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- E-ISSN 2288-8985
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Determination of the Homocysteine, Cysteine and Methionine in Human Plasma by Gas Chromatography with Electron Capture Detector
Analytical Science and Technology / Analytical Science and Technology, (P)1225-0163; (E)2288-8985
1999, v.12 no.5, pp.408-414
Myung,
S., Chang,
Y., Yoo,
E., Park,
J., Min,
H., &
Kim,
M.
(1999). Determination of the Homocysteine, Cysteine and Methionine in Human Plasma by Gas Chromatography with Electron Capture Detector. Analytical Science and Technology, 12(5), 408-414.
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Abstract
A selective and sensitive method for the derivatization of total homocysteine (Hcy) and the related compounds in plasma by gas chromatograph (GC)-electron capture detector (ECD) has been developed. To determine total homocysteine, cysteine (Cys), and methionine (Met) in human plasma using GC-ECD, analytes were reduced and converted into their N(O,S)-ethylearbonyl pentafluoropropyl (PFP) ester by derivatization with ethyl chloroformate and pentafluoropropyl alcohol (PFP-OH) in plasma. The best derivatizing agent N(O,S)-ethyl carbonyl PFP ester, was chosen by comparing the sensitivity of derivatized analysis in GC-ECD. The derivatized analytes in plasma were extracted by chloroform, and subsequently back-extracted with hexane and analyzed by GC-ECD. The calibration carves (<TEX>$R^2$</TEX> > 0.990) were linear over the range <TEX>$5-50{\mu}mol/L$</TEX> of Hcy and Met, <TEX>$40-400{\mu}mol/L$</TEX> of Cys spiked in plasma. The detection limit observed by the established method was below <TEX>$0.5{\mu}mol/L$</TEX>. This method is highly sensitive and specific in the analysis of Hcy, Cys, and Met. Therefore, we suggest that this method is appropriate in the analysis of trace concentration of Hcy, Cys, and Met in biological fluids.
- keywords
- GC-EDD, homocysteine, cysteine, methionine
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