Validated methods for determination of neurotransmitters and metabolites in rodent brain tissue and extracellular fluid by reversed phase UHPLC–MS/MS

Bergh, Marianne Skov-Skov; Bogen, Inger Lise; Lundanes, Elsa; Øiestad, Åse Marit Leere

doi:10.1016/j.jchromb.2016.06.011

Cited by 48 publications

(32 citation statements)

References 28 publications

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“…The distribution tendencies of DA, 5-HT and GABA determined in rat brain were similar to those reported in previous studies (Bergh et al, 2016;González et al, 2011;Wojnicz et al, 2016). Levels of the inhibitory amino acid GABA were the most abundant of the three NTs, and were similar among the regions (around 300 μg/g), except in the hypothalamus (520 μg/g).…”

Section: Determination Of Nts In Rat Brain Samplessupporting

confidence: 87%

“…For the calibration curve preparation of surrogateanalyte approach, 10 μL of mixed working solution at different concentration levels and 150 μL of cold 0.1% FA in ACN (with IS) were spiked into 40 μL brain homogenate. For the calibration curve preparation for the surrogate-matrix approach, Ringer's solution (148 mM NaCl, 2.7 mM KCl, 1.2 mM CaCl 2 and 0.85 mM MgCl 2 ) was used as surrogate matrix for the brain homogenate (Bergh et al, 2016). The final concentrations of DA, 5-HT, DA-d4 and 5-HT-d4 were 0.5, 1, 5, 25, 100 and 200 ng/mL, respectively, while the concentrations of GABA and GABA-d6 were 1, 5, 25, 100, 200 and 500 ng/mL, respectively.…”

Section: Preparation Of Calibration Standards and Quality Control Smentioning

confidence: 99%

“…In the surrogate-matrix approach, an authentic analyte is spiked in a neat solution/artificial matrix/stripped matrix devoid of endogenous analyte to construct a calibration curve (González, Fernández, Vidal, Frenich, & Pérez, 2011;Le, Ng, Kwan, Cusmano-Ozog, & Cowan, 2014). Further, the increased amount of deuterium in the isotope-labeled standards may negatively affect the accuracy of LC/MS analysis by increasing the retention time (Bergh et al, 2016). Moreover, a review reported that there are risks associated with a stripped-matrix approach because efficient removal of endogenous analytes is difficult and time consuming, and the oxidized products might be converted back to the analytes, which might affect the quantification of analytes in study samples (Thakare, Chhonker, Gautam, Alamoudi, & Alnouti, 2016;Zhang et al, 2012;Zhou et al, 2015).…”

mentioning

confidence: 99%

“…In our study, Ringer's solution was used as surrogate matrix, as it simulates the cerebrospinal fluid and contains components that reflect the physiological state of the central nervous system(Bergh et al, 2016;Thakare et al, 2016). (a) Chromatograms of blank Ringer's solution sample for authentic analytes and internal standard (IS).…”

mentioning

confidence: 99%

“…Moreover, it requires the correction of the responses between surrogate analytes and the corresponding authentic analytes before analysis. Further, the increased amount of deuterium in the isotope-labeled standards may negatively affect the accuracy of LC/MS analysis by increasing the retention time (Bergh et al, 2016). Both approaches are indirect means by which to quantify endogenous NTs, and each has its own advantages and limitations.…”

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confidence: 99%

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Development, validation and comparison of surrogate matrix and surrogate analyte approaches with UHPLC–MS/MS to simultaneously quantify dopamine, serotonin and γ‐aminobutyric acid in four rat brain regions

Liu

Xian

et al. 2018

Biomedical Chromatography

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As biomarkers, endogenous neurotransmitters play critical roles in the process of neuropsychiatric diseases, and neurotransmitter levels in different brain regions can contribute to neurological disease diagnosis and treatment. Due to the lack of a blank matrix for endogenous neurotransmitters, surrogate-matrix and surrogate-analyte approaches have been used for the determination of neurotransmitters to solve this problem. In this study, we capitalised on the high accuracy, precision, and throughput of UHPLC-MS/MS and developed new methods based on the two approaches. Both approaches satisfied FDA and EMA validation criterias after an appropriate parallelism assessment, and they were used to further quantify the three endogenous neurotransmitters, including dopamine (DA), serotonin (5-HT) and γ-aminobutyric acid (GABA) in rat brain four regions (cortex, striatum, hypothalamus and hippocampus) which represent the catecholamines, indolamines, and amino acids, respectively. Comparison of the results in the same rats (n = 10) showed there was no significant difference in DA, 5-HT, or GABA levels between the two approaches (P > 0.05). The concentrations of DA and GABA were highest in striatum and hypothalamus, respectively, and the levels of 5-HT were paralleled in striatum and hippocampus almost 2-fold higher than other regions. This is the first study to compare these two approaches in the determination of endogenous neurotransmitter content in the rat brain, and the surrogate-matrix approach proved to be simple, rapid, and reliable, considering cost, matrix similarity, and practicality.

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Section: Determination Of Nts In Rat Brain Samplessupporting

confidence: 87%

Section: Preparation Of Calibration Standards and Quality Control Smentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Development, validation and comparison of surrogate matrix and surrogate analyte approaches with UHPLC–MS/MS to simultaneously quantify dopamine, serotonin and γ‐aminobutyric acid in four rat brain regions

Liu

Xian

et al. 2018

Biomedical Chromatography

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Liquid chromatography–tandem mass spectrometry method for simultaneous quantification of neurotransmitters in rat brain tissue exposed to 4′‐fluoro‐α‐PHP

Viana

Pego

Oliveira

et al. 2022

Biomedical Chromatography

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The combination of different advanced analytical techniques makes it possible to determine the concentrations of neurotransmitters in various biological matrices, providing a complex and comprehensive study of the effects of psychoactive substances. The present study aimed to develop and validate a fast and simple analytical method for the determination of acetylcholine, serotonin, γ-aminobutyric acid, glutamate, dopamine and metabolites in rats brain tissue by liquid chromatography coupled to tandem mass spectrometry. The brain was homogenized and aliquots of the sample, dopamine-d 4 , and acetone were added to a tube and then vortexed and centrifuged. The supernatant was collected and dried. The residue was reconstituted and injected. The LLOQ ranged from 0.001 to 1 μg/g; the intra-run precision ranged from 0.47 to 11.52%; the inter-run precision ranged from 0.68 to 17.54%; and the bias ranged from 89.10 to 109.60%. As proof of concept, the method was applied to animals exposed to the synthetic cathinone 4 0 -fuoro-α-pyrrolidinohexanophenone (300 mg/kg). In addition, the workflow proved to be simple, rapid and useful to estimate the concentration of neurotransmitters. This analytical tool can be used to support the investigation of the changes in the neurochemical profile for the characterization of the mechanism of action of psychoactive substances, as well as both neurological and psychiatric diseases.

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Uncarialins J—M from Uncaria rhynchophylla and Their Anti‐depression Mechanism in Unpredictable Chronic Mild Stress‐Induced Mice via Activating 5‐HT_1A Receptor

Bai

Zhou

et al. 2021

Chin. J. Chem.

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Monoterpene indole alkaloids | Hirsuteine | Depression | 5-HT 1A receptor | Site-directed amino acid mutation Uncaria rhynchophylla has been widely used to treat central nervous system diseases for a long history. After investigation of U. rhynchophylla, eleven monoterpene indole alkaloids, including four new compounds uncarialins J-M (1-4) and seven known analogues (5-11), were isolated and identified. Their structural characterization was conducted using HRESIMS, 1D and 2D NMR, electronic circular dichroism (ECD) spectra, and quantum chemical computations. Compounds 1, 2, 7, and 9-11 displayed significant agonistic effects towards 5-HT 1A receptor, and their EC 50 values were 7.86, 7.32, 2.24, 1.18, 1.52, and 3.75 μmol/L, respectively. Furthermore, in vivo experimental results fully revealed that hirsuteine (7) displayed a significant antidepression effect in unpredictable chronic mild stress (UCMS)-induced depression mice mainly via regulating 5-HT 1A signaling pathway. Molecular docking and site-directed amino acid mutation verified that amino acid residues Asp116 and Asn386 were the binding sites of hirsuteine (7) with 5-HT 1A receptor. In addition, pre-treatment of mice with WAY 100635 also blocked the anti-depression effect of hirsuteine ( 7), which further demonstrated that 5-HT 1A receptor was a potential target of hirsuteine (7) to effectively treat depression. These findings indicated the therapeutic material basis of U. rhynchophylla and the anti-depression underlying mechanism of hirsuteine (7), and further provided the useful guidance for the development of hirsuteine (7) as a potential antidepressant candidate.

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Validated methods for determination of neurotransmitters and metabolites in rodent brain tissue and extracellular fluid by reversed phase UHPLC–MS/MS

Cited by 48 publications

References 28 publications

Development, validation and comparison of surrogate matrix and surrogate analyte approaches with UHPLC–MS/MS to simultaneously quantify dopamine, serotonin and γ‐aminobutyric acid in four rat brain regions

Development, validation and comparison of surrogate matrix and surrogate analyte approaches with UHPLC–MS/MS to simultaneously quantify dopamine, serotonin and γ‐aminobutyric acid in four rat brain regions

Liquid chromatography–tandem mass spectrometry method for simultaneous quantification of neurotransmitters in rat brain tissue exposed to 4′‐fluoro‐α‐PHP

Uncarialins J—M from Uncaria rhynchophylla and Their Anti‐depression Mechanism in Unpredictable Chronic Mild Stress‐Induced Mice via Activating 5‐HT_1A Receptor

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Validated methods for determination of neurotransmitters and metabolites in rodent brain tissue and extracellular fluid by reversed phase UHPLC–MS/MS

Cited by 48 publications

References 28 publications

Development, validation and comparison of surrogate matrix and surrogate analyte approaches with UHPLC–MS/MS to simultaneously quantify dopamine, serotonin and γ‐aminobutyric acid in four rat brain regions

Development, validation and comparison of surrogate matrix and surrogate analyte approaches with UHPLC–MS/MS to simultaneously quantify dopamine, serotonin and γ‐aminobutyric acid in four rat brain regions

Liquid chromatography–tandem mass spectrometry method for simultaneous quantification of neurotransmitters in rat brain tissue exposed to 4′‐fluoro‐α‐PHP

Uncarialins J—M from Uncaria rhynchophylla and Their Anti‐depression Mechanism in Unpredictable Chronic Mild Stress‐Induced Mice via Activating 5‐HT1A Receptor

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Uncarialins J—M from Uncaria rhynchophylla and Their Anti‐depression Mechanism in Unpredictable Chronic Mild Stress‐Induced Mice via Activating 5‐HT_1A Receptor