HPLC Column


  • Overview
  • Product list
  • Instruction manual

CAPCELL CORE instruction manual

Thank you for choosing a CAPCELL CORE column. CAPCELL CORE C18 is provided as a high-performance column of core-shell type packed with 2.7µm spherical silica particle (1.7µm solid core and 0.5µm porous layer), which is coated by a mono-layer silicone polymer and further bonded by octadecyl (C18) as function group.

1. Column Handling

  1. Handle the column with great care. A strong shock may cause damage.
  2. Attach or detach the column when the pressure gage indicates zero.
  3. The maximum of the column operating pressure is 60 MPa.

2. Column Installation

  1. The column joint is of the male nut type for tubing of 1/16 inch OD. Check that the tubing joints of the system fit correctly and that the ferrule tips are deeply inserted into the joints. (See Fig. 1.)
  2. Before attaching the column, replace the liquid in the system with the mobile phase to be used.
    Note the replacement procedure to avoid salting out. The shipment solvent is described in the column report enclosed with the column.
  3. Install the column according to the direction of the arrow.

3. Analysis

3-1. Mobile phase

  1. All solvents applicable for the conventional chemically bonded silica columns can be used.In addition, high temperature and organic solvent poor condition will result in short column lifetime when working at the extremes of pH.
  2. The applicable pH range for CAPCELL CORE C18 is 1.5~10.For continued high performance of the columns, ensure to avoid using pH exceeding the recommended one.
  3. After fully degassing, filtrate the mobile phase using a membrane filter 0.45µm or smaller to remove dust.To prevent foreign matter from clogging the column inlet filter, an online filter connected before the column is recommended.
  4. To prevent column from deterioration, avoid the followings;
    • Frequent change of the mobile phase compositions
    • Direct change to any mobile phase with bad compatibility
    • Rapid change of the pressure in column inlet
    • High column pressure due to the use of a high-viscosity mobile phase
    • Long-term flowing of water

3-2. Sample Preparation

  1. Dissolve the sample in a solvent with the same composition as that used for elution wherever possible.
  2. Using a solvent with strong elution may lead to poor resolution resulting to broad peak or cause the sample to precipitin at the column head.
  3. If there is insoluble matter remaining in the sample solution, filtrate the solution using a filter 0.45µm or smaller.
  4. The pH of the sample solution should be set under the pH range applicable for particles.

3-3. Attention on Analysis

CAPCELL CORE has smaller specific surface than that of totally porous particles because of the base material of core shell type particle.Accordingly when shifting a conventional method with sample loading near the upper limit to CAPCELL CORE column, please keep in mind that it is necessary to reduce the sample loading amount and optimize the injection.

4. Column Storage

  1. Completely seal the column with the accessory plugs and store in a cold place where there is little temperature fluctuation.
  2. For storage within one month, replace the column with solution having the same compositions of organic solvent as that of the mobile phase but without acid and salt included.In addition, replacement by water only is not recommended.
  3. For storage more than one month or longer, replace with acetonitrile after processing the procedure in 2.

5. End Fittings

  1. An analytical column of up to 6-mm ID uses a filter-embedded end fitting as shown in Fig. 1. The filter cannot be changed alone. If the filter is clogged or the column pressure is high, replace the end fitting.
  2. See Fig. 1 for the column connection. If the tubing is inappropriate, especially if a tube for a different type of column is used, the length after the ferrule tip (V in Fig. 1) is often different from the end fitting length L, and a problem may occur.
    If L is greater than V, dead volume may be generated and cause peak broadening or tailing or deterioration of separation performance.
    If L is smaller than V, liquid may leak because of inadequate ferrule adhesion.
    Therefore, we recommend replacing the ferrule together with the column.
    *If the column is replaced frequently, the male nut may crush the ferrule and liquid may leak. Since tightening the nut too much may cause its head to come off, replace the ferrule at an early stage.

Fig. 1  Column connection

【Fig. 1】 Column connection

6. Troubleshooting

Problems in HPLC and UHPLC are attributable to various causes that cannot all be listed up. The table below describes some comparatively common problems related to the column.

Symptom Causes Remedies
1. Column pressure rise. Blocking with foreign matter
1. Dust or insoluble matter in the mobile phase or sample solution.
2. Dirt in the tubing.
3. Plunger seal fragment.
4. Precipitation of sample components.
• Sonicate the filter or replace it.
• Filtrate the mobile phase and sample solution in advance using a membrane filter.
• Attach a line filter.
• Clean the tubing and replace the plunger seal.
• Prepare a sample solution with the mobile phase.
2. Peak splitting, tailing, and broadening. 1. Void in the column head.
2. Dead volume due to inappropriate connections.
3. Inappropriate mobile phase conditions.
4. Column deterioration.
*Deterioration derived from damage in packing bed is irreparable.
• Reconnect the tubing.
• Review the pH, salt concentration, sample amount, and other conditions.
• Review the ion pair agent concentration, pH, sample amount, and other conditions.
• Check the column performance using standard inspection solution.
3. Retention time too long or unstable. 1. Liquid leak (Indicated on the pressure gage of the pump).
2. Inappropriate mobile phase conditions.
3. Inadequate column equilibration time.
• Check the pump and tubing for any leaks.
• Secure adequate equilibration time.
4. Retention time too short. 1. Hydrolysis (deterioration) of a bonded groups by strong acid or base.
2. Inappropriate mobile phase conditions.
3. Inadequate column equilibration time.
• Check the column performance by using standard solution for verfication.
• Secure adequate equilibration time.
• Ensure equilibration time.

CAPCELL CORE is shipped after a strict performance check. However, if you should find any defect, please contact your dealer or Osaka Soda for replacement.
Note that Osaka Soda does not warrant the product against column lifetime or any deterioration caused by the failure to follow the above instructions for care and use.
We assume that the product was delivered in good condition, if there is no any claim ten or more days after reception by the customer, and will not accept a later replacement request.

August 1st 2012

Page top