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
doi:10.1258/acb.2009.008191
© 2010 Association for Clinical Biochemistry

 

This Article
Right arrow
Figures Only
Right arrow
Full Text
Right arrow

Full Text (PDF)

Right arrow
All Versions of this Article:

acb.2009.008191v1

47/1/56

most recent

Right arrow
Alert me when this article is cited
Right arrow
Alert me if a correction is posted
Services
Right arrow
Email this article to a friend
Right arrow

Similar articles in this journal

Right arrow
Similar articles in PubMed
Right arrow
Alert me to new issues of the journal
Right arrow
Download to citation manager
Right arrow
Google Scholar
Right arrow
Articles by Griffith, C A
Right arrow
Articles by Keevil, B G
PubMed
Right arrow
PubMed Citation
Social Bookmarking

What’s this?

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}nhs.net


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.


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What’s this?