I am trying to detect viral capsids using an UltiMate HPLC system. I have a strong peak at the expected retention time using a 260/280nm UV detector, however I am not receiving any signal from the FLD detector (should detect innate tryptophan residues present in the viral capsids). The detector is connected to the HPLC and has a green status/connected light, however the only signal received looks like a binary on/off type waveform. Picture of this waveform is attached. There are no errors on the detector or in the software controlling it.
Some solvents like Methanol will quench or prevent fluoresence. Be sure you do not use Methanol in both your extraction and HPLC solvents.
Are you sure the detector is working? Is there a gain adjustment on the detector? Do you have a positive control you can run, such as tryptophan?
Are the wavelength settings on the detector suitable for detecting tryptophan (excitation 280-290 nm, emission 330-360 nm)?
Is the value being reported equal to zero fluorescence or maximal fluorescence (detector saturation)?
- Badges
- Science method