Analytical and Bioanalytical Chemistry

, Volume 407, Issue 17, pp 5227–5233 | Cite as

LC–MS/MS-based quantification of cholesterol and related metabolites in dried blood for the screening of inborn errors of sterol metabolism

  • S. Becker
  • S. Röhnike
  • S. Empting
  • D. Haas
  • K. Mohnike
  • S. Beblo
  • U. Mütze
  • R. A. Husain
  • J. Thiery
  • U. Ceglarek
Research Paper
Part of the following topical collections:
  1. Lipidomics


Smith–Lemli–Opitz syndrome (SLOS) is an inherited metabolic disease in the cholesterol biosynthesis pathway which is characterised by accumulation of 7- and 8-dehydrocholesterol and by reduced cholesterol concentrations in all tissues and body fluids. With this study, we developed a new, rapid, robust and high-throughput tandem mass spectrometric method as routine application for the selective SLOS screening and therapy monitoring in serum and dried blood. After protein precipitation of 10 μL serum or 4.7 mm dried blood spot, the sum of 7- and 8-dehydrocholesterol (DHC) was analysed by rapid chromatography combined with tandem mass spectrometry. Method comparison with GC–MS was performed for 46 serum samples. A comparison between serum and corresponding dried blood spots for DHC and cholesterol was performed with 40 samples from SLOS patients. Concentrations of DHC and cholesterol were analysed in 2 dried blood samples from newborns with SLOS and 100 unaffected newborns. Intra- and inter-assay variabilities ranged between 3.7 and 17.7 % for serum and dried blood spots. Significant correlations between the new LC–MS/MS method and GC–MS were determined for DHC (r = 0.937, p < 0.001) and for cholesterol (r = 0.946, p < 0.001). Significant coefficients of correlation between serum and dried blood spot samples above 0.8 were calculated for both analytes. A cut-off value of 5.95 for the ratio of DHC/cholesterol (multiplied by 1000) was found to distinguish newborns diagnosed with SLOS from normal newborns in a retrospective analysis after 5 years. The developed method enables a rapid quantification of the sum parameter 7- and 8-DHC in newborns and SLOS patients under therapy in serum as well as dried blood spot samples.


7-Dehydrocholesterol Dried blood spots Smith–Lemli–Opitz syndrome Tandem mass spectrometry 



Atmospheric pressure photo ionisation




Confidence interval






7-Dehydrocholesterol reductase


Electrospray ionisation


Gas chromatography mass spectrometry


Liquid chromatography tandem mass spectrometry


Limit of detection


Lower limit of quantification


Smith–Lemli–Opitz syndrome

Supplementary material

216_2015_8731_MOESM1_ESM.pdf (266 kb)
ESM 1(PDF 265 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • S. Becker
    • 1
    • 2
  • S. Röhnike
    • 3
  • S. Empting
    • 3
  • D. Haas
    • 4
  • K. Mohnike
    • 3
  • S. Beblo
    • 5
  • U. Mütze
    • 5
  • R. A. Husain
    • 6
  • J. Thiery
    • 1
    • 2
  • U. Ceglarek
    • 1
    • 2
  1. 1.Institute of Laboratory Medicine, Clinical Chemistry and Molecular DiagnosticsUniversity Hospital LeipzigLeipzigGermany
  2. 2.LIFE—Leipzig Research Center for Civilization DiseasesUniversity LeipzigLeipzigGermany
  3. 3.Department of PediatricsOtto-von-Guericke University MagdeburgMagdeburgGermany
  4. 4.Division of Inborn Metabolic DiseasesUniversity Children’s HospitalHeidelbergGermany
  5. 5.Hospital for Children and Adolescents, Centre of Paediatric Research (CPR), Department of Women and Child Health, University HospitalsUniversity of LeipzigLeipzigGermany
  6. 6.Center for Inborn Metabolic Disorders, Children’s HospitalJena University HospitalJenaGermany

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