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Plasma L-ergothioneine measurement by high-performance liquid chromatography and capillary electrophoresis after a pre-column derivatization with 5-iodoacetamidofluorescein (5-IAF) and fluorescence detection

Sotgia, Salvatore and Pisanu, Elisabetta and Pintus, Gianfranco and Erre, Gian Luca and Pinna, Gérard Aimé and Deiana, Luca and Carru, Ciriaco and Zinellu, Angelo (2013) Plasma L-ergothioneine measurement by high-performance liquid chromatography and capillary electrophoresis after a pre-column derivatization with 5-iodoacetamidofluorescein (5-IAF) and fluorescence detection. PLoS One, Vol. 8 (7), e70374. ISSN 1932-6203. Article.

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DOI: 10.1371/journal.pone.0070374

Abstract

Two sensitive and reproducible capillary electrophoresis and high-performance liquid chromatography-fluorescence procedures were established for quantitative determination of L-egothioneine in plasma. After derivatization of L-ergothioneine with 5-iodoacetamidofluorescein, the separation was carried out by HPLC on an ODS-2 C-18 sperisorb column by using a linear gradient elution and by HPCE on an uncoated fused silica capillary, 50 µm id, and 60 cm length. The methods were validated and found to be linear in the range of 0.3 to 10 µmol/l. The limit of quantification was 0.27 µmol/l for HPCE and 0.15 µmol/l for HPLC. The variations for intra- and inter-assay precision were around 6 RSD%, and the mean recovery accuracy close to 100% (96.11%).

Item Type:Article
ID Code:9521
Status:Published
Refereed:Yes
Uncontrolled Keywords:Capillary electrophoresis, high-performance liquid chromatography-fluorescence, HPLC
Subjects:Area 05 - Scienze biologiche > BIO/10 Biochimica
Area 05 - Scienze biologiche > BIO/12 Biochimica clinica e biologia molecolare clinica
Area 03 - Scienze chimiche > CHIM/08 Chimica farmaceutica
Divisions:001 Università di Sassari > 01-a Nuovi Dipartimenti dal 2012 > Scienze Biomediche
001 Università di Sassari > 01-a Nuovi Dipartimenti dal 2012 > Medicina Clinica e Sperimentale
Publisher:Public Library of Science
ISSN:1932-6203
Copyright Holders:© 2013 Sotgia et al.
Deposited On:03 Dec 2013 13:41

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