Quantification of endogenous neurotransmitters and related compounds by liquid chromatography coupled to tandem mass spectrometry

Eulàlia Olesti, Jose Rodríguez-Morató, Alex Gomez-Gomez, Johannes G Ramaekers, Rafael de la Torre, Oscar J Pozo

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Neurotransmitters are signaling molecules, playing key roles in neuronal communications in the brain. Drug induced changes in neurotransmitters and other brain metabolite concentration may be used to characterize drugs according to their targeted metabolomics profile. Here, we report the development and validation of a straightforward liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of 16 endogenous small polar compounds in rat plasma and brain homogenates. The method enables the quantification of the neurotransmitters γ-aminobutyric acid, glutamate, acetylcholine and adenosine, as well as choline, glutamine, acetylcarnitine, carnitine, creatine, creatinine, valine, leucine, isoleucine, phenylalanine, tyrosine and tryptophan. After optimizing the sample preparation, chromatographic and spectrometric conditions, the method was successfully validated using the standard addition approach and a hydrophilic interaction chromatography (HILIC) with an amide column. The method was shown to be linear (r > 0.99) as all the compounds were within the ±25% values of intra and inter-day precision and accuracy acceptance. A matrix effect was corrected with the use of 10 isotopically labelled internal standards and the compound stability was evaluated for all compounds. Relevant exaltation of choline (in plasma) and creatinine (in brain) were solved with -20 °C conditions. The applicability of the method was tested by evaluating brain alterations in the concentrations of neurotransmitters and related compounds after the administration of two psychostimulant drugs of abuse (cocaine and methylenedioxypyrovalerone) to rats. A neuro-metabolic fingerprint of each drug was obtained that reflected their pharmacological profile. Altogether, this methodology presents a valuable targeted metabolomics tool for basic and clinical research studies.

Original languageEnglish
Pages (from-to)93-102
Number of pages10
JournalTalanta
Volume192
DOIs
Publication statusPublished - 15 Jan 2019

Keywords

  • 3,4-METHYLENEDIOXYPYROVALERONE MDPV
  • AMINO-ACID
  • BATH SALTS
  • BRAIN-TISSUE
  • Brain neurotransmitters
  • Hydrophilic interaction chromatography
  • Liquid chromatography coupled to mass spectrometry
  • METABOLITES
  • MICROEXTRACTION
  • MONOAMINE NEUROTRANSMITTERS
  • Psychoactive substances
  • Quantitative method
  • RAT-BRAIN
  • Standard addition
  • TOXICOLOGY
  • UHPLC-MS/MS

Cite this

Olesti, Eulàlia ; Rodríguez-Morató, Jose ; Gomez-Gomez, Alex ; Ramaekers, Johannes G ; de la Torre, Rafael ; Pozo, Oscar J. / Quantification of endogenous neurotransmitters and related compounds by liquid chromatography coupled to tandem mass spectrometry. In: Talanta. 2019 ; Vol. 192. pp. 93-102.
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abstract = "Neurotransmitters are signaling molecules, playing key roles in neuronal communications in the brain. Drug induced changes in neurotransmitters and other brain metabolite concentration may be used to characterize drugs according to their targeted metabolomics profile. Here, we report the development and validation of a straightforward liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of 16 endogenous small polar compounds in rat plasma and brain homogenates. The method enables the quantification of the neurotransmitters γ-aminobutyric acid, glutamate, acetylcholine and adenosine, as well as choline, glutamine, acetylcarnitine, carnitine, creatine, creatinine, valine, leucine, isoleucine, phenylalanine, tyrosine and tryptophan. After optimizing the sample preparation, chromatographic and spectrometric conditions, the method was successfully validated using the standard addition approach and a hydrophilic interaction chromatography (HILIC) with an amide column. The method was shown to be linear (r > 0.99) as all the compounds were within the ±25{\%} values of intra and inter-day precision and accuracy acceptance. A matrix effect was corrected with the use of 10 isotopically labelled internal standards and the compound stability was evaluated for all compounds. Relevant exaltation of choline (in plasma) and creatinine (in brain) were solved with -20 °C conditions. The applicability of the method was tested by evaluating brain alterations in the concentrations of neurotransmitters and related compounds after the administration of two psychostimulant drugs of abuse (cocaine and methylenedioxypyrovalerone) to rats. A neuro-metabolic fingerprint of each drug was obtained that reflected their pharmacological profile. Altogether, this methodology presents a valuable targeted metabolomics tool for basic and clinical research studies.",
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Quantification of endogenous neurotransmitters and related compounds by liquid chromatography coupled to tandem mass spectrometry. / Olesti, Eulàlia; Rodríguez-Morató, Jose; Gomez-Gomez, Alex; Ramaekers, Johannes G; de la Torre, Rafael; Pozo, Oscar J.

In: Talanta, Vol. 192, 15.01.2019, p. 93-102.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Quantification of endogenous neurotransmitters and related compounds by liquid chromatography coupled to tandem mass spectrometry

AU - Olesti, Eulàlia

AU - Rodríguez-Morató, Jose

AU - Gomez-Gomez, Alex

AU - Ramaekers, Johannes G

AU - de la Torre, Rafael

AU - Pozo, Oscar J

N1 - Copyright © 2018 Elsevier B.V. All rights reserved.

PY - 2019/1/15

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N2 - Neurotransmitters are signaling molecules, playing key roles in neuronal communications in the brain. Drug induced changes in neurotransmitters and other brain metabolite concentration may be used to characterize drugs according to their targeted metabolomics profile. Here, we report the development and validation of a straightforward liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of 16 endogenous small polar compounds in rat plasma and brain homogenates. The method enables the quantification of the neurotransmitters γ-aminobutyric acid, glutamate, acetylcholine and adenosine, as well as choline, glutamine, acetylcarnitine, carnitine, creatine, creatinine, valine, leucine, isoleucine, phenylalanine, tyrosine and tryptophan. After optimizing the sample preparation, chromatographic and spectrometric conditions, the method was successfully validated using the standard addition approach and a hydrophilic interaction chromatography (HILIC) with an amide column. The method was shown to be linear (r > 0.99) as all the compounds were within the ±25% values of intra and inter-day precision and accuracy acceptance. A matrix effect was corrected with the use of 10 isotopically labelled internal standards and the compound stability was evaluated for all compounds. Relevant exaltation of choline (in plasma) and creatinine (in brain) were solved with -20 °C conditions. The applicability of the method was tested by evaluating brain alterations in the concentrations of neurotransmitters and related compounds after the administration of two psychostimulant drugs of abuse (cocaine and methylenedioxypyrovalerone) to rats. A neuro-metabolic fingerprint of each drug was obtained that reflected their pharmacological profile. Altogether, this methodology presents a valuable targeted metabolomics tool for basic and clinical research studies.

AB - Neurotransmitters are signaling molecules, playing key roles in neuronal communications in the brain. Drug induced changes in neurotransmitters and other brain metabolite concentration may be used to characterize drugs according to their targeted metabolomics profile. Here, we report the development and validation of a straightforward liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of 16 endogenous small polar compounds in rat plasma and brain homogenates. The method enables the quantification of the neurotransmitters γ-aminobutyric acid, glutamate, acetylcholine and adenosine, as well as choline, glutamine, acetylcarnitine, carnitine, creatine, creatinine, valine, leucine, isoleucine, phenylalanine, tyrosine and tryptophan. After optimizing the sample preparation, chromatographic and spectrometric conditions, the method was successfully validated using the standard addition approach and a hydrophilic interaction chromatography (HILIC) with an amide column. The method was shown to be linear (r > 0.99) as all the compounds were within the ±25% values of intra and inter-day precision and accuracy acceptance. A matrix effect was corrected with the use of 10 isotopically labelled internal standards and the compound stability was evaluated for all compounds. Relevant exaltation of choline (in plasma) and creatinine (in brain) were solved with -20 °C conditions. The applicability of the method was tested by evaluating brain alterations in the concentrations of neurotransmitters and related compounds after the administration of two psychostimulant drugs of abuse (cocaine and methylenedioxypyrovalerone) to rats. A neuro-metabolic fingerprint of each drug was obtained that reflected their pharmacological profile. Altogether, this methodology presents a valuable targeted metabolomics tool for basic and clinical research studies.

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KW - Liquid chromatography coupled to mass spectrometry

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KW - MICROEXTRACTION

KW - MONOAMINE NEUROTRANSMITTERS

KW - Psychoactive substances

KW - Quantitative method

KW - RAT-BRAIN

KW - Standard addition

KW - TOXICOLOGY

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