Column Image

  • Triazole bonded stationary phase
  • Enhanced hydrophilic interaction
  • Excellent retention for highly polar analytes
  • Unique anion-exchange mechanism

For information on COSMOSIL 2.5HILIC (Ultra-High Performance Column)

Product Information


Packing Material HILIC
USP Code L104
Silica Gel High Purity Porous Spherical Silica
Average Particle Size 5 µm
Average Pore Size approx. 120Å
Specific Surface Area approx. 300 m2/g
Stationary Phase HILIC
Interactions Hydrophilic Interaction, Anion Exchange
Object Substance Hydrophilic Compound, Acidic Compound
Feature Suitable for non-retaining by C18

Comparison with C18

The hydrophilic interaction chromatography is a variation of normal phase chromatography where a polar stationary phase is used with a mobile phase which contains a high concentration of water miscible organic solvent and a low concentration of aqueous eluent. The main retention mechanism is the partitioning of the polar analytes between the polar stationary and the non-polar mobile phase. As it is also called “aqueous normal phase”, the elution order is similar to that of normal phase and the sample elution is in the order of increasing hydrophilicity. Without using ion-pair reagent COSMOSIL HILIC retains highly polar analytes that would not be retained in reversed phase chromatography. It also shows a weak anion-exchange mechanism with the positively charged stationary phase, thus acidic compound is strongly retained.



Melamine Related Compunds

Column Size 4.6 mm I.D. x 250 mm
Mobile Phase Acetonitrile/ 10mmol/l Ammonium Acetate = 70/30
Flow Rate 1.0 ml/min
Temperature 30°C
Detection UV225nm
  1. Melamine (0.1 µg)
  2. Ammeline (0.07 5µg)
  3. Cyanuric Acid (0.75 µg)
  4. Ammelide (0.05μg)

>> More melamine applications by HPLC and LC/MS/MS using COSMOSIL HILIC.

BCAA (amino acid branhed-chain)

Column Size 4.6 mm I.D. x 250 mm
Mobile Phase Acetonitrile/ 10mmol/l Ammonium Acetate = 85/15
Flow Rate 1.0 ml/min
Temperature 30°C
Detection ELSD
  1. L-Leucine (3.0 µg)
  2. L-Isoleucine (3.0 µg)
  3. L-Valine (3.0 µg)

Optimizing Analytical Conditions

The COSMOSIL HILIC Chromatogram Index, which includes 167 chromatograms using COSMOSIL HILIC, is available. This index is a useful tool for all chromatographers, assisting them to optimize analytical conditions for hydrophilic interaction chromatography.

COSMOSIL HILIC Application Notebook (PDF file : 18.5 MB)

Usage information

Column 5 µm HILIC
I.D. (mm) 2.0 3.0 4.6 10.0 20.0
Washing method 1. Remove buffer, salts and/or acid from column: wash for 10-15 min. using the mobile phase last used, without buffer, salts or acid. For example, if your mobile phase was 50:50 acetonitrile/20 mmol/l phosphate buffer, wash with 50:50 acetonitrile/water.

2. Remove adsorbed compounds and fix unstable baselines: Wash with 50:50 acetonitrile/water or up to 100% water.
Storage conditions Remove buffer, salts and/or acid from column: wash for 10-15 min. using the mobile phase last used, without buffer, salts or acid.
Then, replace the solvent with 90:10 acetonitrile/water.

Tightly plug the column, and store in a cool and dry place.
Usable pH range pH2~pH7.5
Max. pressure 20MPa 15MPa
Temperature range The maximum usable temperature is 60°C. However. for regular use, please use at a constant temperature between 20°C and 50°C.
Usable solvents - Acetonitrile/water mobile phases are recommended.
Other instructions - Retention increases with increased acetonitrile concentration.
- Acetonitrile concentration should be within 0-95% (usually 50-95%).
- Retention will decrease when using methanol/water mobile phases.
- Only use HPLC-grade solvents.
- It is necessary to use salts or buffers for dissociating compounds. However, these have low solubility in the high-organic mobile phases used in HILIC mode. Phosphate buffers, widely used in reversed-phase, are not suitable for HILIC for this reason. When using salts or buffers, the acetonitrile concentration should be 70% or less. Before use, please confirm that the additives are completely soluble in the mobile phase.
- Neutral pH will maximize retention time.




  • Simultaneous separation of pharmaceutical substances and counterions using a serial connection of hydrophilic interaction and reversed phase columns
    (Poster from HPLC 2010, Boston, MA, USA)
    pdf (0.2 MB)


Reference list

Ordering Information


Product Name Size Product No. Price
Guard Column
4.6 mm l.D.×10 mm 07055-61 e-Nacalai
10 mm l.D.×20 mm 07058-31
20 mm l.D.×20 mm 07854-91
20 mm l.D.×50 mm 07873-41
28 mm l.D.×50 mm 07874-31
Packed Column
1.0 mm l.D.×150 mm 07869-11 e-Nacalai
1.0 mm l.D.×250 mm 07870-71
2.0 mm l.D.×50 mm 07052-91
2.0 mm l.D.×100 mm 08569-11
2.0 mm l.D.×150 mm 07054-71
2.0 mm l.D.×250 mm 07489-91
3.0 mm l.D.×150 mm 07871-61
3.0 mm l.D.×250 mm 07872-51
4.6 mm l.D.×150 mm 07056-51
4.6 mm l.D.×250 mm 07057-41
10 mm l.D.×250 mm 07059-21
20 mm l.D.×250 mm 07060-81
28 mm l.D.×250 mm 07875-21

(Storage) RT: Room temperature, A: Cool and dark, R: Refrigerator, F: Freeze