High-performance liquid chromatography/tandem mass spectrometry assay for the rapid high sensitivity measurement of basal acetylcholine from microdialysates

Hows, Mark E. P.; Organ, Andrew J.; Murray, S; Dawson, Lee A.; Foxton, Richard M.; Heidbreder, Christian; Hughes, Zoë A.; Lacroix, Laurent; Shah, Ajit J.

doi:10.1016/s0165-0270(02)00228-5

Cited by 72 publications

(94 citation statements)

References 19 publications

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“…The limit of detection of ACh achieved by using LC-MS 2 was comparable to the detection that can be achieved using LC-ECD methods. However, obvious advantages of LC-MS 2 assays for the detection of ACh can be summarized as follows: (i) there is no enzymatic reactions needed in order to separate choline from ACh; (ii) the assay provides a means of confirming the identity of the analyte using the specific mass transition together with its chromatographic retention time, and (iii) although neostigmine was used in the present study, we recently showed (Hows et al, 2002) that LC-MS/MS allows ACh to be measured in dialysates without the need to add neostigmine or physostigmine to the perfusate.…”

Section: Technical Notementioning

confidence: 96%

“…While the three monoamines, NE, DA, and 5-HT, have been detected simultaneously using LC with electrochemical detection (Heidbreder et al, 2001b), ACh was quantified by using a newly developed analytical method based on LC-MS 2 detection (Hows et al, 2002). In comparison with the most commonly used LC methods coupled with electrochemical detection, the LC-MS assay method used in the present study is very specific, minimizing the need to separate ACh from other components present in dialysates.…”

Section: Technical Notementioning

confidence: 99%

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Selective Antagonism at Dopamine D3 Receptors Enhances Monoaminergic and Cholinergic Neurotransmission in the Rat Anterior Cingulate Cortex

Lacroix

Hows

Shah

et al. 2002

Neuropsychopharmacol

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Recent neuroanatomical and functional investigations focusing on dopamine (DA) D 3 receptors have suggested a potential role of this receptor in psychiatric diseases such as schizophrenia and drug dependence. In line with the key role of the prefrontal cortex in psychiatric disorders, the present study aimed at assessing the effects of the acute systemic administration of the selective DA D 3 receptor antagonist SB-277011-A on the in vivo extracellular levels of monoamines (DA, norepinephrine (NE), and serotonin (5-HT)) and acetylcholine (ACh) in the anterior cingulate subregion of the medial prefrontal cortex. The in vivo neurochemical profile of SB-277011-A (10 mg/kg, i.p.) in the anterior cingulate cortex was compared with both typical and atypical antipsychotics including clozapine (10 mg/kg, s.c.), olanzapine (10 mg/kg, s.c.), sulpiride (10 mg/kg, s.c.), and haloperidol (0.5 mg/kg, s.c.). The acute administration of SB-277011-A, clozapine, and olanzapine produced a significant increase in extracellular levels of DA, NE, and ACh without affecting levels of 5-HT. Sulpiride also significantly increased extracellular DA, but with a delayed onset over SB-277011-A, clozapine, and olanzapine. In contrast, haloperidol failed to alter any of the three monoamines and ACh in the anterior cingulate cortex. These findings add to a growing body of evidence suggesting a differentiation between typical and atypical antipsychotic drugs (APDs) in the anterior cingulate cortex and a role of DA D 3 receptors in desired antipsychotic drug profile. Similar to their effects on DA and NE, SB-277011-A, clozapine, and olanzapine increased extracellular levels of ACh, whereas haloperidol and sulpiride did not alter ACh. The results obtained in the present study provide evidence of the important role of DA D 3 receptors in the effect of pharmacotherapeutic agents that are used for the treatment of psychiatric disorders such as schizophrenia and drug dependence.

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Section: Technical Notementioning

confidence: 96%

Section: Technical Notementioning

confidence: 99%

Selective Antagonism at Dopamine D3 Receptors Enhances Monoaminergic and Cholinergic Neurotransmission in the Rat Anterior Cingulate Cortex

Lacroix

Hows

Shah

et al. 2002

Neuropsychopharmacol

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“…The use of electrospray ionization (ESI)-MS is rapidly expanding into the study of enzyme kinetics [17][18][19][20][21][22][23]. Also, combinations of chromatographic and electrophoretic separation methods with ESI-MS, such as liquid chromatography (LC)/MS [24], LC/tandem mass spectroscopy (MS/MS) [25], gas chromatography (GC)/MS [26], frontal affinity chromatography (FAC)/MS [27], or capillary electrophoresis (CE)/MS [28] have been reported for the determination of acetylcholine or the screening of AChE inhibitors. These methods are quite specific but have longer cycle times.…”

mentioning

confidence: 99%

Monitoring enzyme reaction and screening of inhibitors of acetylcholinesterase by quantitative matrix-assisted laser desorption/ionization fourier transform mass spectrometry

Yao

Wei

et al. 2008

J. Am. Soc. Mass Spectrom.

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A matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI-FTMS)-based assay was developed for kinetic measurements and inhibitor screening of acetylcholinesterase. Here, FTMS coupled to MALDI was applied to quantitative analysis of choline using the ratio of choline/acetylcholine without the use of additional internal standard, which simplified the experiment. The Michaelis constant (K m ) of acetylcholinesterase (AChE) was determined to be 73.9 mol L Ϫ1 by this approach. For Huperzine A, the linear mixed inhibition of AChE reflected the presence of competitive and noncompetitive components. The half maximal inhibitory concentration (IC 50 ) value of galantamine obtained for AChE was 2.39 mol L Ϫ1 . Inhibitory potentials of Rhizoma Coptidis extracts were identified with the present method. In light of the results the referred extracts as a whole showed inhibitory action against AChE. The use of high-resolution FTMS largely eliminated the interference with the determination of ACh and Ch, produced by the low-mass compounds of chemical libraries for inhibitor screening. The excellent correlation with the reported kinetic parameters confirms that the MS-based assay is both accurate and precise for determining kinetic constants and for identifying enzyme inhibitors. The obvious advantages were demonstrated for quantitative analysis and also high-throughput characterization. This study offers a perspective into the utility of MALDI-FTMS as an alternate quantitative tool for inhibitor screening of

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“…NIH-PA Author Manuscript NIH-PA Author Manuscript [7][8][9]. Despite the successful applications of microdialysis sampling, the main limitations for sampling large neuropeptides are a combination of their low relative recoveries and low basal concentrations.…”

Section: Nih-pa Author Manuscriptmentioning

confidence: 99%

“…It is by far the most commonly used method for the collection of low molecular weight hydrophilic neurotransmitters, such as acetylcholine, dopamine, and glutamate from mammalian brain with high relative recovery [7][8][9]. Despite the successful applications of microdialysis sampling, the main limitations for sampling large neuropeptides are a combination of their low relative recoveries and low basal concentrations.…”

Section: Introductionmentioning

confidence: 99%

An in vitro comparison of microdialysis relative recovery of Met- and Leu-enkephalin using cyclodextrins and antibodies as affinity agents

Fletcher

Stenken

2008

Analytica Chimica Acta

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Cyclodextrins and antibodies have been used as affinity agents to improve relative recovery during microdialysis sampling. Two neuropeptides, methionine-enkephalin (ME) and leucine-enkephalin (LE), were chosen to compare the use of cyclodextrins and antibodies as possible affinity agents for improving their relative recovery across polycarbonate and polyethersulfone membranes during in vitro sampling. Cyclodextrins (CD) including β-CD, 2-hydroxypropyl-β-cyclodextrin (2HPβ-CD), and γ-CD gave improvements of relative recovery for both peptides of less than 2-fold as compared to controls. Comparisons of relative recovery between tyrosine-glycine-glycine, tyrosine, and phenylalanine using different cyclodextrins in the perfusion fluid were also obtained. Inclusion of an antibody against met-enkephalin in the microdialysis perfusion fluid resulted in relative recovery increases of up to 2.5-fold. These results show that using antibodies as affinity agents during microdialysis sampling may be more effective agents to improve the relative recovery of these opioid neuropeptides.Keywords microdialysis sampling; neuropeptides; affinity agents; cyclodextrins; antibodies IntroductionNeuropeptides are a class of neurotransmitters that are the most structurally diverse group of neuromodulators [1]. Enkephalins are endogenous opioid neuropeptides that have morphinelike action or pain relief in the body and are readily associated with the neurochemistry of addiction [2]. Enkephalins also play significant roles in mediating body temperature control, food intake, reward mechanisms, and stress [3].Microdialysis sampling has been used for over thirty years for chemical sampling applications in neuroscience, pharmacokinetics, and drug metabolism [4][5][6]. It is by far the most commonly used method for the collection of low molecular weight hydrophilic neurotransmitters, such as acetylcholine, dopamine, and glutamate from mammalian brain with high relative recovery Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access Author ManuscriptAnal Chim Acta. Author manuscript; available in PMC 2009 July 14. Published in final edited form as:Anal Chim Acta. 2008 July 14; 620(1-2): 170-175. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript [7][8][9]. Despite the successful applications of microdialysis sampling, the main limitations for sampling large neuropeptides are a combination of their low relative recoveries and low basal concentrations. A neuropeptide with a high molecular weight will exhibit a smaller aqueous diffusion coefficient, which directly aff...

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High-performance liquid chromatography/tandem mass spectrometry assay for the rapid high sensitivity measurement of basal acetylcholine from microdialysates

Cited by 72 publications

References 19 publications

Selective Antagonism at Dopamine D3 Receptors Enhances Monoaminergic and Cholinergic Neurotransmission in the Rat Anterior Cingulate Cortex

Selective Antagonism at Dopamine D3 Receptors Enhances Monoaminergic and Cholinergic Neurotransmission in the Rat Anterior Cingulate Cortex

Monitoring enzyme reaction and screening of inhibitors of acetylcholinesterase by quantitative matrix-assisted laser desorption/ionization fourier transform mass spectrometry

An in vitro comparison of microdialysis relative recovery of Met- and Leu-enkephalin using cyclodextrins and antibodies as affinity agents

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