silicone bonded phases, Hypercarb has a different structural shape and is stable under any pH conditions, allowing highly polar substances to be ideally retained and separated. The HYPERCARB liquid chromatography column is highly suitable for solving the separation challenges of reverse phase, normal phase high-performance liquid chromatography, and LC/MS reaction species. The retention on the Hypercarb column is the result of the charge induced effect of adsorption and polar substances on the graphite polarized surface. The Hypercarb chromatography column enhances the retention of polar compounds, separates analytes based on their structure, and exhibits extreme stability under pH extremes and high temperature environments.

High temperature HPLC using Hypercarb
• Characteristics of Porous Graphitic Carbon
• Retention mechanism: Shape and polarity of the target analyte
Develop high-temperature HPLC to improve analysis speed and separation efficiency
• “Green” LC
Suitable for analyzing polar compounds
• PH stability, 0-14
• Stereoselective
• Provide another selective chromatographic column

We only list some commonly used specification analysis columns here. If you need more product information, please contact our sales staff.

——100% porous graphitized carbon enhances separation capability
Ideal retention of strongly polar analytes
Separate compounds with similar structures
The pH value remains stable from 0 to 14
Ø Very suitable for high-temperature applications
Porous graphitized carbon (PGC) is a fixed phase in which carbon atoms are tightly bound in a planar hexagonal structure, forming large molecules similar to polycyclic aromatic hydrocarbons. Hypercarb differs from traditional silicone bonds and phases in terms of structure and retention properties, and remains stable at all pH values. Can be used for the retention and separation of highly polar compounds. Hypercarb columns are highly suitable for solving separation challenges in reverse phase, normal phase HPLC, and HPLC/MS (LC/MS) applications.
Retention and Separation

The separation mechanism of Hypercarb mainly depends on the polarity and shape of the target analyte. This special retention mechanism can successfully retain and separate certain compounds that cannot be separated by typical reverse phase high-performance liquid chromatography. By using Hypercarb to analyze polar substances, complex buffer solutions and ion pair reagents can be avoided, and the organic phase elution strength can be correspondingly increased, making the system more compatible with various detectors such as MS.

The retention of Hypercarb's original liquid chromatography column is the result of a combination of two mechanisms:

1. Adsorption: The adsorption effect of Hypercarb on analytes largely depends on its contact area with graphitized carbon molecules, as well as the type and position of functional groups on the contact surface. The strength of the interaction depends on the size and direction of the surface area of the molecules in contact with graphite. In three-dimensional space, molecules with structures closer to a plane have stronger retention.

2. The second mechanism is the induction effect of polar compounds and graphite polarized surfaces. The charge induced dipole effect results in strong retention of polar substances. When polar substances approach the surface of graphite, dipole effects are induced, enhancing the mutual attraction between the sample and the graphite surface. These charges will not be disturbed by molecular ionization charges, such as ions produced by alkaline compound ions under acidic conditions. The polarity induced dipole mechanism is attributed to the interaction between graphite surface and molecular polarized charges. Due to the strong interaction of Hypercarb, shorter chromatographic columns can be used during the method development process. In most cases, 100mm or shorter is sufficient for separation.

Increased retention of polar analytes in Hypercarb original liquid chromatography columns

In typical reverse phase chromatography, the retention of analytes is directly related to their hydrophobicity: analytes with stronger hydrophobicity have longer retention times; On the contrary, as the polarity of the analyte increases, the interaction between the analyte and solvent begins to dominate and retention decreases. This rule is correct for most inverse systems. Hypercarb is an exception to this rule, and at times, retention increases as the polarity of the analyte increases. This phenomenon is called "Polarity Retention Effect on Graphite" (PREG). This characteristic makes the Hypercarb column particularly useful for the separation of highly polar compounds (logP as low as 4), which are often difficult to retain and separate on alkyl bonds and silica gel stationary phases. Compounds with strong polarity can be retained and separated on a Hypercarb column without the need for ion pair reagents or complex mobile phases.

Hypercarb original liquid chromatography column width and pH stability

Another major advantage of Hypercarb columns is their stability in any chemical and physical environment. Due to this characteristic, it can remain stable in the pH range of 0-14 and can be analyzed at pH values that classical silica gel bonds and columns cannot use. Under high temperature and high pressure conditions, the Hypercarb column is suitable for various buffer salt mobile phases.

Separation of Compounds with Similar Structures by Hypercarb Original Liquid Chromatography Column

Due to the influence of fixed phase surface and analyte shape on retention, Hypercarb columns are capable of separating analytes with similar structures, such as isomers and homologs. Traditional C18 columns are difficult to separate methylene and methyl groups, while using Hypercarb columns can achieve good separation efficiency. The isomers of antibiotic Axetil, due to their structural similarity, exhibit a spatial structure that is distinct from the graphite surface. Hypercarb columns not only have significant improvements in separation compared to silica gel columns, but also have changes in elution sequence.

Hypercarb's original liquid chromatography column is highly suitable for separating polar compounds
The analysis of highly polar compounds is a significant challenge for RPLC/MS, as these compounds cannot be well separated on traditional hydrophobic stationary phases. Hypercarb overcomes these challenges because it:
Use "MS compatible" mobile phase,; For example, 0.1% formic acid or acetic acid and low concentration volatile buffer solutions (such as ammonium acetate or formic acid) can preserve and separate highly polar complexes
Ø can be used for mobile phases containing high concentrations of organic solvents, making them easier to atomize during atmospheric pressure ionization and improving the sensitivity of analysis.
When using shorter and smaller diameter chromatography columns, the peak capacity will not decrease. When using narrow pore and capillary columns, lower flow rates are more compatible with MS technology
In any mobile phase, it is extremely stable and does not cause loss of stationary phase, as the Hypercarb porous graphitized surface does not have any bonds or phases.