Analysis of acetylcholine and choline in microdialysis samples by liquid chromatography/tandem mass spectrometry

Päivi Uutela, Ruut Reinilä, Petteri Piepponen, Raimo A Ketola, Risto Kostiainen, Raimo Ketola

    Research output: Contribution to journalArticleScientificpeer-review

    Abstract

    "A sensitive liquid chromatography/electrospray ionisation tandem mass spectrometric (LC/ESI-MS/ MS) method was developed for the analysis of acetylcholine and choline in microdialysis samples. A Ringer's solution that contains high (150 mM) concentrations of inorganic salts was used to extract acetylcholine and choline from a rat or mouse brain. The separation of acetylcholine, choline, an internal standard acetyl-p-methylcholine, endogenous compounds and inorganic cations was achieved with hydrophilic interaction chromatography using a diol column. The eluent consisted of 20 mM ammonium formate (pH 3.3) and acetonitrile (20:80) which is favourable for the ESI process. Limits of detection (signal-to-noise (S/N) ratio = 3) of 0.02 nM (0.2 fmol) for acetylcholine and 1 nM (10 fmol) for choline were observed using standards diluted in Ringer's solution. A good linearity was obtained from the limit of quantitation: 0.1 nM (S/N ratio =10) to 50 nM (r = 0.999) for acetylcholine and within the concentration range of 100-3500 nM (r = 0.998) for choline. The between-day repeatability of the method was good; RSD was 3.1% at 1 nM level of acetylcholine and 3.5% at 1000 nM level of choline. The recoveries for addition of 1 or 2.5 nM acetylcholine and 0.2 or 1 mu M choline in microdialysis balancing samples were between 93 and 101% indicating that no suppressing endogenous compounds were co-eluting with acetylcholine or choline. The developed method was applied to the analysis of microdialysis balancing samples collected from rat and mouse brains. Copyright (C)2005 John Wiley & Sons, Ltd."
    Original languageEnglish
    JournalRapid Communications in Mass Spectrometry
    Volume19
    Issue number20
    Pages (from-to)2950-2956
    Number of pages7
    ISSN0951-4198
    DOIs
    Publication statusPublished - 2005
    MoE publication typeA1 Journal article-refereed

    Fields of Science

    • asetyylikoliini
    • koliini

    Cite this

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    title = "Analysis of acetylcholine and choline in microdialysis samples by liquid chromatography/tandem mass spectrometry",
    abstract = "{"}A sensitive liquid chromatography/electrospray ionisation tandem mass spectrometric (LC/ESI-MS/ MS) method was developed for the analysis of acetylcholine and choline in microdialysis samples. A Ringer's solution that contains high (150 mM) concentrations of inorganic salts was used to extract acetylcholine and choline from a rat or mouse brain. The separation of acetylcholine, choline, an internal standard acetyl-p-methylcholine, endogenous compounds and inorganic cations was achieved with hydrophilic interaction chromatography using a diol column. The eluent consisted of 20 mM ammonium formate (pH 3.3) and acetonitrile (20:80) which is favourable for the ESI process. Limits of detection (signal-to-noise (S/N) ratio = 3) of 0.02 nM (0.2 fmol) for acetylcholine and 1 nM (10 fmol) for choline were observed using standards diluted in Ringer's solution. A good linearity was obtained from the limit of quantitation: 0.1 nM (S/N ratio =10) to 50 nM (r = 0.999) for acetylcholine and within the concentration range of 100-3500 nM (r = 0.998) for choline. The between-day repeatability of the method was good; RSD was 3.1{\%} at 1 nM level of acetylcholine and 3.5{\%} at 1000 nM level of choline. The recoveries for addition of 1 or 2.5 nM acetylcholine and 0.2 or 1 mu M choline in microdialysis balancing samples were between 93 and 101{\%} indicating that no suppressing endogenous compounds were co-eluting with acetylcholine or choline. The developed method was applied to the analysis of microdialysis balancing samples collected from rat and mouse brains. Copyright (C)2005 John Wiley & Sons, Ltd.{"}",
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    author = "P{\"a}ivi Uutela and Ruut Reinil{\"a} and Petteri Piepponen and Ketola, {Raimo A} and Risto Kostiainen and Raimo Ketola",
    year = "2005",
    doi = "10.1002/rcm.2160",
    language = "English",
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    journal = "Rapid Communications in Mass Spectrometry",
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    Analysis of acetylcholine and choline in microdialysis samples by liquid chromatography/tandem mass spectrometry. / Uutela, Päivi; Reinilä, Ruut; Piepponen, Petteri; Ketola, Raimo A; Kostiainen, Risto; Ketola, Raimo.

    In: Rapid Communications in Mass Spectrometry, Vol. 19, No. 20, 2005, p. 2950-2956.

    Research output: Contribution to journalArticleScientificpeer-review

    TY - JOUR

    T1 - Analysis of acetylcholine and choline in microdialysis samples by liquid chromatography/tandem mass spectrometry

    AU - Uutela, Päivi

    AU - Reinilä, Ruut

    AU - Piepponen, Petteri

    AU - Ketola, Raimo A

    AU - Kostiainen, Risto

    AU - Ketola, Raimo

    PY - 2005

    Y1 - 2005

    N2 - "A sensitive liquid chromatography/electrospray ionisation tandem mass spectrometric (LC/ESI-MS/ MS) method was developed for the analysis of acetylcholine and choline in microdialysis samples. A Ringer's solution that contains high (150 mM) concentrations of inorganic salts was used to extract acetylcholine and choline from a rat or mouse brain. The separation of acetylcholine, choline, an internal standard acetyl-p-methylcholine, endogenous compounds and inorganic cations was achieved with hydrophilic interaction chromatography using a diol column. The eluent consisted of 20 mM ammonium formate (pH 3.3) and acetonitrile (20:80) which is favourable for the ESI process. Limits of detection (signal-to-noise (S/N) ratio = 3) of 0.02 nM (0.2 fmol) for acetylcholine and 1 nM (10 fmol) for choline were observed using standards diluted in Ringer's solution. A good linearity was obtained from the limit of quantitation: 0.1 nM (S/N ratio =10) to 50 nM (r = 0.999) for acetylcholine and within the concentration range of 100-3500 nM (r = 0.998) for choline. The between-day repeatability of the method was good; RSD was 3.1% at 1 nM level of acetylcholine and 3.5% at 1000 nM level of choline. The recoveries for addition of 1 or 2.5 nM acetylcholine and 0.2 or 1 mu M choline in microdialysis balancing samples were between 93 and 101% indicating that no suppressing endogenous compounds were co-eluting with acetylcholine or choline. The developed method was applied to the analysis of microdialysis balancing samples collected from rat and mouse brains. Copyright (C)2005 John Wiley & Sons, Ltd."

    AB - "A sensitive liquid chromatography/electrospray ionisation tandem mass spectrometric (LC/ESI-MS/ MS) method was developed for the analysis of acetylcholine and choline in microdialysis samples. A Ringer's solution that contains high (150 mM) concentrations of inorganic salts was used to extract acetylcholine and choline from a rat or mouse brain. The separation of acetylcholine, choline, an internal standard acetyl-p-methylcholine, endogenous compounds and inorganic cations was achieved with hydrophilic interaction chromatography using a diol column. The eluent consisted of 20 mM ammonium formate (pH 3.3) and acetonitrile (20:80) which is favourable for the ESI process. Limits of detection (signal-to-noise (S/N) ratio = 3) of 0.02 nM (0.2 fmol) for acetylcholine and 1 nM (10 fmol) for choline were observed using standards diluted in Ringer's solution. A good linearity was obtained from the limit of quantitation: 0.1 nM (S/N ratio =10) to 50 nM (r = 0.999) for acetylcholine and within the concentration range of 100-3500 nM (r = 0.998) for choline. The between-day repeatability of the method was good; RSD was 3.1% at 1 nM level of acetylcholine and 3.5% at 1000 nM level of choline. The recoveries for addition of 1 or 2.5 nM acetylcholine and 0.2 or 1 mu M choline in microdialysis balancing samples were between 93 and 101% indicating that no suppressing endogenous compounds were co-eluting with acetylcholine or choline. The developed method was applied to the analysis of microdialysis balancing samples collected from rat and mouse brains. Copyright (C)2005 John Wiley & Sons, Ltd."

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    KW - koliini

    KW - asetyylikoliini

    KW - koliini

    KW - asetyylikoliini

    KW - koliini

    U2 - 10.1002/rcm.2160

    DO - 10.1002/rcm.2160

    M3 - Article

    VL - 19

    SP - 2950

    EP - 2956

    JO - Rapid Communications in Mass Spectrometry

    JF - Rapid Communications in Mass Spectrometry

    SN - 0951-4198

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    ER -