LC/ESI-MS/MS analysis of urinary 3β-sulfooxy-7β-N- acetylglucosaminyl- 5-cholen-24-oic acid and its amides: New biomarkers for the detection of Niemann-Pick type C disease

Masamitsu Maekawa, Yasushi Misawa, Ayako Sotoura, Hiroaki Yamaguchi, Masami Togawa, Kousaku Ohno, Hiroshi Nittono, Genta Kakiyama, Takashi Iida, Alan F. Hofmann, Junichi Goto, Miki Shimada, Nariyasu Mano

Research output: Contribution to journalArticlepeer-review

34 Citations (Scopus)

Abstract

We developed a sensitive, reliable, and accurate LC/ESI-MS/MS method for measurement of 3β-sulfooxy- 7β-N-acetylglucosaminyl-5-cholen-24-oic acid and its glycine and taurine amides in urine. This atypical C24 bile acid has been reported previously to be present in the urine of patients with Niemann-Pick Type C (NPC) disease. In the method, targeted analytes are concentrated at the front edge of a trapping column, Shim-pack MAYI-C8, which permits elimination of contaminating molecules in the urinary matrix. The trapped analytes are then eluted, separated on a YMC-Pack Pro C18, and quantified with MS/MS using selected reaction monitoring. The method could detect (as amount injected) 2 pg of nonamidated 3b-sulfooxy- 7b-N-acetylglucosaminyl-5-cholen-24-oic acid, 2 pg of its glycine-amide, and 0.6 pg of its taurineamide, and is linear up to 300 pg. The method was then used to measure the three analytes in the urine of NPC patients (N = 2), 3b-hydroxysteroid dehydrogenase deficiency patients (N = 2), and healthy volunteers (N = 8). Measurable concentrations of all three analytes were present in all subjects. The urinary concentration of the sum of all three analytes was four hundred times greater in the 3 month NPC patient and 40 times greater in the adult patient than that of healthy volunteers. The markedly elevated urinary concentration of 3b-sulfooxy-7b-N-acetylglucosaminyl-5-cholen-24-oic acid and its amides in NPC patients suggests that these compounds may be valuable biomarkers for detection of NPC disease.

Original languageEnglish
Pages (from-to)967-972
Number of pages6
JournalSteroids
Volume78
Issue number10
DOIs
Publication statusPublished - 2013

Keywords

  • Biomarker
  • LC/ESI-MS/MS
  • Multiple conjugate
  • Niemann-Pick type C disease

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Endocrinology
  • Pharmacology
  • Clinical Biochemistry
  • Organic Chemistry

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