Columns for the Octave^™
Chromatography System

The Semba Octave™ Chromatography System can be used for SMBC purification in a variety of applications, depending on the separation medium and chemistry. The Octave System is an "open platform" permitting the use of a wide range of chromatography columns and solid phases, provided they meet some critical requirements.

An important consideration for the type of column to be used with the Semba Octave System is the expected back pressure that will be generated during operation. The total fluid back pressure limit for the instrument is 270 psi (18.6 bar, 1.86 MPa), which is due to the pressure limit of the pneumatic valve system (300 psi). Note that back pressures are additive. During typical applications no more than 3 columns are in any one zone, so in general if the back pressure generated by a single column with a given solvent and flow rate is less than 90 psi, it should be compatible with the system.

Bioseparation

The system has a non-metallic flow path making it suitable for use with biological samples including:

• crude cell lysates
• tissue culture fluids
• serum samples

In all cases, as with any high performance chromatography system, samples should be free of particulate matter. Filtration of samples through a 0.45 micron membrane is usually sufficient.

The system accommodates a variety of column sizes. In general, columns up to 2 cm x 10 cm packed with any polymeric resin (e.g. cross-linked agarose, methacrylate, polyacrylamide) having a particle size greater than 30 microns in diameter are compatible with the Octave System. Convenient 1-ml and 5-ml biochromatography columns for many applications are available for use with the Octave System.

Chemical Separation

The increased productivity of SMBC allows the use of smaller columns with larger particle size and dramatically decreased plate numbers in the Octave System versus equivalent separations performed by HPLC (Perrin and Nicoud, 2001). These conditions generate lower back pressures and enable higher flow rates for preparative SMBC. In general, columns having a particle size of 10 microns or greater (ideally 15 – 30 microns) are suitable for efficient performance on the Octave System. Columns having a particle size less than 10 microns in diameter usually generate back pressures exceeding 270 psi under SMBC conditions and are not recommended for use with the Octave System.

The flow path in the Octave System is compatible with most organic solvents normally used in HPLC applications. Unsaturated halogenated hydrocarbons, e.g. methylene dichloride, trichloroethane, chloroform should be avoided.