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Stable isotope dilution analysis of N-acetylaspartic acid in urine by liquid chromatography electrospray ionization tandem mass spectrometry.
Biomed Chromatogr. 2007 Sep; 21(9):898-902.BC

Abstract

N-acetylaspartic acid (NAA) is a specific urinary marker for Canavan disease, an autosomal recessive leukodystrophy. We developed a 'dilute and shoot' stable isotope dilution liquid chromatography tandem mass spectrometry (LC-MS/MS) method for determination of NAA in urine. Deuterated internal standard d(3)-NAA was added to untreated urine and the mixture was injected into the LC-MS/MS system operated in the negative ion mode. Chromatography was carried out on a C(8) minibore column using 50% acetonitrile solution containing 0.05% formic acid at a flow rate of 0.25 mL/min. The retention time was 1.6 min and the turnaround time was 2.2 min. NAA and d(3)-NAA were analyzed in multiple reaction monitoring mode. Calibrators and quality control samples were prepared in pooled control urine. The assay was linear up to 2000 micromol/L with limit of quantification at 1 micromol/L (S/N = 12). Interassay and intraassay coefficients of variation were less than 7% and recovery at three different concentrations was 98.9-102.5%. The LC-MS/MS method for NAA as described involves no extraction and no derivatization, showed no interference and gave excellent recovery with low variability and short analytical time. The method was successfully applied for the retrospective analysis of urine from 21 Canavan disease cases.

Authors+Show Affiliations

Al-Dirbashi OY
National Laboratory for Newborn Screening, King Faisal Specialist Hospital and Research Centre, PO Box 3354, Riyadh 11211, Saudi Arabia.
Rashed MS
No affiliation info available
Al-Mokhadab MA
No affiliation info available
Al-Qahtani K
No affiliation info available
Al-Sayed MA
No affiliation info available
Kurdi W
No affiliation info available

MeSH

Aspartic AcidChromatography, High Pressure LiquidHumansIsotopesReference StandardsReproducibility of ResultsSpectrometry, Mass, Electrospray IonizationTandem Mass Spectrometry

Pub Type(s)

Journal Article
Validation Study

Language

eng

PubMed ID

17441217

Citation

Al-Dirbashi, Osama Y., et al. "Stable Isotope Dilution Analysis of N-acetylaspartic Acid in Urine By Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry." Biomedical Chromatography : BMC, vol. 21, no. 9, 2007, pp. 898-902.
Al-Dirbashi OY, Rashed MS, Al-Mokhadab MA, et al. Stable isotope dilution analysis of N-acetylaspartic acid in urine by liquid chromatography electrospray ionization tandem mass spectrometry. Biomed Chromatogr. 2007;21(9):898-902.
Al-Dirbashi, O. Y., Rashed, M. S., Al-Mokhadab, M. A., Al-Qahtani, K., Al-Sayed, M. A., & Kurdi, W. (2007). Stable isotope dilution analysis of N-acetylaspartic acid in urine by liquid chromatography electrospray ionization tandem mass spectrometry. Biomedical Chromatography : BMC, 21(9), 898-902.
Al-Dirbashi OY, et al. Stable Isotope Dilution Analysis of N-acetylaspartic Acid in Urine By Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry. Biomed Chromatogr. 2007;21(9):898-902. PubMed PMID: 17441217.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Stable isotope dilution analysis of N-acetylaspartic acid in urine by liquid chromatography electrospray ionization tandem mass spectrometry. AU - Al-Dirbashi,Osama Y, AU - Rashed,Mohamed S, AU - Al-Mokhadab,Manhal A, AU - Al-Qahtani,Khalid, AU - Al-Sayed,Moeen A A, AU - Kurdi,Wesam, PY - 2007/4/19/pubmed PY - 2007/11/8/medline PY - 2007/4/19/entrez SP - 898 EP - 902 JF - Biomedical chromatography : BMC JO - Biomed Chromatogr VL - 21 IS - 9 N2 - N-acetylaspartic acid (NAA) is a specific urinary marker for Canavan disease, an autosomal recessive leukodystrophy. We developed a 'dilute and shoot' stable isotope dilution liquid chromatography tandem mass spectrometry (LC-MS/MS) method for determination of NAA in urine. Deuterated internal standard d(3)-NAA was added to untreated urine and the mixture was injected into the LC-MS/MS system operated in the negative ion mode. Chromatography was carried out on a C(8) minibore column using 50% acetonitrile solution containing 0.05% formic acid at a flow rate of 0.25 mL/min. The retention time was 1.6 min and the turnaround time was 2.2 min. NAA and d(3)-NAA were analyzed in multiple reaction monitoring mode. Calibrators and quality control samples were prepared in pooled control urine. The assay was linear up to 2000 micromol/L with limit of quantification at 1 micromol/L (S/N = 12). Interassay and intraassay coefficients of variation were less than 7% and recovery at three different concentrations was 98.9-102.5%. The LC-MS/MS method for NAA as described involves no extraction and no derivatization, showed no interference and gave excellent recovery with low variability and short analytical time. The method was successfully applied for the retrospective analysis of urine from 21 Canavan disease cases. SN - 0269-3879 UR - https://www.unboundmedicine.com/medline/citation/17441217/Stable_isotope_dilution_analysis_of_N_acetylaspartic_acid_in_urine_by_liquid_chromatography_electrospray_ionization_tandem_mass_spectrometry_ L2 - https://doi.org/10.1002/bmc.815 DB - PRIME DP - Unbound Medicine ER -
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