Psi-screen, an in vitro toxicity test system: applications in the bioassay of perfumes and fragrance chemicals.
The effects of 65 perfume formulations (perfume oils, perfumes, eau de parfum, eau de toilette) on mitochondrial membrane potential (Psim) and mitochondrial respiration have been investigated using a mitochondria-based assay for (Psim, termed Psi-Screen. All the perfume formulations tested are highly active in the Psi-Screen assay, and the major site of inhibition in all cases is NADH-ubiquinone reductase (Complex I). This is confirmed in studies on the inhibition of NADH oxidase and NADH-ubiquinone reductase. Some formulations also inhibit succinate oxidation at either Complex II or Complex III. Evidence for the inhibition of mitochondrial ATPase is presented, as well as for the induction of reactive oxygen species production by perfume inhibition of Complex I. Thus, perfume formulations are multiple inhibitor mixtures which inhibit multiple bioenergetic functions at high dilutions. The implications of these findings are discussed with respect to cell toxicity via necrosis and/or apoptosis. Twenty candidate fragrance chemicals were investigated and all inhibited Complex I (5 at <35 microM). Mass screening strategies and high-throughput screening assays are discussed.
Griffiths
David E
DE
Department of Chemistry, University of Warwick, Coventry, CV4 7AL, UK. d.griffiths@warwick.ac.uk
D000818:Animals; D002417:Cattle; D016208:Databases, Factual; D042967:Electron Transport Complex I / Q000032:analysis / Q000187:drug effects; D042963:Electron Transport Complex II / Q000187:drug effects; D014450:Electron Transport Complex III / Q000187:drug effects; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000187:drug effects; D008929:Mitochondria, Heart / Q000187:drug effects*; D051336:Mitochondrial Membranes / Q000187:drug effects; D025261:Mitochondrial Proton-Translocating ATPases / Q000037:antagonists & inhibitors; D009097:Multienzyme Complexes / Q000032:analysis / Q000187:drug effects; D009247:NADH, NADPH Oxidoreductases / Q000032:analysis / Q000187:drug effects; D010476:Perfume / Q000633:toxicity*; D051381:Rats; D017382:Reactive Oxygen Species / Q000032:analysis; D018675:Toxicity Tests / Q000379:methods*
D009097:Multienzyme Complexes; D010476:Perfume; D017382:Reactive Oxygen Species; D042963:Electron Transport Complex II; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; D025261:Mitochondrial Proton-Translocating ATPases; D042967:Electron Transport Complex I; D014450:Electron Transport Complex III