Quantitation of cytidine-5'-monophospho-N-acetylneuraminic acid in human leukocytes using LC-MS/MS: method development and validation.
LC-MS/MS
cytidine-5′-monophospho-N-acetylneuraminic acid
human leukocytes
hydrophilic interaction chromatography
Journal
Biomedical chromatography : BMC
ISSN: 1099-0801
Titre abrégé: Biomed Chromatogr
Pays: England
ID NLM: 8610241
Informations de publication
Date de publication:
Feb 2020
Feb 2020
Historique:
received:
26
08
2019
revised:
18
10
2019
accepted:
21
10
2019
pubmed:
7
11
2019
medline:
26
2
2020
entrez:
7
11
2019
Statut:
ppublish
Résumé
The biosynthesis of sialic acid (Neu5Ac) leads to the intracellular production of cytidine-5'-monophospho-N-acetylneuraminic acid (CMP-Neu5Ac), the active sialic acid donor to nascent glycans (glycoproteins and glycolipids) in the Golgi. UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase myopathy is a rare autosomal recessive muscular disease characterized by progressive muscle weakness and atrophy. To quantify the intracellular levels of CMP-Neu5Ac as well as N-acetylmannosamine (ManNAc) and Neu5Ac in human leukocytes, we developed and validated robust liquid chromatography-tandem mass spectrometry methods. A fit-for-purpose approach was implemented for method validation. Hydrophilic interaction chromatography was used to retain three hydrophilic analytes. The human leukocyte pellets were lysed and extracted in a methanol-water mixture and the leukocyte extract was used for LC-MS/MS analysis. The lower limits of quantitation for ManNAc, Neu5Ac and CMP-Neu5Ac were 25.0, 25.0 and 10.0 ng/ml, respectively. These validated methods were applied to a clinical study.
Identifiants
pubmed: 31691999
doi: 10.1002/bmc.4735
pmc: PMC8336284
mid: NIHMS1728435
doi:
Substances chimiques
Sialic Acids
0
cytidine-5'-monophosphosialic acid
0
Cytidine Monophosphate
F469818O25
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e4735Subventions
Organisme : Intramural NIH HHS
ID : Z01 HG000215
Pays : United States
Organisme : Intramural NIH HHS
ID : ZIB TR000002
Pays : United States
Organisme : NCATS NIH HHS
Pays : United States
Organisme : NHGRI NIH HHS
Pays : United States
Informations de copyright
© 2019 John Wiley & Sons, Ltd.
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