Isolation, separation and analysis in neurochemistry: Trace amines and acids as an illustrative example

Abstract

In this brief overview various neurochemical isolation procedures that can be adopted for the analysis of several monoamine neurotransmitters/neuromodulators and their principal oxidatively deaminated metabolites are outlined. With respect to the trace amines, they can be identified and quantitated as their so-called dansyl derivatives after thin-layer chromatographic separation by mass spectrometric (MS) electron-impact (EI) ionisation followed by selected-ion monitoring (SIM) of their molecular ions. Deuterated homologues are added as internal standards at the start of the analytical procedure. The MS-EI-SIM procedure offers a tissue extract or releasate sensitivity of about 100 pg/g of tissue or fluid. In the case of tryptamine or phenylethylamine, by utilising different derivatives (N-acetylpentafluoropropionyl or N-acetylpentafluorobenzoyl), which cyclise to form perfluorinated spirocyclic compounds, it is possible using MS negative chemical ionisation techniques coupled with monitoring of the (M — HF) ions to achieve sensitivities for tissue extracts of 1 pg/g or less. Acidic and neutral metabolites (up to twelve of them can be assayed simultaneously) can be detected and quantitated in tissue extracts, releasates or biological fluids as their methyl-pentafluoropropionyl or trifluoroethyl-pentafluoropropionyl derivatives in the 100–1000 pg range using gas chromatographic-MS-SIM procedures.