Monitoring System Functions; Wavelength Calibration And Verification - Thermo Scientific Dionex UltiMate 3000 Series Operating Instructions Manual

UltiMate 3000 Series:
Fluorescence Detectors FLD-3100 and FLD-3400RS

5.8 Monitoring System Functions

This section provides a short overview of some special functions for qualification,
documentation and monitoring the system that Chromeleon supports for the detector. Some
functions are available also on the control panel for the detector. For additional information
about a function, see the Chromeleon Help.
For information about how to monitor the remaining lamp lifetime, refer to section 7.2
(→ page 108).

5.8.1 Wavelength Calibration and Verification

A simple calibration procedure is performed after power-up of the detector. This requires
that a flow cell is installed in the detector. During the calibration process, ensure that the
eluent composition in the flow cell does not change and no air bubbles are present in the
flow cell. If the message "Not calibrated" appears when the detector is turned on, verify
that these conditions are met and execute the Calibrate command in Chromeleon.
In addition, you can verify the wavelength calibration using water (Raman measurement). If
verification fails, that is, if the calibration shows a significant deviation, you can perform a
wavelength calibration using water (Raman measurement). During wavelength calibration,
the light spectrum of the xenon flash lamp is used to calibrate the excitation wavelength.
Afterward, the emission monochromator is adjusted with the help of the maximum of the
Raman emission spectrum (379 mm).
The following conditions must be met for both wavelength verification and Raman
wavelength calibration:
• The system is equilibrated and environmental conditions are stable.
• There are no air bubbles in the flow cell. We recommend degassing the water. Set a flow
rate of 0.5 mL/min and wait about 15 minutes before you begin, until the baseline is
stable.
• The water has no impurities. Always use fluorescence-grade water and consider using a
filter or a suitable column before the detector.
Page 90 Operating Instructions