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Detection of Δ4-3-oxo-steroid 5β-reductase deficiency by LC-ESI-MS/MS measurement of urinary bile acids


, : Detection of Δ4-3-oxo-steroid 5β-reductase deficiency by LC-ESI-MS/MS measurement of urinary bile acids. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences 900: 24-31

The synthesis of bile salts from cholesterol is a complex biochemical pathway involving at least 16 enzymes. Most inborn errors of bile acid biosynthesis result in excessive formation of intermediates and/or their metabolites that accumulate in blood and are excreted in part in urine. Early detection is important as oral therapy with bile acids results in improvement. In the past, these intermediates in bile acid biosynthesis have been detected in neonatal blood or urine by screening with FAB-MS followed by detailed characterization using GC-MS. Both methods have proved difficult to automate, and currently most laboratories screen candidate samples using LC-MS/MS. Here, we describe a new, simple and sensitive analytical method for the identification and characterization of 39 conjugated and unconjugated bile acids, including Δ(4)-3-oxo- and Δ(4,6)-3-oxo-bile acids (markers for Δ(4)-3-oxo-steroid 5β-reductase deficiency), using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). In this procedure a concentrated, desalted urinary sample (diluted with ethanol) is injected directly into the LC-ESI-MS/MS, operated with ESI and in the negative ion mode; quantification is obtained by selected reaction monitoring (SRM). To evaluate the performance of our new method, we compared it to a validated method using GC-MS, in the analysis of urine from two patients with genetically confirmed Δ(4)-3-oxo-steroid 5β-reductase deficiency as well as a third patient with an elevated concentration of abnormal conjugated and unconjugated Δ(4)-3-oxo-bile acids. The Δ(4)-3-oxo-bile acids concentration recovered in three patients with 5β-reductase deficiency were 48.8, 58.9, and 49.4 μmol/mmol creatinine, respectively by LC-ESI-MS/MS.

US$19.90

PMID: 22695323

DOI: 10.1016/j.jchromb.2012.05.023


Other references

Zorn, K.K., 2007: Advancing our understanding of endometrial cancer histologic subtypes: should we lump or split?. Gynecologic Oncology 106(1): 6-7

Hanley T.R.Jr; Young J.T.; John J.A.; Rao K.S., 1984: Ethylene glycol monomethyl ether and propylene glycol monomethyl ether inhalation fertility and teratogenicity studies in rats mice and rabbits. Environmental Health Perspectives 57: 7-12

Hope, E.W., 1926: The Influence of the Universities upon the Advancement of the Public Health. Proceedings of the Royal Society of Medicine 19(Sect Epidemiol State Med): 22-30

Lafay, M.; Lidove, O.; Chanson, P.; Ziza, J-Marc., 2016: Thick heel pads. Joint, Bone, Spine 82(6): 463-463

Michon, J.; Arnaud, R., 1962: Rapid colorimetric determination of blood urea. Clinica Chimica Acta; International Journal of Clinical Chemistry 7: 739-739

Greenman D.L.; Cernosek S.F.; Gutierrez Cernosek R.M., 1977: Uterine vaginal and prolactin responses to dietary di ethyl stilbestrol in the mouse. Federation Proceedings 36(3): 912

Flynn, V., 1974: Some aspects of silage making and feeding. Silage yield, quality and additives, efficiency of conservation and wilting are discussed.

Clark, B.B., 1974: Too many questionnaires--and some solutions. Hospitals 48(12): 83-86

Dubey, JP.; Speer, CA.; Munday, BL.; Lipscomb, TP., 1989: Ovine sporozoan encephalomyelitis linked to Sarcocystis infection. The unidentified sporozoan causing encephalomyelitis in sheep described by Hartley and Blakemore (1974) was restudied, and the parasite was identified as a Sarcocystis species based on its location and structure. The parasite was located free in t...

Johnson S.B., 1988: Chronic illness and pain. Mash, Ed L. G. Terdal. (ed.). Behavioral Assessment Of Childhood Disorders, Second Edition. Ix 824p. The Guilford Press: New York, New York, Usa; London, England, Uk. Illus. 491-527