Analytical and Bioanalytical Chemistry

, Volume 405, Issue 4, pp 1345–1351 | Cite as

Intracellular in vitro probe acylcarnitine assay for identifying deficiencies of carnitine transporter and carnitine palmitoyltransferase-1

  • Jamiyan Purevsuren
  • Hironori Kobayashi
  • Yuki Hasegawa
  • Kenji Yamada
  • Tomoo Takahashi
  • Masaki Takayanagi
  • Toshiyuki Fukao
  • Seiji Fukuda
  • Seiji Yamaguchi
Original Paper


Mitochondrial fatty acid oxidation (FAO) disorders are caused by defects in one of the FAO enzymes that regulates cellular uptake of fatty acids and free carnitine. An in vitro probe acylcarnitine (IVP) assay using cultured cells and tandem mass spectrometry is a tool to diagnose enzyme defects linked to most FAO disorders. Extracellular acylcarnitine (AC) profiling detects carnitine palmitoyltransferase-2, carnitine acylcarnitine translocase, and other FAO deficiencies. However, the diagnosis of primary carnitine deficiency (PCD) or carnitine palmitoyltransferase-1 (CPT1) deficiency using the conventional IVP assay has been hampered by the presence of a large amount of free carnitine (C0), a key molecule deregulated by these deficiencies. In the present study, we developed a novel IVP assay for the diagnosis of PCD and CPT1 deficiency by analyzing intracellular ACs. When exogenous C0 was reduced, intracellular C0 and total AC in these deficiencies showed specific profiles clearly distinguishable from other FAO disorders and control cells. Also, the ratio of intracellular to extracellular C0 levels showed a significant difference in cells with these deficiencies compared with control. Hence, intracellular AC profiling using the IVP assay under reduced C0 conditions is a useful method for diagnosing PCD or CPT1 deficiency.


Fatty acid oxidation Carnitine cycle disorder Acylcarnitine profile ESI-MS/MS 


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Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Jamiyan Purevsuren
    • 1
    • 5
  • Hironori Kobayashi
    • 1
  • Yuki Hasegawa
    • 1
  • Kenji Yamada
    • 1
  • Tomoo Takahashi
    • 1
  • Masaki Takayanagi
    • 2
  • Toshiyuki Fukao
    • 3
    • 4
  • Seiji Fukuda
    • 1
  • Seiji Yamaguchi
    • 1
  1. 1.Department of PediatricsShimane University School of MedicineIzumoJapan
  2. 2.Division of MetabolismChiba Children’s HospitalChibaJapan
  3. 3.Department of Pediatrics, Graduate School of MedicineGifu UniversityGifuJapan
  4. 4.Medical Information Sciences Division, United Graduate School of Drug Discovery and Medical Information SciencesGifu UniversityGifuJapan
  5. 5.Medical Genetics LaboratoryNational Center for Maternal and Child HealthUlaanbaatarMongolia

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