A sensitive and specific LC-MS/MS method for rapid diagnosis of Niemann-Pick C1 disease from human plasma

作者：Jiang Xuntian; Sidhu Rohini; Porter Forbes D; Yanjanin Nicole M; Speak Anneliese O; Vruchte Danielle Taylor Te; Platt Frances M; Fujiwara Hideji; Scherrer David E; Zhang Jessie; Dietzen Dennis J; Schaffer Jean E; Ory Daniel S^*

来源：The Journal of Lipid Research, 2011, 52(7): 1435-1445.

DOI：10.1194/jlr.D015735

• cholesterol
• diagnostic tools
• liquid chromatography/tandem mass spectrometry
• Niemann-Pick disease
• oxysterols
• neurodegeneration

Niemann-Pick type C1 (NPC1) disease is a rare, progressively fatal neurodegenerative disease for which there are no FDA-approved therapies. A major barrier to developing new therapies for this disorder has been the lack of a sensitive and noninvasive diagnostic test. Recently, we demonstrated that two cholesterol oxidation products, specifically cholestane-3 beta,5 alpha,6 beta-triol (3 beta,5 alpha,6 beta-triol) and 7-ketocholesterol (7-KC), were markedly increased in the plasma of human NPC1 subjects, suggesting a role for these oxysterols in diagnosis of NPC1 disease and evaluation of therapeutics in clinical trials. In the present study, we describe the development of a sensitive and specific LC-MS/MS method for quantifying 3 beta,5 alpha,6 beta-triol and 7-KC human plasma after derivatization with N,N-dimethylglycine. We show that dimethylglycine derivatization successfully enhanced the ionization and fragmentation of 3 beta,5 alpha,6 beta-triol and 7-KC for mass spectrometric detection of the oxysterol species in human plasma. The oxysterol dimethylglycinates were resolved with high sensitivity and selectivity, and enabled accurate quantification of 3 beta,5 alpha,6 beta-triol and 7-KC concentrations in human plasma. The LC-MS/MS assay was able to discriminate with high sensitivity and specificity between control and NPC1 subjects, and offers for the first time a noninvasive, rapid, and highly sensitive method for diagnosis of NPC1 disease.-Jiang, X., R. Sidhu, F. D. Porter, N. M. Yanjanin, A. O. Speak, D. Taylor te Vruchte, F. M. Platt, H. Fujiwara, D. E. Scherrer, J. Zhang, D. J. Dietzen, J. E. Schaffer, and D. S. Ory. A sensitive and specifi c LC-MS/MS method for rapid diagnosis of Niemann-Pick C1 disease from human plasma. J. Lipid Res. 2011. 52: 1435-1445.

• 出版日期2011-7