Ultra-high pressure HPLC can be one method to
combat the high sample complexity of shot gun proteomics mixtures. During the
years there have been an o lot of effort in optimizing the hardware setup. For
example using solid core particles or longer columns.
The Coon Lab tried to reduce the van Deemter A-Term
by creating an really tight and homogeneous packing bed of C18 particle utilizing
pressures of over 2000bar for column packing. The packed columns turned out of
display reduced backpressure and improved sequence coverage of +23%.
How this has been achieved?
Well, here a special type of pump was used. An air
driven liquid pump based on pneumatic principle from heskel. This company
usually operates in the hydrogen-gas fueled car industry but there multi head
pump system which are able to create outlet pressures of up to 7000bar shown to
be beneficial for column loading. The fittings and values used have been
manufactured by HiP. Especially the connection from the slurry reservoir,
having a ID of 1mm to the capillary with and OD of 0.3mm seems to be crucial to
me (an modified femal to male fitting was used).
Keine Kommentare:
Kommentar veröffentlichen