Characterization of thioether compounds formed from alkaline degradation products of enflurane.
BACKGROUND
Renal toxicity has occasionally been observed after enflurane anesthesia. Although originally attributed to its oxidative metabolism to inorganic fluoride, serum levels of inorganic fluoride appear to be small to explain these renal effects. Formation of potentially nephrotoxic halogenated alkenes during alkaline degradation in carbon dioxide absorbers and subsequent bioactivation via the glutathione conjugation pathway may be considered as an alternative mechanism for renal toxicity. The aim of this study was to characterize the thioethers formed chemically and biosynthetically.


METHODS
Alkaline degradation of enflurane was achieved by stirring with pulverized potassium hydroxide. Volatile degradation products were analyzed by 19F nuclear magnetic resonance (NMR) analysis of head space gasses trapped in dimethyl sulfoxide (DMSO). Thioethers were generated chemically by trapping head space gasses in DMSO containing N-acetyl-L-cysteine or 2-mercaptoacetic acid as model thiol compounds. Glutathione conjugates were generated biosynthetically by passing head space through rat liver fractions in presence of glutathione. Products formed were analyzed by gas chromatography-mass spectroscopy and 19F-NMR.


RESULTS
Direct analysis of head space gasses showed formation of 1-chloro-1,2-difluorovinyl difluoromethyl ether and two unidentified fluorine-containing products as alkaline degradation products of enflurane. When trapped in DMSO-N-acetyl-L-cysteine-triethylamine, N-acetyl-S-(2-chloro-1,2-difluoro-1-(difluoromethoxy)ethyl)-L-cysteine was identified as the major product. Another N-acetyl-L-cysteine S-conjugate formed was N-acetyl-S-(2-chloro-1,1,2-trifluoroethyl)-L-cysteine, a potent nephrotoxin in rats. 19F-NMR analysis of glutathione conjugates formed after incubation with rat liver fractions resulted in formation of corresponding S-conjugates.


CONCLUSIONS
The current study demonstrates that alkaline degradation products of enflurane can be conjugated to thiol compounds, forming S-conjugates that could theoretically contribute to adverse renal effects observed occasionally with enflurane anesthesia. The N-acetyl-L-cysteine S-conjugates identified may be biomarkers to assess exposure of humans to alkaline degradation products of enflurane.
Orhan
H
H
Division of Molecular Toxicology, Vrije Universiteit, Amsterdam, The Netherlands.
Vermeulen
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NP
Sahin
G
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Commandeur
J N
JN
D000111:Acetylcysteine / Q000737:chemistry; D000468:Alkalies; D018685:Anesthetics, Inhalation / Q000737:chemistry*; D000818:Animals; D003600:Cytosol / Q000378:metabolism; D004737:Enflurane / Q000737:chemistry*; D008401:Gas Chromatography-Mass Spectrometry; D005978:Glutathione / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008862:Microsomes, Liver / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013440:Sulfides / Q000737:chemistry*; D013864:Thioglycolates / Q000737:chemistry
D000468:Alkalies; D018685:Anesthetics, Inhalation; D013440:Sulfides; D013864:Thioglycolates; C017487:2-mercaptoacetate; D004737:Enflurane; D005978:Glutathione; D000111:Acetylcysteine