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Articles |
1
Metabolic Unit, Department of Clinical Chemistry, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
a Author for correspondence. Fax 31-20-4440305; e-mail C.Jakobs{at}AZVU.NL
Background: Available methods for the determination of nanomolar concentrations of S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) in plasma and cerebrospinal fluid (CSF) are time-consuming. We wished to develop a method for their rapid and simultaneous measurement.
Methods: We used tandem mass spectrometry (MS/MS) for the simultaneous determination of SAM and SAH, with stable-isotope-labeled internal standards. The 13C5-SAH internal standard was enzymatically prepared using SAH-hydrolase and [13C5]adenosine. The method comprises a weak anion-exchange solid-phase extraction procedure serving as clean-up step for the deproteinized plasma and CSF samples. After clean-up, samples were injected on a C18 HPLC column, which was connected directly to the tandem mass spectrometer, operating in MS/MS mode.
Results: In plasma samples, the intraassay CVs for SAM and SAH were 4.2% and 4.0%, respectively, and the interassay CVs were 7.6% and 5.9%, respectively. In CSF, the intraassay CVs for SAM and SAH were 6.8% and 6.9%, respectively, and the interassay CVs were 4.2% and 5.5%, respectively. Mean recovery of SAM and SAH for both matrices at two concentrations was 93%. Detection limits for SAM and SAH in samples were 7.5 and 2.5 nmol/L, respectively. Concentrations of SAM and SAH in plasma from healthy subjects were within the previously reported ranges. In 10 CSF samples, the mean concentrations (range) were 248 (137–385) nmol/L for SAM and 11.3 (8.9–14.1) nmol/L for SAH.
Conclusions: SAM and SAH can be analyzed by MS/MS, taking optimal advantage of the speed and high sensitivity and specificity of this relatively new analytical technique.
The following articles in journals at HighWire Press have cited this article:
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 R. G. V. Smolders, K. de Meer, P. Kenemans, C. Jakobs, W. Kulik, and M. J. van der Mooren Oral Estradiol Decreases Plasma Homocysteine, Vitamin B6, and Albumin in Postmenopausal Women But Does Not Change the Whole-Body Homocysteine Remethylation and Transmethylation Flux J. Clin. Endocrinol. Metab., April 1, 2005; 90(4): 2218 - 2224. [Abstract] [Full Text] [PDF]
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 L. Anderson Candidate-based proteomics in the search for biomarkers of cardiovascular disease J. Physiol., February 15, 2005; 563(1): 23 - 60. [Abstract] [Full Text] [PDF]
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 N. P. Riksen, G. A. Rongen, G. H.J. Boers, H. J. Blom, P. H.H. van den Broek, and P. Smits Enhanced Cellular Adenosine Uptake Limits Adenosine Receptor Stimulation in Patients With Hyperhomocysteinemia Arterioscler Thromb Vasc Biol, January 1, 2005; 25(1): 109 - 114. [Abstract] [Full Text] [PDF]
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 F. Stam, C. van Guldener, P. M. ter Wee, W. Kulik, D. E. C. Smith, C. Jakobs, C. D. A. Stehouwer, and K. de Meer Homocysteine clearance and methylation flux rates in health and end-stage renal disease: association with S-adenosylhomocysteine Am J Physiol Renal Physiol, August 1, 2004; 287(2): F215 - F223. [Abstract] [Full Text] [PDF]
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S. P. Stabler and R. H. Allen Quantification of Serum and Urinary S-Adenosylmethionine and S-Adenosylhomocysteine by Stable-Isotope-Dilution Liquid Chromatography-Mass Spectrometry Clin. Chem., February 1, 2004; 50(2): 365 - 372. [Abstract] [Full Text] [PDF]
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R. Castro, I. Rivera, E. A. Struys, E. E.W. Jansen, P. Ravasco, M. E. Camilo, H. J. Blom, C. Jakobs, and I. Tavares de Almeida Increased Homocysteine and S-Adenosylhomocysteine Concentrations and DNA Hypomethylation in Vascular Disease Clin. Chem., August 1, 2003; 49(8): 1292 - 1296. [Abstract] [Full Text] [PDF]
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 K. Herbig, E.-P. Chiang, L.-R. Lee, J. Hills, B. Shane, and P. J. Stover Cytoplasmic Serine Hydroxymethyltransferase Mediates Competition between Folate-dependent Deoxyribonucleotide and S-Adenosylmethionine Biosyntheses J. Biol. Chem., October 4, 2002; 277(41): 38381 - 38389. [Abstract] [Full Text] [PDF]
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