Controlling detrimental effects of metal cations in the quantification of energy metabolites via ultrahigh pressure-liquid chromatography-electrospray-tandem mass spectrometry by employing acetylacetone as a volatile eluent modifier.

摘要: The majority of energy metabolites involved in glycolysis, the citric acid cycle and phosphate pentose pathway shows a high affinity for metal cations. In this paper, we demonstrate negative effect cations on UHPLC-ESI-MS/MS analysis suggest acetylacetone (acac) as volatile, metal-chelating eluent modifier with potential to remediate these effects. For 52 analytes considered, modification acac led significant improvements almost all analytical figures merit. Using aqueous standards, peak intensities were increased 2.5-fold average. biological matrices (yeast, murine tissue), 5-fold increase was observed, whereas number detectable by 26% from 31 39 Limits detection improved 4-fold average (equaling two 1:1 dilution steps), most seen efficient chelators like citrate (256-fold), salicylate or coenzyme A acetyl-coenzyme (16-fold each). beneficial resulted mainly an averaged column efficiency (number theoretical plates, N) 57%. To determine species scavenged acac, quantified metal-acetylacetonate complexes UHPLC ESI-MS/MS. Our results indicate that Al(3+) (342±34pM/min leakage into effluent) Fe(3+) (22±1pM/min effluent), presumably due mild corrosion inside column, are responsible detrimental effects alleviated acac.