4.7 Article

Two mass-spectrometric techniques for quantifying serine enantiomers and glycine in cerebrospinal fluid: Potential confounders and age-dependent ranges

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CLINICAL CHEMISTRY
卷 54, 期 9, 页码 1443-1450

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AMER ASSOC CLINICAL CHEMISTRY
DOI: 10.1373/clinchem.2007.100412

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  1. Department of Metabolic and Endocrine Diseases
  2. University Medical Center Utrecht
  3. Laboratory for Medical Microbiology
  4. Laboratory of Metabolic and Endocrine Diseases
  5. Department of Neonatology of the University Medical Center Utrecht

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BACKGROUND: The recent discovery and specific functions of D-amino acids in humans are bound to lead to the revelation Of D-amino acid abnormalities in human disorders. Therefore, high-throughput analysis techniques are warranted to determine D-amino acids in biological fluids in a routine laboratory setting. METHODS: We developed 2 chromatographic techniques, a nonchiral derivatization with chiral (chirasil-L-val column) separation in a GC-MS system and a chiral derivatization with Marfey's reagent and LC-MS analysis. We validated the techniques for D-serine, L-serine, and glycine determination in cerebrospinal fluid (CSF), evaluated several confounders, and determined age-dependent human concentration ranges. RESULTS: Quantification limits for D-serine, L-serine, and glycine in cerebrospinal fluid were 0.14, 0.44, and 0.14 mu mol/L, respectively, for GC-MS and 0.20, 0.41, and 0.14 mu mol/L for LC-MS. Within-run imprecision was <3% for both methods, and between-run imprecision was <13%. Comparison of both techniques with Deming regression yielded coefficients of 0.90 (D-serine), 0.92 (L-serine), and 0.96 (glycine). Sample collection, handling, and transport is uncomplicated - there is no rostrocaudal CSF gradient, no effect of storage at 4 C for 1 week before storage at -80 degrees C, and no effect of up to 3 freeze/thaw cycles. Conversely, contatnination with erythrocytes increased D-serine, L-serine, and glycine concentrations. CSF concentrations for 145 apparently healthy controls demonstrated markedly and specifically increased (5 to 9 times) D-serine concentrations during early central nervous System development. CONCLUSIONS: These 2 clinically applicable analysis techniques will help to unravel pathophysiologic, diagnostic, and therapeutic issues for disorders associated with central nervous system abnormalities, NMDA-receptor dysfunction, and other pathology associated with p-amino acids. (C) 2008 American Association for Clinical Chemistry.

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