Development and validation of a sample stabilization strategy and a UPLC–MS/MS method for the simultaneous quantitation of acetylcholine (ACh), histamine (HA), and its metabolites in rat cerebrospinal fluid (CSF)

A UPLC–MS/MS assay was developed and validated for simultaneous quantification of acetylcholine (ACh), histamine (HA), tele-methylhistamine (t-mHA), and tele-methylimidazolacetic acid (t-MIAA) in rat cerebrospinal fluid (CSF). The biological stability of ACh in rat CSF was investigated. Following fi...

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Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2011-07, Vol.879 (22), p.2023-2033
Main Authors: Zhang, Yanhua, Tingley, F. David, Tseng, Elaine, Tella, Max, Yang, Xin, Groeber, Elizabeth, Liu, Jianhua, Li, Wenlin, Schmidt, Christopher J., Steenwyk, Rick
Format: Article
Language:English
Subjects:
Acetylcholine
Acetylcholine - cerebrospinal fluid
Acetylcholine - metabolism
Acetylcholine - pharmacokinetics
Acetylcholinesterase
Analysis
Analytical, structural and metabolic biochemistry
Animals
Benzofurans - cerebrospinal fluid
Benzofurans - chemistry
Benzofurans - pharmacology
Biological and medical sciences
Biological stability
Biomarker
biomarkers
Cerebrospinal fluid
Cholinesterase Inhibitors - pharmacology
Chromatography, High Pressure Liquid - methods
derivatization
Donepezil
Drug discovery
Drug Stability
Fundamental and applied biological sciences. Psychology
General pharmacology
half life
High-performance liquid chromatography
Histamine
Histamine - cerebrospinal fluid
Histamine - metabolism
Histamine - pharmacokinetics
hydrophilic interaction chromatography
Hydrophobic and Hydrophilic Interactions
Imidazoles - cerebrospinal fluid
Imidazoles - pharmacokinetics
Indans - pharmacology
isotope labeling
Male
Medical sciences
Metabolites
Methylhistamines - cerebrospinal fluid
Methylhistamines - pharmacokinetics
Neurotransmitters
Pharmacology. Drug treatments
Piperidines - pharmacology
Prucalopride
Quantitation
quantitative analysis
Rat cerebrospinal fluid
Rats
Rats, Sprague-Dawley
Reproducibility of Results
Sensitivity and Specificity
Tandem Mass Spectrometry - methods
tele-Methylhistamine
tele-Methylimidazolacetic acid
UPLC–MS/MS
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Summary:A UPLC–MS/MS assay was developed and validated for simultaneous quantification of acetylcholine (ACh), histamine (HA), tele-methylhistamine (t-mHA), and tele-methylimidazolacetic acid (t-MIAA) in rat cerebrospinal fluid (CSF). The biological stability of ACh in rat CSF was investigated. Following fit-for-purpose validation, the method was applied to monitor the drug-induced changes in ACh, HA, t-mHA, and t-MIAA in rat CSF following administration of donepezil or prucalopride. The quantitative method utilizes hydrophilic interaction chromatography (HILIC) Core–Shell HPLC column technology and a UPLC system to achieve separation with detection by positive ESI LC–MS/MS. This UPLC–MS/MS method does not require extraction or derivatization, utilizes a stable isotopically labeled internal standard (IS) for each analyte, and allows for rapid throughput with a 4 min run time. Without an acetylcholinesterase (AChE) inhibitor present, ACh was found to have 1.9 ± 0.4 min in vitro half life in rat CSF. Stability studies and processing modification, including the use of AChE inhibitor eserine, extended this half life to more than 60 min. The UPLC–MS/MS method, including stabilization procedure, was validated over a linear concentration range of 0.025–5 ng/mL for ACh and 0.05–10 ng/mL for HA, t-mHA, and t-MIAA. The intra-run precision and accuracy for all analytes were 1.9–12.3% CV and −10.2 to 9.4% RE, respectively, while inter-run precision and accuracy were 4.0–16.0% CV and −5.3 to 13.4% RE, respectively. By using this developed and validated method, donepezil caused increases in ACh levels at 0.5, 1, 2, and 4 h post dose as compared to the corresponding vehicle group, while prucalopride produced approximately 1.6- and 3.1-fold increases in the concentrations of ACh and t-mHA at 1 h post dose, respectively, compared to the vehicle control. Overall, this methodology enables investigations into the use of CSF ACh and HA as biomarkers in the study of these neurotransmitter systems and related drug discovery efforts.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2011.05.030