Quantitative analysis of adenosine using liquid chromatography/atmospheric pressure chemical ionization-tandem mass spectrometry (LC/APCI-MS/MS).
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.
Van Dycke
Annelies
A
Laboratory for Clinical and Experimental Neurophysiology, Department of Neurology, Ghent University Hospital, Ghent, Belgium. Annelies.VanDycke@UGent.be
Verstraete
Alain
A
Pil
Kristof
K
Raedt
Robrecht
R
Vonck
Kristl
K
Boison
Detlev
D
Boon
Paul
P
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*
D003470:Culture Media; D015762:2-Chloroadenosine; D000241:Adenosine
J Pain Palliat Care Pharmacother. 2015 Jun;29(2):133-9
26095483
J Chromatogr B Biomed Sci Appl. 1998 Nov 20;719(1-2):55-61
9869364
J Pharm Biomed Anal. 2004 Nov 19;36(4):877-82
15533683
J Chromatogr A. 1997 Feb 28;763(1-2):193-7
9129323
Zhongguo Zhong Yao Za Zhi. 2007 Oct;32(19):2018-21
18161295
J Pharm Biomed Anal. 2001 Mar;24(5-6):1143-6
11248511
Rheumatology (Oxford). 2005 Jan;44(1):74-9
15454631
Eur J Pharmacol. 1994 Mar 21;254(3):277-82
8013563
J Chromatogr A. 1998 Dec 18;828(1-2):489-96
9916327
J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Jan 18;830(2):293-300
16310419
Adv Neurol. 1999;79:1001-10
10514882
Biochem Biophys Res Commun. 2005 Jul 22;333(1):88-94
15935989
Epilepsy Res. 2009 Aug;85(2-3):131-41
19428218
Forensic Sci Int. 2007 Jan 17;165(2-3):216-24
16781833
J Pharm Biomed Anal. 2001 Mar;24(5-6):1131-6
11248509
J Chromatogr. 1990 Aug 3;529(2):437-41
2172267
Rapid Commun Mass Spectrom. 2004;18(22):2730-8
15499664
J Chromatogr A. 2004 Nov 26;1058(1-2):61-6
15595652
J Chromatogr B Biomed Sci Appl. 1999 Mar 19;724(2):231-8
10219663
J Neurochem. 1982 Jul;39(1):280-2
6806443
Brain Res Brain Res Protoc. 2003 Feb;10(3):168-74
12565687
J Pharm Biomed Anal. 2001 Dec;26(5-6):967-73
11600309
J Neurochem. 1987 Jul;49(1):227-31
3585332
Int Rev Neurobiol. 2005;63:191-270
15797469
Exp Neurol. 1999 Nov;160(1):164-74
10630201
Anal Chem. 2003 Jul 1;75(13):3019-30
12964746
Curr Opin Pharmacol. 2008 Feb;8(1):2-7
17942368
J Pharmacol Exp Ther. 2008 Jun;325(3):920-6
18322147
J Neurochem. 2004 Mar;88(5):1305-12
15009686
Neurodegener Dis. 2007;4(1):28-33
17429216
Anal Biochem. 1996 Jun 15;238(1):34-9
8660582
J Chromatogr. 1991 Jan 4;536(1-2):265-72
2050767