Method Development - Agilent Technologies 1100 Series Reference Manual

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.
Table 5 Steps for thorough method development
Fluorescence scan
Signal mode
Spectral
mode/multi-wavelength
detection
1100 Series FD Reference Manual
Step 1:
Check system
Find impurities (for example,
in solvents and reagents)
First Steps with the Fluorescence Detector
Step 2:
Optimize limits of detection
and selectivity
Determine simultaneously
the excitation and emission
spectra of a pure
compound
Perform wavelength
switching
Determine Ex/Em spectra for
all separated compounds in a
single run
Activate up to four
wavelength simultaneously
Step 3:
Set up routine methods
Use for lowest limits of
detection
Collect online spectra,
perform library search,
determine peak purity
Deactivate wavelength
switching
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