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SeQuant® ZIC®-HILIC HPLC Columns

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Excellent Separations of Polar Hydrophilic Compounds

Merck:/Freestyle/LE-Lab-Essentials/Chromatography/LE-SeQuant ZIC-HILIC HPLC columns-150x248-03032014.jpgSeQuant® ZIC®-HILIC HPLC columns have documented excellent selectivity for separations of polar hydrophilic compounds in HPLC and LC-MS methods. These columns carry densely bonded, truly zwitterionic functional groups with a charge balance of 1:1. Separation is achieved by hydrophilic partitioning combined with weak ionic interactions resulting in maximum selectivity, high loadability and easy optimization of methods. The comprehensive SeQuant® ZIC®-HILIC product range includes classical analytical and capillary columns, as well as semi-preparative formats. All columns are available with 3 or 5 µm particles, and pore sizes of 100 or 200 Å.

Most Popular HILIC Columns

Its favorable retention and robust performance have made SeQuant® ZIC®-HILIC one of the most popular HILIC columns throughout the world. The columns have been used in numerous methods for a variety of compounds, such as acids and bases, anions and cations, carbohydrates, metabolites, metal complexes, amino acids, peptides, protein digests and many more. Alternative choices for separation of polar hydrophilic compounds include SeQuant® ZIC®-cHILIC with complementary selectivity, and SeQuant® ZIC®-pHILIC for higher pH stability.

Independent Comparison of HILIC Columns

A group of distinguished Japanese scientists performed comprehensive screening to compare the performance of 15 different HILIC columns. Their conclusion was that the selectivity of SeQuant® ZIC®-HILIC was higher and more versatile than that of all other stationary phases.

“In particular, the zwitterionic phase ZIC®-HILIC showed better selectivity for the test samples than the other phases.”
Kawachi et al., Journal of Chromatography A, Vol. 1218, Issue 35, 2 September 2011, Pages 5903–5919

Read the Article

What is HILIC?

HILIC or hydrophilic interaction liquid chromatography is a high-performance liquid chromatographic (HPLC) technique for separation of polar and hydrophilic compounds. Simply put, HILIC is a normal-phase type of separation but uses reversed-phase type eluents. Thus, it unites the benefits of aqueous mobile phases with separation based on normal phase principles in which more polar compounds respond with higher retention. This results in more efficient separation. Furthermore, HILIC is easy to combine with a wide range of detection methods, and boasts improved sensitivity with some of these techniques, such as electrospray ionization mass spectrometry (ESI-MS).

Application Examples

Among numerous other applications, the use of SeQuant® ZIC®-HILIC columns has been recommended by the US FDA (United States Food and Drug Administration) in its methods for the analysis of melamine ad related potential contaminants. Learn more about melamine analysis in the pharmaceutical industry.

Separation of Cyanuric Acid Melamine Food Contaminants

GradientTimeA: 95% acetonitrile in 0.1% aqueous formic acid
B: 50% acetonitrile in 20 mM aqueous ammonium formateFlow rate
0 100 0 0.4
4.2 100 0 0.4
8.0 65 35 0.4
8.5 65 35 0.4
9.0 25 75 0.4
11.0 25 75 0.4
11.2 100 0 0.6
13.0 100 0 0.6
14.0 100 0 0.4
ZIC®-HILIC 150 x 2.1 mm, 5 μm, 200 Å 
Column ZIC®-HILIC 150 x 2.1 mm, 5 μm, 200 Å 
Detection MS/MS, -ESI and +ESI
Temperature 30°C
Inj. volume 5 μL
Sample Standard in mobile phase
(7 ng/mL) equivalent to 1 μg/g
Courtesy of David N. Heller, FDA Center for Veterinary Medicine, Laurel, MD 20708 USA
Reference David N. Heller and Cristina B. Nochetto, Rapid Commun. Mass Spectrom., 22 (2008)
3624-3632. DOI: 10.1002/rcm.3779
Separation of cyanuric acid melamine food contaminants on SeQuant® ZIC®-HILIC

Separation of Amino Acids

Column ZIC®-HILIC 150 x 4.6 mm, 3.5 μm, 100 Å 
Mobile Phase 80% acetonitrile
20% ammonium acetate in water, 50 mM pH adjusted to 4.5 with formic acid
Flow rate 0.75 mL/min
Detection Refractive index, cell 9 μL, 40°C
Temperature 40°C
Inj. volume 50 μL
Gradient 1. 2-Amino-5-Hydroxy benzoic acid
2. Tryptophan
3. Isoleucine
4. Methionine
5. Lycine
6. Threonine
100 ppm
100 ppm
100 ppm
100 ppm
100 ppm
100 ppm
Courtesy of Gora Sharangi, MilliporeSigma India Application Lab
Separation of amino acids on SeQuant® ZIC®-HILIC

Bioanalysis of Methyl Malonic Acid

Column ZIC®-HILIC 100 x 2.1 mm, 3.5 μm,100 Å
Mobile Phase A: 80% (v/v) acetonitrile
B: 20% (v/v) 100 mM NH4Ac buffer pH 4.5
Flow rate MS-ESI- with SIM at 117.2 m/z           
Drying-gas flow rate
Drying-gas temperature
Capillary voltage

10 L/min
3.0 kV
Detection Refractive index, cell 9 μL, 40°C
Inj. volume 4 μL
Sample Precipitation-treated plasma sample containing 0.137 mol/L MMA
Reference H-A Lakso, P. Appelblad, J. Schneede
Clin. Chem., 54 (2008) 2028
DOI: 10.1373/clinchem.2007.101253
Merck:/Freestyle/LE-Lab-Essentials/Chromatography/Diagrams/LE-SeQuant ZIC-Bioanalysis of methyl malonic acid-480x384-03042014.jpg

Functional groupFeaturesBase particleParticle sizePore sizeColumn types
Well-known for high-performance selectivity and robustness silica 3.5 µm,
5 µm
100 Å, 200 Å analytical,
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