Re: HPLC: 1 mm columns

Kevin Howland (k.howland@ukc.ac.uk)
Wed, 13 May 1998 16:49:00 +0100

I used to run a 140B based LC system with 1mm columns (but with a different
detector). The dynamic mixer needs to be replaced with a 75 ul mixer -
because this was developed for the 140c you need to swop the motor off the
old mixer keep the electrical connections the same. Its a good idea to use
0.005" tubing from the mixer and to drill through the casing of the pump to
allow the shortest possible connection to the injector.

When programming a gradient, you need to pressurise with at least 5% B or
you get cross flow of buffer A into piston B. You also need to allow the
system to equilibrate for a minimum of 15 min before you inject. If you
have 50 or 75 um i.d. fused silica capillary as the flow cell outlet this
should reduce/eliminate bubbles in the flow cell.

Kevin

>ABRF colleagues,
>
> I would like to know if it is possible to use an Applied
>Biosystems Model 140B Solvent Delivery system to run 1 mm
>i.d. columns. The detector is a Model 785A absorbance
>detector but I do not know the size of the flow cell.
> Similarly, can the ABI Model 120 HPLC system be used for
>1 mm columns? I would assume it would be necessary to
>change or replace the relavtively large volume dynamic
>mixers on each.
> Thanks in advance for your input.
>-Mark
>
>Mark O. Lively, Ph.D.
>Professor of Biochemistry
>Wake Forest University School of Medicine
>Medical Center Blvd.
>Winston-Salem, North Carolina 27157
>Phone: 336-716-2969
>Fax: 336-716-7200
>email: mlively@wfubmc.edu

_______________________________________
Kevin Howland

Protein Science Facility Manager
Research School of Biosciences
University of Kent
Canterbury CT2 7NJ
Tel: +44-1227-764000 ext 7987
Fax: +44-1227-763912
email: k.howland@ukc.ac.uk