High-performance liquid chromatography/tandem mass spectrometry assay for the determination of 1-methyl-4-phenyl pyridinium (MPP+) in brain tissue homogenates.
A high-throughput liquid chromatography/tandem mass spectrometry method has been developed for the quantitative assessment of 1-methyl-4-phenylpyridinium (MPP+) in brain tissue samples. This separation is based on reversed phase chromatography using formic acid and acetonitrile as the mobile phase. Using gradient separation conditions, MPP+ was resolved within 5 min and detected using tandem mass spectrometry in the positive ion electrospray mode. The limit of detection for MPP+ was found to be 1 fmol on column with a signal to noise ratio of 3:1. The assay has been used routinely in our laboratory for the measurement of MPP+ levels in brain tissue from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice, and can be used to distinguish neuroprotective efficacy and monoamine oxidase inhibition.
Hows
Mark E P
ME
Computational Analytical and Structural Sciences, GlaxoSmithKline, New Frontiers Science Park, Third Avenue, Harlow CM19 5AW, Essex, UK. mark.e.hows@gsk.com
Ashmeade
Tracey E
TE
Billinton
Andrew
A
Perren
Marion J
MJ
Austin
Angela A
AA
Virley
David J
DJ
Organ
Andrew J
AJ
Shah
Ajit J
AJ
D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine / Q000032:analysis / Q000737:chemistry; D015655:1-Methyl-4-phenylpyridinium / Q000032:analysis* / Q000737:chemistry / Q000633:toxicity; D000818:Animals; D000978:Antiparkinson Agents / Q000494:pharmacology; D001921:Brain / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D015203:Reproducibility of Results; D012642:Selegiline / Q000494:pharmacology; D012680:Sensitivity and Specificity; D013997:Time Factors; D014018:Tissue Distribution
D000978:Antiparkinson Agents; D012642:Selegiline; D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D015655:1-Methyl-4-phenylpyridinium