Method Development for the Profiling Analysis of Urine Globotriaosylceramide (Gb3) for the Screening of Fabry Disease by Tandem Mass Spectrometry

ESI-MS/MS를 이용한 소변 중 Globotriaosylceramide(Gb3)의 정량 및 임상 응용; 패브리병(Fabry) 진단

  • Yoon, Hye-Ran (Dept. of Biomedical & Pharmaceutical Analysis, Duksung Women's University) ;
  • Cho, Kyung-Hee (Dept. of Analytical Research, BioCore Co., LTD) ;
  • Kang, Seung-Woo (Dept. of Analytical Research, BioCore Co., LTD) ;
  • Kwon, Young-Joo (Dept. of Internal Medicine, College of Medicine, Korea University) ;
  • Jeong, Choon-Sik (Dept. of Pabathophysiology & Pharmacology, College of Pharmacy, Duksung Women's University) ;
  • Lee, Yong-Soo (Dept. of Pabathophysiology & Pharmacology, College of Pharmacy, Duksung Women's University)
  • 윤혜란 (덕성여자대학교 약학대학 생의학분석실) ;
  • 조경희 (바이오코아 분석연구팀) ;
  • 강승우 (바이오코아 분석연구팀) ;
  • 권영주 (고려대학교 의과대학 내과학 교실) ;
  • 정춘식 (덕성여자대학교 병태생리 및 약물학실) ;
  • 이용수 (덕성여자대학교 병태생리 및 약물학실)
  • Published : 2007.04.30

Abstract

Measurement of globotriaosylceramide (Gb3, ceramide trihexoside) in urine has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ${\alpha}$-galactosidase A (${\alpha}$-Gal A). The lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. A simple, rapid, and highly sensitive analytical method for Gb3 in urine was developed without labor-extensive pre-treatment by electrospray ionization MS/MS (ESI-MS/MS). Only simple 5-fold dilution of urine is necessary for the extraction and isolation of Gb3 in urine. Gb3 in diluted urine was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Eight isoforms of Gb3 were completely resolved from urine matrix. C24:0 Gb3 occupied 50% of total Gb3 as a major component in urine. Linear relationship for Gb3 isoforms was found in the range of 0.005${\sim}$5.0 ${\mu}$g/ml. The limit of detection (S/N=5) was 0.005 ${\mu}$g/ml and limit of quantification was 0.05 ${\mu}$g/ml for C24:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9598 to 0.9975. This method could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.

Keywords

References

  1. Desnick, R. J., Brady, R., Barranger, J., Collins, A. J., Germain, D. P., Goldman, M. G., Packman, S., Wilcox, W. R. and Grabowski, J. : Fabry disease, an under-recognized multisystemic disorder : expert recommendations for diagnosis, management, and enzyme replacement therapy. Ann. Intern. Med. 138(4), 338 (2003) https://doi.org/10.7326/0003-4819-138-4-200302180-00014
  2. Barbey, F., Hayoz, D., Widmer, U. and Burnier, M. : Efficacy of enzyme replacement therapy in Fabry disease. Curr. Med. Chem. Cardiovasc. Hematol. Agents. 2(4), 277 (2004) https://doi.org/10.2174/1568016043356192
  3. Eng, C. M., Guffon, N., Wilcox, W. R., Germain, D. P., Lee, P., Waldek, S., Caplan, L., Linthorst, G. E. and Desnick, R. J. : Safety and efficacy of recombinant human alpha-galactosidase A--replacement therapy in Fabry's disease. N. Engl. J. Med. 345(1), 9 (2001)
  4. Thurberg, B. L., Rennke, H., Colvin, R. B., Dikman, S., Gordon, R. E., Collins, A. B., Desnick, R. J. and O'Callaghan, M. : Globotriaosylceramide accumulation in the Fabry kidney is cleared from multiple cell types after enzyme replacement therapy. Kidney Int. 62, 1933 (2002) https://doi.org/10.1046/j.1523-1755.62.s82.2.x
  5. Desnick, R. J., Brady, R., Barranger, J., Collins, A. J., Germain, D. P., Goldman, M., Grabowski, G., Packman, S. and Wilcox, W. R. : Fabry disease, an under-recognized multisystemic disorder: expert recommendations for diagnosis, management, and enzyme replacement therapy. Ann. Int. Med. 138, 338 (2003) https://doi.org/10.7326/0003-4819-138-4-200302180-00014
  6. Frustaci, A., Chimenti, C., Ricci, R., Natale, L., Russo, M. A., Pieroni, M., Eng, C. M. and Desnick, R. J. : Improvement in cardiac function in the cardiac variant of Fabry's disease with galactose-infusion therapy. N. Engl. J. Med. 345(1), 25 (2001)
  7. Schiffmann, R., Murray, G. J., Trec, D., Daniel, P., Sellos-Moura, M., Myers, M., Quirk, J. M., Zirzow, G. C., Borowski, M., Loveday, K., Anderson, T., Gillespie, F., Oliver, K. L., Jeffries, N. O., Doo, E., Liang, T. J., Kreps, Gunter, C. K., Frei, K., Crutchfield, K., Selden, R. F. and O'Brady, R. O. : Infusion of alpha-galactosidase A reduces tissue globotriaosylceramide storage in patients with Fabry disease. Proc. Natl. Acad. Sci. 97, 365 (2000)
  8. Nelson, B. C., Roddy, T., Araghi, S., Wilkens, D., Thomas, J. J., Zhang, K., Sung, C. C. and Richards, S. M. : Globotriaosylceramide isoform profiles in human plasma by liquid chromatography-tandem mass spectrometry. J. Chromatogr. B 805(1), 127 (2004) https://doi.org/10.1016/j.jchromb.2004.02.032
  9. Berna, L., Asfaw, B., Conzelmann, E., Cerny, B. and Ledvinova, J. : Determination of urinary sulfatides and other lipids by combination of reversed-phase and thin-layer chromatographies. Anal Biochem. 269(2), 304 (1999)
  10. Zeidner, K. M., Desnick, R. J. and Ioannou, Y. A. : Quantitative determination of globotriaosylceramide by immunodetection of glycolipid-bound recombinant verotoxin B subunit. Anal. Biochem. 267, 104 (1999)
  11. Schiffmann, R., Murray, G. J., Treco, D., Daniel, P., Sellos-Moura, M., Myers, M., Quirk, J. M., Zirzow, G. C., Borowski, M., Loveday, K., Anderson, T., Gillespie, F., Oliver, K. L., Jeffries, N. O., Doo, E., Liang, T. J., Kreps, C., Gunter, K., Frei, K., Crutchfield, K., Selden, R. F. and O'Brady, R. O. : Infusion of alpha-galactosidase A reduces tissue globotriaosylceramide storage in patients with Fabry disease. Proc. Natl. Acad. Sci. 97, 365 (2000)
  12. Gross, S. K. and McCluer, R. H. : High-performance liquid chromatographic analysis of neutral glycosphingolipids as their per-O-benzoyl derivatives. Anal. Biochem. 102, 429 (1980)
  13. Ullman, M. D. and McCluer, R. : Quantitative microanalysis of perbenzoylated neutral glycosphingolipids by highperformance liquid chromatography with detection at 230 nm. J. Lipid Res. 19, 910 (1978)
  14. Lee, W. M. F., Westrick, M. A. and Macher, B. A. : Highperformance liquid chromatography of long-chain neutral glycosphingolipids and gangliosides. Biochim. Biophys. Acta. 712, 498 (1982)
  15. Strasberg, P. M., Warren, I., Skomorowski, M. A. and Lowden, J. A. : HPLC analysis of neutral glycolipids: an aid in the diagnosis of lysosomal storage disease. Clin. Chim. Acta. 132, 29 (1983)
  16. Yoo, H. H., Son, J. and Kim, D. H. : Liquid chromatographytandem mass spectrometric determination of ceramides and related lipid species in cellular extracts. J Chromatogr. B 843(2), 327 (2006) https://doi.org/10.1016/j.jchromb.2006.06.025
  17. Fauler, G., Rechberger, G. N., Devrnja, D., Erwa, W., Plecko, B., Kotanko, P., Breunig, F. and Paschke, E. : Rapid determination of urinary globotriaosylceramide isoform profiles by electrospray ionization mass spectrometry usingstearoyl-d35-globotriaosylceramide as internal standard. Rapid. Commun. Mass. Spectrom. 19(11), 1499 (2005) https://doi.org/10.1002/rcm.1948
  18. Boscaro, F., Pieraccini, G., la Marca, G., Bartolucci, G., Luceri, C., Luceri, F. and Moneti, G. : Rapid quantitation of globotriaosylceramide in human plasma and urine: a potential application for monitoring enzyme replacement therapy in Anderson-Fabry disease. Rapid. Commun. Mass. Spectrom. 16(16), 1507 (2002)
  19. Nelson, B. C., Roddy, T., Araghi, S., Wilkens, D., Thomas, J. J., Zhang, K., Sung, C. C. and Richards, S. M. : Globotriaosylceramide isoform profiles in human plasma by liquid chromatography-tandem mass spectrometry. J. Chromatogr. B 805(1), 127 (2004) https://doi.org/10.1016/j.jchromb.2004.02.032