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. Steps for thorough method development gives an overview of how to benefit from the operation modes during these steps.
| 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 |
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