Analysis of thiols by microchip capillary electrophoresis for in situ planetary investigations

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.