First HPLC-UV method for the determination of homocysteine thiolactone in human urine after derivatization with 1-benzyl-2-chloropyridinium bromide.

ABSTRACT: The report presents first high performance liquid chromatography coupled with ultraviolet detection (HPLC-UV) based method for the determination of homocysteine thiolactone (HTL) in human urine. Sample preparation procedure chiefly involves chloroform liquid-liquid extraction of HTL and its re-extraction with hydrochloric acid, followed by evaporation to dryness and treatment of the residue with 1-benzyl-2-chloropyridinium bromide and perchloric acid. The chromatographic separation of obtained UV-absorbing 2-S-pyridinium derivative is achieved within 6 min at 25 °C on Zorbax SB-C18 (150 × 4.6 mm, 5 μm) column using gradient elution. The eluent consists of 0.1% acetic acid in a mixture with acetonitrile delivered at a flow rate of 1 mL/min. The analyte is quantified by monitoring at 316 nm.

SUBMITTER: Gawel M 

PROVIDER: S-EPMC11873195 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature