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I'm running EEMS on some water samples on a Horiba Fluoromax 4 spectrofluorometer. Method below.

  • $\pu{240-580nm}$ emission, $\pu{5nm}$ interval, $\pu{1nm}$ width.

  • $\pu{240-480nm}$ excitation, $\pu{5nm}$ interval, $\pu{1nm}$ width.

  • $\pu{0.01 mg/L}$ quinine sulfate standard in $\pu{0.5M}~\ce{H2SO4}$ + water.

  • Raman EEMS to use as references for my unknowns.

I use factory correction files for both detectors and then use the 3D profile button on my output charts. My data is only letting me export it as ASCII, and my quinine sulfate EEMS looks like the water Raman EEMS.

My questions are:

  • Is it possible my quinine sulfate is expired? I don't know how old it is (at least 2+ years, but it's been in the fridge in amber plastic). There's also a chance I set it up wrong which is why the signal just looks like water showing some Rayleigh scatter.

  • How can I export the images of my EEMs intensity profiles? I get an error saying I need to convert my matrix to a worksheet first, but I don't know how to do that.

  • Finally, how do I get an intensity map of my EEMs (heat map)? Does that happen in Matlab/R? There's a button in FluorEscence to get a 2D intensity map, but it doesn't seem to do anything. Maybe I'm only supposed to export ASCII and the Matlab/R package makes an intensity map. I'm not sure, but the manuals and help for the Horiba software aren't helpful in the least.

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    $\begingroup$ You have three questions and need rather detailed and specific information. But you can easily check the quinine fluorescence issue since quinine water fluoresces very well: see here. An inexpensive UV flashlight, such as is used for scorpion hunting, counterfeit specie examination or pet accident detection, works fine if you just want to see if your old quinine solution has gone south. $\endgroup$
    – Ed V
    Commented May 12, 2021 at 21:30
  • $\begingroup$ @EdV Thank you; I will test the flashlight tomorrow - I believe we have one in the lab. Do you know anything about EEMs processing or Horiba software? $\endgroup$
    – dogman
    Commented May 12, 2021 at 21:38
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    $\begingroup$ I know nothing about Horiba software and probably very little about EEM processing. I know a little about what we used to call synchronous luminescence spectroscopy, where you scan both the excitation and emission and get fluorescence intensity as a function of the excitation and emission wavelengths. But this is not useful: you have a fancy instrument, presumably, and the adventure of mastering it! Best of success! $\endgroup$
    – Ed V
    Commented May 12, 2021 at 22:55

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