Simulation of Scanning Fluorescence Spectrometer.docVIP

Simulation of Scanning Fluorescence Spectrometer This is a simulation of a scanning spectrofluorometer. The simulation displays excitation spectra, emission spectra, and synchronous spectra, relative fluorescence intensity, and absorbance at the excitation wavelength. Operating instructions are contained in the scrolling text field in the upper right of the screen. Answer the following questions. Set A=1 ppm and B=0. Determine the wavelengths of maximum excitation and emission for component A. What is its Stokes shift? Dose Vavilov’s Law*seem to hold for compound A, that is, is the shape of the emission spectra independent of the excitation wavelength, and vice versa, except for the scatter peaks? Is there any sign of Rayleigh or Raman scatter? How could you distinguish these from genuine fluorescence? Check the blank (click on “Insert Blank”).Increase the sensitivity setting as necessary. Is there any sign of dark current or background fluorescence? What are the main features of the excitation and emission spectra of the blank? Estimate the spectral bandpass of the monochromators. Dose the wavelength separation between the Rayleigh and Raman scatter peaks in the emission spectrum vary with excitation wavelength? What is the frequency, in cm-1, of the vibration causing the Raman peak? What vibration is most likely the cause? Find the combination of excitation and emission wavelength that gives the best precision of measurement of low concentrations of component A. Estimate the detection limit of component A in ppm. Is the detection limit lower by fluorescence or by absorption measurement? By approximately what factor? Over most of the concentration range, what is the source of noise in the intensity readings and in the spectra? How could you prove this? Is there evidence of non-linearity in the relationship between concentration and intensity at high concentrations? What is the most likely source of the non-linearity? Vary the wavelength offset and observe the sync