beautypg.com

Care and use manual, Band spreading minimization – Waters XBridge Columns User Manual

Page 7

background image

[ Care and Use ManUal ]

XBridge

Columns

7

Figure 4: Waters Ferrule in a Parker Style Endfitting

There are two ways to fix the problem:

1. Tighten the screw a bit more. The ferrule moves forward, and reaches

the sealing surface. Do not overtighten since this may end in break-
ing the screw.

2. Cut the tubing, replace the ferrule and make a new connection.

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 5: Waters Part Numbers for SLIPFREE® Connectors

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


Band Spreading Minimization

Figure 6 shows the influence of tubing internal diameter on system band
spreading and peak shape. As can be seen, the larger tubing diameter
causes excessive peak broadening and lower sensitivity.


Figure 6:
Effect of Connecting Tubing on System

b. Measuring System Bandspreading Volume
and System Variance

This test should be performed on an HPLC system with a single wavelength
UV detector (not a Photodiode Array (PDA)).

1. Disconnect column from system and replace with a zero dead volume

union.

2. Set flow rate to 1 mL/min.

3. Dilute a test mix in mobile phase to give a detector sensitivity

of 0.5 - 1.0 AUFS (system start up test mix can be used which
contains uracil, ethyl and propyl parabens; Waters part number
WAT034544).

4. Inject 2 to 5 µL of this solution.

Gap

Diluted/Distorted Sample Band

0.005 inches

0.020 inches

0.040 inches