Waters AccQ-Tag Amino Acid Analysis Column - Manual (Page 2)

[ CARE AND USE MANUAL ] [ CARE AND USE MANUAL ]
2
II. INSTALLING THE COLUMN
a. Preparing the System
This procedure requires a zero dead-volume union. Before installing
the column in the flow path:
1. Directly connect the injector to the detector by replacing the old
column with a zero dead-volume union.
2. Flush the HPLC system with acetonitrile/water (60%/40%) at a
flow rate of 1.0 ml/min.
3. Remove the union.
4. Install the column in the column heater (see Section 2.2).
b. Installing the Columns
To install the column:
1. Remove the end plugs from the column and save them for use when
you store the column.
2. Screw the inlet and outlet fittings into the column until finger-
tight. An arrow on the label indicates the direction of flow.
If you are using a stainless steel system, use two 5/16 inch wrenches
to tighten the fittings 1/4 to 1/2 turn. Do not over tighten. Over tight-
ening damages the connection.
Preparing a new PEEK ™ connection
Prepare a new tubing-ferrule connection (Figure 2-1) if the existing
connection is damaged or worn.
To prepare a new PEEK ™ tubing-ferrule connection:
1. Use a single-edge razor to make a straight, square cut in front of
the compression screw on the worn fitting.
2. Slide a compression screw and washer over the end of the
cut tubing.
Properly bottom the tubing in the fitting seat. Dead volume results in
sample bandspreading.
To prepare a new stainless steel tubing-ferrule connection:
1. Use a file with a cutting edge or a tube cutter to scribe the
circumference of the steel tubing at the desired break point.
2. Grasp the tubing on both sides of the scribe mark with cloth-
covered pliers (to prevent marring the tube surface), and gently
work the tube back and forth until it separates.
3. Ensure that the tubing end is straight, open, and free of burrs.
4. Slide the compression fitting, followed by the ferrule (large end of
the toper first) over the tube.
5. Properly bottom the tubing in the fitting seat. Dead volume results
in sample bandspreading.
Figure 1. Ferrule and Compression Screw Assembly
III. USING THE COLUMN
a. Chromatography Guidelines
Before running the first amino acid analysis on the new column,
perform the efficiency test in Section IV. b. Efficiency Testing.
General Considerations
The following operating guidelines will help you obtain the best
performance from your AccQ•Tag ™ column:
• Do not exceed an operating pressure of 26 MPa /260 atm or 4000 psi).
• Filter all eluents with a 0.22 pm membrane filter. Never use turbid
or cloudy eluents.
• Protect the column from vibration and mechanical shock.
Ferrule 0.009-inch I.D. Tubing (0.25 mm)
0.130-inch gap (3.3 mm)