Abstract
Microchip capillary electrophoresis with laser-induced fluorescence detection (μCE-LIF) enables sensitive analyses of a wide range of analytes employing small volumes of sample and reagent (nL to μL) on an instrument platform with minimal mass, volume, and power requirements. This technique has been used previously in the analysis of amino acids and other organic molecules of interest in the fields of astrobiology and planetary science. Here, we present a protocol for the analysis of thiols using μCE-LIF. This protocol utilizes Pacific Blue C5-maleimide for fluorescent derivatization of thiols, enabling limits of detection in the low nM range (1.4–15 nM). Separations are conducted in micellar electrokinetic chromatography mode with 25 mM sodium dodecyl sulfate in 15 mM tetraborate, pH 9.2. This method allows analysis of 12 thiols in less than 2 min following a labeling step of 2 h. A step-by-step protocol, including tips on microchip capillary electrophoresis, is described here.
Copyright 2012 California Institute of Technology. Government sponsorship acknowledged.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Pilcher CB (2003) Biosignatures of early earths. Astrobiology 3(3):471–486
Vance S, Christensen LE, Webster CR, Sung K (2011) Volatile organic sulfur compounds as biomarkers complementary to methane: Infrared absorption spectroscopy of CH3SH enables in situ measurements on Earth and Mars. Planet Space Sci 59(2–3):299–303
Brown RH, Lebreton J-P, Waite JH (eds) (2009) Titan from Cassini-Huygens. Springer, New York
Toyo’oka T (2009) Recent advances in separation and detection methods for thiol compounds in biological samples. J Chromatogr B 877(28):3318–3330
Zhou J, O'Shea TJ, Lunte SM (1994) Simultaneous detection of thiols and disulfides by capillary electrophoresis-electrochemical detection using a mixed-valence ruthenium cyanide-modified microelectrode. J Chromatogr A 680(1):271–277
Inoue T, Kirchhoff JR (2000) Electrochemical detection of thiols with a coenzyme pyrroloquinoline quinone modified electrode. Anal Chem 72(23):5755–5760
Stenken JA, Puckett DL, Lunte SM, Lunte CE (1990) Detection of N-acetylcysteine, cysteine and their disulfides in urine by liquid chromatography with a dual-electrode amperometric detector. J Pharm Biomed Anal 8(1):85–89
Kuśmierek K, Chwatko G, Głowacki R, Bald E (2009) Determination of endogenous thiols and thiol drugs in urine by HPLC with ultraviolet detection. J Chromatogr B 877(28):3300–3308
McDermott GP, Terry JM, Conlan XA, Barnett NW, Francis PS (2011) Direct detection of biologically significant thiols and disulfides with manganese(IV) chemiluminescence. Anal Chem 83(15):6034–6039
Seiwert B, Karst U (2007) Simultaneous LC/MS/MS determination of thiols and disulfides in urine samples based on differential labeling with ferrocene-based maleimides. Anal Chem 79(18):7131–7138
Mora MF, Greer F, Stockton AM, Bryant S, Willis PA (2011) Toward total automation of microfluidics for extraterrestrial in situ analysis. Anal Chem 83(22):8636–8641
Mora MF, Stockton AM, Willis PA (2012) Microchip capillary electrophoresis instrumentation for in situ analysis in the search for extraterrestrial life. Electrophoresis 33:2624–2638
Skelley AM, Scherer JR, Aubrey AD, Grover WH, Ivester RHC, Ehrenfreund P, Grunthaner FJ, Bada JL, Mathies RA (2005) Development and evaluation of a microdevice for amino acid biomarker detection and analysis on Mars. Proc Natl Acad Sci U S A 102(4):1041–1046
Benhabib M, Chiesl TN, Stockton AM, Scherer JR, Mathies RA (2010) Multichannel capillary electrophoresis microdevice and instrumentation for in situ planetary analysis of organic molecules and biomarkers. Anal Chem 82(6):2372–2379
Mora MF, Stockton AM, Willis PA (2013) Analysis of thiols by microchip capillary electrophoresis for in situ planetary investigations. Electrophoresis 34:309–316
Willis PA, Jiao H, Greer F, Fisher AM, Mora MF, Stockton AM, Cable ML (2012) The chemical laptop. U.S. Patent CIT-5905-P
Acknowledgments
The research described here was carried out at the Jet Propulsion Laboratory, California Institute of Technology, under a contract with the National Aeronautics and Space Administration. Financial support for this project was provided by ASTID (Project #104320) and the NASA Postdoctoral Program (NPP) at the Jet Propulsion Laboratory, administered by Oak Ridge Associated Universities through a contract with NASA.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Mora, M.F., Stockton, A.M., Willis, P.A. (2015). Analysis of Thiols by Microchip Capillary Electrophoresis for In Situ Planetary Investigations. In: Van Schepdael, A. (eds) Microchip Capillary Electrophoresis Protocols. Methods in Molecular Biology, vol 1274. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2353-3_4
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2353-3_4
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2352-6
Online ISBN: 978-1-4939-2353-3
eBook Packages: Springer Protocols