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Analysis of intact glucuronides and sulfates of serotonin, dopamine, and their phase I metabolites in rat brain microdialysates by liquid chromatography-tandem mass spectrometry.
Anal Chem. 2009 Oct 15; 81(20):8417-25.AC

Abstract

A method for the analysis of intact glucuronides and sulfates of common neurotransmitters serotonin (5-HT) and dopamine (DA) as well as of 5-hydroxy-3-indoleacetic acid (5-HIAA), 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA) in rat brain microdialysates by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. Enzyme-assisted synthesis using rat liver microsomes as a biocatalyst was employed for the production of 5-HT-, 5-HIAA-, DOPAC-, and HVA-glucuronides for reference compounds. The sulfate conjugates were synthesized either chemically or enzymatically using a rat liver S9 fraction. The LC-MS/MS method was validated by determining the limits of detection and quantitation, linearity, and repeatability for the quantitative analysis of 5-HT and DA and their glucuronides, as well as of 5-HIAA, DOPAC, and HVA and their sulfate-conjugates. In this study, 5-HT-glucuronide was for the first time detected in rat brain. The concentration of 5-HT-glucuronide (1.0-1.7 nM) was up to 2.5 times higher than that of free 5-HT (0.4-2.1 nM) in rat brain microdialysates, whereas the concentration of DA-glucuronide (1.0-1.4 nM) was at the same level or lower than the free DA (1.2-2.4 nM). The acidic metabolites of neurotransmitters, 5-HIAA, HVA, and DOPAC, were found in free and sulfated form, whereas their glucuronidation was not observed.

Authors+Show Affiliations

Division of Pharmaceutical Chemistry, Faculty of Pharmacy, P.O. Box 56, FI-00014 University of Helsinki, Helsinki, Finland.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19772284

Citation

Uutela, Päivi, et al. "Analysis of Intact Glucuronides and Sulfates of Serotonin, Dopamine, and Their Phase I Metabolites in Rat Brain Microdialysates By Liquid Chromatography-tandem Mass Spectrometry." Analytical Chemistry, vol. 81, no. 20, 2009, pp. 8417-25.
Uutela P, Reinilä R, Harju K, et al. Analysis of intact glucuronides and sulfates of serotonin, dopamine, and their phase I metabolites in rat brain microdialysates by liquid chromatography-tandem mass spectrometry. Anal Chem. 2009;81(20):8417-25.
Uutela, P., Reinilä, R., Harju, K., Piepponen, P., Ketola, R. A., & Kostiainen, R. (2009). Analysis of intact glucuronides and sulfates of serotonin, dopamine, and their phase I metabolites in rat brain microdialysates by liquid chromatography-tandem mass spectrometry. Analytical Chemistry, 81(20), 8417-25. https://doi.org/10.1021/ac901320z
Uutela P, et al. Analysis of Intact Glucuronides and Sulfates of Serotonin, Dopamine, and Their Phase I Metabolites in Rat Brain Microdialysates By Liquid Chromatography-tandem Mass Spectrometry. Anal Chem. 2009 Oct 15;81(20):8417-25. PubMed PMID: 19772284.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analysis of intact glucuronides and sulfates of serotonin, dopamine, and their phase I metabolites in rat brain microdialysates by liquid chromatography-tandem mass spectrometry. AU - Uutela,Päivi, AU - Reinilä,Ruut, AU - Harju,Kirsi, AU - Piepponen,Petteri, AU - Ketola,Raimo A, AU - Kostiainen,Risto, PY - 2009/9/24/entrez PY - 2009/9/24/pubmed PY - 2010/1/19/medline SP - 8417 EP - 25 JF - Analytical chemistry JO - Anal Chem VL - 81 IS - 20 N2 - A method for the analysis of intact glucuronides and sulfates of common neurotransmitters serotonin (5-HT) and dopamine (DA) as well as of 5-hydroxy-3-indoleacetic acid (5-HIAA), 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA) in rat brain microdialysates by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. Enzyme-assisted synthesis using rat liver microsomes as a biocatalyst was employed for the production of 5-HT-, 5-HIAA-, DOPAC-, and HVA-glucuronides for reference compounds. The sulfate conjugates were synthesized either chemically or enzymatically using a rat liver S9 fraction. The LC-MS/MS method was validated by determining the limits of detection and quantitation, linearity, and repeatability for the quantitative analysis of 5-HT and DA and their glucuronides, as well as of 5-HIAA, DOPAC, and HVA and their sulfate-conjugates. In this study, 5-HT-glucuronide was for the first time detected in rat brain. The concentration of 5-HT-glucuronide (1.0-1.7 nM) was up to 2.5 times higher than that of free 5-HT (0.4-2.1 nM) in rat brain microdialysates, whereas the concentration of DA-glucuronide (1.0-1.4 nM) was at the same level or lower than the free DA (1.2-2.4 nM). The acidic metabolites of neurotransmitters, 5-HIAA, HVA, and DOPAC, were found in free and sulfated form, whereas their glucuronidation was not observed. SN - 1520-6882 UR - https://www.unboundmedicine.com/medline/citation/19772284/Analysis_of_intact_glucuronides_and_sulfates_of_serotonin_dopamine_and_their_phase_I_metabolites_in_rat_brain_microdialysates_by_liquid_chromatography_tandem_mass_spectrometry_ L2 - https://doi.org/10.1021/ac901320z DB - PRIME DP - Unbound Medicine ER -