Waters nanoACQUITY Columns 5000 psi - Manual (Page 2)

NanoACQUITY ™ COLUMNS CARE AND USE:
NanoACQUITY ™ columns are manufactured to exacting specifications,
which provide unmatched peak symmetry for maximum sensitivity
and accurate quantitation. As the columns are fritted at both ends,
they are able to withstand pressure changes during injection and
thus provide superior longevity. In order to achieve maximum
performance, it is important to ensure that proper connections are
made to minimize peak tailing and poor efficiency.
CONTENTS:
• NanoACQUITY ™ Column
• Care and Use Instruction Sheet – This instruction sheet contains
important information about using and maximizing performance
from your NanoACQUITY
™ Column.
• Certificate of Batch Analysis – Demonstrates the results of the
physical and chemical characterization tests performed on the
batch of chromatographic packing in your NanoACQUITY
™
column.
• Certificate of Column Performance – Demonstrates a
chromatographic separation of a peptide mixture from a
representative column.
COLUMN DESCRIPTIONS:
The NanoACQUITY ™ column you have purchased contains several
innovative features designed to provide maximum performance,
ease-of use and longevity:
75 µm, 100 µm, and 150 µm Column
1) Optimized column packing protocol to provide superior
column efficiency.
2) Diamond-polished column ends and connecting tubing to
provide minimum extra-column peak broadening.
3) Proprietary fritting technology to provide stable, long-lasting
column (patent pending).
4) Special fitting to connect to a NanoACQUITY ™ system.
5) Protective PTFE oversleeve for easier handling and more
rugged column design (patent pending).
6) 500 cm of 20 µm ID connection tubing coupled to column
outlet with zero-dead-volume PTFE coupler to maintain
chromatographic performance while simplifying column set-up.
7) Zero-dead volume PTFE protector/coupler for joining column
to electrospray emitter top or other detector.
300 µm Column
1) Optimized column packing protocol to provides superior
column efficiency
2) Low volume Valco style end fitting for minimum band-broadening
and ease of use
GENERAL COLUMN INSTALLATION:
NOTE: Take care in handling NanoACQUITY ™ columns to prevent
inadvertent damage. With the 75 µm, 100 µm, and 150 µm
columns, do not bend the PTFE oversleeve, and do not pull the
capillary connection tubing away from the column coupling.
Flow mobile phase only in the direction indicated by the arrow
on the column label. Connect the column with the direction of
the flow arrow on the label pointing to the detector or Mass
Spectrometer.
75 µm, 100 µm, and 150 µm Installation Column:
1) Remove the protective tube from the inlet end of the column. Connect
the column inlet to the NanoACQUITY
™ system. Do not overtighten.
2) For connecting the NanoACQUITY ™ column to your electrospray
emitter or other detector connection tubing, Waters recommends
that you use the supplied PTFE zero-dead volume coupler to connect
your column to the detector. Firmly grasp the coupler that is attached
to the connecting tubing, and slide it down the tubing until the tube
end is positioned approximately halfway inside the coupler. If you
have access to a low power microscope, use it to examine the end
of the emitter to be inserted in the coupler to ensure that there is
no particulate contamination. Remove contaminants by wiping
carefully with a Kim-wipe moistened with methanol. While grasping
the coupler, carefully insert and slide your electrospray emitter in
the coupler until the emitter and connecting tubing are butted
against each other. Inspect the union to ensure that both ends are
butted together properly. If you observe shavings of PTFE between
the connections, remove the emitter and slide the connecting tubing
through the coupler to remove the pieces of PTFE. Repeat the procedure
described above to re-connect the connector to the emitter. This
approach will provide the lowest dead volume connection and
will result in the best chromatographic peak shape.
Alternatively, you can remove the coupler and connect your column to
the detector using commercially available microtight connectors. However,
this approach can result in poor chromatographic performance because
there can be significant unswept dead volume.
300 µm Installation Column:
1) Connect the column inlet to the injector utilizing a Valco style
compression screw.
2) Connect the column outlet to the detector or Mass Spectrometer
using a Valco style compression screw.
COLUMN MAINTENANCE:
Caring for the column
To ensure continued high performance of your NanoACQUITY ™
column, follow these guidelines:
1) Always use filtered mobile phases.
2) Filter or centrifuge particulate-laden samples.
3) Do not subject the column to sudden changes of pressure or flow.
Cleaning and regenerating the column
A shift in retention or resolution may indicate contamination of the
column. Flushing with a neat organic solvent is usually sufficient to
remove the contaminant. If system backpressure increases with usage,
the column inlet is most likely becoming plugged with particulates
from the mobile phase or sample. Use an appropriate solvent to
dissolve the precipitate. For example, proteinaceous material can
be removed with repeated injections of neat DMSO.
Column storage
For overnight storage, continuously flush the column with the mobile
phase at 10 - 20% of the maximum recommended flow rate. Store
the column in the HPLC grade water when it will be used within
24 hrs or in 100% acetonitrile for long term storage. (Caution:
Rinse column in HPLC grade water before switching to 100%
organic solvent to avoid potential salt precipitation in column.)
SAFETY:
Use appropriate safety glasses or goggles meeting ANSI Z87.1-
1989 requirements when handling fused silica tubing.