First Steps with the Fluorescence Detector 2
1100 Series FD Reference Manual 37
Method Development
Fluorescence detectors are used in liquid chromatography when superior
limits of detection and selectivity are required. Thorough method
development, including spectra acquisition, is fundamental to achieve good
results. This chapter describes three different steps that can be taken with the
Agilent 1100 Series fluorescence detector. Table 5 gives an overview of how to
benefit from the operation modes during these steps.
Tabl e 5 Steps for thorough method development
Step 1:
Check system
Step 2:
Optimize limits of detection
and selectivity
Step 3:
Set up routine methods
Fluorescence scan Find impurities (for example,
in solvents and reagents)
Determine simultaneously
the excitation and emission
spectra of a pure
compound
Signal mode Perform wavelength
switching
Use for lowest limits of
detection
Spectral
mode/multi-wavelength
detection
Determine Ex/Em spectra for
all separated compounds in a
single run
Activate up to four
wavelength simultaneously
Collect online spectra,
perform library search,
determine peak purity
Deactivate wavelength
switching
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