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Clinical Chemistry, Vol 39, 263-271, Copyright © 1993 by American Association for Clinical Chemistry
T Fiskerstrand, H Refsum, G Kvalheim and PM Ueland
Department of Pharmacology and Toxicology, University of Bergen, Norway.
We have developed a modified version of our fully automated column- switching HPLC method for determining total plasma homocysteine based on single-column (reversed-phase) separation. Homocysteine, cysteine, and cysteinylglycine in plasma (total concentrations), acid- precipitated plasma (non-protein-bound concentrations), and urine can be determined. The derivatization and chromatography were performed automatically by a sample processor. The successful separation of all thiol species (within 15 min) was accomplished by accurate adjustment of the pH of the mobile phase to 3.65 (plasma) or 3.50 (acid- precipitated plasma, urine). Maximal fluorescence yield of cysteine, cysteinylglycine, and, to a lesser degree, homocysteine was dependent on optimal concentrations of EDTA and dithioerythritol during reduction (with NaBH4) and derivatization (with monobromobimane). The method is sensitive (detection limit approximately 0.05 pmol) and has a high degree of precision (CV < 5%). The sample output is approximately 70 samples in 24 h. Serum and heparin plasma can also be analyzed. Hemolysis up to approximately 2.0 g/L of hemoglobin did not interfere with the analytical recovery of homocysteine or cysteine. Collection of blood, separation of plasma from whole blood, and acid precipitation must be standardized to obtain reproducible thiol results. Our modifications and the standardization of blood-sampling procedures have substantially improved the method and broadened its applications.
The following articles in journals at HighWire Press have cited this article:
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  A. D. Ricart, J. D. Berlin, K. P. Papadopoulos, S. Syed, D. W. Drolet, C. Quaratino-Baker, J. Horan, J. Chick, W. Vermeulen, A. W. Tolcher, et al. Phase I, Pharmacokinetic and Biological Correlative Study of OSI-7904L, a Novel Liposomal Thymidylate Synthase Inhibitor, and Cisplatin in Patients with Solid Tumors Clin. Cancer Res., December 1, 2008; 14(23): 7947 - 7955. [Abstract] [Full Text] [PDF]
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 A. K Elshorbagy, E. Nurk, C. G. Gjesdal, G. S Tell, P. M Ueland, O. Nygard, A. Tverdal, S. E Vollset, and H. Refsum Homocysteine, cysteine, and body composition in the Hordaland Homocysteine Study: does cysteine link amino acid and lipid metabolism? Am. J. Clinical Nutrition, September 1, 2008; 88(3): 738 - 746. [Abstract] [Full Text] [PDF]
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 A. Ulvik, S. E. Vollset, G. Hoff, and P. M. Ueland Coffee Consumption and Circulating B-Vitamins in Healthy Middle-Aged Men and Women Clin. Chem., September 1, 2008; 54(9): 1489 - 1496. [Abstract] [Full Text] [PDF]
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P. Detopoulou, D. B Panagiotakos, S. Antonopoulou, C. Pitsavos, and C. Stefanadis Dietary choline and betaine intakes in relation to concentrations of inflammatory markers in healthy adults: the ATTICA study Am. J. Clinical Nutrition, February 1, 2008; 87(2): 424 - 430. [Abstract] [Full Text] [PDF]
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E. Nurk, C. A Drevon, H. Refsum, K. Solvoll, S. E Vollset, O. Nygard, H. A Nygaard, K. Engedal, G. S Tell, and A D. Smith Cognitive performance among the elderly and dietary fish intake: the Hordaland Health Study Am. J. Clinical Nutrition, November 1, 2007; 86(5): 1470 - 1478. [Abstract] [Full Text] [PDF]
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C. H Halsted, D. H Wong, J. M Peerson, C. H Warden, H. Refsum, A D. Smith, O. K Nygard, P. M Ueland, S. E Vollset, and G. S Tell Relations of glutamate carboxypeptidase II (GCPII) polymorphisms to folate and homocysteine concentrations and to scores of cognition, anxiety, and depression in a homogeneous Norwegian population: the Hordaland Homocysteine Study Am. J. Clinical Nutrition, August 1, 2007; 86(2): 514 - 521. [Abstract] [Full Text] [PDF]
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P. Berstad, S. V Konstantinova, H. Refsum, E. Nurk, S. E. Vollset, G. S Tell, P. M Ueland, C. A Drevon, and G. Ursin Dietary fat and plasma total homocysteine concentrations in 2 adult age groups: the Hordaland Homocysteine Study Am. J. Clinical Nutrition, June 1, 2007; 85(6): 1598 - 1605. [Abstract] [Full Text] [PDF]
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 M. den Heijer, H. P. J. Willems, H. J. Blom, W. B. J. Gerrits, M. Cattaneo, S. Eichinger, F. R. Rosendaal, and G. M. J. Bos Homocysteine lowering by B vitamins and the secondary prevention of deep vein thrombosis and pulmonary embolism: a randomized, placebo-controlled, double-blind trial Blood, January 1, 2007; 109(1): 139 - 144. [Abstract] [Full Text] [PDF]
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 E. Nurk, G.S. Tell, H. Refsum, P.M. Ueland, and S.E. Vollset Factor V Leiden, pregnancy complications and adverse outcomes: the Hordaland Homocysteine Study QJM, May 1, 2006; 99(5): 289 - 298. [Abstract] [Full Text] [PDF]
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 C. G. Gjesdal, S. E. Vollset, P. M. Ueland, H. Refsum, C. A. Drevon, H. K. Gjessing, and G. S. Tell Plasma Total Homocysteine Level and Bone Mineral Density: The Hordaland Homocysteine Study Arch Intern Med, January 9, 2006; 166(1): 88 - 94. [Abstract] [Full Text] [PDF]
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A. Windelberg, O. Arseth, G. Kvalheim, and P. M. Ueland Automated Assay for the Determination of Methylmalonic Acid, Total Homocysteine, and Related Amino Acids in Human Serum or Plasma by Means of Methylchloroformate Derivatization and Gas Chromatography-Mass Spectrometry Clin. Chem., November 1, 2005; 51(11): 2103 - 2109. [Abstract] [Full Text] [PDF]
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G. Beutel, H. Glen, P. Schoffski, J. Chick, S. Gill, J. Cassidy, and C. Twelves Phase I Study of OSI-7904L, a Novel Liposomal Thymidylate Synthase Inhibitor in Patients with Refractory Solid Tumors Clin. Cancer Res., August 1, 2005; 11(15): 5487 - 5495. [Abstract] [Full Text] [PDF]
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I. M van Beynum, M. den Heijer, C. M. Thomas, L. Afman, D. Oppenraay-van Emmerzaal, and H. J Blom Total homocysteine and its predictors in Dutch children Am. J. Clinical Nutrition, May 1, 2005; 81(5): 1110 - 1116. [Abstract] [Full Text] [PDF]
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 E.-C. Chan, P.-Y. Chang, T.-L. Wu, and J. T. Wu Enzymatic Assay of Homocysteine on Microtiter Plates or a TECAN Analyzer Using Crude Lysate Containing Recombinant Methionine {gamma}-Lyase Ann. Clin. Lab. Sci., April 1, 2005; 35(2): 155 - 160. [Abstract] [Full Text] [PDF]
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A. Brevik, S. E. Vollset, G. S Tell, H. Refsum, P. M. Ueland, E. B. Loeken, C. A Drevon, and L. F. Andersen Plasma concentration of folate as a biomarker for the intake of fruit and vegetables: the Hordaland Homocysteine Study Am. J. Clinical Nutrition, February 1, 2005; 81(2): 434 - 439. [Abstract] [Full Text] [PDF]
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 P. I. Holm, P. M. Ueland, S. E. Vollset, O. Midttun, H. J. Blom, M. B.A.J. Keijzer, and M. den Heijer Betaine and Folate Status as Cooperative Determinants of Plasma Homocysteine in Humans Arterioscler. Thromb. Vasc. Biol., February 1, 2005; 25(2): 379 - 385. [Abstract] [Full Text] [PDF]
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A. Bowron, A. Barton, J. Scott, and D. Stansbie Blood Spot Homocysteine: A Feasibility and Stability Study Clin. Chem., January 1, 2005; 51(1): 257 - 258. [Full Text] [PDF]
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 H A W Neil, V Seagroatt, D J Betteridge, M P Cooper, P N Durrington, J P Miller, M Seed, R P Naoumova, G R Thompson, R Huxley, et al. Established and emerging coronary risk factors in patients with heterozygous familial hypercholesterolaemia Heart, December 1, 2004; 90(12): 1431 - 1437. [Abstract] [Full Text] [PDF]
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G. V Dedoussis, D. B Panagiotakos, C. Chrysohoou, C. Pitsavos, A. Zampelas, D. Choumerianou, and C. Stefanadis Effect of interaction between adherence to a Mediterranean diet and the methylenetetrahydrofolate reductase 677C->T mutation on homocysteine concentrations in healthy adults: the ATTICA Study Am. J. Clinical Nutrition, October 1, 2004; 80(4): 849 - 854. [Abstract] [Full Text] [PDF]
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S. Hustad, B. G Nedrebo, P. M. Ueland, J. Schneede, S. E. Vollset, A. Ulvik, and E. A Lien Phenotypic expression of the methylenetetrahydrofolate reductase 677C->T polymorphism and flavin cofactor availability in thyroid dysfunction Am. J. Clinical Nutrition, October 1, 2004; 80(4): 1050 - 1057. [Abstract] [Full Text] [PDF]
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H. P.J. Willems, M. den Heijer, J. Lindemans, H. W.A. Berenschot, W. B.J. Gerrits, G. M.J. Bos, and H. J. Blom Measurement of Total Homocysteine Concentrations in Acidic Citrate- and EDTA-Containing Tubes by Different Methods Clin. Chem., October 1, 2004; 50(10): 1881 - 1883. [Full Text] [PDF]
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Q. Han, X. Sun, M. Xu, N. Zhang, L. Tang, Y. Tan, and R. M. Hoffman 3-Deazaadenosine, a Stabilizer of Whole-Blood Homocysteine Content, Does Not Interfere with the Single-Enzyme Homocysteine Assay while Totally Inhibiting the Enzyme Conversion Homocysteine Immunoassay Clin. Chem., September 1, 2004; 50(9): 1703 - 1704. [Full Text] [PDF]
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E. Nurk, G. S Tell, S. E Vollset, O. Nygard, H. Refsum, R. M Nilsen, and P. M Ueland Changes in lifestyle and plasma total homocysteine: the Hordaland Homocysteine Study Am. J. Clinical Nutrition, May 1, 2004; 79(5): 812 - 819. [Abstract] [Full Text] [PDF]
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 C. Chrysohoou, D. B Panagiotakos, C. Pitsavos, A. Zeimbekis, A. Zampelas, L. Papademetriou, C. Masoura, and C. Stefanadis The associations between smoking, physical activity, dietary habits and plasma homocysteine levels in cardiovascular disease-free people: the 'ATTICA' study Vascular Medicine, May 1, 2004; 9(2): 117 - 123. [Abstract] [PDF]
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 E. Bozkurt, S. Keles, M. Acikel, M. Islek, and S. Atesal Plasma Homocysteine Level and the Angiographic Extent of Coronary Artery Disease Angiology, May 1, 2004; 55(3): 265 - 270. [Abstract] [PDF]
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A. B Guttormsen, E. Solheim, and H. Refsum Variation in plasma cystathionine and its relation to changes in plasma concentrations of homocysteine and methionine in healthy subjects during a 24-h observation period Am. J. Clinical Nutrition, January 1, 2004; 79(1): 76 - 79. [Abstract] [Full Text] [PDF]
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H. Refsum, A. D. Smith, P. M. Ueland, E. Nexo, R. Clarke, J. McPartlin, C. Johnston, F. Engbaek, J. Schneede, C. McPartlin, et al. Facts and Recommendations about Total Homocysteine Determinations: An Expert Opinion Clin. Chem., January 1, 2004; 50(1): 3 - 32. [Abstract] [Full Text] [PDF]
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 T.-M. Lu, Y.-A. Ding, S.-J. Lin, W.-S. Lee, and H.-C. Tai Plasma levels of asymmetrical dimethylarginine and adverse cardiovascular events after percutaneous coronary intervention Eur. Heart J., November 1, 2003; 24(21): 1912 - 1919. [Abstract] [Full Text] [PDF]
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 M. S. Post, M. O. Verhoeven, M. J. van der Mooren, P. Kenemans, C. D. A. Stehouwer, and T. Teerlink Effect of Hormone Replacement Therapy on Plasma Levels of the Cardiovascular Risk Factor Asymmetric Dimethylarginine: A Randomized, Placebo-Controlled 12-Week Study in Healthy Early Postmenopausal Women J. Clin. Endocrinol. Metab., September 1, 2003; 88(9): 4221 - 4226. [Abstract] [Full Text] [PDF]
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 H Refsum and A D Smith Low vitamin B-12 status in confirmed Alzheimer's disease as revealed by serum holotranscobalamin J. Neurol. Neurosurg. Psychiatry, July 1, 2003; 74(7): 959 - 961. [Abstract] [Full Text] [PDF]
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H. Zetterberg, E. Nexo, B. Regland, L. Minthon, R. Boson, M. Palmer, L. Rymo, and K. Blennow The Transcobalamin (TC) Codon 259 Genetic Polymorphism Influences Holo-TC Concentration in Cerebrospinal Fluid from Patients with Alzheimer Disease Clin. Chem., July 1, 2003; 49(7): 1195 - 1198. [Full Text] [PDF]
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 I. Bjelland, G. S. Tell, S. E. Vollset, H. Refsum, and P. M. Ueland Folate, Vitamin B12, Homocysteine, and the MTHFR 677C->T Polymorphism in Anxiety and Depression: The Hordaland Homocysteine Study Arch Gen Psychiatry, June 1, 2003; 60(6): 618 - 626. [Abstract] [Full Text] [PDF]
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L. El-Khairy, S. E. Vollset, H. Refsum, and P. M. Ueland Plasma Total Cysteine, Mortality, and Cardiovascular Disease Hospitalizations: The Hordaland Homocysteine Study Clin. Chem., June 1, 2003; 49(6): 895 - 900. [Abstract] [Full Text] [PDF]
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 M.-J. J. Pouwels, M. den Heijer, H. J. Blom, C. J. Tack, and A. R. Hermus Improved Insulin Sensitivity and Metabolic Control in Type 2 Diabetes Does Not Influence Plasma Homocysteine Diabetes Care, May 1, 2003; 26(5): 1637 - 1639. [Full Text] [PDF]
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A. de Bree, W. M. Verschuren, A.-L. Bjorke-Monsen, N. M. van der Put, S. G Heil, F. J. Trijbels, and H. J Blom Effect of the methylenetetrahydrofolate reductase 677C->T mutation on the relations among folate intake and plasma folate and homocysteine concentrations in a general population sample Am. J. Clinical Nutrition, March 1, 2003; 77(3): 687 - 693. [Abstract] [Full Text] [PDF]
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L. El-Khairy, S. E Vollset, H. Refsum, and P. M Ueland Plasma total cysteine, pregnancy complications, and adverse pregnancy outcomes: the Hordaland Homocysteine Study Am. J. Clinical Nutrition, February 1, 2003; 77(2): 467 - 472. [Abstract] [Full Text] [PDF]
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R. Meleady, P. M Ueland, H. Blom, A. S Whitehead, H. Refsum, L. E Daly, S. E. Vollset, C. Donohue, B. Giesendorf, I. M Graham, et al. Thermolabile methylenetetrahydrofolate reductase, homocysteine, and cardiovascular disease risk: the European Concerted Action Project Am. J. Clinical Nutrition, January 1, 2003; 77(1): 63 - 70. [Abstract] [Full Text] [PDF]
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L. El-Khairy, S. E. Vollset, H. Refsum, and P. M. Ueland Predictors of Change in Plasma Total Cysteine: Longitudinal Findings from the Hordaland Homocysteine Study Clin. Chem., January 1, 2003; 49(1): 113 - 120. [Abstract] [Full Text] [PDF]
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 J. Y. Jeremy, N. Shukla, G. D. Angelini, A. Day, I. Y.P. Wan, S. P. Talpahewa, and R. Ascione Sustained increases of plasma homocysteine, copper, and serum ceruloplasmin after coronary artery bypass grafting Ann. Thorac. Surg., November 1, 2002; 74(5): 1553 - 1557. [Abstract] [Full Text] [PDF]
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D. M. Hill, L. J. Johnson, P. J. Burns, A. M. Neale, D. M. Harmening, and A. C. Kenney Effects of Temperature on Stability of Blood Homocysteine in Collection Tubes Containing 3-Deazaadenosine Clin. Chem., November 1, 2002; 48(11): 2017 - 2022. [Abstract] [Full Text] [PDF]
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P. O'Callaghan, R. Meleady, T. Fitzgerald, I. Graham, and European COMAC group Smoking and plasma homocysteine Eur. Heart J., October 2, 2002; 23(20): 1580 - 1586. [Abstract] [Full Text] [PDF]
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E. Nurk, G. S. Tell, S. E. Vollset, O. Nygard, H. Refsum, and P. M. Ueland Plasma Total Homocysteine and Hospitalizations for Cardiovascular Disease: The Hordaland Homocysteine Study Arch Intern Med, June 24, 2002; 162(12): 1374 - 1381. [Abstract] [Full Text] [PDF]
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B. G. Nedrebo, O. Nygard, P. M. Ueland, and E. A. Lien Plasma Total Homocysteine in Hyper- and Hypothyroid Patients before and during 12 Months of Treatment Clin. Chem., September 1, 2001; 47(9): 1738 - 1741. [Full Text] [PDF]
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 A.-L. B. Monsen, P. M. Ueland, S. E. Vollset, A. B. Guttormsen, T. Markestad, E. Solheim, and H. Refsum Determinants of Cobalamin Status in Newborns Pediatrics, September 1, 2001; 108(3): 624 - 630. [Abstract] [Full Text] [PDF]
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H. Refsum, C. S Yajnik, M. Gadkari, J. Schneede, S. E Vollset, L. Orning, A. B Guttormsen, A. Joglekar, M. G Sayyad, A. Ulvik, et al. Hyperhomocysteinemia and elevated methylmalonic acid indicate a high prevalence of cobalamin deficiency in Asian Indians Am. J. Clinical Nutrition, August 1, 2001; 74(2): 233 - 241. [Abstract] [Full Text] [PDF]
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 A. d. Bree, W. M. M. Verschuren, H. J. Blom, and D. Kromhout Lifestyle Factors and Plasma Homocysteine Concentrations in a General Population Sample Am. J. Epidemiol., July 15, 2001; 154(2): 150 - 154. [Abstract] [Full Text] [PDF]
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S. E. Vollset, H. Refsum, A. Tverdal, O. Nygard, J. E. Nordrehaug, G. S Tell, and P. M. Ueland Plasma total homocysteine and cardiovascular and noncardiovascular mortality: the Hordaland Homocysteine Study Am. J. Clinical Nutrition, July 1, 2001; 74(1): 130 - 136. [Abstract] [Full Text] [PDF]
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A. de Bree, W. M. Verschuren, H. J Blom, and D. Kromhout Association between B vitamin intake and plasma homocysteine concentration in the general Dutch population aged 20-65 y Am. J. Clinical Nutrition, June 1, 2001; 73(6): 1027 - 1033. [Abstract] [Full Text] [PDF]
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R. H. Williams, J. A. Maggiore, R. D. Reynolds, and C. M. Helgason Novel Approach for the Determination of the Redox Status of Homocysteine and Other Aminothiols in Plasma from Healthy Subjects and Patients with Ischemic Stroke Clin. Chem., June 1, 2001; 47(6): 1031 - 1039. [Abstract] [Full Text] [PDF]
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G. L. Hortin, P. Sullivan, and G. Csako Relationships among Plasma Homocysteine, Cysteine, and Albumin Concentrations: Potential Utility of Assessing the Cysteine/Homocysteine Ratio Clin. Chem., June 1, 2001; 47(6): 1121 - 1124. [Full Text] [PDF]
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 L. El-Khairy, P. M. Ueland, H. Refsum, I. M. Graham, and S. E. Vollset Plasma Total Cysteine as a Risk Factor for Vascular Disease : The European Concerted Action Project Circulation, May 29, 2001; 103(21): 2544 - 2549. [Abstract] [Full Text] [PDF]
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M K Al-Obaidi, P J Stubbs, R Amersey, and M I M Noble Acute and convalescent changes in plasma homocysteine concentrations in acute coronary syndromes Heart, April 1, 2001; 85(4): 380 - 384. [Abstract] [Full Text]
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 E. Nurk, G. S. Tell, O. Nygård, H. Refsum, P. M. Ueland, and S. E. Vollset Plasma Total Homocysteine Is Influenced by Prandial Status in Humans: The Hordaland Homocysteine Study J. Nutr., April 1, 2001; 131(4): 1214 - 1216. [Abstract] [Full Text]
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 J. C. Chambers, L. Fusi, I. S. Malik, D. O. Haskard, M. De Swiet, and J. S. Kooner Association of Maternal Endothelial Dysfunction With Preeclampsia JAMA, March 28, 2001; 285(12): 1607 - 1612. [Abstract] [Full Text] [PDF]
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 M Cahill, M Karabatzaki, C Donoghue, R Meleady, L A Mynett-Johnson, D Mooney, I M Graham, A S Whitehead, and D C Shields Thermolabile MTHFR genotype and retinal vascular occlusive disease Br. J. Ophthalmol., January 1, 2001; 85(1): 88 - 90. [Abstract] [Full Text]
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 C. M. Tribouilloy, M. Peltier, M. C. Iannetta Peltier, F. Trojette, M. Andrejak, and J.-P. M. Lesbre Plasma Homocysteine and Severity of Thoracic Aortic Atherosclerosis Chest, December 1, 2000; 118(6): 1685 - 1689. [Abstract] [Full Text] [PDF]
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J. C. Chambers, P. M. Ueland, O. A. Obeid, J. Wrigley, H. Refsum, and J. S. Kooner Improved Vascular Endothelial Function After Oral B Vitamins : An Effect Mediated Through Reduced Concentrations of Free Plasma Homocysteine Circulation, November 14, 2000; 102(20): 2479 - 2483. [Abstract] [Full Text] [PDF]
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J. C. Chambers, H. Ireland, E. Thompson, P. Reilly, O. A. Obeid, H. Refsum, P. Ueland, D. A. Lane, and J. S. Kooner Methylenetetrahydrofolate Reductase 677 C->T Mutation and Coronary Heart Disease Risk in UK Indian Asians Arterioscler. Thromb. Vasc. Biol., November 1, 2000; 20(11): 2448 - 2452. [Abstract] [Full Text] [PDF]
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B. J. Bolann, J. D. Solli, J. Schneede, K. A. Grottum, A. Loraas, M. Stokkeland, A. Stallemo, A. Schjoth, R. B. Bie, H. Refsum, et al. Evaluation of Indicators of Cobalamin Deficiency Defined as Cobalamin-induced Reduction in Increased Serum Methylmalonic Acid Clin. Chem., November 1, 2000; 46(11): 1744 - 1750. [Abstract] [Full Text] [PDF]
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C. M. Pfeiffer, S. P. Caudill, E. W. Gunter, B. A. Bowman, P. F. Jacques, J. Selhub, C. L. Johnson, D. T. Miller, and E. J. Sampson Analysis of Factors Influencing the Comparison of Homocysteine Values between the Third National Health and Nutrition Examination Surve |
