Quantitative amino acid analysis of individual snail neurons by open tubular liquid chromatography

Abstract

A method is described for the determination of amino acids in individual cells. The amino acids are derivatized with naphthalene-2,3-dicarboxaldehyde and then analyzed by open tubular liquid chromatography with amperometric detection. The total volume present after derivatization is approximately 25 nL. It was possible to quantitatively determine 17 amino acids in three different neurons of the land snail Helix aspersa. Quantitation was accomplished through the use of two internal standards and a calibration curve. Alanine was found to be the most abundant amino acid by about a factor of 2 over glutamine in all three types of neurons. This method has the advantages of sensitivity in the attomole range (5 X 10(-9) M) and selectivity for a specific class of compounds and is at least as reliable as other methods used for single-cell analysis.

Variation in biochemical composition during gonad maturation of the tropical abaloneHaliotis variaLinnaeus 1758 (Vetigastropoda: Haliotidae)

“…In contrast to the voluminous studies on oysters and other lamellibranchs, biochemical studies on gastropods are limited. Some of the available literature elucidates various aspects of the biochemical composition of limpets (Barry & Munday 1959;Blackmore 1969), snails (Giese 1967;Oates et al 1990;Carasco et al 2006) and abalone (Mercer et al 1993;Knauer et al 1994;Chiou et al 2001Chiou et al , 2002Litaay & De Silva 2003). Chemical investigations of abalone body components, especially the biochemical composition of foot muscle and its texture properties (Hatae et al 1995) are of great significance because of their high commercial value in the international market (Guo et al 1999).…”

Analysis of amino acids in individual human erythrocytes by capillary electrophoresis with electroporation for intracellular derivatization and laser‐induced fluorescence detection

“…Jorgenson and co-workers [8,9] described a derivatization method in single-cell analysis by open-tubular liquid chromatography (OT-LC). One giant neuron with ca.…”

Analytical chemistry in microenvironments: single nerve cells

“…This method has been used to demonstrate the coexistence of dopamine and serotonin in a single neuron in Helix aspersa (3,37). Additionally, whole cells have been removed from Helix aspersa and the amino acids derivatized with naphthalene-2,3-dicarboxaldehyde followed by OTLC separation with amp-erometric detection (39). The methods described here have also been used with this added derivatization step to apply both the OTLC and capillary electrophoresis techniques with laser fluorescence detection to whole cell profiling of amines (3).…”