Reversed-Phase Chromatography: General Introduction For Improved Method Development
Today, more than 80% of all analytical chromatographic separations are performed using reversed-phase sorbents which have shown higher versatility compared to normal-phase chromatography sorbents. Reversed-phase sorbents have found their use in a wide range of applications such as process purification, isolation of active biomolecules, analytical separation of drugs and metabolites as well as extraction of various contaminants in environmental samples.
However, reversed-phase chromatography includes a large number of different phases that differ significantly in both chemical and physical properties which will have a significant impact on their chromatographic behaviours. Thus, one chromatographer will have to consider all of the following stationary phase properties while gathering information on the sample to be analyzed or purified:
- Hydrophobicity
- Particle and Pore Diameter
- Bonding Chemistry (Surface Coverage, Polymeric vs Monomeric)
- Particle Geometry (Irregular vs Spherical)
- Silica Purity
- Analyte pKa
- Molecular Weight
- Sample Matrix (pH, salts, concentration, interferents)
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SiliaBond® C18 (23%)
SiliaBond® C18 (or C18) is the traditional matrix for reversed-phase chromatography. The high loading provides the highest degree of hydrophobicity commercially available. Considered as the least selective phase since it interacts with a wide range of organic molecules.
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SiliaBond® C18 nec (23%)
SiliaBond® C18 (or C18) is the traditional matrix for reversed-phase chromatography. The high loading provides the highest degree of hydrophobicity commercially available. Considered as the least selective phase since it interacts with a wide range of organic molecules. Also used in ion-pairing chromatography (non-endcapped) for the separation of biomolecules.
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SiliaBond® C18 (17%)
The SiliaBond® C18 17% BDS is the most efficient deactivated C18 reversed-phase. This versatile support is the best choice for your applications in reversed-phase chromatography. This chromatographic phase has been compared with different competitors and presents a better separation for basic compounds.
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SiliaBond® C18 nec (17%)
This non-endcapped version of the SiliaBond® C18 17% BDS, presents a perfect monolayer of dimethyloctadecylsilane and silanol functions. The hydrophilic/hydrophobic balance is permitting to use a mobile phase with a higher concentration of water and ion pairing conditions for separation of biomolecules.
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SiliaBond® C18 (11%)
SiliaBond® C18 11% is used as a reversed-phase chromatographic matrix when large hydrophobic molecules are too strongly retained on SiliaBond C18 with higher carbon content.
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SiliaBond® C8
SiliaBond® C8 (or C8) is a reversed-phase matrix with a moderate degree of hydrophobicity that works well for separating a wide range of compounds. It can be used instead of C18 when shorter retention times are desired.
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SiliaBond® C8 nec
SiliaBond® C8 (or C8) is a reversed-phase matrix with a moderate degree of hydrophobicity that works well for separating a wide range of compounds. It can be used instead of C18 when shorter retention times are desired.
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SiliaBond® C4
SiliaBond® C4 (or C4) provides less retention towards non-polar compounds than C18 and C8, and is useful in ion-pairing chromatography or to separate large biomolecules. It works best for molecules with a large hydrophilic region or in case where the hydrophobic region is buried within the structure.
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SiliaBond® C4 nec
SiliaBond® C4 (or C4) provides less retention towards non-polar compounds than C18 and C8, and is useful in ion-pairing chromatography or to separate large biomolecules. It works best for molecules with a large hydrophilic region or in case where the hydrophobic region is buried within the structure.
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SiliaBond® C2
SiliaBond® C2 nec is a non-polar sorbent and presents a moderate hydrophobicity. This phase is used to extract extremely non-polar compounds.
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SiliaBond® C2 nec
SiliaBond® C2 nec is a non-polar sorbent and presents a moderate hydrophobicity. This phase is used to extract extremely non-polar compounds.
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SiliaBond® C1
SiliaBond® C1 (or C1) has the lowest degree of hydrophobicity of all reversed stationary phases, useful in the separation of large biomolecules that have extensive hydrophobic regions.
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SiliaBond® Phenyl
SiliaBond® Phenyl (Si-PHE) is a sorbent of medium polarity used in reversed-phase chromatography. It has similar retention than C8 but with different selectivity especially for molecules containing aromatics and fatty acids.
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SiliaBond® Phenyl nec
SiliaBond® Phenyl (Si-PHE) is a sorbent of medium polarity used in reversed-phase chromatography. It has similar retention than C8 but with different selectivity especially for molecules containing aromatics and fatty acids.
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SiliaBond® Cyano
SiliaBond® Cyano (Si-CN) can be used both in normal and reversed-phase chromatography as its polarity marks the separation between the polar and non-polar phases. SiliaBond Cyano is the least retentive polar sorbent in normal phase chromatography and the least retentive of the non-polar sorbents in reversed-phase chromatography.
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SiliaBond® C12 (Adamantyl)
SiliaBond® C12 (or C12, Si-Adamantyl) has a polarity similar to C18 with additional steric bulk to provide additional separation characteristics.
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SiliaBond® Allyl
SiliaBond® Allyl (Si-Allyl) can be used as solid support for further derivatization.
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