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Analysis of urine for cysteine, cysteinylglycine, and homocysteine by high-performance liquid chromatography

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Abstract

A new analytical method is proposed for simultaneous determination, by liquid chromatography, of the three main urinary thiols–cysteine, cysteinylglycine, and homocysteine. To measure the total amount of these thiols urine is reduced with sodium borohydride, to convert disulfides to thiols which are then derivatized with 2-chloro-1-methylquinolinium tetrafluoroborate. Separation and quantitation of the 2-S-quinolinium thiol derivatives formed were achieved by high-performance liquid chromatography with detection at 355 nm. Validation showed the method enabled reliable simultaneous determination of these aminothiols in urine. The calibration graphs for each analyte, obtained by use of normal urine spiked with increasing amounts of cysteine, cysteinylglycine, and homocysteine, were linear (R 2≥0.997) over the range covering most practical situations. The recovery of the assay was 98–100% and sensitivity was 0.12–0.25 μmol L−1. The method was applied to 91 different samples of normal urine to establish reference values for the aminothiols, normalized on creatinine.

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Acknowledgements

The authors wish to thank the University of Lodz for financial support of this research.

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Authors and Affiliations

  1. Department of Environmental Chemistry, University of Lodz, ul. Narutowicza 65, Lodz, 90-131, Poland

    K. Kuśmierek, R. Głowacki & E. Bald

Authors
  1. K. Kuśmierek
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  2. R. Głowacki
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  3. E. Bald
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Correspondence to E. Bald.

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Kuśmierek, K., Głowacki, R. & Bald, E. Analysis of urine for cysteine, cysteinylglycine, and homocysteine by high-performance liquid chromatography. Anal Bioanal Chem 385, 855–860 (2006). https://doi.org/10.1007/s00216-006-0454-x

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  • DOI: https://doi.org/10.1007/s00216-006-0454-x

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