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Hilic HPLC Column

Hilic Column Wholesale and OEM Supplier

uHPLCs is one of the best HILIC Column Wholesale and OEM suppliers, specializing in providing top-tier HILIC columns designed to meet the diverse needs of your laboratory. Our columns are crafted with precision and high-quality materials, ensuring reliable and reproducible results for your chromatographic applications.

Why Choose uHPLCs HILIC Columns

1. High Efficiency and Resolution: 
Our HILIC columns are engineered to deliver sharp, well-resolved peaks, enhancing the accuracy and reliability of your analyses.

2. Superior Reproducibility: 
Each column is manufactured to stringent quality standards, guaranteeing consistent performance across different batches.

3. Wide Range of Selectivities: 
We offer a variety of stationary phases, allowing you to choose the most suitable column for your specific separation needs.

4. Enhanced Durability: 
Constructed with robust materials, our columns withstand high pressures and extended use, providing excellent longevity and value for your investment.

5. Optimized Particle Size: 
Our columns feature optimized particle sizes that improve separation efficiency and reduce analysis time.

6. Excellent Chemical Compatibility: 
Compatible with a broad range of solvents and samples, our columns are versatile for various applications.

7. Custom Solutions Available: 
We provide tailored solutions to meet unique separation requirements, ensuring you achieve the best possible results.

Choose uHPLCs for your HILIC columns and experience superior performance, reliability, and expert support. Contact us today to learn more about our products and how we can assist with your chromatographic needs.

Hilic HPLC Column

3.0 Empty HPLC Column Hardware

USHD HILIC-SiO2
Hilic HPLC Column

2.1mm Empty HPLC Columns Normal Pressure for Lab Equipment

USHD HILIC-Amide
Hilic HPLC Column

2.1mm High Pressure Empty UHPLC Columns

USHD HILIC-Zil
Hilic HPLC Column

Hilic HPLC Column Specification

Item USP Pore Size Option Surface area (m²/g) Carbon Load (%) PH Tolerance Range Column Specifications Features and Application
USHD HILIC-SiO2
L3
5μm, 120A; 3μm, 120A; 1.8μm, 120A;
320
#
2-8
4.6X250mm; 4.6X150mm; 3.0X100mm; 3.0X50mm; 2.1X100mm; 2.1X50mm;
1.) Hydrophilic interaction silica gel column, special treatment of silica gel matrix, can better retain water-soluble compounds separated by polarity differences, suitable for challenging polar analysis such as separation of highly polar substances in high water phase.
2.) It has strong stability and long column life.
3.) HILIC mode can obtain better selectivity, strong retention performance and separation effect, and can separate strongly basic polar compounds under HILIC conditions. It can also be used in reverse phase mode.
USHD HILIC-Amide
L68
5μm, 120A
320
5
2-9
4.6X250mm; 4.6X150mm;
1.) Amide chimeric HILIC hydrophilic interaction chromatography column, with separation ability similar to HILIC amino column, but different selectivity and longer column life.
2.) Under HILIC conditions, it has better retention ability for highly polar ionic compounds dissolved in highly organic mobile phase.
3.) It has a strong separation effect on acidic and highly polar compounds, and can separate some mixed molecules that are often separated by amino phase in low pH buffer.
USHD HILIC-ZIL
#
5μm, 100A
400
6
2-9
4.6X250mm; 4.6X150mm;
1.) HILIC hydrophilic interaction ion exchange chromatography column, bonded with zwitterionic functional groups can separate solutes containing acidic, basic and/or zwitterionic substances.
2.) It is extremely hydrophilic and can use 100% aqueous mobile phase, which can effectively retain highly polar ionic compounds and basic drug molecules that are weakly or not retained in RP-LC.
HPLC Colun in the HPLC System Connect Diagram by uhplcs

In addition to custom columns, uHPLCs also offers a wide range of standard columns for various applications such as reversed-phase, normal-phase, ion exchange, size exclusion, and HILIC. We also offers prepacked columns and accessories, such as frits, end-fittings, and Guards Columns.

uHPLCs uses state-of-the-art manufacturing techniques to produce high-quality columns that are consistent, reliable, and provide excellent performance. we have a strict quality control program to ensure that all columns meet the highest CE , SGS and UL standards for performance and reproducibility.

Contact Us For Excellent HPLC Columns

Experience the Precision and Reliability of Our High-Quality C18 HPLC Columns Today

WHY uHPLCs Hilic HPLC Column ?

Some Advantage You Should Choose uHPLCs Hilic Column:

1. High Efficiency and Resolution: uHPLCs HILIC columns are designed to deliver sharp, well-resolved peaks, ensuring precise and accurate chromatographic analyses.

2. Consistent Reproducibility: Each column is manufactured under stringent quality control standards, ensuring consistent performance and reliable results across different batches.

3. Versatile Selectivity Options: We offer a wide range of stationary phases, allowing you to choose the most suitable column for your specific separation needs, enhancing your method development flexibility.

4. Exceptional Durability: Constructed from robust, high-quality materials, our HILIC columns are built to withstand high pressures and prolonged use, providing long-lasting performance and value.

5. Optimized Particle Sizes: Featuring optimized particle sizes, our columns improve separation efficiency, reduce analysis time, and enhance overall productivity.

6. Broad Chemical Compatibility: Our HILIC columns are compatible with a wide range of solvents and sample types, making them versatile for various applications in different industries.

7. Customizable Solutions: We offer customized solutions to meet your unique separation requirements, ensuring you achieve the best possible results for your specific applications.

8. Expert Support: At uHPLCs, we provide comprehensive support and expertise to help you optimize your chromatographic processes and achieve superior performance.

Choose uHPLCs HILIC columns for their superior performance, reliability, and expert support. Contact us today to learn more about our products and how we can assist with your chromatographic needs.

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Hilic HPLC Column FAQ

Main Features of Hilic Column?

HILIC (Hydrophilic Interaction Liquid Chromatography) columns are a type of HPLC column that is specifically designed for the separation of polar compounds. Polar compounds are those that have a strong affinity for water. Traditional reversed-phase HPLC columns are not very effective at separating polar compounds, as they tend to elute (come out of the column) too quickly.

Here are some of the main features of HILIC columns:

  • Hydrophilic stationary phase: HILIC columns have a stationary phase that is hydrophilic, or water-loving. This stationary phase can be made from a variety of materials, such as silica gel or bonded phases.
  • High organic mobile phase: The mobile phase in HILIC chromatography is typically high in organic solvent (e.g., acetonitrile) and low in water. This is opposite to reversed-phase chromatography, where the mobile phase is typically high in water and low in organic solvent.
  • Reversed elution order: Compared to reversed-phase chromatography, HILIC columns elute analytes in the opposite order. More polar analytes will elute later than less polar analytes.
  • Strong retention of polar compounds: HILIC columns are able to strongly retain polar compounds that would be poorly retained by reversed-phase columns. This makes them ideal for the separation of a wide variety of polar compounds, including pharmaceuticals, natural products, and metabolites.

Here are some of the advantages of using HILIC columns:

  • Improved separation of polar compounds: HILIC columns can provide better separation of polar compounds than reversed-phase columns.
  • Reduced ion suppression: HILIC columns can help to reduce ion suppression, which can be a problem with some mass spectrometry techniques.
  • Increased compatibility with mass spectrometry: HILIC columns are often more compatible with mass spectrometry than reversed-phase columns.

Here are some of the disadvantages of using HILIC columns:

  • Higher cost: HILIC columns can be more expensive than reversed-phase columns.
  • Limited range of stationary phases: There is a more limited range of stationary phases available for HILIC columns than for reversed-phase columns.
  • Slower analysis times: HILIC columns can sometimes result in slower analysis times than reversed-phase columns.

Some Applications of Hilic Column

Hydrophilic interaction liquid chromatography (HILIC) is a powerful separation technique that has found numerous applications, particularly for the analysis of polar and hydrophilic compounds.

Here are some key applications of HILIC columns:

1.Separation of Biomolecules

Glycans and Glycoconjugates: HILIC is widely used for the separation and analysis of glycans, glycopeptides, and glycoproteins. It provides excellent resolution and selectivity for these highly polar compounds.
Peptides and Proteins: HILIC is effective for separating polar and hydrophilic peptides and proteins, including membrane proteins, histones, and their variants with different modifications (acetylation, methylation, phosphorylation).

2. Oligonucleotides: 

HILIC shows potential for improved resolution in the analysis of complex oligonucleotides due to its ability to separate polar compounds.

Pharmaceutical and Biomedical Applications
1. Polar Drugs and Metabolites:
HILIC is suitable for analyzing polar pharmaceuticals, drug metabolites, and other small polar molecules that are poorly retained in reversed-phase LC.
2. Metabolomics and Biomarker Analysis:
HILIC is valuable for the analysis of polar metabolites, biomarkers, and other small molecules present in biological fluids and tissues.
Complementary to Reversed-Phase LC
3. Two-Dimensional LC (2D-LC):
HILIC is highly orthogonal to reversed-phase LC, making it an ideal technique for comprehensive 2D-LC separations, especially when coupled with mass spectrometry (LC-MS).
4. Fractionation and Sample Cleanup:
HILIC can be used in combination with solid-phase extraction (SPE) or other techniques to fractionate and clean up complex samples before separation by reversed-phase LC or other methods.

So, As we know, HILIC has proven to be a powerful and versatile technique, complementing traditional reversed-phase LC and expanding the range of separations possible for polar and hydrophilic compounds in various fields, including biotechnology, pharmaceuticals, and biomedical research.

Frequently Asked Questions

A HILIC (Hydrophilic Interaction Liquid Chromatography) column is a type of analytical column used in liquid chromatography that separates analytes based on their hydrophilicity. It contains a polar stationary phase that attracts and retains hydrophilic (water-loving) compounds through mechanisms like hydrogen bonding, dipole-dipole interactions, and electrostatic forces. HILIC is complementary to reversed-phase chromatography and is particularly useful for separating highly polar and ionizable compounds that are poorly retained by non-polar stationary phases.

1. Excellent selectivity for polar and hydrophilic compounds:
HILIC provides superior separation of highly polar analytes like glycans, peptides, nucleic acids, metabolites, and polar drugs/metabolites that are poorly retained in reversed-phase LC.
2. Orthogonal selectivity to reversed-phase LC:
HILIC offers an orthogonal separation mechanism, making it ideal for 2D-LC setups and comprehensive analysis when combined with reversed-phase separations.
3. Compatible with mass spectrometry:
HILIC mobile phases containing high organic content are MS-friendly and facilitate efficient ionization and detection of analytes.
4. Improved peak shapes:
For ionizable compounds, HILIC can provide sharper peaks compared to ion-exchange chromatography by reducing secondary interactions.

There are several types of HILIC stationary phases available, each with its own advantages and disadvantages. The most common types include:

1.Bare silica:

This is the simplest type of HILIC stationary phase.

It is made from silica gel, which is a polar material that can interact with hydrogen bonding groups on analytes.

Bare silica is not very selective, but it can be a good choice for separating polar compounds that are difficult

to retain on reversed-phase columns.

Image of Bare silica HILIC stationary phase 

 

2.Bonded phases:

These stationary phases have a layer of bonded organic molecules on the surface of the silica gel.

The type of bonded molecule can have a significant impact on the selectivity of the column.

Some common bonded phases for HILIC include:

2.1 Diol phases:

These phases contain bonded diol groups, which can form hydrogen bonds with analytes.

Diol phases are a good choice for separating polar compounds such as carbohydrates,

peptides, and oligonucleotides.

Image of Diol phases HILIC stationary phase 

 

2.2 Amino phases:

These phases contain bonded amino groups, which can interact with analytes through hydrogen bonding and ion exchange.

Amino phases are a good choice for separating polar compounds such as amino acids and peptides.

Image of Amino phases HILIC stationary phase

 

2.3 Zwitterionic phases:

These phases contain both positively and negatively charged groups.

Zwitterionic phases can be a good choice for separating polar compounds that are difficult

to retain on other types of HILIC columns.

Image of Zwitterionic phases HILIC stationary phase

 

3.Hydride phases:

These phases are made from a special type of silica gel that is capped with a layer of hydride groups.

Hydride phases are very hydrophilic and can be a good choice for separating very polar compounds.

Image of Hydride phases HILIC stationary phase

The choice of HILIC stationary phase will depend on the specific needs of the separation.

It is important to consider the polarity of the analytes, the desired selectivity, and the compatibility of

the stationary phase with the mobile phase.

 

Optimizing a HILIC separation can be an iterative process, but there are several key factors you can adjust to achieve the best results for your analytes. Here’s a breakdown of the key points:

1. Mobile Phase:

  • Organic Modifier: This is the major factor influencing retention in HILIC. Unlike reversed-phase chromatography, increasing the organic content (like acetonitrile) strengthens interactions and increases retention time. Explore a range of organic modifier percentages (e.g., acetonitrile, methanol) to find the sweet spot for your analytes.
  • Buffer and pH: Buffers like ammonium formate or acetate help control mobile phase pH and mitigate unwanted ionic interactions. Consider buffer type, concentration, and pH to fine-tune selectivity and peak shape.
  • Additives: For specific analyte functionalities, additives like formic acid or modifiers with specific functional groups can enhance resolution or ionization for MS compatibility.

Stationary Phase:

  • Column Selection: Different HILIC phases (bare silica, amide, diol, etc.) offer varying selectivity based on their surface chemistry. Experimenting with different column types might be necessary to achieve optimal separation for your analytes.


2. Chromatographic Conditions:

  • Flow Rate: Adjusting flow rate can impact resolution and analysis time. Start with a moderate flow rate and explore a range to find the balance between peak efficiency and analysis speed.
  • Gradient: HILIC often utilizes gradient elution with increasing organic content to elute retained analytes. Experiment with gradient slope and time to optimize peak separation while maintaining reasonable analysis time.
  • Column Temperature: Temperature can influence analyte retention and peak shape. Consider a slightly elevated temperature (e.g., 30-40°C) compared to room temperature for some separations.


Additional Tips:

  • Sample Preparation: Ensure proper sample cleanup to minimize matrix effects that can interfere with peak separation.
  • System Equilibration: HILIC columns rely on a stable water layer for interaction. Allow sufficient equilibration time after injections and between runs for consistent retention times.
  • Method Development Tools: Consider using software or statistical methods to design experiments and optimize multiple parameters simultaneously.

But You can Remember, optimization is often an iterative process. Start by making small adjustments to one factor at a time, monitor the effects on your separation, and refine further based on your observations. 

Consulting application notes or technical resources from column manufacturers can also provide valuable insights for specific HILIC stationary phases.

HILIC mobile phases are quite unique compared to other chromatography techniques like reversed-phase HPLC. Here’s the breakdown:

Key Components:

  • High Organic Content (60-70%+): The bulk of the mobile phase in HILIC is a strong organic solvent, typically acetonitrile (ACN). This creates a weak interaction with the polar stationary phase.
  • Low Aqueous Content (3-10%+): A small percentage of water or another polar solvent like methanol is added. This forms a water-rich layer on the stationary phase for analyte partitioning.
  • Buffers (Optional): Buffers can be included to control pH and influence analyte ionization and retention. Popular choices include ammonium formate and ammonium acetate due to their compatibility with MS detection and solubility in high-ACN environments.

Important Points:

  • Water is the Strong Solvent: Unlike reversed-phase, where water is the weak solvent, water acts as the stronger solvent in HILIC, eluting more polar analytes later.
  • Methanol Not Ideal: While methanol is a polar solvent, it’s generally not recommended in HILIC because it can disrupt the formation of the water layer.
  • Mobile Phase Sensitivity: HILIC separations can be sensitive to small changes in mobile phase composition, especially the organic solvent percentage.

Need Help ?

Contact uHPLCs Today for Any Questions for HPLC / UHPLC 

+(86) 0755-28502380

sales@uhplcs.com

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