Ion Exchange HPLC Columns

IonSep™ - SCX, WCX, SAX, and WAX are non-porous resin phases coated with a hydrophilic layer and further bonded with SCX, WCX, SAX, and WAX functional groups, respectively. The chemistry feature provides high resolution, efficiency and recovery separations of proteins, oligonucleotides and peptides. The packing materials are made of 1.8µm, 3µm, 5µm, and 10µm highly cross-linked polystyrene divinylbenzene (PS/DVB) non-porous particle.

Selection Guide

Order Information

Application Examples

Application Note:

1. IonSep-SAX and -WAX columns are shipped in 20 mM Tris at pH 8.0 and IonSep-SCX and -WCX columns in 20 mM phosphate at pH 6.0.
2. All samples and solvents including buffers should be filtered through 0.45 μm or 0.2 μm filters to avoid column clogging.
3. The column compatible with aqueous mobile phase or a mixture of organic and water, such as methanol or acetonitrile and water. Please be noted that (3.1) The SAX, WAX, SCX, and WCX are compatibles with nonionic and zwitterionic detergents; (3.2) SAX and WAX are not compatible with anionic detergents; (3.3) SCX and WCX are not compatible with cationic detergents; and (3.4) Typical buffers include phosphate, tris, and acetate.
4. Always degas the mobile phase. A simple way for degassing is to sonicate it for 5 minutes under water pumped vacuum.
5. Normal flow rate: 0.1 - 1.0 mL/min for 4.6mm I.D. columns.
6. Flush the column with gradual increasing the mobile phase flow rate from 0.1 mL/min to the target until the baseline is stable.
7. The air bubbles trapped inside the column may cause the column backpressure and baseline to fluctuate, please continue flush the column with higher flow rate for 2-5 minutes, for example 0.5 mL/min for a 4.6x300 mm column.
8. A sudden increase in backpressure may be due to plugging of the inlet frit, which could be resolved by flushing column with reverse flow.
9. The optimum operation temperature is 10-50^oC to reach longest operating life (maximum 80^oC) at pH 2 - 12.
10. Keep SAX and WAX columns in 20 mM Tris at pH 8.0/0.1% NaN₃, SCX and WCX columns in 20 mM phosphate buffer at pH 6.0/0.1% NaN₃.

Column Cleaning:

1. Clean the pre-column filter or the guard column first by flushing the filter or the guard column in reverse flow direction using washing solutions for 20 min. Recommended washing solutions are 150 mM KNO₃ in 75% ACN at pH 2 (adjusted by HCl) for SAX and WAX columns and 50 mM phosphate buffer in 1.0 M NaCl at pH 10 for SCX and WCX columns.
2. Disconnect the column from the detector and clean column in the reverse flow direction by flowing cleaning solution.
3. Flow rate of cleaning solution typically should be less than 50% of the maximum recommended flow rate
4. After about 10 column volumes (CVs) of cleaning solution are used, rinse column 3 CVs of distilled/deionized water between each rinse.
5. Cleaning solutions: (4.1) 150 mM KNO₃ in 75% ACN at pH 2 (adjusted by HCl) for SAX and WAX columns and (4.2) 50 mM phosphate buffer in 1.0 M NaCl at pH 10 for SCX and WCX columns.
6. A guard column is highly recommended to effectively trap adsorptive sample components and residual particulates in the sample and mobile phase.