Document Detail


Quantification of free and total sialic acid excretion by LC-MS/MS.
MedLine Citation:
PMID:  17123874     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: The main purpose for measuring urinary free sialic acid (FSA) is to diagnose sialic acid (SA) storage diseases. Elevated amounts of conjugated sialic acid (CSA) are observed in several diseases indicating the need to quantify CSA as well. A LC-MS/MS method for quantification of FSA and total sialic acid (TSA) in urine is developed and validated. METHODS: FSA is analyzed directly after filtration of urine samples. For determination of TSA an enzymatic (neuraminidase) and a chemical (acid) hydrolysis were compared. 13C3-sialic acid was used as internal standard. LC-MS/MS was performed in negative electrospray ionisation mode with multiple reaction monitoring of transitions m/z 308.2-->87.0 (SA) and m/z 311.2-->90.0 (13C3-SA). CSA was calculated by subtracting FSA from TSA. RESULTS: Limit of detection for FSA and TSA was 0.3 and 1.7 micromol/L, respectively. Limit of quantification for FSA and TSA was 1.0 and 5.0 micromol/L. Intra- and inter-assay variations of FSA were 4.6% and 6.6% (n=10) for FSA and 6.5% and 3.6% (n=10) for TSA. Linearity was tested till 7800 micromol/L (r2=0.9998). Values of SA analyzed after neuraminidase- or acid hydrolysis treatment were comparable. Urine samples from patients with inborn errors of SA (related) metabolism were analyzed and compared with age-related reference values. CONCLUSION: A method has been developed for routine determination of urinary FSA and TSA. The method is rapid, specific, robust and sensitive. Age-related reference values for FSA, TSA and CSA were determined and improved diagnostic efficacy.
Authors:
Maria van der Ham; Berthil H C M T Prinsen; Jan G M Huijmans; Nicolaas G G M Abeling; Bert Dorland; Ruud Berger; Tom J de Koning; Monique G M de Sain-van der Velden
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Publication Detail:
Type:  Journal Article     Date:  2006-11-22
Journal Detail:
Title:  Journal of chromatography. B, Analytical technologies in the biomedical and life sciences     Volume:  848     ISSN:  1570-0232     ISO Abbreviation:  J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.     Publication Date:  2007 Apr 
Date Detail:
Created Date:  2007-03-27     Completed Date:  2007-06-11     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101139554     Medline TA:  J Chromatogr B Analyt Technol Biomed Life Sci     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  251-7     Citation Subset:  IM    
Affiliation:
Department of Metabolic and Endocrine Diseases, University Medical Centre Utrecht, The Netherlands.
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MeSH Terms
Descriptor/Qualifier:
Adolescent
Adult
Aged
Child
Child, Preschool
Chromatography, Liquid / methods*
Chromatography, Thin Layer
Humans
Infant
Infant, Newborn
Middle Aged
N-Acetylneuraminic Acid / analysis,  urine*
Reproducibility of Results
Sialic Acid Storage Disease / urine
Tandem Mass Spectrometry / methods*
Chemical
Reg. No./Substance:
131-48-6/N-Acetylneuraminic Acid

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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