Chromatography Forum

Hi HPLC experts,

Is there any relationship between fluorescence and UV detector for vitamin E determination in normal phase HPLC.
This problem arise because the fluorescence HPLC is currenctly using by another researcher (may take few weeks for her to complete) while UV HPLC is unused.So I planned to evaluate the efffect of few variables e.g. temp and time to the tocols in my sample using HPLC- UV/Vis. After that, I wish to use fluorescence HPLC for method optimization.
Can I follow this way??.
Will there be a difference in amount regardless the tocols concentration

PLease advise me.
Thank You

The first entry in a google search (fluorescence of tocopherols) gave this (the URL was ridiculously long):

Journal of Chromatography A
Volume 178, Issue 2, 31 October 1979, Pages 525-531
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doi:10.1016/S0021-9673(00)92511-1 | How to Cite or Link Using DOI
Copyright © 1979 Published by Elsevier B.V. Cited By in Scopus (50)
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Application of a high-performance liquid chromatographic fluorescence method for the rapid determination of α-tocopherol in the plasma of cattle and pigs and its comparison with direct fluorescence and high-performance liquid chromatography—ultraviolet detection methods.






C.H. McMurraya and W.J. Blanchflowera

Veterinary Research Laboratories, Stoney Road, Stormont, Belfast Northern Ireland

Received 2 July 1979. Available online 30 October 2001.

Abstract
High-performance liquid chromatography is used to develop a sensitive method for the determination of tocopherol levels in the plasma of cattle and pigs. This method is compared with a similar method using UV detection and one using direct fluorescence determination of tocopherol.

Finally a double injection technique used in conjunction with fluorescence detection is shown to enhance the rate of analysis of the tocopherol levels in bovine plasma extracts.

Fluorescent detection can be 100 – 10000 times more sensitive than UV detection. The main reason to use fluorescence detection is low concentration of an analyte in a sample, otherwise I would start with a UV detector.

The complexity of the sample/matrix also comes in to play. Fluorescence is more selective than UV (2 wavelengths involved, and more compounds absorb than fluoresce).