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Quantification of free and total sialic acid excretion by LC-MS/MS

van der Ham, Maria ; Prinsen, Berthil H.C.M.T. ; Huijmans, Jan G.M. ; Abeling, Nicolaas G.G.M. ; Dorland, Bert ; Berger, Ruud ; de Koning, Tom J. LU and de Sain-van der Velden, Monique G.M. (2007) In Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences 848(2). p.251-257
Abstract

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... (More)

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 μmol/L, respectively. Limit of quantification for FSA and TSA was 1.0 and 5.0 μmol/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 μmol/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.

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author
; ; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Galactosialidosis, Hemolytic uremic syndrome, Salla disease, Sialic acid, Sialidosis, Sialuria
in
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
volume
848
issue
2
pages
7 pages
publisher
Elsevier
external identifiers
  • pmid:17123874
  • scopus:33947533124
ISSN
1570-0232
DOI
10.1016/j.jchromb.2006.10.066
language
English
LU publication?
no
id
2424b82f-e633-4d38-a59c-09273e569f50
date added to LUP
2020-02-28 13:57:47
date last changed
2024-06-26 12:51:02
@article{2424b82f-e633-4d38-a59c-09273e569f50,
  abstract     = {{<p>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. <sup>13</sup>C<sub>3</sub>-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 (<sup>13</sup>C<sub>3</sub>-SA). CSA was calculated by subtracting FSA from TSA. Results: Limit of detection for FSA and TSA was 0.3 and 1.7 μmol/L, respectively. Limit of quantification for FSA and TSA was 1.0 and 5.0 μmol/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 μmol/L (r<sup>2</sup> = 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.</p>}},
  author       = {{van der Ham, Maria and Prinsen, Berthil H.C.M.T. and Huijmans, Jan G.M. and Abeling, Nicolaas G.G.M. and Dorland, Bert and Berger, Ruud and de Koning, Tom J. and de Sain-van der Velden, Monique G.M.}},
  issn         = {{1570-0232}},
  keywords     = {{Galactosialidosis; Hemolytic uremic syndrome; Salla disease; Sialic acid; Sialidosis; Sialuria}},
  language     = {{eng}},
  month        = {{04}},
  number       = {{2}},
  pages        = {{251--257}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences}},
  title        = {{Quantification of free and total sialic acid excretion by LC-MS/MS}},
  url          = {{http://dx.doi.org/10.1016/j.jchromb.2006.10.066}},
  doi          = {{10.1016/j.jchromb.2006.10.066}},
  volume       = {{848}},
  year         = {{2007}},
}