Interaction Kinetics of Sulfadiazine and N-Acetyl-sulfadiazine with Soil Humic Acid: ESR Investigations with Nitroxide Spin Label
- 53 Downloads
The interaction of sulfadiazine (SDZ) and its main metabolite N-acetyl-SDZ (N-ac-SDZ) with model humic acid was investigated with stable paramagnetic nitroxide spin probes. Leonardite humic acid (LHA) was mixed with laccase to enhance the amount of reactive quinone groups of LHA and then incubated with nitroxide spin-labelled analogs of SDZ and N-ac-SDZ. The labeling at the pyrimidine moiety of SDZ leaves the aniline moiety susceptible to covalent binding to LHA, which is blocked by the N-acetylation. A broadened electron spin resonance (ESR) signal was observed for SDZ, which increased immediately after incubation and indicates strong restriction of the re-orientational motion of the spin probe, i.e., immobilization due covalent binding of the aniline moiety of SDZ to reactive quinone sites of LHA. A fast first-order reaction with a time constant of 17.6 ± 3.4 h of covalent binding was determined. The broadened ESR signal of N-ac-SDZ declined immediately after incubation with LHA and is caused by unspecific sorption to LHA, not by covalent binding. Short time constants of the bound and free SDZ were found for the reduction by the antioxidant sodium ascorbate demonstrating that SDZ and N-ac-SDZ are not physically entrapped by LHA.
The study was partly financed by the Higher Education Institutional Excellence Programme of the Ministry of Human Capacities in Hungary, within the framework of the 20765-3/2018/FEKUTSTRAT Innovation for sustainable and healthy living and environment thematic programme of the University of Pécs.
Compliance with Ethical Standards
Conflict of Interest
The authors declare no competing financial interest.
- 1.Sales of veterinary antimicrobial agents in 30 European countries in 2015 (EMA/184855/2017). (European Medicines Agency, London, 2017), http://www.ema.europa.eu/docs/en_GB/document_library/Report/2017/10/WC500236750.pdf. Accessed 18 Jun 2018
- 2.Critically important antimicrobials for human medicine—5th rev. (World Health Organization, Geneva, 2017), http://who.int/foodsafety/publications/antimicrobials-fifth/en/. Accessed 18 Jun 2018
- 10.F. Führ, H. Ophoff, P. Burauel, U. Wanner, K. Haider, Modification of Definition of Bound Residues, in Pesticide Bound Residues in Soil, ed. by F. Führ, H. Ophoff (Wiley-VCH, Weinheim, 1998), pp. 175–176Google Scholar
- 12.Understanding the relationship between extraction technique and bioavailability. (European Centre for Ecotoxicology and Toxicology of Chemicals, Brussels, 2013), http://www.ecetoc.org/wp-content/uploads/2014/08/ECETOC-TR-117-Understanding-the-relationship-between-extraction-technique-and-bioavailability.pdf. Accessed 18 Jun 2018
- 14.A. Eschenbach, Characterization of non extractable residues for their risk assessment in soil with special regard to pharmaceuticals. (Umweltbundesamt, Dessau-Rosslau, 2013), https://www.umweltbundesamt.de/sites/default/files/medien/376/dokumente/eschenbach_presentation.pdf. Accessed 18 Jun 2018
- 21.J.P. Klare, H.J. Steinhoff, Structural Information from Spin-Labels and Intrinsic Paramagnetic Centres in the Biosciences, in Structure and Bonding, 152nd edn., ed. by C.R. Timmel, J.R. Harmer (Springer, Berlin, 2013), pp. 205–248Google Scholar
- 30.A.A. Bobko, I.A. Kirilyuk, I.A. Grigor’ev, J.L. Zweier, V.V. Khramtsov, Free Rad. Biol. Med. 42, 404 (2007)Google Scholar
- 31.J.H. Freed, in Spin Labeling: Theory and Applications, ed. by L.J. Berliner (Academic Press, New York, 1976), pp. 53–132Google Scholar
- 33.L.J. Berliner (ed.), Spin Labeling: Theory and Applications (Academic Press, New York, 1976)Google Scholar
- 36.M. Loos, M. Krauss, K. Fenner, Environ. Sci. Technol. 46, 9830 (2012)Google Scholar