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

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• 作者：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
• 关键词：CE ; collision voltage ; C.V. ; coefficient of variation ; ESI ; electrospray ionization ; GlcNAc ; N-acetylglucosamine ; 3¦Â-HSD ; 3¦Â-hydroxy-¦¤5-C27-steroid dehydrogenase/isomerase ; IS ; internal standard ; LC/MS/MS ; liquid chromatography/tandem mass spectrometry
• 刊名：Steroids
• 出版年：2013
• 出版时间：October, 2013
• 年：2013
• 卷：78
• 期：10
• 页码：967-972
• 全文大小：1048 K

文摘

We developed a sensitive, reliable, and accurate LC/ESI-MS/MS method for measurement of 3¦Â-sulfooxy-7¦Â-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 3¦Â-sulfooxy-7¦Â-N-acetylglucosaminyl-5-cholen-24-oic acid, 2 pg of its glycine-amide, and 0.6 pg of its taurine-amide, 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), 3¦Â-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 3¦Â-sulfooxy-7¦Â-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.