Hydrophilic Interaction Liquid Chromatography (HILIC) is a polar chromatographic mode designed for the retention and separation of compounds that are poorly retained in reversed phase systems. It combines features of normal phase chromatography with the convenience and versatility of aqueous mobile phases, making it ideal for the analysis of polar metabolites, sugars, nucleotides, peptides, and pharmaceuticals.
The stationary phase is typically polar or zwitterionic, while the mobile phase consists of a high organic content (commonly acetonitrile) mixed with a smaller portion of water. This composition creates a stable, water-rich layer on the stationary phase surface, within which analytes partition according to their polarity and hydrogen bonding capacity.
HILIC offers excellent separation selectivity for highly polar analytes, fast equilibration times, and full compatibility with mass spectrometry, making it a key technique in metabolomics, bioanalysis, and pharmaceutical research.
Hydrophilic Interaction Liquid Chromatography has become an essential complement to reversed phase HPLC, extending chromatographic capability to compounds with high polarity or limited solubility in nonpolar solvents.
It provides reproducible retention behavior, sharp peak shapes, and high sensitivity for analytes ranging from small polar molecules to biomolecules. HILIC is especially valuable in applications where traditional reversed phase methods offer insufficient retention or selectivity.
The stationary phase in HILIC is polar or zwitterionic, often based on silica or polymeric supports modified with functional groups such as amide, diol, amino, or cyano. These surface chemistries create a thin, adsorbed layer of water when exposed to high-organic mobile phases.
Analytes partition between this water layer and the surrounding solvent, while additional interactions such as hydrogen bonding, dipole–dipole forces, and electrostatic attraction contribute to selectivity. The choice of stationary phase determines the balance between hydrophilic interaction and ionic contribution, allowing precise tuning for specific analyte classes.
The mobile phase in HILIC typically contains a high proportion of acetonitrile (60–95 %) mixed with a smaller aqueous fraction containing salts or volatile buffers such as ammonium acetate or ammonium formate.
This composition promotes analyte retention by maintaining a polar stationary layer while ensuring full compatibility with LC-MS and ESI detection. Adjusting the water content, buffer concentration, or pH allows fine control over retention and selectivity without compromising reproducibility or peak symmetry.
Retention in HILIC is governed by a combination of partitioning and adsorption mechanisms. Polar analytes partition into the water-enriched layer on the stationary phase surface, while additional hydrogen bonding and electrostatic interactions modulate selectivity.
Highly polar molecules exhibit stronger retention, whereas less polar compounds elute earlier as the organic solvent content decreases. This tunable balance between hydrophilic and electrostatic forces provides excellent flexibility for separating complex mixtures of polar compounds.
| Name | Particle Sizes | Carbon Load | Pore Size |
|---|---|---|---|
| Adsorbosil CN | 5 – 10 µm | 5 % | 60 Å |
| Adsorbosil NH2 | 5 – 10 µm | 3 % | 60 Å |
| Adsorbosil Si | 5 – 10 µm | 0 % | 60 Å |
| Adsorbosphere CN | 3 – 10 µm | 6 % | 100 Å |
| Adsorbosphere NH2 | 3 – 10 µm | 6 % | 100 Å |
| Adsorbosphere PAC | 3 – 10 µm | 6 % | 100 Å |
| Adsorbosphere Si | 3 – 10 µm | 0 % | 100 Å |
| Adsorbosphere HS Si | 5 µm | 0 % | 100 Å |
| Adsorbosphere XL NH2 | 3 – 10 µm | 5 % | 100 Å |
| Adsorbosphere XL Si | 3 – 10 µm | 0 % | 100 Å |
| Allsphere Amino | 3 – 10 µm | 5.5 % | 80 Å |
| Allsphere Cyano | 3 – 10 µm | 7.5 % | 80 Å |
| Allsphere Silica | 3 – 10 µm | 0 % | 80 Å |
| Brava Amino | 3 – 10 µm | 4 % | 130 Å |
| Brava CN-BDS | 3 – 10 µm | 5 % | 145 Å |
| Brava Cyano | 3 – 10 µm | 5 % | 130 Å |
| Brava Silica | 3 – 10 µm | 0 % | 130 Å |
| CT-Sil CN | 5 µm | 7 % | 100 Å |
| CT-Sil Diol | 5 µm | 0 % | 100 Å |
| Econosil CN | 5 µm | 5 % | 60 Å |
| Econosil NH2 | 5 µm | 3 % | 60 Å |
| Econosil Silica | 5 – 10 µm | 0 % | 60 Å |
| Econosphere Amino | 3 – 10 µm | 3 % | 100 Å |
| Econosphere Cyano | 3 – 10 µm | 6 % | 100 Å |
| Econosphere Silica | 3 – 10 µm | 0 % | 100 Å |
| EcoPrep 60 Si | 10 µm | 0 % | 60 Å |
| EcoPrep 120 Si | 10 – 50 µm | 0 % | 120 Å |
| Equisil APS | 3 – 5 µm | 4 % | 120 Å |
| Equisil APS-2 | 5 µm | 4 % | 120 Å |
| Equisil CPS | 5 µm | 5 % | 120 Å |
| Equisil CPS-2 | 3 µm | 5 % | 120 Å |
| Equisil Si | 3 – 5 µm | 0 % | 120 Å |
| Equisil BDS CPS | 5 µm | 5 % | 130 Å |
| Equisorb CN | 3 – 10 µm | 7.5 % | 80 Å |
| Equisorb NH2 | 3 – 10 µm | 5.5 % | 80 Å |
| Equisorb Si | 3 – 10 µm | 0 % | 80 Å |
| Exsil 80 Amino | 3 – 10 µm | 80 Å | |
| Exsil 80 Nitrile EC | 5 µm | 80 Å | |
| Exsil 80 Nitrile HL | 3 – 5 µm | 80 Å | |
| Exsil 100 Amino | 1.5 – 10 µm | 100 Å | |
| Exsil 100 Diol | 5 µm | 100 Å | |
| Exsil 100 HAAX | 5 µm | 100 Å | |
| Exsil 100 Nitrile | 1.5 – 10 µm | 100 Å | |
| Exsil 100 Nitrile HL | 5 µm | 100 Å | |
| Exsil 100 Si | 1.5 – 10 µm | 0 % | 100 Å |
| Exsil Pure 120 ALKAM | 1.5 – 10 µm | 120 Å | |
| Exsil Pure 120 Amino | 1.5 – 10 µm | 120 Å | |
| Exsil Pure 120 Nitrile | 1.5 – 10 µm | 120 Å | |
| Exsil Pure 120 Phenyl-Amino | 1.5 – 5 µm | 120 Å | |
| Exsil Pure 120 Si | 1.5 – 10 µm | 0 % | 120 Å |
| Exsil Pure 120 ZIK-HILIC | 1.5 – 10 µm | 120 Å | |
| Exsil 300 PAX | 3 – 7 µm | 300 Å | |
| Exsil Avanti CN (1) | 3 – 5 µm | 130 Å | |
| Exsil Avanti NH2 (2) | 3 – 5 µm | 130 Å | |
| Exsil Avanti BDS ALKAM | 1.5 – 10 µm | 145 Å | |
| Exsil Avanti BDS CN | 3 – 5 µm | 145 Å | |
| Exsil Clasico CN | 3 – 10 µm | 90 Å | |
| Exsil Clasico NH2 | 8 µm | 90 Å | |
| Gold-Turbo Amid-C12 | 1.5 µm | ||
| GreatSmart NH2 | 3 – 10 µm | 5 % | 120 Å |
| GreatSmart Silica | 3 – 10 µm | 0 % | 120 Å |
| Grom Sapphire 65 Si | 5 µm | 65 Å | |
| Grom Sapphire 110 Si | 5 µm | 110 Å | |
| GromSil Amino-3 CP | 5 µm | 100 Å | |
| GromSil 60 Diol | 5 µm | 60 Å | |
| GromSil 60 NP-2 | 3 – 5 µm | 60 Å | |
| GromSil 80 Amino-1 PR | 3 – 10 µm | 80 Å | |
| GromSil 80 Cyan-2 PR | 3 – 5 µm | 80 Å | |
| GromSil 80 NP-1 | 3 – 5 µm | 80 Å | |
| GromSil 100 Amino-1 PR | 5 – 10 µm | 100 Å | |
| GromSil 100 NP-1 | 3 – 10 µm | 100 Å | |
| GromSil 100 NP-4 | 5 µm | 100 Å | |
| GromSil 120 Cyan-1 ST | 3 – 10 µm | 120 Å | |
| GromSil 120 Diol | 3 – 10 µm | 120 Å | |
| GromSil 120 NP-2 | 3 – 20 µm | 120 Å | |
| GromSil 120 ODS-4 HE | 3 – 10 µm | 120 Å | |
| GromSil 120 SEC | 3 – 5 µm | 120 Å | |
| GromSil 200 Diol | 5 µm | 200 Å | |
| GromSil 200 ODS-4 HE | 3 – 10 µm | 200 Å | |
| GromSil 200 SEC | 5 µm | 200 Å | |
| GromSil 300 Amino-4 PR | 3 – 7 µm | 300 Å | |
| GromSil 300 Cyan-1 ST | 3 – 10 µm | 300 Å | |
| GromSil 300 Diol | 3 – 5 µm | 300 Å | |
| GromSil 300 NP-1 | 5 µm | 300 Å | |
| GromSil 300 SEC | 5 µm | 300 Å | |
| Spherigrom CN | 5 µm | 80 Å | |
| ReproSil Mixed-Mode Amino-C8 | 3 – 5 µm | 11 % | 100 Å |
| ReproSil Mixed-Mode Amino-C18 | 3 – 5 µm | 17 % | 100 Å |
| ReproSil Mixed-Mode Cyano-C18 | 5 µm | 12 % | 100 Å |
| Platinum Amino | 3 – 10 µm | 100 Å | |
| Platinum C18-PFP | 1.5 – 10 µm | 7.5 % | 100 Å |
| Platinum Cyano | 3 – 10 µm | 100 Å | |
| Platinum Si | 3 – 10 µm | 0 % | 100 Å |
| Platinum 300 CN | 5 µm | 300 Å | |
| Platinum 300 Si | 3 – 15 µm | 0 % | 300 Å |
| Prosphere HP 300 C18-AQ | 3 – 10 µm | 14 % | 300 Å |
| Reprobond CN | 10 µm | 7 % | 120 Å |
| Reprobond NH2 | 10 µm | 4 % | 120 Å |
| Reprobond Si | 10 – 45 µm | 0 % | 120 Å |
| ReproKrom Si | 5 – 10 µm | 0 % | 100 Å |
| ReproShell PFP | 2.7 – 5 µm | 4 % | 90 Å |
| ReproShell Si | 2.7 – 5 µm | 0 % | 90 Å |
| ReproSil-Si, Saturator column | N/A | ||
| ReproSil 70 CN | 5 µm | 10 % | 70 Å |
| ReproSil 70 Diol | 5 µm | 4 % | 70 Å |
| ReproSil 70 NH2 | 5 µm | 5 % | 70 Å |
| ReproSil 70 Si | 5 – 50 µm | 0 % | 70 Å |
| ReproSil 80 CN | 3 – 10 µm | 3.5 % | 80 Å |
| ReproSil 80 Diol | 5 µm | 0 % | 80 Å |
| ReproSil 80 NH2 | 3 – 5 µm | 2 % | 80 Å |
| ReproSil 80 Si | 3 – 5 µm | 0 % | 80 Å |
| ReproSil 100 CN | 3 – 10 µm | 2 % | 100 Å |
| ReproSil 100 Diol | 5 – 10 µm | 3 % | 100 Å |
| ReproSil 100 DNH | 5 – 50 µm | 5 % | 100 Å |
| ReproSil 100 NH2 | 3 – 10 µm | 3 % | 100 Å |
| ReproSil 100 ODS-AQ | 3 – 10 µm | 13 % | 100 Å |
| ReproSil 100 Polyamin-2 | 3 – 5 µm | 8 % | 100 Å |
| ReproSil 100 Si | 3 – 20 µm | 0 % | 100 Å |
| ReproSil Amid-C12 | 1.5 – 10 µm | 9 % | 120 Å |
| ReproSil Amid-C16 | 5 µm | 100 Å | |
| ReproSil Amid-C18 | 3 – 10 µm | 100 Å | |
| ReproSil Amid-C18 ABZ | 3 – 5 µm | 12.5 % | 200 Å |
| ReproSil Amid-C18-e | 5 µm | 100 Å | |
| ReproSil 150 CN | 3 – 5 µm | 5 % | 150 Å |
| ReproSil 150 Si | 5 µm | 0 % | 150 Å |
| ReproSil 200 Diol | 5 µm | 200 Å | |
| ReproSil 200 Si | 5 µm | 0 % | 200 Å |
| ReproSil 300 DEAE | 5 µm | 300 Å | |
| ReproSil 300 Polyamin | 3 – 7 µm | 300 Å | |
| ReproSil DIBS-C4 | 20 µm | 120 Å | |
| ReproSil DIBS-C8 | 20 µm | 120 Å | |
| ReproSil DIBS-C18 | 20 µm | 120 Å | |
| ReproSil DIBS-RP-C4 | 10 µm | 120 Å | |
| ReproSil Fluosil 60 PFP | 3 – 5 µm | 9 % | 60 Å |
| ReproSil Fluosil 100 PFP | 1.8 – 10 µm | 12 % | 100 Å |
| ReproSil Fluosil 120 PFP | 2.2 µm | 10 % | 120 Å |
| ReproSil Fluosil C8 | 3 – 5 µm | 15 % | 145 Å |
| ReproSil Fluosil C8-NE | 5 µm | 14 % | 145 Å |
| ReproSil Fluosil RP8F | 5 µm | 15 % | 145 Å |
| ReproSil-Pur 60 Si | 3 – 5 µm | 0 % | 60 Å |
| ReproSil-Pur 120-C18 ace-EPS | 3 – 5 µm | 12.5 % | 120 Å |
| ReproSil-Pur 120 CN | 3 – 10 µm | 7 % | 120 Å |
| ReproSil-Pur 120 Diol | 3 – 10 µm | 7 % | 120 Å |
| ReproSil-Pur 120 NH2 | 3 – 10 µm | 4 % | 120 Å |
| ReproSil-Pur 120 Si | 1.9 – 10 µm | 0 % | 120 Å |
| ReproSil-Pur 200 C18-AQ | 1.9 – 50 µm | 11 % | 200 Å |
| ReproSil-Pur 200 Diol | 1.9 – 5 µm | 5 % | 200 Å |
| ReproSil-Pur 200 Si | 3 – 5 µm | 0 % | 200 Å |
| ReproSil-Pur 300 CN | 3 µm | 300 Å | |
| ReproSil-Pur 300 Diol | 3 – 5 µm | 3.5 % | 300 Å |
| ReproSil-Pur 300 Si | 3 – 10 µm | 0 % | 300 Å |
| ReproSil-Pur 1000 NH2 | 3 – 5 µm | 1 % | 1 000 Å |
| ReproSil-Pur 1000 Si | 5 µm | 0 % | 1 000 Å |
| ReproSil-Pur Basic 100 Si | 3 – 5 µm | 0 % | 100 Å |
| ReproSil Saphir 100 NH2 | 5 µm | 4 % | 100 Å |
| ReproSil Star NH2 | 5 µm | 3.5 % | 120 Å |
| ReproSil Star Si | 5 µm | 120 Å | |
| ReproSil Star ZIK HILIC | 1.5 – 5 µm | 5 % | 120 Å |
| ReproSil-XR 120 CN | 3 – 5 µm | 5 % | 120 Å |
| ReproSil-XR 120 NH2 | 3 – 10 µm | 5 % | 120 Å |
| ReproSil-XR 120 NH2-e | 5 µm | 5 % | 120 Å |
| ReproSil-XR 120 Si | 3 – 10 µm | 0 % | 120 Å |
| Reprospher 100 C4-Aqua | 5 – 30 µm | 6 % | 100 Å |
| Reprospher 100 C8-Aqua | 1.8 – 10 µm | 8 % | 100 Å |
| Reprospher 100 C8-Aqua-DE | 3 – 5 µm | 8 % | 100 Å |
| Reprospher 100 C18-Aqua | 1.7 – 30 µm | 12 % | 100 Å |
| Reprospher 100 C18-Aqua-DE | 3 – 5 µm | 12 % | 100 Å |
| Reprospher 100 CN | 1.8 – 10 µm | 7 % | 100 Å |
| Reprospher 100 CN-DE | 1.7 – 10 µm | 7 % | 100 Å |
| Reprospher 100 Diol | 3 – 10 µm | 7 % | 100 Å |
| Reprospher 100 Diol-DE | 5 µm | 7 % | 100 Å |
| Reprospher 100 DNH | 3 – 10 µm | 5 % | 100 Å |
| Reprospher 100 NH2 | 3 – 5 µm | 4 % | 100 Å |
| Reprospher 100 NH2-DE | 1.8 – 5 µm | 4 % | 100 Å |
| Reprospher 100 PEI | 3 – 50 µm | 13 % | 100 Å |
| Reprospher 100 Si | 1.7 – 10 µm | 0 % | 100 Å |
| Reprospher Carbohydrate | 5 µm | 4.5 % | 100 Å |
| Reprospher HILIC-A | 3 – 10 µm | 9 % | 100 Å |
| Reprospher HILIC-ARG | 3 – 5 µm | 15 % | 100 Å |
| Reprospher HILIC-P | 3 – 10 µm | 9 % | 100 Å |
| Reprospher 200 Si | 5 µm | 0 % | 200 Å |
| Reprospher 300 C4-Aqua | 1.8 – 5 µm | 3 % | 300 Å |
| Reprospher 300 C18-Aqua | 3 – 5 µm | 4 % | 300 Å |
| Reprospher 300 PEI | 3 – 10 µm | 4 % | 300 Å |
| Stability ABZ-Amid-C18 | 3 – 5 µm | ||
| Stability Polyamin | 3 – 5 µm | 300 Å | |
| Stability 100 Amid-C16 | 3 – 5 µm | 100 Å | |
| Stability 100 Amid-C18 | 3 – 5 µm | 100 Å | |
| Stability 100 Amid-C25 | 5 µm | 100 Å | |
| Stability 100 Polyamin-2 | 3 – 10 µm | 100 Å | |
| Stability 120 Amid-C12 | 1.5 – 5 µm | 120 Å | |
| Stability 300 Polyamin-2 | 3 – 10 µm | 300 Å | |
| Techsphere NH2 | 3 – 5 µm | 100 Å | |
| Ultrasep 100 NH2-DE | 5 µm | 100 Å | |
| Ultrasep ES 100 CN | 5 µm | 100 Å | |
| Ultrasep ES 100 NH2 | 5 µm | 100 Å | |
| Ultrasep ES 100 OH | 5 µm | 100 Å | |
| Ultrasep ES 100 Si | 5 µm | 100 Å | |
| VisionHT C18 Polar | 1.5 – 10 µm | 100 Å | |
| VisionHT HILIC | 1.5 – 10 µm | 120 Å | |
| VisionHT Silica | 1.5 – 10 µm | 120 Å |