Abstract
This chapter describes two different methods using reversed-phase HPLC with electrochemical detection on a boron-doped diamond (BDD) working electrode for the direct, routine, sensitive and simultaneous measurement of a number of aminothiols, disulfides, and thioethers, in either plasma or tissue homogenates.
Key words
- Thiol
- Disulfide
- Glutathione
- Glutathione disulfide
- Cysteine
- Cystine
- S-nitrosoglutathione
- Liver
- Brain
- Plasma
- Boron-doped diamond
- Electrochemical detection
- HPLC
This is a preview of subscription content, access via your institution.
Buying options
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Acworth IN (2003) The handbook of redox biochemistry. CD-ROM. ESA Biosciences. Part number: 70-6090
Fariss MW, Reed DJ (1987) High-performance liquid chromatography of thiols and disulfides: Dinitrophenol derivatives. Methods Enzymol 143:101–109
Keller DA, Menzel DB (1985) Picomole analysis of glutathione, glutathione disulfide, glutathione S-sulfonate, and cysteine S-sulfonate by high-performance liquid chromatography. Anal Biochem 151:418–423
Michelet F, Gueguen R, Leroy P, Wellman M, Nicolas A, Siest G (1995) Blood and plasma glutathione measured in healthy subjects by HPLC: Relation to sex, aging, biological variables, and life habits. Clin Chem 41:1509–1517
Fahey RC, Newton GL (1987) Determination of low-molecular-weight thiols using monobromobimane fluorescent labeling and high-performance liquid chromatography. Methods Enzymol 143:85–96
Martin J, White IN (1991) Fluorimetric determination of oxidised and reduced glutathione in cells and tissues by high-performance liquid chromatography following derivatization with dansyl chloride. J Chromatogr 568:219–225
Acworth IN, Bowers M (1997) An introduction to HPLC-based electrochemical detection: From single electrode to multi-electrode arrays. In: Acworth IN, Naoi M, Parvez H, Parvez S (eds) Coulometric electrode array detectors for HPLC. Progress in HPLC-HPCE, vol 6. VSP, Utrecht, The Netherlands, pp 3–50
Flanagan RJ, Perrett D, Whelpton R (eds) (2005) Thiols, disulfides and related compounds. In: Smith RM (Series ed) Electrochemical detection in HPLC: Analysis of drugs and poisons. RSC chromatography monographs, Athenaeum, UK, pp79–103
Xu J, Granger M, Chen Q, Strojek JW, Lister TE, Swain GM (1997) Boron-Doped Diamond Thin-Film Electrodes: Diamond thin films could be an electrochemist’s best friend. Anal Chem 69:591A–597A
Rao T, Fujishima A, Angus J (2005) In: Fujishima A, Einaga Y, Rao T, Tryk D (eds) Historical survey of diamond electrodes in diamond electrochemistry. Elsevier, Amsterdam, pp 1–10
Waraska J, Acworth IN (2007) A novel electrode material that extends the utility of HPLC-electrochemical detection. American Lab 39(14):38–41
Jones DP, Carlson JL, Samiec PS, Sternberg P, Mody VC, Reed RL, Brown LA (1998) Glutathione measurement in human plasma. Evaluation of sample collection, storage and derivatization conditions for analysis of derivatives by HPLC. Clin Chim Acta 275:175–184
Zhang Y, Hogg N (2005) S-Nitrosothiols: Cellular formation and transport. Free Radic Biol Med 38:831–838
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Bailey, B., Waraska, J., Acworth, I. (2010). Direct Determination of Tissue Aminothiol, Disulfide, and Thioether Levels Using HPLC-ECD with a Novel Stable Boron-Doped Diamond Working Electrode. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress II. Methods in Molecular Biology, vol 594. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-411-1_23
Download citation
DOI: https://doi.org/10.1007/978-1-60761-411-1_23
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-410-4
Online ISBN: 978-1-60761-411-1
eBook Packages: Springer Protocols