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Hitachi PRIMAIDE 1440 - Notes on Fluorescence Analysis

Hitachi PRIMAIDE 1440
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2 - 11
2.3.3 Notes on fluorescence analysis
(1) Raman Scattering
When fluorescence is measured, two additional peaks may
appear in the spectrum. The Rayleigh peak appears at the
excitation wavelength and is due to scattered light, while
the Raman peak appears at longer wavelength than the
excitation. The position of the Raman band is dependent
on the excitation wavelength, while the position of the
fluorescence is independent of the excitation wavelength
though its peak height changes. Both Rayleigh and
Raman scatterings occur due to a solvent. Take care not
to mistake them for fluorescence peaks.
Table 3-2 presents the position of the Raman peak for a
variety of excitation wavelengths.
In liquid chromatography, the Raman scattering appears as
a background rise. This means that baseline noise
increases to disturb high-sensitivity measurement.
Table 2-5 Raman Spectral Peak Position at Each Excitation Wavelength
(Excitation
Wavelength)
Water
Ethanol
Cyclohexane
Carbon
Tetrachloride
Chloroform
Excitation
wavelength, and
Raman peak
position (nm)
248
271
267
267
313
350
344
344
320
346
365
416
405
408
375
410
405
469
459
458
418
461
436
511
500
499
450
502

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