title: Quantitative analysis of s-adenosylmethionine and s-adenosylhomocysteine in neurulation-stage mouse embryos by liquid chromatography tandem mass spectrometry.
abstract: The potential importance of the methylation cycle during embryonic development necessitates the establishment of methodology to detect alterations in the relative abundance of s-adenosylmethionine (SAM) and s-adenosylhomocysteine (SAH) in an embryonic experimental system. We have developed a precise and sensitive method for measurement of SAM and SAH based on liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in single neurulation-stage mouse embryos. Use of a penta-fluorinated high-performance liquid chromatography (HPLC) stationary phase gave enhanced sensitivity due to optimal ionisation in organic mobile phase and increased retention time compared to standard reversed-phase separation. Calibration curves suitable for the analysis of neurulation-stage mouse embryos (SAM 0.02-25.0microM, SAH 0.01-10.0microM) were linear (r(2)>0.997) with limits of detection for SAM and SAH of 10 and 2.5nmol/L, respectively.
authors.affiliation: Neural Development Unit, Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, United Kingdom.
authors.forename: Katie A
authors.initials: KA
authors.lastname: Burren
authors.forename: Kevin
authors.initials: K
authors.lastname: Mills
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Copp
authors.forename: Nicholas D E
authors.initials: ND
authors.lastname: Greene
mesh_terms: D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004622:Embryo, Mammalian / Q000737:chemistry*; D005260:Female; D008297:Male; D051379:Mice; D011247:Pregnancy; D015203:Reproducibility of Results; D012435:S-Adenosylhomocysteine / Q000032:analysis*; D012436:S-Adenosylmethionine / Q000032:analysis*; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine