Identification and quantification of gamma-glutamyl conjugates of biogenic amines in the nervous system of the snail, Helix aspersa, by gas chromatography-negative-ion chemical ionisation mass spectrometry.

The gamma-glutamyl conjugates of p-octopamine and dopamine were identified unambiguously for the first time and quantified in a single cerebral ganglion or pleural plus pedal ganglia of the snail, Helix aspersa, by gas chromatography-negative-ion chemical ionisation mass spectrometry. A new method was used for synthesis of gamma-glutamylamine standards. The concentration of gamma-glutamyltyramine was found to be low in the tissues, therefore it was used as an internal standard. The gamma-glutamylamines were extracted with water and derivatised with pentafluoropropionic anhydride and trifluoroethanol. Under negative-ion chemical ionisation conditions, the trifluoroethyl and pentafluoropropionyl derivatives produced significant ions which were sufficiently abundant to be suitable for selective ion monitoring. The method had a limit of detection of ca. 80 pg of gamma-glutamyl conjugate per tissue and calibration curves were linear over the range examined.