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Determination of selenomethionine and seleno-methyl-selenocysteine in biota by ultrasonic-assisted enzymatic digestion and multi-shot stir bar sorptive extraction-thermal desorption-gas chromatography-mass spectrometry



Determination of selenomethionine and seleno-methyl-selenocysteine in biota by ultrasonic-assisted enzymatic digestion and multi-shot stir bar sorptive extraction-thermal desorption-gas chromatography-mass spectrometry



Journal of Chromatography. a 1300: 151-158



A method based on stir bar sorptive extraction (SBSE) and thermal desorption (TD)-gas chromatography-mass spectrometry (GC-MS) has been optimized for the determination of seleno-methyl-selenocysteine (SeMetSeCys) and selenomethionine (SeMet) in biota samples. Aliquots of freeze-dried tissue, a mixture of protease XIV-lipase and water were sonicated for 2min. After extraction, the extract was separated by centrifugation and subjected to derivatization and SBSE-TD-GC-MS. The parameters affecting derivatization, absorption and desorption steps were investigated. The optimized conditions consist of a derivatization with 40μL of ethyl chloroformate (ECF) in 400μL of a water:ethanol:pyridine (60:32:8) mixture, followed by dilution to 1.5mL of 70g NaClL(-1) in water at neutral pH and an extraction step using 10mm×1mm PDMS stir bar, stirring at 800rpm for 20min at room temperature (23±1°C). Three stir bars were used for the extraction of three different aliquots of the same sample and then placed in a single glass desorption liner and simultaneously desorbed for GC-MS analysis. The desorption step required the following conditions: 300°C (desorption temperature), 6min (desorption time), 50mLmin(-1) (vent flow) and -5°C (cryotrapping temperature). The method provided precise (8.1%) and accurate results in the mgSekg(-1) range (using the selected-ion monitoring-SIM mode) against certified reference material SELM-1 yeast, with recoveries higher than 80% for spiked algae and clams samples.

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Accession: 052535124

Download citation: RISBibTeXText

PMID: 23497851

DOI: 10.1016/j.chroma.2013.02.029


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