Annals of Laboratory Medicine

Korean Society for Laboratory Medicine (대한진단검사의학회)
권호 표지
DOI QR코드

DOI QR Code

Analysis of Acid Sphingomyelinase Activity in Dried Blood Spots Using Tandem Mass Spectrometry

  • Legnini, Elisa (Women’s and Children’s Health Department, University of Padua) ;
  • Orsini, Joe J. (New York State Dept. of Health, Wadsworth Center) ;
  • Muhl, Adolf (Centogene GmbH) ;
  • Johnson, Britt (Dr. John T. Macdonald Foundation Department of Human Genetics, University of Miami Miller School of Medicine) ;
  • Dajnoki, Angela (Dr. John T. Macdonald Foundation Department of Human Genetics, University of Miami Miller School of Medicine) ;
  • Bodamer, Olaf A (Dr. John T. Macdonald Foundation Department of Human Genetics, University of Miami Miller School of Medicine)
  • Published : 2012.09.01
⟨ Previous Next ⟩

Abstract

Background: Niemann Pick disease (NP) is a rare, lysosomal storage disorder due to deficiency of the intra-lysosomal enzyme acid sphingomyelinase (ASM) resulting in intracellular accumulation of sphingomyelin. We evaluated a tandem mass spectrometry (MS/MS) method to analyze ASM activity in dried blood spots (DBS) that may be suitable for laboratory diagnosis of NP including high throughput screening of at-risk populations and potentially for newborn screening. Methods: ASM activity was measured in 3.2 mm punches from DBS. The eluate was incubated with the ASM substrate (N-Hexanoyl-D-erythro-sphingosylphosphorylcholine [C6-sphingomyelin ($C_{29}H_{59}N_{2}O_{6}P$)]) and an internal standard (N-butyroyl-D-erythro-sphingosine [C4-ceramide ($C_{22}H_{43}NO_{3}$)]). ASM product and IS were analyzed using MS/MS in multiple reaction monitoring mode for transitions m/z 370.6>264.3 (ASM internal standard) and m/z 398.6>264.3 (ASM product). Results: ASM activities were stable for up to 2 months at or below $4^{\circ}C$. Position of the punch in the DBS and/or hematocrit of the DBS had a limited effect on ASM activities. Both intraand inter-assay variability were below 10%. There was no carry-over. The median ASM activity in 2,085 newborn infants was 9.5 ${\mu}mol$/h/L (mean 10.6) with a SD of 5.06 ${\mu}mol$/h/L. Six of 2,085 (0.3%) infants were found to have ASM activities below the cut-off of 2.5 ${\mu}mol$/h/L. ASM activities were below the cut-off level in all 10 previously diagnosed cases with NP (range: 0.16 to 2.08 ${\mu}mol$/h/L). Conclusions: This MS/MS method for the measurement of ASM activity in DBS is robust and suitable for laboratory diagnosis of NP.

Keywords

References

  1. Schuchman EH and Desnick RJ. Niemann-Pick Disease Types A and B: acid sphingomyelinase deficiencies. In: Scriver CR, Beaudet AL, Sly WS, Valle D, Childs B, Kinzler KW, Vogelstein B, eds. The metabolic and molecular bases of inherited disease. 8th ed. New York: McGraw-Hill, 2001:3589-610.
  2. Simonaro CM, Desnick RJ, McGovern MM, Wasserstein MP, Schuchman EH. The demographics and distribution of type B Niemann-Pick disease: novel mutations lead to new genotype/phenotype correlations. Am J Hum Genet 2002;71:1413-9. https://doi.org/10.1086/345074
  3. Vanier MT. Niemann-Pick disease type C. Orphanet J Rare Dis 2010;5:16. https://doi.org/10.1186/1750-1172-5-16
  4. Schuchman EH. The pathogenesis and treatment of acid sphingomyelinase-deficient Niemann-Pick disease. Int J Clin Pharmacol Ther 2009;47(S1):S48-57.
  5. He X, Chen F, Dagan A, Gatt S, Schuchman EH. A fluorescence-based, high-performance liquid chromatographic assay to determine acid sphingomyelinase activity and diagnose types A and B Niemann-Pick disease. Anal Biochem 2003;314:116-20. https://doi.org/10.1016/S0003-2697(02)00629-2
  6. Gelb MH, Turecek F, Scott CR, Chamoles NA. Direct multiplex assay of enzymes in dried blood spots by tandem mass spectrometry for the newborn screening of lysosomal storage disorders. J Inherit Metab Dis 2006;29:397-404. https://doi.org/10.1007/s10545-006-0265-4
  7. Mechtler TP, Stary S, Metz TF, DeJesus VR, Greber-Platzer S, Pollak A, et al. Neonatal screening for lysosomal storage disorders: feasibility and incidence from a nationwide study in Austria. Lancet 2012;379:335-41. https://doi.org/10.1016/S0140-6736(11)61266-X
  8. Wittmann J, Karg E, Turi S, Legnini E, Wittmann G, Giese AK, et al. Newborn screening for lysosomal storage disorders in Hungary. J Inherit Metab Dis 2012 (in press).

Cited by

  1. Inborn errors of metabolism and expanded newborn screening: review and update vol.50, pp.6, 2012, https://doi.org/10.3109/10408363.2013.847896
  2. A column‐switching HPLC‐MS/MS method for mucopolysaccharidosis type I analysis in a multiplex assay for the simultaneous newborn screening of six lysosomal storage disorders vol.28, pp.8, 2012, https://doi.org/10.1002/bmc.3133
  3. Rapid evaluation of 25 key sphingolipids and phosphosphingolipids in human plasma by LC-MS/MS vol.407, pp.17, 2012, https://doi.org/10.1007/s00216-015-8585-6
  4. The use of mass spectrometry to analyze dried blood spots vol.35, pp.3, 2012, https://doi.org/10.1002/mas.21441
  5. Multiplex Assay of Second-Line Anti-Tuberculosis Drugs in Dried Blood Spots Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry vol.36, pp.5, 2012, https://doi.org/10.3343/alm.2016.36.5.489
  6. A Robust Liposomal Platform for Direct Colorimetric Detection of Sphingomyelinase Enzyme and Inhibitors vol.12, pp.8, 2018, https://doi.org/10.1021/acsnano.8b03308