Add time:08/23/2019 Source:sciencedirect.com
The assay of the ethyl chloride metabolite S-ethyl-N-acetyl-l-cysteine in human urine by HPLC is described. The compound is enriched by adsorption on a non-polar adsorbent of graphitized non-porous carbon, and then stripped from positively charged compounds by application onto a strong acid cation-exchanger. Subsequently, an enzymatic deacetylation is carried out and the acylase is removed by centrifugal ultrafiltration. Separation of the sample is performed by cation-exchange chromatography applying an eluent of a very low elution strength (diluted formic acid). In the column effluent S-ethyl-l-cysteine is derivatized by o-phthaldialdehyde and the reaction product is detected by fluorescence measurement. In human urine a detection limit in the low ppb range is achieved.
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