The toxicity of opiates and their metabolites in HepG2 cells

Mark Jairaj, D.G. Watson, M. Helen Grant, G.G. Skellern

Research output: Contribution to journalArticle

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Abstract

The toxicity of codeine (C), codeinone (CO), morphine (M), oxycodone (OC), pholcodine (P) and pholcodine-N-oxide (P-NOX) was assessed in HepG2 cells by determining cell viability via the measurement of lactate dehydrogenase (LDH) leakage through the membrane, depletion of reduced glutathione (GSH) and measurement of total protein content. Incubation of C, M, OC, P or P-NOX with HepG2 cells resulted in no significant loss of cell viability, depletion of GSH or decreased total protein content. In contrast, with CO there was a marked depletion of GSH with significant differences from control cells (P<0.05) being detected after as little as 5 min. This effect preceded the loss of cell viability and the decrease in total protein content. To identify the cause of GSH depletion during incubations with CO, the incubation solutions were analysed by liquid chromatography/tandem mass spectrometry (LC/MS/MS). Analysis showed that a codeinone-glutathione conjugate (CO-SG) had been formed. This adduct was synthesised and characterised by LC/MS/MS and by nuclear magnetic resonance spectroscopy (NMR). CO-SG was quantified in the incubation solutions using the synthesised standard substance. Results obtained in this study support the hypothesis that the toxicity of CO may be partly due to GSH depletion. The absence of LDH leakage and GSH depletion in the incubations containing C or OC suggests, that the presence of both a double bond at - 7 and an adjoining keto-group in the 6-position are necessary to elicit the toxicity of M analogues with regard to GSH depletion.
LanguageEnglish
Pages121-129
Number of pages9
JournalChemico-Biological Interactions
Volume146
Issue number2
DOIs
Publication statusPublished - 2003

Fingerprint

Opiate Alkaloids
Hep G2 Cells
Metabolites
Toxicity
Oxycodone
Cell Survival
Cells
L-Lactate Dehydrogenase
Oxides
Glutathione
Codeine
Proteins
Liquid chromatography
Tandem Mass Spectrometry
6-codeinone
Liquid Chromatography
Morphine
Nuclear magnetic resonance spectroscopy
Mass spectrometry
Magnetic Resonance Spectroscopy

Keywords

  • codeine
  • codeinone–glutathione conjugate
  • HepG2 cell
  • opiates

Cite this

Jairaj, Mark ; Watson, D.G. ; Grant, M. Helen ; Skellern, G.G. / The toxicity of opiates and their metabolites in HepG2 cells. In: Chemico-Biological Interactions. 2003 ; Vol. 146, No. 2. pp. 121-129.
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The toxicity of opiates and their metabolites in HepG2 cells. / Jairaj, Mark; Watson, D.G.; Grant, M. Helen; Skellern, G.G.

In: Chemico-Biological Interactions, Vol. 146, No. 2, 2003, p. 121-129.

Research output: Contribution to journalArticle

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