Studies on Poisoning Characteristics of Phosphylated Oxime on AChE and Efficacy of Reactivators Based on Quantitative Determination of Effect Biomarkers by Liquid Chromatograph-mass Spectrometry†

doi:10.7503/cjcu20160945

Abstract

Abstract:

Organophosphorus compounds(OPs) have been extensively utilized throughout the world for many years, however, the security problems are always concerned due to the high toxicity. OPs primarily inhibit acetylcholinesterase(AChE) in nerve system and will cause cholinergic crisis in the synaptic cleft. Enzyme reactivatorsd representing oxime derivatives of quarternary pyridinium compounds are considered as specific medications. Recently, the exact clinical effectiveness of oxime reactivators is a matter of controversy. Although oxime reactivators can recover the activity of AChE and improve neuromuscular transmission, some study reports found oximes could also react with OPs and formmore potent inhibitors of AChE during the process of reactivation, which may intoxicate the reactivated AChE once again, even worsen the poisoning of enzyme due to the high reactivity. In this paper, paraoxon as an example of OPs and obidoxime representing a typical oxime were chosen to demonstrate the reactivation efficacy of oximes in the treatment of OPs poisoning. The reaction product diethylphosphoryl obidoxime(DEP-obidoxime) was first prepared. Then, the poisoning characteristics of phosphylated oxime on AChE and efficacy of reactivators were studied via quantitative determination of the effect biomarkers formed between paraoxon and AChE. The experimental results confirmed that DEP-obidoxime can intoxicate AChE in extremely short time, fortunately, the poisoned AChE by DEP-obidoxime can still be reactivated by typical oximes[pralidoxime, pyramidoxime monohydrate(HI-6) and obidoxime]. EC₅₀ values of each oxime were determined and indicated that the reactivation effectiveness of the above oximes as the following ranking: pralidoxime>HI-6>obidoxime.

Key words: Organophosphorus compound, Oxime, Phosphoryl oxime, Biomarker, Liquid chromatograph-mass spectrometry(LC-MS)

CLC Number:

O627.51

TrendMD:

XIE Yan, CHEN Jia, XU Bin, YAN Long, TANG Jijun, XIE Jianwei. Studies on Poisoning Characteristics of Phosphylated Oxime on AChE and Efficacy of Reactivators Based on Quantitative Determination of Effect Biomarkers by Liquid Chromatograph-mass Spectrometry^†[J]. Chem. J. Chinese Universities, 2017, 38(5): 758.

Figures/Tables 5

Scheme 1 Formation mechanism of DEP-obidoxime by reaction of paraoxon with obidoxime

Fig.1 HPLC separation chromatogram of the mixture of obidoxime and paraoxon(λ=260 nm)(A) and QTOF-MS spectrum of DEP-obidoxime(B)

Fig.2 Product ion spectrum of the diethylphosphoryl adducted peptides containing active site serine residue(X: diethylphosphoryl)

Fig.3 Inhibition degree of AChE by 200 nmol/L paraoxon(a) and 20 nmol/L DEP-obidoxime(b) over time

Table 1 EC50 values of oximes in reactivating paraoxon-inhibited AChE

Oxime	EC₅₀/(μmol·L^-1)	95% Confidence intervals of EC₅₀/(μmol·L^-1)
Pralidoxime	7.94	6.817—9.245
HI-6	37.39	31.65—44.17
Obidoxime	61.13	49.64—75.27

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