Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual
93
Optimizing the Detector
5
Method Development
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 fluorescence detector.
on page 93 gives an overview of how to
benefit from the operation modes during these steps.
Table 7
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
wavelengths simultaneously
Collect online spectra,
perform library search,
determine peak purity
Deactivate wavelength
switching
