Search for


search for

Measurement of Succinylacetone Using HPLC-Tandem Mass Spectrometry and Establishment of a Cut-off Value
J Lab Med Qual Assur 2018;40:149-154
Published online September 30, 2018
© 2018 Korean Association of External Quality Assessment Service.

Sun Hee Jun1,*, Jong Do Seo2,*, Kyunghoon Lee1,2, and Junghan Song1,2

1Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam; 2Department of Laboratory Medicine, Seoul NationalUniversity College of Medicine, Seoul, Korea
Correspondence to: Junghan Song
Department of Laboratory Medicine, Seoul National University Bundang Hospital, 82 Gumi-ro 173beon-gil, Bundanggu, Seongnam 13620, Korea
Tel: +82-31-787-7691
Fax: +82-31-787-4015
*These authors contributed equally to this work.
Received April 11, 2018; Revised July 9, 2018; Accepted July 9, 2018.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Background: Newborn screening of tyrosinemia type 1 is important for identifying infants at risk for developing this disease before life-threatening symptoms occur. It is difficult to differentiate between tyrosinemia type 1 and transient neonatal tyrosinemia (TNT) by analyzing tyrosine alone. Thus, succinylacetone must be analyzed. In this study, we measured succinylacetone in dried blood spot (DBS) by HPLC-tandem mass spectrometry (HPLC-MS/MS) and established cut-off values.
Methods: We used the hydrazine derivatization method to measure succinylacetone in 127 DBSs showing normal results in the newborn screening test and 93 DBSs showing increased tyrosine levels. We established cut-off values using the 99.9th percentile value or median+5 standard deviation value.
Results: Succinylacetone levels determined by our method were well-correlated with the results recommended by the Centers for Disease Control and Prevention for proficiency testing (r =0.9968). The succinylacetone levels in normal newborn DBSs were significantly lower than those in DBSs with high tyrosine levels (P 竊0.001). The cut-off values were calculated to be 1.3 關M from the results of 127 normal DBS samples and 2.2 關M from 220 DBSs, including in 93 newborns with TNT.
Conclusions: Measurement of succinylacetone in DBSs by HPLC-MS/MS is useful in individuals with increased tyrosine concentrations and can be used for rapid differential diagnosis of tyrosinemia when an appropriate cut-off value is established.
Keywords : Succinylacetone, Tyrosinemias, Tandem mass spectrometry
  1. Mitchell GA, Grompe M, Lambert M, Tanguay RM. Hypertyrosinemia. In: Scriver CR, Beaudet AL, Sly WS, Valle DV, editors. The metabolic and molecular bases of inherited disease. 8th ed. New York (NY): McGraw-Hill, 2001:1777-805.
  2. Chinsky JM, Singh R, Ficicioglu C, van Karnebeek CDM, Grompe M, Mitchell G, et al. Diagnosis and treatment of tyrosinemia type I: a US and Canadian consensus group review and recommendations. Genet Med 2017;19.
  3. Berger R, van Faassen H, Smith GP. Biochemical studies on the enzymatic deficiencies in hereditary tyrosinemia. Clin Chim Acta 1983;134:129-41.
  4. Kvittingen EA. Tyrosinaemia type I: an update. J Inherit Metab Dis 1991;14:554-62.
    Pubmed CrossRef
  5. Holme E, Lindstedt S. Nontransplant treatment of tyrosinemia. Clin Liver Dis 2000;4:805-14.
  6. Grenier A, Lescault A, Laberge C, Gagne R, Mamer O. Detection of succinylacetone and the use of its measurement in mass screening for hereditary tyrosinemia. Clin Chim Acta 1982;123:93-9.
  7. Jakobs C, Dorland L, Wikkerink B, Kok RM, de Jong AP, Wadman SK. Stable isotope dilution analysis of succinylacetone using electron capture negative ion mass fragmentography: an accurate approach to the pre- and neonatal diagnosis of hereditary tyrosinemia type I. Clin Chim Acta 1988;171:223-31.
  8. La Marca G, Malvagia S, Pasquini E, Innocenti M, Fernandez MR, Donati MA, et al. The inclusion of succinylacetone as marker for tyrosinemia type I in expanded newborn screening programs. Rapid Commun Mass Spectrom 2008;22:812-8.
    Pubmed CrossRef
  9. Metz TF, Mechtler TP, Merk M, Gottschalk A, Lukacin R, Herkner KR, et al. Evaluation of a novel, commercially available mass spectrometry kit for newborn screening including succinylacetone without hydrazine. Clin Chim Acta 2012;413:1259-64.
    Pubmed CrossRef
  10. Clinical and Laboratory Standards Institute. Defining, establishing, and verifying reference intervals in the clinical laboratory: approved guideline: C28-A3C. 3rd ed. Wayne (PA): Clinical and Laboratory Standards Institute, 2010.
  11. Stinton C, Geppert J, Freeman K, Clarke A, Johnson S, Fraser H, et al. Newborn screening for Tyrosinemia type 1 using succinylacetone: a systematic review of test accuracy. Orphanet J Rare Dis 2017;12:48.
    Pubmed KoreaMed CrossRef