The Most-Used HILIC Columns in Metabolomics: A 2026 Guide

If you work in metabolomics or polar-compound analysis, choosing a HILIC column can feel overwhelming — dozens of phases, each claiming to be the best. This guide cuts through the noise by looking at which HILIC columns actually appear in the recent scientific literature, what analytes they are used for, and in which sample matrices. For each column we link to real, published methods so you can see the evidence for yourself.

New to HILIC? Start with our HILIC column guide for polar metabolites, which explains the underlying principles, and our article on how HILIC retention works for the mechanisms behind the selectivity differences.

A note on “most used”

An honest caveat up front: there is no single, definitive public database that ranks HILIC columns by exact publication count, and any specific number would be misleading. What we can do — and what is more useful — is show which columns recur again and again in current peer-reviewed methods, and give you real examples with the analytes and matrices they were applied to. The ranking below reflects how frequently each phase appears in the recent metabolomics and bioanalytical literature, grouped into broad tiers rather than false-precision counts.

1. Waters ACQUITY BEH Amide

Usage: very high — arguably the most-cited HILIC phase in metabolomics. A bridged-ethylene-hybrid amide phase prized for robustness and broad metabolite coverage.

• Analytes: amino acids, biogenic amines, sugars, nucleotides, acylcarnitines, sugar phosphates, organic acids.
• Matrix: human and animal plasma, cell extracts, tumours, media.
• Recent evidence: a 2026 study optimised a low-temperature BEH-based Z-HILIC/amide method for plasma and K562 cell extracts [1]; a 2025 method used BEH Amide together with reversed phase to quantify 235 metabolites in porcine plasma [2]. Manufacturer page.

2. Merck SeQuant ZIC-HILIC

Usage: very high. A silica-based zwitterionic (sulfobetaine) phase, one of the most widely adopted HILIC columns worldwide, with a permanent 1:1 charge balance.

• Analytes: amino acids, organic acids, nucleotides, carbohydrates, metabolites, peptides.
• Matrix: plasma, serum, urine, cell and tissue extracts.
• Evidence: documented as a benchmark zwitterionic phase for polar metabolomics by the manufacturer [9], with operating conditions specified in the user guide [10]; a systematic HILIC-phase evaluation for human plasma metabolomics compares it against other phases [3].

3. Merck SeQuant ZIC-pHILIC

Usage: very high in untargeted metabolomics. The polymeric sibling of ZIC-HILIC, with an extended pH range that suits high-pH buffers — ideal for organic acids and phosphates of central metabolism [11].

• Analytes: organic acids, sugar phosphates, nucleotides, central-carbon metabolites.
• Matrix: serum, cell extracts, biological fluids.
• Recent evidence: directly compared against Z-HILIC in the 2026 low-temperature study [1]; used with ion-mobility MS on chicken serum extracts in the RHIMMS method [4].

4. Agilent InfinityLab Poroshell 120 HILIC-Z

Usage: high and growing. A superficially porous (core-shell) zwitterionic phase with hybrid, high-pH-resistant particles and inert hardware to reduce metal-analyte losses.

• Analytes: amino acids (incl. isobaric leucine/isoleucine), organic acids, polyamines, polar metabolites.
• Matrix: plasma, serum, cell media, plant extracts.
• Recent evidence: a July 2026 article details amino acid and metabolite analysis on Poroshell 120 HILIC-Z in K562 cells and media [5]; the phase is also used in the RHIMMS ion-mobility method [4]. Manufacturer page.

5. Merck SeQuant ZIC-cHILIC

Usage: high. A zwitterionic phase with the charges reversed relative to ZIC-HILIC, giving complementary selectivity — useful when standard zwitterionic phases do not resolve a target set.

• Analytes: polar metabolites, amino acids, isomeric species.
• Matrix: human plasma, biological fluids.
• Evidence: identified as offering superior isomer separation and metabolic coverage at neutral pH in the systematic plasma evaluation [3].

6. Waters ACQUITY Premier BEH Amide

Usage: high and rising. The next-generation BEH Amide with MaxPeak High Performance Surfaces to reduce metal-analyte interactions — important for phosphates and carboxylic acids, which otherwise adsorb to metal surfaces and tail [8].

• Analytes: phosphorylated metabolites, organic acids, nucleotides, amino acids.
• Matrix: plasma, cell extracts.
• Evidence: recommended for metabolomics studies by the manufacturer [12]; the BEH Amide chemistry underpins the 2026 low-temperature method [1].

7. Imtakt Intrada Amino Acid

Usage: high in amino acid analysis specifically. A dedicated LC-MS phase engineered for underivatized (intact) amino acids, notable for resolving leucine and isoleucine [13].

• Analytes: underivatized amino acids, branched-chain amino acids, dipeptides, β/γ-amino acid isomers.
• Matrix: plasma, clinical biomarkers, fermentation and beverage samples.
• Evidence: used in a validated mixed-mode assay for branched-chain amino acids in maple syrup urine disease [6]. More on this application in our article on underivatized amino acid analysis.

8. Agilent InfinityLab Poroshell 120 HILIC (bare silica)

Usage: moderate to high. The bare-silica core-shell HILIC option, valued for high efficiency at moderate pressure.

• Analytes: free amino acids (incl. isobaric leucine/isoleucine), polar small molecules.
• Matrix: standards, biological samples.
• Evidence: baseline resolution of 12 free amino acids including leucine and isoleucine is documented in an application note [7]. Manufacturer page.

9. Phenomenex Kinetex HILIC

Usage: moderate. A core-shell HILIC phase offering high efficiency at lower back-pressure — a practical middle ground between HPLC and UHPLC.

• Analytes: polar metabolites, hydrophilic small molecules.
• Matrix: biological and food samples.
• Note on evidence: unlike the phases above, we did not identify a recent metabolomics publication built specifically on this column during this review. It is a well-established general-purpose HILIC phase, and the manufacturer specifies roughly 10–30 column volumes for equilibration [14]. Manufacturer page.

10. Amino (NH₂) and bare-silica HILIC phases (general)

Usage: moderate, often application-specific. Classic HILIC chemistries that remain in use, especially for carbohydrates (amino) and simple polar separations (bare silica).

• Analytes: sugars and carbohydrates (amino phases), polar acids and small molecules (silica).
• Matrix: food, beverages, biological extracts.
• Evidence: bare silica, amino, amide and zwitterionic remain the four core HILIC surface chemistries described in the current mechanistic literature [8], and amino phases are still the classic choice for carbohydrate work — with the caveat that they are chemically less stable than alkyl or amide phases.

What the recent literature tells us

Two clear patterns emerge from current (2025–2026) publications:

• Zwitterionic and amide phases dominate. BEH Amide, ZIC-HILIC/pHILIC and the newer Z-HILIC (Poroshell HILIC-Z) account for the large majority of polar-metabolite methods.
• Hardware and conditions matter as much as chemistry. Recent work focuses on metal-inert hardware, high-pH stability and even column temperature — the 2026 study found that cooling BEH-based columns sharpened peak shapes [1].

How to use this list

Treat this as a shortlist, not a verdict. The best column for you depends on your target analytes, your matrix and your existing methods — the same selection logic covered in our guide on how to choose the right HPLC column. Bear in mind that nominally similar phases are not interchangeable: a 2025 characterisation of five zwitterionic HILIC columns found large differences in their residual ion-exchange character [15]. For most polar-metabolite work, starting with a BEH Amide or a zwitterionic phase is a well-supported, defensible choice.

Key takeaways

• BEH Amide and zwitterionic phases (ZIC-HILIC, ZIC-pHILIC, Poroshell HILIC-Z) are the most frequently used HILIC columns in current metabolomics.
• Common analytes across these columns: amino acids, organic acids, sugars, nucleotides, sugar phosphates.
• Common matrices: plasma, serum, urine, cell extracts.
• Recent research emphasises metal-inert hardware, pH stability and temperature control alongside phase chemistry.

References

1. Low-temperature HILIC method development for plasma and K562 cell extracts. Journal of Proteome Research. 2026. doi:10.1021/acs.jproteome.5c01216 — preprint version: bioRxiv (PDF)
2. Combined HILIC and reversed-phase quantification of 235 metabolites in porcine plasma. Molecules. 2025. Open-access article (PMC11820780)
3. Systematic evaluation of HILIC stationary phases for human plasma metabolomics. Metabolites. Open-access article (PMC8875576)
4. RHIMMS: reproducible HILIC ion-mobility mass spectrometry method applied to serum extracts. Open-access article (PMC8885480)
5. Use of a HILIC zwitterionic phase with superficially porous particles for metabolomics analysis. LCGC International. 2026. chromatographyonline.com
6. Rapid mixed-mode LC-MS/MS assay for branched-chain amino acids in maple syrup urine disease. Open-access article (PMC9120951)
7. Analysis of free amino acids on InfinityLab Poroshell 120 HILIC — application note. Agilent. agilent.com (PDF)
8. Guo Y, Muscatiello D. An update on the progress in fundamental understanding of hydrophilic interaction liquid chromatography. Journal of Chromatography A. 2026;1765:466520. doi:10.1016/j.chroma.2025.466520
9. SeQuant HILIC technical article. Merck. sigmaaldrich.com
10. SeQuant ZIC-HILIC user guide — buffer range, water content, sample solvent, equilibration. Merck. sigmaaldrich.com (PDF)
11. SeQuant ZIC-pHILIC user guide — extended pH range of the polymeric phase. Merck. sigmaaldrich.com (PDF)
12. Which column is a good choice for metabolomics studies? Waters knowledge base. support.waters.com
13. Intrada Amino Acid column — product information. Imtakt. imtakt.com
14. Column care and use guide — equilibration volumes and HILIC storage. Phenomenex. phenomenex.com (PDF)
15. Vosáhlová Z, Poláchová J, Svoboda J, Kohout M, Kalíková K. Interaction properties and separation potential of zwitterionic stationary phases in HILIC. Journal of Chromatography Open. 2025;8. Open-access article