Use of stable isotopes and gas chromatography-mass spectrometry in the study of different pools of neurotransmitter amino acids in brain slices

GC/MS Profiling of γ-Hydroxybutyrate and Precursors in Various Animal Tissues Using Automatic Solid-Phase Extraction. Preliminary Investigations of Its Potential Interest in Postmortem Interval Determination

“…Different protocols of derivatization of target compounds were studied to improve the sensitivity of MS detection. Derivatization by silylation with MTBSTFA, initially developed for the assay of GABA, was successfully employed for both compounds. − This derivatization method made it possible to obtain the ions of quantifications for each molecule with the largest m / z ratios, making it possible to decrease the detection thresholds.…”

GC/MS Profiling of γ-Hydroxybutyrate and Precursors in Various Animal Tissues Using Automatic Solid-Phase Extraction. Preliminary Investigations of Its Potential Interest in Postmortem Interval Determination

“…Different protocols of derivatization of target compounds were studied to improve the sensitivity of MS detection. Derivatization by silylation with MTBSTFA, initially developed for the assay of GABA, was successfully employed for both compounds. − This derivatization method made it possible to obtain the ions of quantifications for each molecule with the largest m / z ratios, making it possible to decrease the detection thresholds.…”

Compartmentation of cerebral glutamate in situ as detected by 1H/13C n.m.r

“…spectra (interpulse interval 6.5 s, spectral width 11.6 kHz, 32768 data points) were acquired with a continuous broad-band irradiation accomplished by composite pulses at the 'H frequency for both decoupling and nuclear Overhauser enhancement. (Kapetanovic et al, 1990). Briefly, the residues were derivatized with 200,u of acetonitrile/N-methyl-N(t-butyldimethylsilyl)trifluoroacetamide (3: 1, v/v) at 80°C for 30 min.…”

Mass spectrometric studies on brain metabolism, using stable isotopes

“…To alleviate that complexity, simple experimental designs may be adopted in the first instance, before progressing to in vivo studies. Therefore, in vitro systems that use cultured neural cells or brain slices have frequently been employed for neurochemical studies, including the application of stable isotopes (Kapetanovic, Yonekawa, & Kupferberg, 1990; Hertz et al, 2000; Waagepetersen et al, 2000).…”

“…Brain slices were employed as an in vitro brain model in which cellular architecture and neuronal connections were, in part, functionally preserved as in vivo , but in which the BBB was lost. GC/MS was applied to the simultaneous determination of the endogenous and newly synthesized neurotransmitter amino acids (glutamate, GABA, and aspartate) and glutamine (Kapetanovic, Yonekawa, & Kupferberg, 1990). Hippocampal slices were incubated for 30 min in artificial cerebrospinal fluid in the presence of [ 13 C 6 ]glucose, [2,3‐ 13 C 2 ]pyruvate, and [1,2‐ 13 C 2 ]acetate.…”

GC/MS Profiling of γ-Hydroxybutyrate and Precursors in Various Animal Tissues Using Automatic Solid-Phase Extraction. Preliminary Investigations of Its Potential Interest in Postmortem Interval Determination

“…Different protocols of derivatization of target compounds were studied to improve the sensitivity of MS detection. Derivatization by silylation with MTBSTFA, initially developed for the assay of GABA, was successfully employed for both compounds. − This derivatization method made it possible to obtain the ions of quantifications for each molecule with the largest m / z ratios, making it possible to decrease the detection thresholds.…”

Compartmentation of cerebral glutamate in situ as detected by 1H/13C n.m.r

“…spectra (interpulse interval 6.5 s, spectral width 11.6 kHz, 32768 data points) were acquired with a continuous broad-band irradiation accomplished by composite pulses at the 'H frequency for both decoupling and nuclear Overhauser enhancement. (Kapetanovic et al, 1990). Briefly, the residues were derivatized with 200,u of acetonitrile/N-methyl-N(t-butyldimethylsilyl)trifluoroacetamide (3: 1, v/v) at 80°C for 30 min.…”

Mass spectrometric studies on brain metabolism, using stable isotopes

“…To alleviate that complexity, simple experimental designs may be adopted in the first instance, before progressing to in vivo studies. Therefore, in vitro systems that use cultured neural cells or brain slices have frequently been employed for neurochemical studies, including the application of stable isotopes (Kapetanovic, Yonekawa, & Kupferberg, 1990; Hertz et al, 2000; Waagepetersen et al, 2000).…”

“…Brain slices were employed as an in vitro brain model in which cellular architecture and neuronal connections were, in part, functionally preserved as in vivo , but in which the BBB was lost. GC/MS was applied to the simultaneous determination of the endogenous and newly synthesized neurotransmitter amino acids (glutamate, GABA, and aspartate) and glutamine (Kapetanovic, Yonekawa, & Kupferberg, 1990). Hippocampal slices were incubated for 30 min in artificial cerebrospinal fluid in the presence of [ 13 C 6 ]glucose, [2,3‐ 13 C 2 ]pyruvate, and [1,2‐ 13 C 2 ]acetate.…”

GC/MS Profiling of γ-Hydroxybutyrate and Precursors in Various Animal Tissues Using Automatic Solid-Phase Extraction. Preliminary Investigations of Its Potential Interest in Postmortem Interval Determination

“…Different protocols of derivatization of target compounds were studied to improve the sensitivity of MS detection. Derivatization by silylation with MTBSTFA, initially developed for the assay of GABA, was successfully employed for both compounds. − This derivatization method made it possible to obtain the ions of quantifications for each molecule with the largest m / z ratios, making it possible to decrease the detection thresholds.…”

Compartmentation of cerebral glutamate in situ as detected by 1H/13C n.m.r

“…spectra (interpulse interval 6.5 s, spectral width 11.6 kHz, 32768 data points) were acquired with a continuous broad-band irradiation accomplished by composite pulses at the 'H frequency for both decoupling and nuclear Overhauser enhancement. (Kapetanovic et al, 1990). Briefly, the residues were derivatized with 200,u of acetonitrile/N-methyl-N(t-butyldimethylsilyl)trifluoroacetamide (3: 1, v/v) at 80°C for 30 min.…”

Mass spectrometric studies on brain metabolism, using stable isotopes

“…To alleviate that complexity, simple experimental designs may be adopted in the first instance, before progressing to in vivo studies. Therefore, in vitro systems that use cultured neural cells or brain slices have frequently been employed for neurochemical studies, including the application of stable isotopes (Kapetanovic, Yonekawa, & Kupferberg, 1990; Hertz et al, 2000; Waagepetersen et al, 2000).…”

“…Brain slices were employed as an in vitro brain model in which cellular architecture and neuronal connections were, in part, functionally preserved as in vivo , but in which the BBB was lost. GC/MS was applied to the simultaneous determination of the endogenous and newly synthesized neurotransmitter amino acids (glutamate, GABA, and aspartate) and glutamine (Kapetanovic, Yonekawa, & Kupferberg, 1990). Hippocampal slices were incubated for 30 min in artificial cerebrospinal fluid in the presence of [ 13 C 6 ]glucose, [2,3‐ 13 C 2 ]pyruvate, and [1,2‐ 13 C 2 ]acetate.…”