Waters Symmetry Columns - Manual (Page 5)

[ Care and Use ManUal ]
Symmetry Columns 5
Guard columns need to be replaced at regular intervals, as
determined by sample contamination. When system backpressure
steadily increases above a set pressure limit, it is usually an
indication that the guard column should be replaced. A sudden
appearance of split peaks is also indicative of a need to replace the
guard column.
b. Storage
For periods longer than four days at room temperature, store the
column in 100% acetonitrile. For elevated temperature applications,
store immediately after use in 100% acetonitrile for the best column
lifetime. Do not store columns in buffered eluents. If the mobile
phase contained a buffer salt, flush the column with 10 column
volumes of HPLC grade water (see Table 1 for common column
volumes) and replace with 100% acetonitrile for storage. Failure to
perform this intermediate step could result in precipitation of the
buffer salt in the column when 100% acetonitrile is introduced.
Completely seal column to avoid evaporation and drying out of the
bed.
Note: If a column has been run with a mobile phase that contains
formate (e.g., ammonium formate, formic acid, etc.) and is then
flushed with 100% acetonitrile, slightly longer equilibration times
may be necessary when the column is re-installed and run again with
a formate-containing mobile phase.
Vii. connectinG the column to the hplc
a. Column Connectors and System Tubing Considerations
Tools needed:
3/8 inch wrench
5/16 inch wrench
Handle the column with care. Do not drop or hit the column on a hard
surface as it may disturb the bed and affect its performance.
1. Correct connection of 1/16 inch outer diameter stainless
steel tubing leading to and from the column is essential for
high quality chromatographic results.
2. When using standard stainless steel compression screw fittings,
it is important to ensure proper fit of the 1/16 inch outer
diameter stainless steel tubing. When tightening or loosening
the compression screw, place a 5/16 inch wrench on the
compression screw and a 3/8 inch wrench on the hex head of the
column endfitting.
Note: If one of the wrenches is placed on the column tube flat during
this process, the endfitting will be loosened and leak.
3. If a leak occurs between the stainless steel compression screw
fitting and the column endfitting, a new compression screw
fitting, tubing and ferrule must be assembled.
4. An arrow on the column identification label indicates correct
direction of solvent flow.
Correct connection of 1/16 inch outer diameter stainless steel
tubing leading to and from the column is essential for high-quality
chromatographic results. Tubing touches the bottom of the column
endfitting, with no void between them. It is important to realize that
extra column peak broadening can destroy a successful separation.
The choice of appropriate column connectors and system tubing is
discussed in detail below.
Due to the absence of an industry standard, various column
manufacturers have employed different types of chromatographic
column connectors. The chromatographic performance of the
separation can be negatively affected if the style of the column
endfittings does not match the existing tubing ferrule settings. This
section explains the differences between Waters style and Parker
style ferrules and endfittings (Figure 1). Each endfitting style varies
in the required length of the tubing protruding from the ferrule.
The Symmetry column is equipped with Waters style endfittings
that require a 0.130 inch ferrule. If a non-Waters style column is
presently being used, it is critical that ferrule depth be reset for
optimal performance prior to installing a Symmetry column.
Figure 1: Waters and Parker Style Ferrule Types
0.130 inches 0.0 0 inches
Parker Style Ferrule Settings Waters Ferrule Settings