Development of a new standard reference material: SRM 1955 (homocysteine and folate in human serum)

  • Mary B. Satterfield
  • Lorna T. Sniegoski
  • Katherine E. Sharpless
  • Michael J. Welch
  • Adriana Hornikova
  • Nien-Fan Zhang
  • Christine M. Pfeiffer
  • Zia Fazili
  • Mindy Zhang
  • Bryant C. Nelson
Original Paper

Abstract

Total homocysteine (tHCY) and folate are interrelated biomarkers for arteriosclerosis and coronary heart disease. Although many different methods for both tHCY and folate are clinically available, the intermethod and interlaboratory results are often poor, resulting in the need for a matrix reference material and reference methods. The National Institute of Standards and Technology (NIST) has developed isotope dilution liquid chromatography/mass spectrometry (LC/MS) and liquid chromatography/ tandem mass spectrometry (LC/MS/MS) methods for determination of tHCY and several folate forms including 5-methyltetrahydrofolic acid (5MT) and folic acid (FA). Additionally, a method for simultaneous measurement of tHCY, 5MT, and FA has been developed and validated. In collaboration with the Centers for Disease Control and Prevention (CDC), mass spectrometric methods and methods used in clinical laboratories have been applied to characterize a new Standard Reference Material (SRM), SRM 1955, “Homocysteine and Folate in Human Serum,” containing low, medium, and high levels of tHCY and 5MT. Additionally, FA, 5-formyltetrahydrofolic acid (5FT), vitamin B12, and total folate values are provided. Use of the new SRM should improve clinical measurements and will permit traceability to internationally recognized certified reference materials, as described by European Directive 98/79/EC on in vitro diagnostic medical devices.

Keywords

Isotope dilution mass spectrometry Standard reference material Homocysteine Folate 5-Methyltetrahydrofolic acid 5-Formyltetrahydrofolic acid Folic acid 

Notes

Acknowledgements

Sonya Strider (FPIA) and Lily Jia (BioRad).

References

  1. 1.
    Boushey CJ, Beresford SA, Omenn GS, Motulsky AG (1995) A quantitative assessment of plasma homocysteine as a risk factor for vascular disease. Probable benefits of increasing folic acid intakes. JAMA 274:1049–1057CrossRefPubMedGoogle Scholar
  2. 2.
    Refsum H, Ueland PM, Nygard O, Vollset SE (1998) Homocysteine and cardiovascular disease. Annu Rev Med 49:31–62CrossRefPubMedGoogle Scholar
  3. 3.
    Weiss N, Hilge R, Hoffmann U (2004) Mild hyperhomocysteinemia: risk factor or just risk predictor for cardiovascular diseases? Vasa 33:191–203CrossRefPubMedGoogle Scholar
  4. 4.
    MRC Vitamin Study Research Group (1991) Prevention of neural tube defects: results of the Medical Research Council Vitamin Study. Lancet 338:131–137CrossRefGoogle Scholar
  5. 5.
    Pfeiffer CM, Fazili Z, McCoy L, Zhang M, Gunter EW (2004) Determination of folate vitamers in human serum by stable-isotope-dilution tandem mass spectrometry and comparison with radioassay and microbiologic assay. Clin Chem 50:423–432CrossRefPubMedGoogle Scholar
  6. 6.
    Honein MA, Paulozzi LJ, Mathews TJ, Erickson JD, Wong LY (2001) Impact of folic acid fortification of the US food supply on the occurrence of neural tube defects. JAMA 285:2981–2986CrossRefPubMedGoogle Scholar
  7. 7.
    Schnyder G, Roffi M, Pin R, Flammer Y, Lange H, Eberli FR et al (2001) Decreased rate of coronary restenosis after lowering of plasma homocysteine levels. N Engl J Med 345:1593–1600CrossRefPubMedGoogle Scholar
  8. 8.
    Choi SW, Mason JB (2002) Folate status: effects on pathways of colorectal carcinogenesis. J Nutr 132:2413S–2418SPubMedGoogle Scholar
  9. 9.
    D’Eramo JL, Finkelstein AE, Boccazzi FO, Fridman O (1998) Total homocysteine levels in plasma: high-performance liquid chromatographic determination with electrochemical detection and glassy carbon electrode. J Chromatogr B 720(1–2):205–210Google Scholar
  10. 10.
    Frantzen F, Faaren AL, Alfheim I, Nordhei AK (1998) Enzyme conversion immunoassay for determining total homocysteine in plasma or serum. Clin Chem 44:311–316PubMedGoogle Scholar
  11. 11.
    Marangon K, O’Byrne D, Devaraj S, Jialal I (1999) Validation of an immunoassay for measurement of plasma total homocysteine. Am J Clin Path 112:757–762PubMedGoogle Scholar
  12. 12.
    Pfeiffer CM, Huff DL, Gunter EW (1999) Rapid and accurate HPLC assay for plasma total homocysteine and cysteine in a clinical laboratory setting. Clin Chem 45:290–293PubMedGoogle Scholar
  13. 13.
    Stabler SP, Lindenbaum J, Savage DG, Allen RH (1993) Elevation of serum cystathionine levels in patients with cobalamin and folate deficiency. Blood 81:3404–3413PubMedGoogle Scholar
  14. 14.
    Gempel K, Gerbitz K-D, Casetta B, Bauer MF (2000) Rapid determination of total homocysteine in blood spots by liquid chromatography-electrospray ionization-tandem mass spectrometry. Clin Chem 46:122–123PubMedGoogle Scholar
  15. 15.
    Magera MJ, Lacey JM, Casetta B, Rinaldo P (1999) Method for the determination of total homocysteine in plasma and urine by stable isotope dilution and electrospray tandem mass spectrometry. Clin Chem 45:1517–1522PubMedGoogle Scholar
  16. 16.
    Nelson B, Pfeiffer CM, Sniegoski LT, Satterfield MB (2003) Development and evaluation of an isotope dilution LC/MS method for the determination of total homocysteine in human plasma. Anal Chem 75:775–784CrossRefPubMedGoogle Scholar
  17. 17.
    Nexo E, Engbaek F, Ueland PM, Westby C, O’Gorman P, Johnston C et al (2000) Evaluation of novel assays in clinical chemistry: quantification of plasma total homocysteine. Clin Chem 46:1150–1156PubMedGoogle Scholar
  18. 18.
    Tripodi A, Chantarangkul V, Lombardi R, Lecchi A, Mannucci PM, Cattaneo M (2001) Multicenter study of homocysteine measurement-performance characteristics of different methods, influence of standards on interlaboratory agreement of results. Thromb Haemost 85:291–295PubMedGoogle Scholar
  19. 19.
    Molloy AM, Scott JM (1997) Microbiological assay for serum, plasma, and red cell folate using cryopreserved, microtiter plate method. Methods Enzymol 281:43–53, 43–53PubMedCrossRefGoogle Scholar
  20. 20.
    O’Broin S, Kelleher B (1992) Microbiological assay on microtitre plates of folate in serum and red cells. J Clin Pathol 45:344–347PubMedCrossRefGoogle Scholar
  21. 21.
    Bio-Rad Laboratories (1993) Instruction manual, bio-rad quantaphase II folate/B12 radioassay kit. Bio-Rad Laboratories, Hercules, CAGoogle Scholar
  22. 22.
    Gunter EW, Bowman BA, Caudill SP, Twite DB, Adams MJ, Sampson EJ (1996) Results of an international round robin for serum and whole-blood folate. Clin Chem 42:1689–1694PubMedGoogle Scholar
  23. 23.
    Bagley PJ, Selhub J (2000) Analysis of folate form distribution by affinity followed by reversed-phase chromatography with electrical detection. Clin Chem 46:404–411PubMedGoogle Scholar
  24. 24.
    Lucock MD, Hartley R, Smithells RW (1989) A rapid and specific HPLC-electrochemical method for the determination of endogenous 5-methyltetrahydrofolic acid in plasma using solid phase sample preparation with internal standardization. Biomed Chromatogr 3:58–63CrossRefPubMedGoogle Scholar
  25. 25.
    Hart DJ, Finglas PM, Wolfe CA, Mellon F, Wright AJ, Southon S (2002) Determination of 5-methyltetrahydrofolate (13C-labeled and unlabeled) in human plasma and urine by combined liquid chromatography mass spectrometry. Anal Biochem 305:206–213CrossRefPubMedGoogle Scholar
  26. 26.
    Nelson BC, Dalluge JJ, Margolis SA (2001) Preliminary application of liquid chromatography-electrospray-ionization mass spectrometry to the detection of 5-methyltetrahydrofolic acid monoglutamate in human plasma. J Chromatogr B Biomed Sci Appl 765:141–150CrossRefPubMedGoogle Scholar
  27. 27.
    Nelson BC, Pfeiffer CM, Margolis SA, Nelson CP (2003) Affinity extraction combined with stable isotope dilution LC/MS for the determination of 5-methyltetrahydrofolate in human plasma. Anal Biochem 313:117–127CrossRefPubMedGoogle Scholar
  28. 28.
    Nelson BC, Pfeiffer CM, Margolis SA, Nelson CP (2004) Solid-phase extraction-electrospray ionization mass spectrometry for the quantification of folate in human plasma or serum. Anal Biochem 325:41–51CrossRefPubMedGoogle Scholar
  29. 29.
    Pawlosky RJ, Flanagan VP, Pfeiffer CM (2001) Determination of 5-methyltetrahydrofolic acid in human serum by stable-isotope dilution high-performance liquid chromatography-mass spectrometry. Anal Biochem 298:299–305CrossRefPubMedGoogle Scholar
  30. 30.
    Nelson BC, Satterfield MB, Sniegoski LT, Welch MJ (2005) Simultaneous quantification of homocysteine and folate in human serum or plasma using liquid chromatography/tandem mass spectrometry. Anal Chem 77:3586–3593CrossRefPubMedGoogle Scholar
  31. 31.
    Guide to the expression of uncertainty in measurement, 1st edn. Geneva, Switzerland: ISO, 1993Google Scholar
  32. 32.
    Levenson MS, Banks DL, Eberhardt KR, Gill LM, Guthrie WF, Liu HK et al (2000) An approach to combining results from multiple methods motivated by the ISO GUM. J Res Natl Inst Stand Technol 105:571–579Google Scholar
  33. 33.
    Liu H-K, Zhang NF (2001) Bayesian approach to combining results from multiple methods. Proceedings of the Section of Bayesian Statistical Science of American Statistical SocietyGoogle Scholar
  34. 34.
    Satterfield MB, Sniegoski LT, Welch MJ, Nelson BC, Pfeiffer CM (2003) Comparison of isotope dilution mass spectrometry methods for the determination of total homocysteine in plasma and serum. Anal Chem 75:4631–4638CrossRefPubMedGoogle Scholar
  35. 35.
    Pfeiffer CM, Twite D, Shih J, Holets-McCormack SR, Gunter EW (1999) Method comparison for total plasma homocysteine between the Abbott IMx analyzer and an HPLC assay with internal standardization. Clin Chem 45:152–153PubMedGoogle Scholar
  36. 36.
    Shipchandler MT, Moore EG (1995) Rapid, fully automated measurement of plasma homocyst(e)ine with the Abbott IMx analyzer. Clin Chem 41:991–994PubMedGoogle Scholar
  37. 37.
    Hornikova A, Zhang NF, Welch MJ, Tai S (2006) An application of combining results from multiple methods-statistical evaluation of uncertainty for NIST SRM 1508a. Metrologia 43:205–212CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Mary B. Satterfield
    • 1
  • Lorna T. Sniegoski
    • 1
  • Katherine E. Sharpless
    • 1
  • Michael J. Welch
    • 1
  • Adriana Hornikova
    • 2
  • Nien-Fan Zhang
    • 2
  • Christine M. Pfeiffer
    • 3
  • Zia Fazili
    • 3
  • Mindy Zhang
    • 3
  • Bryant C. Nelson
    • 1
  1. 1.Analytical Chemistry DivisionNational Institute of Standards and TechnologyGaithersburgUSA
  2. 2.Statistical Engineering DivisionNational Institute of Standards and TechnologyGaithersburgUSA
  3. 3.Division of Laboratory SciencesCenters for Disease Control and PreventionAtlantaUSA

Personalised recommendations