HPLC-assay with electrochemical detection for the neurotoxin MPTP, its metabolite MPP+ and MPTP-analogues in biological samples after purification over Sephadex G10.
A sensitive and selective method for assaying the neurotoxin MPTP and some MPTP-analogues in mouse brain and serum is described. The method is based on isolation of the compounds from biological samples on small Sephadex G10 columns followed by reverse phase HPLC with amperometric detection. HPLC separation was performed at pH 3, after which the pH was increased to 6.8 by mixing the column effluent with 0.5 M phosphate pH 9, to provide the conditions required for electrochemical detection. A metabolite of MPTP, MPP+, was determined as MPTP after reduction with NaBH4. This assay allows the determination of brain and serum concentrations in the pmol/g range of administered MPTP and MPTP-analogues and the effects of these substances on dopamine and its metabolites in the same tissue sample.
D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D015655:1-Methyl-4-phenylpyridinium; D000818:Animals; D001923:Brain Chemistry; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006863:Hydrogen-Ion Concentration; D008297:Male; D051379:Mice; D011725:Pyridines / Q000032:analysis* / Q000378:metabolism; D011726:Pyridinium Compounds / Q000032:analysis*
D011725:Pyridines; D011726:Pyridinium Compounds; D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D015655:1-Methyl-4-phenylpyridinium