Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche ''Togo Rosati''

Authors    Pucciarini L, Saluti G, Galarini R, Carotti A, Macchiarulo A, Rudaz S, Sardella R

Abstract   Cysteine is a sulfur-containing amino acid which plays an outstanding role in many biological pathways in mammals. The analysis and quantification of native cysteine remains a critical issue due to its highly reactive thiol group evolving to the disulfide cystine derivative through oxidation reaction. Aimed at improving the derivative stability, cysteine was labelled with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), which reacts with both amino and thiol groups. The derivatization was optimized and the chemical identity of the reaction product was assessed via high-resolution mass spectrometry. The NBD-cysteine derivative resulted stable for 10 days. This derivative was enantioresolved (α and R_s equal to 1.25 and 2.70, respectively) thanks to a (R,R)-Whelk-O1 phase with the following chromatographic setting: eluent, MeOH/water-90/10 (v/v) with 15 mM ammonium formate (pwsH 6.0); column temperature, 35 °C; flow rate, 1.0 mL/min. The developed method was validated following the ICH guidelines and applied for the quality control of a L-cysteine containing dietary supplement.

Publish Date  2020

Volume    180

ISSN         1873-264X

DOI           doi.org/10.1016/j.jpba.2019.1130660731-7085

URL           https://www.sciencedirect.com/science/article/pii/S0731708519323842?via%3Dihub

Journal     Journal of Pharmaceutical and Biomedical Analysis

Pages       art. n. 113066

PMID        31891875