User guide

[ Care and Use ManUal ]
Carbamate Analysis Column 2
Follow generally accepted procedures for quality control and methods
development when using this column.
If you observe a change in peak shape, retention of a particular
compound, or resolution between two compounds, take immediate
steps to determine the reason for the changes. Until the cause of the
change is determined, do not rely on the results of the analyses.
II. InstallatIon
a. Attaching the Column
1. If this column is to be used with an LC system previously used
for a purpose other than carbamate analysis, then before
installing the column in the flow path connect the column
inlet and outlet lines to each other with a union and flush the
lines, as well as the sample injector and injector loops, free of
previous solvents. Make certain that the final flushing solvent
is miscible with water. Remove the union.
2. Remove the end plugs from your column with a 5/16-inch
wrench.
NOTE: Be sure to replace and tighten the end plugs when the
column is removed from the system for storage.
3. The column outlet is indicated by an arrow on the label
showing the direction solvent should flow. Thread the inlet and
outlet tubing fittings into the column until finger tight and then
tighten the fittings with a wrench turning each - turn.
CAUTION: Do not over-tighten. Over-tightening will damage
the connection. Follow the next three steps of this procedure
if you must remove a damaged compression screw or worn
ferrule.
4. Using a miniature tubing cutter, scribe deeply the
circumference of the tubing at the desired break point. Or,
alternatively, using a three-cornered file with a cutting edge,
cut 1/3 of the way through the tubing at the desired break
point.
5. Grasp the tubing on both sides of the scribe mark with cloth-
covered pliers (to prevent marring the tube surface) and gently
work the tub back and forth until it breaks cleanly. Check that
the end is straight and smooth with no burrs. Flush the tube
from the opposite end with mobile phase to remove any metal
particles that may have lodged in the interior of the tube.
6. Slide the compression screw head first followed by the ferrule
(large end of the taper first) over the tube. Insert the end of
the tube into the fitting seat to which it will be connected.
Tighten the compression screw in the fitting seat as in Step 3.
Assembly details are shown in Figure 2.
CAUTION: Properly bottom the tubing in the fitting seat
while tightening the compression screw to the column.
Tubing not completely seated will result in dead volume
which could cause excessive sample band spreading.
Figure 2: Ferrule and Compression Screw Assembly
b. Equilibration
The Waters Carbamate Analysis Column is shipped in the mobile
phase used for column storage: 50/50 v/v methanol/acetonitrile.
For equilibration, flush the column with the initial mobile phase
used for the gradient analysis. This can be conveniently done while
the post-column reaction system, column oven, and detector are
warming up to stable operating conditions (about 30 minutes at 1.5
mL/min). Refer to the Waters Carbamate Analysis System Manual
for details.
Compression Screw or Nut
Ferrule
End must be straight
and smooth to achieve
maximum column efficiency
Tu be
Critical Distance to be determined by
the union or column fitting, etc.