Waters Delta-Pak High Pressure Insert HPLC Column - Manual (Page 2)

[ Care and Use ManUal ]
2
II. InstallatIon
a. Installing the column
Use only plastic tubing and plastic finger-tight endfittings with the High-
Pressure Inert column. The Waters endfitting design is shown in Figure 1.
Over-tightening the endfittings can cause irreversible damage to the filter
housing.
Figure 1: Finger-tight Endfitting
To install a new fitting or replace a worn fitting:
1) Using a single-edge razor, make a straight and square cut on the
plastic tubing. Make this cut in front of the compression screw on the
worn endfitting.
2) Slide a compression screw fitting and a washer over the end of the cut
tubing.
3) Place the gripper ring in the top depression of the tool (see Figure 2).
Insert the end of the tubing into the gripper ring tool as far as it will
go. This seats the gripper ring at the proper length of the tubing.
4) Remove the tubing from the tool and slide the other washer and ferrule
over the end of the tubing. Insert a union over the endfitting assembly
and tighten. This seats the assembly.
Figure 2: Gripper Ring Installation
Make sure that any mobile phase used for startup is miscible with the ship-
ping solvent. Before placing the column in the flow path:
1) Attach a union between the column inlet and outlet lines.
2) Flush the lines to remove any microparticulates and old solvents. Flush
the injector loop if applicable.
3) Remove the union.
Remove the finger-tight end plugs from our column and save them to store
the column when it is removed from the system. Attach the column so that
flow follows the direction of the arrow on the column label. To install the
column, read the inlet and outlet fittings into the column until finger-tight.
b. Equilibration
The column is delivered in the mobile phase indicated in Table 1. Prior to
use, the column should be equilibrated with the mobile phase that will be
used in a analysis. Check mobile phase/shipping solvent miscibility.
Delta-Pak material is highly hydrophobic. If mobile phases with high concen-
trations of water are to be used, it is necessary to thoroughly solvate the
packing with the non-aqueous component before starting. To do so, pure the
column with 5 to 10 column volumes of the non-aqueous solvent to be used
in the mobile phase prior to equilibrating the column with the mobile phase.
Should the mobile phase contain a buffer salt, flush the column with 30
mL of water prior to equilibrating the column with the buffer. With PIC
®
ion- pairing reagents, whose concentration in the mobile phase is very
low (typically 5 mmol/l), 100 to 300 mL of mobile phase is required for
equilibration.
III. MoBIle PHase and saMPle GuIdelInes
Liquid chromatography columns have a finite lifetime directly related to the
care and use they receive. Column life is reduced by:
• contamination from the mobile phase or sample
• improper storage and handling
• frequent solvent switching
• exposure to high or low pH eluents (for example, less than
2.0, greater than 8.0)