Waters Spherisorb Columns - Manual (Page 7)

[ Care and Use ManUal ]
Waters Spherisorb Columns 7
buffer salt in the column when 100% acetonitrile is introduced.
Note: The exception of the storage recommendations above are cyano (CN)
columns used for reversed-phase methods (CN-RP). These columns should never
be stored in 100% intermediate polarity solvents (acetonitrile, methanol, IPA).
Store CN-RP columns in 0.1 M ammonium acetate/acetonitrile 50/50.
2. Normal-Phase Columns
For rapid equilibration upon start-up, store your normal-phase
column in the mobile phase that is commonly used. Completely seal
column to avoid evaporation and drying out of the bed.
vi. connectinG the column to the hplc
a. Column Connectors and System Tubing Considerations
All Spherisorb column and cartridges have Parker style endfittings.
Tools needed for Spherisorb analytical column:
1/2 inch wrench
9/16 inch wrench
Tools needed for Spherisorb analytical cartridge column:
1/4 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 chromato-
graphic 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 1/2 inch
wrench on the hex head of the column endfitting.
Note: If one of the wrenches is improperly placed on the inner column hex head (or the
cartridge 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 direc-
tion 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
Spherisorb column is equipped with Parker style endfittings that require
a 0.090 inch ferrule. If a Waters style column is presently being used,
it is critical that the ferrule depth be reset for optimal performance prior
to installing a Spherisorb column.
Figure 1: Waters and Parker Style Ferrule Types
In a proper tubing/column connection (Figure 2), the tubing touches
the bottom of the column endfitting, with no void between them.
Figure 2: Proper Tubing/Column Connection
The presence of a void in the flow stream reduces column perfor-
mance. This can occur if a Parker style ferrule is connected to a
Waters endfitting (Figure 3).
0.090 inches 0.130 inches
Waters Ferrule Settings Parker Style Ferrule Settings