PRODUCTS > CIM® Industrial Columns

CIM® Epoxy-8f mL Tube Monolithic Column

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About CIM® Tube Monolithic Column

Thank-you for purchasing your CIM® Tube Monolithic Column. The CIM® Tube Monolithic Column is an ideal choice for efficient  laboratory and industrial scale purification of large moleculessuch proteins or DNA and also smaller molecules such as peptides and oligonucleotides as well as for the bioconversion of of various molecules. This monolithic column operates at extremely high flow rates, low back pressures, and is extremely easy to use. The advantageous features of CIM® Tube Monolithic Column include:

  • Use with any HPLC, LC, peristaltic pump or syringe with appropriate adapters
  • Fast large scale separations of biomolecules
  • Flow unaffected dynamic binding capacity
  • High resolution separations even with increased sample loadings
  • Low back pressures even at very high flow rates
  • Extremely fast column equilibration
  • pH stability

Before You Begin

Unpacking

Carefully inspect the purchased products for any damage that may have occurred during shipping. Immediately report any such damage to your vendor and the transport company. CIM® Tube Monolithic Column is shipped in a 20 % ethanol solution and need to be stored in the refrigerator at +4 °C (39 °F) to +8 °C (46 °F) until ready to use.
WARNING: Do not store the CIM® Tube Monolithic Column below 0 °C (32 °F).

Preparation of the CIM® Tube Monolithic Column for Use

CIM® Tube Monolithic Column consist of a CIM® monolith in a polypropylene housing (Figure 1). The chemistry of the monolith and the flow direction are marked on the label.


Connecting the CIM® Tube Monolithic Column

  1. Remove both of the column blind fittings
  2. Attach the CIM® Tube Monolithic Column to the pump system according to the flow direction using standard 1/8” OD fittings 5/16-24 coned (Valco type). If you are using HPLC with capillaries 1/16”, please use appropriate adapters (male 5/16-24, female 10-32).
  3. Avoid contact with the housing and retaining fittings with organic solvents or strong acids.

Equilibrating the CIM® Tube Monolithic Column

Before you begin working with the CIM® Tube Monolithic Column the product should be equilibrated and the flow characteristics and back pressure checked in accordance with the following procedure:
  1. Wash the CIM® Tube Monolithic Column with at least 5 column volumes of the binding mobile phase (e. g. for ion-exchange chromatography a low ionic strength buffer) at one-half of the working flow rate.
    Note:     Allow 2 column volumes to flow into a waste container. This will remove any small particles or air bubbles that may affect the detector cell.
  2. Wash the CIM® Tube Monolithic Column with at least 5 column volumes of the eluting mobile phase (e.g. for ion-exchange chromatography a buffer containing 1–2 M NaCl) at one-half of the working flow rate in order to elute any bound components and impurities.
  3. Finally, wash the CIM® Tube Monolithic Column again with 10 column volumes of starting (binding) mobile phase at a working flow rate.
  4. Your CIM® Tube Monolithic Column is now ready for use.

Column Charecteristics

Monolith Dimensions

Outer diameter, Do: 15 mm
Inner diameter, Di: 6.5 mm
Length, L: 56.0 mm
Bed volume, V: 8 ml
Flow direction: In radial direction from outer to inner surface, see Figure 2.

Calculation of Linear Velocities (Outer, Inner and Average)

Outer surface, Ao, = π Do L = 26.4 cm2
Inner surface, Ai, = π Di< L = 11.4 cm2
Linear velocity at the outer surface, uo (cm/min) = F (cm3/min)/Ao
Linear velocity at the inner surface, ui (cm/min) = F (cm3/min)/Ai

 

Table 1: Outer, inner and average linear velocities (cm/min) at different flow rates

F (cm3/min) 20 40 80 120 240 400
uo (cm/min) 0.76 1.52 3.03 4.55 9.10 15.17
ui (cm/min) 1.75 3.50 7.00 10.50 21.00 35.00
uav (cm/min) 1.12 2.24 4.48 6.71 13.43 22.38

Operating Conditions

  1. Operating Flow Rates: up to 400 mL/min for mobile phases with viscosities close to the viscosity of water (0.001 Pa · s).
    WARNING:     Reversing the flow direction can seriously damage your column!
  2. Operating Back Pressure: up to 20 bar (2 MPa)
    WARNING: Exceeding 2 MPa can seriously damage your column!
  3. Mobile Phases and Organic Solvents: pH: 1–13, salt concentration: up to 8 M, do not use concentrated organic solvents.
  4. Operating Temperature: up to 50 °C (122 °F).
    WARNING    : exceeding 50 °C (122 °F) can seriously damage your column.

Caring for the Product

To extend the life of your CIM® Tube Monolithic Column, please observe the following guidelines:

  • Always use freshly prepared mobile phases (buffers).
  • Always filter your mobile phases (buffers) and samples through a 0.22 μm filter.
  • Protect the tube monolithic column by using high-pressure in-line filters
  • We also recommend using high-pressure in-line filters between the column outlet and the detector.
  • Never remove the caps from the CIM® Tube Monolithic Column.

Cleaning and Regeneration of the CIM® Tube Monolithic Column

Please refer to the Product Specific Information Sheet which accompanied these instructions.

Storing the CIM® Tube Monolithic Column

Up to 2 days of planned storage after column use:
Equilibrate the CIM® Tube Monolithic Column according to the procedure described in Equilibrating the CIM® Tube Monolithic Columns. Than:

  • Stop the pumps and disconnect the column from the HPLC/LC system
  • Carefully seal the ends of the column with blind fittings to prevent drying
  • Store the column at +4 °C (39 °F) to +8 °C to (46 °F).
    WARNING    : Do not store the CIM® Tube Monolithic Column below 0°C (32 °F).

3 or more days of planned storage after column use:
Before long term storage of the CIM® Tube Monolithic Columns perform the equilibration/regeneration procedure as described on Page 2 and then wash the column with 10 column volumes of 20 % ethanol at a working flow rate. The column filled with 20 % ethanol should be carefully sealed with blind fittings and stored at +4 °C (39 °F) to +8 °C to (46 °F).
WARNING: Never let the CIM® Tube(s) dry out!

Troubleshooting

Problem

Possible source

Action
Increased back pressure Blocked tube monolithic column, mixing tee, capillaries, in-line high pressure filter, or detector cell.
  • Perform regeneration of the Tube Monolithic Column as described in the Product Speficic Information Sheet
  • Exchange capillary, in-line filter, or mixing tee
  • Clear out the detector cell
Poor resolution (i.e. Wider peaks)

 

Various
  • Perform regeneration and/or CIP procedure described in the Product Specific Information Sheet
No apparent separation Various
  • Check the pH and the composition of the mobile phase
  • Verify that the tube chemistry is appropriate for your application
Poor or no baseline Various
  • Check the pH and the composition of the mobile phase
  • Make the gradient flatter
  • Increase the flow rate

LOOK-UP CHEMISTRY
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