Abstract
The use of 1,4-naphthoquinone as an advantageous pre-column reagent for liquid chromatography analysis of aliphatic thiol compounds is proposed. The compound reacts selectively in mild conditions (5 min at room temperature; pH 7.5) with thiol function. The resulting adducts were separated under isocratic conditions by using a reversed-phase column (C-12n) with a mobile phase corresponding to methanol/triethylammonium phosphate buffer (pH 3; 0.05 mol L−1) 65:35, v/v, at a flow rate of 0.4 mL min−1 in presence of quercetin as internal standard. Detection was set at a wavelength of 420 nm. The effect of the derivatization reaction conditions on the N-acetylcysteine (NAC) reaction yield was investigated by a series of experiments. The yield of NAC derivative was found to be quantitative at a reagent thiol molar ratio of about 3 by comparison with an authentic specimen of synthesized NAC adduct, which was characterized by 1 H NMR, IR, and UV. Similar linear responses were observed by standard and placebo solutions (determination coefficient, 0.9998). The within- and between-day standard deviations (RSD) were ≤0.47 %. Recovery studies showed good results (100.03 %) with RSD 0.76 %. The limit of detection was about 20 pmol. The utility of the validated method for the determination of NAC in a new dietary supplement and commercial formulations is demonstrated.
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Acknowledgments
We are grateful to Doctor Debora Fanti for her valuable technical assistance. This work was supported by a grant from MIUR (“Cofinanziamento PRIN” 2009, Rome, Italy).
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Published in the special issue Analytical Science in Italy with guest editor Aldo Roda.
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Gatti, R., Andreatta, P. & Boschetti, S. Study of 1,4-naphthoquinone as a new useful derivatization reagent for LC analysis of aliphatic thiols in dietary supplements and pharmaceuticals. Anal Bioanal Chem 405, 817–825 (2013). https://doi.org/10.1007/s00216-012-6250-x
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DOI: https://doi.org/10.1007/s00216-012-6250-x