Monitoring the hydrolysis of toxic organophosphonate nerve agents in aqueous buffer and in bicontinuous microemulsions by use of diisopropyl fluorophosphatase (DFPase) with H-1-P-31 HSQC NMR spectroscopy

Gaeb J, Melzer M, Kehe K, Wellert S, Hellweg T, Blum M-M (2010)
Analytical and Bioanalytical Chemistry 396(3): 1213-1221.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Abstract / Bemerkung
The enzyme diisopropyl fluorophosphatase (DFPase, EC 3.1.8.2) from the squid Loligo vulgaris effectively catalyzes the hydrolysis of diisopropyl fluorophosphate (DFP) and a number of organophosphorus nerve agents, including sarin, soman, cyclosarin, and tabun. Until now, determination of kinetic data has been achieved by use of techniques such as pH-stat titration, ion-selective electrodes, and a recently introduced method based on in situ Fourier-transform infrared (FTIR) spectroscopy. We report the use of 1D H-1-P-31 HSQC NMR spectroscopy as a new method for real-time quantification of the hydrolysis of toxic organophosphonates by DFPase. The method is demonstrated for the agents sarin (GB), soman (GD), and cyclosarin (GD) but can also be used for V-type nerve agents, for example VX. Besides buffered aqueous solutions the method was used to determine enzymatic activities in a biodiesel-based bicontinuous microemulsion that serves as an example of complex decontamination media, for which other established techniques often fail. The method is non-invasive and requires only limited manual handling of small volumes of liquid (700 mu L), which adds to work safety when handling highly toxic organophosphorus compounds. Limits of detection are slightly below 100 mu mol L-1 on a 400 MHz spectrometer with 16 FIDs added for a single time frame. The method is not restricted to DFPase but can be used with other phosphotriesterases, for example paraxonase (PON), and even reactive chemicals, for example oximes and other nucleophiles, as long as the reaction components are compatible with the NMR experiment.
Stichworte
Enzymes; Phosphotriesterases; Nerve agents; NMR; DFPase; Hydrolases
Erscheinungsjahr
2010
Zeitschriftentitel
Analytical and Bioanalytical Chemistry
Band
396
Ausgabe
3
Seite(n)
1213-1221
ISSN
1618-2642
eISSN
1618-2650
Page URI
https://pub.uni-bielefeld.de/record/1999947

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Gaeb J, Melzer M, Kehe K, Wellert S, Hellweg T, Blum M-M. Monitoring the hydrolysis of toxic organophosphonate nerve agents in aqueous buffer and in bicontinuous microemulsions by use of diisopropyl fluorophosphatase (DFPase) with H-1-P-31 HSQC NMR spectroscopy. Analytical and Bioanalytical Chemistry. 2010;396(3):1213-1221.
Gaeb, J., Melzer, M., Kehe, K., Wellert, S., Hellweg, T., & Blum, M. - M. (2010). Monitoring the hydrolysis of toxic organophosphonate nerve agents in aqueous buffer and in bicontinuous microemulsions by use of diisopropyl fluorophosphatase (DFPase) with H-1-P-31 HSQC NMR spectroscopy. Analytical and Bioanalytical Chemistry, 396(3), 1213-1221. doi:10.1007/s00216-009-3299-2
Gaeb, J., Melzer, M., Kehe, K., Wellert, S., Hellweg, T., and Blum, M. - M. (2010). Monitoring the hydrolysis of toxic organophosphonate nerve agents in aqueous buffer and in bicontinuous microemulsions by use of diisopropyl fluorophosphatase (DFPase) with H-1-P-31 HSQC NMR spectroscopy. Analytical and Bioanalytical Chemistry 396, 1213-1221.
Gaeb, J., et al., 2010. Monitoring the hydrolysis of toxic organophosphonate nerve agents in aqueous buffer and in bicontinuous microemulsions by use of diisopropyl fluorophosphatase (DFPase) with H-1-P-31 HSQC NMR spectroscopy. Analytical and Bioanalytical Chemistry, 396(3), p 1213-1221.
J. Gaeb, et al., “Monitoring the hydrolysis of toxic organophosphonate nerve agents in aqueous buffer and in bicontinuous microemulsions by use of diisopropyl fluorophosphatase (DFPase) with H-1-P-31 HSQC NMR spectroscopy”, Analytical and Bioanalytical Chemistry, vol. 396, 2010, pp. 1213-1221.
Gaeb, J., Melzer, M., Kehe, K., Wellert, S., Hellweg, T., Blum, M.-M.: Monitoring the hydrolysis of toxic organophosphonate nerve agents in aqueous buffer and in bicontinuous microemulsions by use of diisopropyl fluorophosphatase (DFPase) with H-1-P-31 HSQC NMR spectroscopy. Analytical and Bioanalytical Chemistry. 396, 1213-1221 (2010).
Gaeb, Juergen, Melzer, Marco, Kehe, Kai, Wellert, Stefan, Hellweg, Thomas, and Blum, Marc-Michael. “Monitoring the hydrolysis of toxic organophosphonate nerve agents in aqueous buffer and in bicontinuous microemulsions by use of diisopropyl fluorophosphatase (DFPase) with H-1-P-31 HSQC NMR spectroscopy”. Analytical and Bioanalytical Chemistry 396.3 (2010): 1213-1221.

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