Issue 7, 2013

Quantification of proteinthiols using ThioGlo 1 fluorescent derivatives and HPLC separation

Abstract

A method for quantification of total soluble protein-derived thiols in beer was developed based on the formation of fluorescent adducts with the maleimide compound ThioGlo 1. The problem of interference from fluorescent adducts of sulfite and ThioGlo 1 was solved by HPLC separation of the adducts followed by fluorescence detection. Using standard addition of GSH, a detection limit of 0.028 μM thiols was achieved. The application and validation of the method was demonstrated for beers with different color intensities, and the application range is in principle for any biological system containing thiols. However, the quantification of cysteine was complicated by a lower fluorescence response of its ThioGlo 1 adducts. Based on the studies of the responses of a series of cysteine-derived thiols and 1H NMR studies of the structures of ThioGlo 1 adducts with GSH and cysteine, it was concluded that thiols with a neighboring free amino group yield ThioGlo 1 adducts with a reduced fluorescence intensity.

Graphical abstract: Quantification of protein thiols using ThioGlo 1 fluorescent derivatives and HPLC separation

Supplementary files

Article information

Article type
Paper
Submitted
04 Sep 2012
Accepted
01 Feb 2013
First published
01 Feb 2013

Analyst, 2013,138, 2096-2103

Quantification of protein thiols using ThioGlo 1 fluorescent derivatives and HPLC separation

S. Hoff, F. H. Larsen, M. L. Andersen and M. N. Lund, Analyst, 2013, 138, 2096 DOI: 10.1039/C3AN36672C

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements