Waters Oligonucleotide Separation Technology XBridge OST C18 Columns - Manual (Page 3)

[ CARE AND USE MANUAL ]
3
Figure 4: Waters Ferrule in a Parker Style Endfitting
The best way to fix the problem is to cut the tubing, replace the
ferrule and make a new connection.
1. Alternatively, replace the conventional compression screw fitting with
an all-in-one PEEK ™ fitting (Waters part number PSL613315) that
allows resetting of the ferrule depth. Another approach is to use a
Keystone, Inc. SLIPFREE
® connector to always ensure the correct fit.
The fingertight SLIPFREE connectors automatically adjust to fit all
compression screw type fittings without the use of tools (Figure 5).
Figure 5: Single and Double SLIPFREE Connectors
SLIPFREE Connector Features:
• Tubing pushed into endfitting, thereby guaranteeing
a void-free connection
• Connector(s) come(s) installed on tubing
• Various tubing IDs and lengths available
• Fingertight to 10,000 psi – never needs wrenches
• Readjusts to all column endfittings
• Compatible with all commercially available endfittings
• Unique design separates tube-holding function from sealing function
Table 1: Waters Part Numbers for SLIPFREE Connectors
2. Flush column with 100% organic mobile phase (acetonitrile with
TEAA or methanol for TEA-HFIP ion-pairing method) by setting
the pump flow rate to 0.1 mL/min and increase the flow rate to
0.5 mL/min over 5 minutes.
3. When the mobile phase is flowing freely from the column outlet,
stop the flow and attach the column outlet to the detector. This
prevents entry of air into the detection system and gives more
rapid baseline equilibration.
4. Gradually increase the flow rate as described in step 3.
5. Once a steady backpressure and baseline at 260nm have been
achieved, proceed to the next section.
b. Column Equilibration
XBridge OST C
18
columns are shipped in 100% acetonitrile. It is
important to ensure mobile phase compatibility before changing
to a different mobile phase system. Equilibrate the column with a
minimum of 10 column volumes of the mobile phase to be used for
the oligonucleotide separation.
Note: When mobile phase additives are present in low concentrations
(e.g., TEAA or TEA-HFIP ion-pairing reagents), 100 to 200 column
volumes may be required for complete XBridge OST C
18
column
equilibration.
Gap
SLIPFREE Type Tubing Internal Diameter
Tubing Length 0.005” 0.010” 0.020”
Single 6 cm PSL 618000 PSL 618006 PSL 618012
Single 10 cm PSL 618002 PSL 618008 PSL 618014
Single 20 cm PSL 618004 PSL 618010 PSL 618016
Double 6 cm PSL 618001 PSL 618007 PSL 618013
Double 10 cm PSL 618003 PSL 618009 PSL 618015
Double 20 cm PSL 618005 PSL 618001 PSL 618017