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Simultaneous determination of homocysteine and other sulphur compounds in human plasma by HPLC/ED
Year: 2004
Type of publication: ostatní - přednáška nebo poster
Name of source: Vitamins 2004 Targeted nutrional therapy
Publisher name: Univerzita Pardubice
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Language Name Abstract Keywords
cze Simultaneous determination of homocysteine and other sulphur compounds in human plasma by HPLC/ED homocystein; kapalinová chromatografie; elektrochemická detekce; glutathion; cystein; síru obsahující aminokyseliny; cystathionin; methionin
eng Simultaneous determination of homocysteine and other sulphur compounds in human plasma by HPLC/ED Homocysteine is a sulphur-containing amino acid and is formed in the metabolism of methionine. Homocysteine is metabolized through two major pathways: the methylation cycle and the formation of cystathionine. Cystathionine is cleaved into cysteine and alpha-ketobutyrate. Cysteine is a precursor of glutathione (GSH), the major cellular redox buffer. The aim of this work was simultaneous determination of compounds containing sulphur. It allowed understand of failure in transsulphuration and methylation pathways at individual patients. We injected 20 ul of the supernatant onto 80 x 4.6 mm RP-C18 column (HR-80, particle size, 3 um; ESA, Inc.). The mobile phase (flow rate, 1 ml/min) consisted 95% sodium dihydrogen phosphate and 5% acetonitrile. The mobile phase was filtered through a 0.45 um cellulose membrane before use. Penicillamine is appropriate internal standard for this method. We detected reduced sulphur compounds with ESA Coulochem II 5100A electrochemical detector (USA), using ESA 5010 coulometric cell. An ESA 5020 Guard cell was installed between the pump and ESA autosampler. We developed a sensitive and inexpensive method with rapid sample preparation for determination of total plasma or serum homocysteine and other sulphur compounds (methionine, glutathione, cystathionine). homocysteine; liquid chromatography; electrochemical detection; glutathione; cysteine; sulphur-containing amino acids; cystathionine, methionine

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