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SeQuant® ZIC®-pHILIC Polymeric HPLC Columns

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Use SeQuant® ZIC®-pHILIC HPLC Columns for More Difficult Separations of Polar Hydrophilic Compounds.

Merck:/Freestyle/LE-Lab-Essentials/Chromatography/LE-SeQuant pHILIC-HILIC-150x225-03042014.jpgSeQuant® ZIC®-pHILIC HPLC columns are the preferred choice when extreme eluent conditions are needed to overcome challenging separations of polar hydrophilic compounds. Offering excellent pH stability from pH 2 to 10, these polymer-based columns carry densely bonded, truly zwitterionic functional groups with a charge balance of 1:1. This results in straightforward and reproducible separations with wide-spectrum pH tuning for optimal separation selectivity and detection sensitivity. Furthermore, SeQuant® ZIC®-pHILIC columns minimize background noise in evaporative detection techniques with high sensitivity to ions, such as charged aerosol detectors (CAD). SeQuant® ZIC®-pHILIC columns are available with 5 µm polymer particles in various formats of analytical dimensions. Alternative choices for the separation of polar hydrophilic compounds include silica-based SeQuant® ZIC®-HILIC columns with identical functional groups, and SeQuant® ZIC®-cHILIC columns with complementary selectivity.

Independent Column Comparison

Researchers of an American pharmaceutical company compared several HILIC columns of different brands for the analysis of counter ions using a charged aerosol detector. According to their results, ZIC®-pHILIC offered favorable selectivity and peak shapes with minimum background noise – a combination not achieved with the other columns tested. “When operating on ZIC®-pHILIC columns, the baseline noise was negligible (<3 pA) and not affected by increasing the aqueous percentage of the eluent.” (Huang et al., J. Pharm. Biomed. Anal. 50 (2009) 809-814)

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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

SeQuant® ZIC®-pHILIC columns have been used for a wide range of applications, and are often preferred for difficult separations in which high mobile phase pH is required for enhanced selectivity, stability or sensitivity of analytes.

Separation of Gentisic Acid, Protocatechuic Acid, and Isophtalic Acid

ZIC®-pHILIC (5 µm, polymer) PEEK 100x4.6 mm
Mobile phase
75:25 Acetonitrile / aqueous buffer pumped at 0.5 mL/min
Buffer salt
Ammonium acetate 17 mM, pH 6.8
Ammonium carbonate 17 mM, pH 9.6
Separation of gentisic acid, protocatechuic acid, and isophtalic acid on SeQuant® ZIC®-pHILIC
Separation of gentisic acid, protocatechuic acid, and isophtalic acid on SeQuant® ZIC®-pHILIC

0 20% 80%
15 60% 40%
20 60% 40%
35 20% 80%
Column ZIC®-pHILIC (5µm, polymer) PEEK 150x2.1 mm
Mobile phase A: Acetonitrile
B: 10 mM (NH4)2CO3 + 0.2% NH4OH
Sample Pantothenate (2, structure), Acetyl-CoA (3), AMP (4),
CoA (5), ADP (6), and ATP (7).
Courtesy of T. Pluskal, K. Nagao,
M. Yanagida, GO Cell Unit,
Initial Research Project Okinawa Institute of Science and Technology,
Okinawa 904-2234, Japan.
Separation of gentisic acid, protocatechuic acid, and isophtalic acid on SeQuant® ZIC®-pHILIC

Functional groupFeaturesBase particleParticle sizePore sizeColumn types
High pH-stability and low noise with ion detectors polymer 5 µm analytical,

Technical Support for SeQuant® HILIC Columns

Merck offers several technical support services to help you achieve optimal separations and reliable analytical results with SeQuant® HILIC products.

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