Automated high-performance liquid chromatographic determination of 5-S-cysteinyl-3,4-dihydroxyphenylalanine in urine

Journal of Chromatography
B KågedalC Hansson

Abstract

An automated high-performance liquid chromatographic (HPLC) method has been developed for measurement of 5-S-cysteinyl-DOPA in urine (DOPA = 3,4-dihydroxyphenylalanine). The urinary sample was injected into an HPLC boronate column. With a mobile phase of 0.1 M phosphate buffer containing 0.2 mM disodium ethylenediaminetetraacetate (Na2EDTA) (pH 6.0) mixed with methanol (9:1), 5-S-cysteinyl-DOPA was adsorbed while most other compounds were washed away. By column switching, the column flow was reversed and 5-S-cysteinyl-DOPA was desorbed by a mobile phase of 0.1 M formic acid and 0.2 mM Na2EDTA at pH 3.0 and chromatographed on a reversed-phase column. The precision, as estimated from repeated analysis of an urinary sample and from duplicate analysis of a number of samples, ranged from 1.4 to 5.2% (coefficient of variation), and the analytical recovery was 93 +/- 4.1%. The method is suitable for use in the clinical laboratory.

References

Jan 1, 1977·Analytical Biochemistry·S HigaY Yamamura
Feb 11, 1983·Journal of Chromatography·B Kågedal, A Pettersson
Dec 12, 1984·Journal of Chromatography·K KemperE Schlimme
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Citations

Sep 3, 2003·Pigment Cell Research·Akihiko TakasakiBertil Kågedal
Mar 10, 2006·Pigment Cell Research·Kazumasa WakamatsuShosuke Ito
Sep 17, 2004·Scandinavian Journal of Clinical and Laboratory Investigation·K ArstrandB Kågedal
Sep 14, 2000·Melanoma Research·R KärnellU Ringborg
Jan 31, 2002·Journal of Chromatography. B, Biomedical Sciences and Applications·J Hartleb, R Arndt

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