Skip to main content
SpringerLink
Log in

Development of a new HPLC diagnostic method for galactosemia using 8-amino-2-naphthalenesulfonic acid

  • Originals
  • Column Liquid Chromatography
  • Published:
Chromatographia Aims and scope Submit manuscript

Summary

In a metabolic pathway from galactose to glucose-1-phosphate, there are three major enzymes, galactokinase, galactose-1-phosphate uridyl transferase (GALT) and UDP-galactose-4-epimerase. The deficiency of one of these enzymes causes accumulation of galactose in blood, which provides a pathognomonic marker. A new reversed-phase HPLC method with fluorescence detector has been developed for the measurement of galactose in 50μL of blood on Guthrie filter paper using 8-amino-2-naphthalenesulfonic acid (8,2-ANS) as derivatization reagent for the diagnosis of Galactosemia. Galactose was extracted from blood spotted on filter paper and derivatized with 8,2-ANS to produce Schiff bases, and reduced with sodium cyanoborohydride. Linear range was from 2 mg dL−1 to 20 mg dL−1 (r=0.9998). Detection limit (S/N=3) of this method was 90 ng dL−1. The mean recovery of galactose was 102.7% (SD =0.3%, n=14). The normal range of blood galactose in Korean neonates (specimen collected within 7 days after birth) was below 6 mg dL−1 (n=5 for each gender) without any gender difference. When applied to 11 anonymous blood spots of heterogeneous genotypes of GALT deficiency all of the patients' blood samples showed abnormal elevation of galactose.

The results indicate that the new method using 8,2-ANS yields consistent and correct galactose determination that is simple and practical as a rapid first screening tool for patients with galactosemia.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Roth, H.; Segal, S.; Bertoli, D.Anal. Biochem. 1952,10, 32–52.

    Article  Google Scholar 

  2. Hjelm, M.; Derverdier, C.H.In Methods of Enzymatic Analysis, Bermeyer, H.U., Ed.;Verlag Chemie International, Deerfield Beach, FL,1974, 1282–1289.

    Google Scholar 

  3. Kurz, G.; Wallenfels, K.In Methods of Enzymatic Analysis; Bergmeyer, H.U., Ed.;Verlag Chemie International; Deer-field Beach, FL,1974, 1279–1282.

    Google Scholar 

  4. Henderson, J.M.; Fales, F.W.Clin. Chem. 1980,26, 282–285.

    CAS  Google Scholar 

  5. Grun, B.R.; Berger, U.F.; Oberdorfer, W.E.; Hull, H.; Ostertag, Keppler, D.Adv. Enzyme. Regul. 1990,30, 231–242.

    Article  CAS  Google Scholar 

  6. D'Amboise, M.; Hanai, T.; Noel, D.Clin. Chem. 1980,26, 1348–1350.

    Google Scholar 

  7. Michael, P.; Crabbe, M.J.C.J. Chromatogr. 1975,107, 180–184.

    Google Scholar 

  8. Lloyd, P.; Crabbe, M.J.C.,J. Chromatogr. 1985,343, 402–406.

    CAS  Google Scholar 

  9. Thompson, R.M.J. Chromatogr. 1978,166, 201–212.

    Article  CAS  Google Scholar 

  10. Nachtmann, F.; Budha, W.K.J. Chromatogr. 1977,136, 279–287.

    Article  CAS  Google Scholar 

  11. Miwa, I.; Kanbara, M.; Wakazone, H.; Okuda, J.Anal. Biochem. 1986,173(1), 39–44.

    Article  Google Scholar 

  12. Jakobe, C.; Kleijer, W.J.; Allen, J.; Holton, J.B.Eur. J. Pediatr. 1995,154 (7 Suppl 2), S33–36.

    Article  Google Scholar 

  13. Berry, G.T.; Palmieri, M.J.; Heales, S.J.; Lenard, V.; Segal, S.Metabolism.1992,41(7), 783–787.

    Article  CAS  Google Scholar 

  14. Xu, Y.K.; Kaufman, F.R.; Donnell, G.N.; Gludici, T.; Alfl, O.; Ng, W.G.Clin. Chem. 1995,240(1), 21–33.

    Article  CAS  Google Scholar 

  15. Gibson, J.B.; Reynold, R.A.; Palmieri, M.J.; Berry, G.T.; Elsas, L.J. 2nd; Levy, H.L.; Segal, S.Metabolism. 1995,44(5), 597–604.

    Article  CAS  Google Scholar 

  16. Keevlil, N.J.; Holton, J.B.; Allen, J.T.J. Inherit. Metab. Dis. 1994,17(1), 23–26.

    Article  Google Scholar 

  17. Keevlil, N.J.; Holton, J.B.; Allen, J.T.Clin. Chim. Acta. 1993,221(1), 135–142.

    Article  Google Scholar 

  18. Gibson, J.B.; Reynold, R.A.; Rogers, S.; Palmieri, M.J.; Segal, S.J. Pediatr. 1993,123(6), 906–914.

    Article  CAS  Google Scholar 

  19. Hase, S.; Ibuki, T.; Ikenaka, T.J.Biochem. 1984,95, 197–203.

    CAS  Google Scholar 

  20. Takemoto, H.; Hase, S.; Ikenaka, T.Anal. Biochem. 1985,145, 245–250.

    Article  CAS  Google Scholar 

  21. Takeda, M.; Maeda, M.; Tsuji, A.J. Chromatogr. 1982,244, 347.

    Article  CAS  Google Scholar 

  22. Mopper, K.; Johnson, L.J. Chromatogr. 1983,256, 27–38.

    Article  CAS  Google Scholar 

  23. Mopper, K.Anal. Chem. 1980,52, 2018.

    Article  CAS  Google Scholar 

  24. Kato, T.; Kinoshita, T.Anal. Biochem. 1980,106, 238.

    Article  CAS  Google Scholar 

  25. Honda, S.; Takahashi, M.; Kakehi, K.; Ganno, S.Anal. Biochem. 1981,113, 130.

    Article  CAS  Google Scholar 

  26. Kato, T.; Kinoshita, T.Bunseki Kagaku 1982,31, 615.

    CAS  Google Scholar 

  27. Mikami, H.; Ishida, Y.Bunseki Kagaku 1983,32, E207.

    CAS  Google Scholar 

  28. Kai, M.; Tamura, K.; Watanabe, H.; Ohkura, Y.Bunseki Kagaku 1989,38, 568–572.

    CAS  Google Scholar 

  29. Hong, S.P.; Nakamura, H.; Nakajima, T.Anal. Sci. 1994,10, 647–648.

    CAS  Google Scholar 

  30. Hong, S.P.; Sano, A.; Nakamura, H.Anal. Sci. 1996,12, 491–493.

    CAS  Google Scholar 

  31. Hong, S.P.; Sano, A.; Nakamura, H.Anal. Sci. 1997,13, 473–478.

    CAS  Google Scholar 

  32. Hong, S.P.; Lee, Y.M.; Nakamura, H.Arch. Pharm. Res. 1997,20(2), 184–190.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Oriental Pharmaceutical Science, Kyung Hee University, Dongdaemoon-gu Hoegi-dong #1, 130-701, Seoul, Korea

    S. P. Hong & M. K. Kim

  2. Metabolic Disease Detection Laboratory, Seoul Medical Science Institute, Seoul Clinical Laboratory, 7-14 Dong-Binggo-dong Yongsan-gu, 140-230, Seoul, Korea

    H. R. Yoon

Authors
  1. S. P. Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H. R. Yoon
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. K. Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hong, S.P., Yoon, H.R. & Kim, M.K. Development of a new HPLC diagnostic method for galactosemia using 8-amino-2-naphthalenesulfonic acid. Chromatographia 54, 83–86 (2001). https://doi.org/10.1007/BF02491838

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02491838

Key Words

Search

Navigation