ExcsepTM NH2 Column User Manual

Introduction

ExcsepTM NH2 columns are made of ultra-high purity, ultra-high mechanical strength spherical chromatography bare silica gel, prepared by the unique surface modification technology of Onetwo Chromatography Technology. Bonded to aminopropyl, it is suitable for the separation of polar and hydrophilic compounds, such as monosaccharides, oligosaccharides and sugar alcohols.

Column Activation

ExcsepTM NH2 columns are stored in hexane:isopropanol (99.5:0.5) solvent after production.

Reversed-phase mode use: When the mobile phase is a polar solvent such as methanol, acetonitrile, water, etc., use 100% isopropanol-100% acetonitrile-do the sample mobile phase in sequence, respectively, at 0.3mL/min, 1.0mL/min, 1.0mL/min, rinse for 12h, 2h, and 40min. If buffer salts or ion-pairing reagents are used in the mobile phase, be sure to use the same proportion of the aqueous phase for the transition.

Normal phase mode of use: when the flow is a weakly polar solvent such as n-hexane, isopropanol, etc., in turn, 100% isopropanol - do the sample mobile phase, respectively, at 0.3mL/min, 1.0mL/min, rinsed 12h, 2h.

Storage

Reversed-phase mode is used:

The mobile phases are polar solvents such as methanol, acetonitrile, and water. After use, rinse the column with pure acetonitrile at a flow rate of 1.0 mL/min for more than 30 min and store. If buffer salts or ion-pairing reagents are used in the mobile phase, be sure to use the same proportion of aqueous phase for transition.

Normal phase mode is used:

The flow is weak polar solvents such as n-hexane and isopropanol. After use, flush the column with n-hexane:isopropanol (95:5) at a flow rate of 1.0 mL/min for more than 30 min and store.

Solution for Column Efficiency Reduction

When the chromatographic column shows abnormal phenomena such as increased column pressure, decreased column efficiency, abnormal peak shape, and decreased separation, it is necessary to maintain the column:

Reversed-phase mode is used:

The mobile phase is a polar solvent such as methanol, acetonitrile, water, etc. Reverse the connection of the column, and use the sampling flow rate to rinse the column sequentially with 100% acetonitrile-100% methanol-100% isopropanol-100% acetonitrile (if buffer salts or ion-pairing reagents are used in the mobile phase, be sure to use the same proportion of the aqueous phase to make a transition first, and then start to rinse with 100% acetonitrile). , rinse each solvent for more than 40min. Isopropanol has a high viscosity and high pressure, so please pay attention to adjusting the appropriate flow rate. After the rinsing is completed, connect the column positively for use.

Normal phase mode is used:

The flow rate is weak polar solvents such as n-hexane, isopropanol, etc. Reverse the column and use the sample flow rate to rinse the column with 100% isopropanol - 100% methanol - 100% isopropanol - 100% n-hexane in sequence, and rinse for more than 40min for each solvent. Isopropanol has a high viscosity and high pressure, please pay attention to adjust the appropriate flow rate. After rinsing, the column should be connected to the normal use.