Development of a method to measure plasma and whole blood choline by liquid chromatography tandem mass spectrometry

This version was published on 1 January 2010

Ann Clin Biochem 2010;47:56-61
© 2010 Association for Clinical Biochemistry


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Original Articles

C A Griffith1,
L J Owen1,
R Body2,
G McDowell2 and
B G Keevil1

1 University Hospital of South Manchester, Southmoor Road, Wythenshawe, Manchester M23 9LT;
2 Manchester Royal Infirmary, Oxford Road, Manchester M13 9WL, UK

Corresponding author: C A Griffith, Department of Clinical Biochemistry, University Hospital of South Manchester, Southmoor Road, Wythenshawe, Manchester M23 9LT, UK. Email: caroline.griffith{at}

Background: Current gold standard markers for myocardial damage are troponinsI and T, which are both sensitive and specific for the detectionof myocardial infarction, but require up to 6 h to become reliablyelevated in serum. Investigation into markers with potentialto identify patients with early ischaemic changes is thereforeintense. Choline is reported to be prognostic in patients presentingwith acute coronary syndromes via its release from ischaemiccell membranes.

Methods: Liquid chromatography tandem mass spectrometry was used to developa method to quantitate choline in plasma and blood. The methodinvolves addition of a deuterated internal standard to an aliquotof plasma or blood followed by organic solvent addition, whichprecipitates the proteins in the sample. Preparation was carriedout directly into a 96-deep-well plate. Chromatography of cholineused a strong cation exchange column and separation used a WatersAtlantis dC18 analytical column positioned directly before themass spectrometer source, allowing on-line preanalytical cleanup of the sample.

Results: The lower limit of quantitation was 0.38 µmol/L, linearitywas observed up to 754 µmol/L, with a working concentrationrange of 0.38–224 µmol/L, inter- and intra-assaycoefficients of variation were <6% and <4%, respectively.Samples were stable throughout five freeze–thaw cyclesand recovery was between 94% and 114%.

Conclusions: The assay was successfully validated in accordance with FDAguidelines and is suitable for quantitation of choline in researchand clinical settings.

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