title: Quantitative analysis of adenosine using liquid chromatography/atmospheric pressure chemical ionization-tandem mass spectrometry (LC/APCI-MS/MS).
abstract: Adenosine-secreting cellular brain implants constitute a promising therapeutic approach for the treatment of epilepsy. To engineer neural stem cells for therapeutic adenosine delivery, a reliable and fast analytical method is necessary to quantify cell-based adenosine release. Here we describe the development, optimization and validation of adenosine measurement using liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry (LC-APCI-MS/MS). LC-MS/MS in positive ion mode used selected reaction monitoring at m/z of 268.2/136.1 and 302.2/170.0 for adenosine and the internal standard, respectively. The bias was within 15% of the nominal value and evaluation of precision showed a relative standard deviation lower than 15% for all measured concentrations. The lower limit of quantification of adenosine was 15.6 ng/ml. Freeze and thaw stability and processed sample stability also fulfilled the acceptance criteria. Evaluation of the matrix effect showed that the method is not affected by relative matrix effects. The major advantages of this method are the absence of an extraction phase and the combination of the high selectivity and sensitivity characteristic for the LC-MS/MS technique, with a short run time of 4.5 min. These results demonstrate that this method is a useful tool to measure adenosine concentrations in culture medium released from stem cells in vitro.
authors.affiliation: Laboratory for Clinical and Experimental Neurophysiology, Department of Neurology, Ghent University Hospital, Ghent, Belgium. Annelies.VanDycke@UGent.be
authors.forename: Annelies
authors.initials: A
authors.lastname: Van Dycke
authors.forename: Alain
authors.initials: A
authors.lastname: Verstraete
authors.forename: Kristof
authors.initials: K
authors.lastname: Pil
authors.forename: Robrecht
authors.initials: R
authors.lastname: Raedt
authors.forename: Kristl
authors.initials: K
authors.lastname: Vonck
authors.forename: Detlev
authors.initials: D
authors.lastname: Boison
authors.forename: Paul
authors.initials: P
authors.lastname: Boon
mesh_terms: D015762:2-Chloroadenosine / Q000032:analysis; D000241:Adenosine / Q000032:analysis* / Q000378:metabolism; D002478:Cells, Cultured; D002853:Chromatography, Liquid / Q000379:methods*; D003470:Culture Media / Q000737:chemistry* / Q000378:metabolism; D004355:Drug Stability; D006801:Humans; D009474:Neurons / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013234:Stem Cells / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D003470:Culture Media; D015762:2-Chloroadenosine; D000241:Adenosine
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