I need information, possibly backed up with literature, on what excitation and emission wavelengths that are suitable for detecting and quantifying phenolics using HPLC fitted with flourimetric detector. I am aware that there are some detectors like UV, MS, DAD,ESI-MS, MALDI-TOF/MS, NMR etc that are better. But my inquiry is specific to fluorescence detector.
Hi Olorunfemi,
It depends on the type of phenolics. Some phenolic compound does fluoresce but others can not be detected at all. Furthermore, you have to change both emission and detection wavelenghts during the run because almost each compound possess specific wavelenghts. In my lab,for example, I use FLuorometric detector for catechins, by setting detector at 278 (emission) and 345 (absorption). Other wavelenghts could be the following: (ex-em)
Gallic acid 278 366
(+)-Catechine 278 350
Caffeic acid 262 426
(−)-Epicatechin 278 360
Syringaldehyde 260 422
p-Coumaric acid 260 422
Ferulic acid 260 422
trans-Resveratrol 330 374
Quercetin 264 420
Kaempferol 268 422
Hope this will help.
Hops, Something get wrong....
Excitation Emission
Gallic acid 278 366
-Catechin 278 360
Caffeic acid 262 426
(−)-Epicatechin 278 360
Syringaldehyde 260 422
p-Coumaric acid 260 422
Ferulic acid 260 422
trans-Resveratrol 330 374
Quercetin 264 420
Kaempferol 268 422
Let me try in this way.......
Gallic acid 278-366; (+)-Catechin (278-360); Caffeic acid (262-426); (−)-Epicatechin (278-360); p-Coumaric acid (260-422) Ferulic acid (260-422); trans-Resveratrol (330- 374); Quercetin (264-420) Kaempferol (268-422).
Fabio,
thanks so much for the information. But what pH range I can use 278-366 for gallic acid
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