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 micromol/L, respectively, for GC-MS and 0.20, 0.41, and 0.14 micromol/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 degrees C for 1 week before storage at -80 degrees C, and no effect of up to 3 freeze/thaw cycles. Conversely, contamination 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 D-amino acids.
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 micromol/L, respectively, for GC-MS and 0.20, 0.41, and 0.14 micromol/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 degrees C for 1 week before storage at -80 degrees C, and no effect of up to 3 freeze/thaw cycles. Conversely, contamination 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 D-amino acids.
Authors: Robert B Clark; Jorge L Cervantes; Mark W Maciejewski; Vahid Farrokhi; Reza Nemati; Xudong Yao; Emily Anstadt; Mai Fujiwara; Kyle T Wright; Caroline Riddle; Carson J La Vake; Juan C Salazar; Sydney Finegold; Frank C Nichols Journal: Infect Immun Date: 2013-07-08 Impact factor: 3.441
Authors: Frank C Nichols; Robert B Clark; Mark W Maciejewski; Anthony A Provatas; Jeremy L Balsbaugh; Floyd E Dewhirst; Michael B Smith; Amanda Rahmlow Journal: J Lipid Res Date: 2020-09-10 Impact factor: 5.922
Authors: M E Wimmer; L A Briand; B Fant; L A Guercio; A C Arreola; H D Schmidt; S Sidoli; Y Han; B A Garcia; R C Pierce Journal: Mol Psychiatry Date: 2017-02-21 Impact factor: 15.992
Authors: Sabine A Fuchs; Cacha M P C D Peeters-Scholte; Martina M J de Barse; Martin W Roeleveld; Leo W J Klomp; Ruud Berger; Tom J de Koning Journal: Amino Acids Date: 2011-09-23 Impact factor: 3.520
Authors: Hanneke A Haijes; Eline A J Willemse; Johan Gerrits; Wiesje M van der Flier; Charlotte E Teunissen; Nanda M Verhoeven-Duif; Judith J M Jans Journal: Metabolites Date: 2019-10-18
Authors: Michael A Swanson; Kristen Miller; Sarah P Young; Suhong Tong; Lina Ghaloul-Gonzalez; Juanita Neira-Fresneda; Lisa Schlichting; Cheryl Peck; Linda Gabel; Marisa W Friederich; Johan L K Van Hove Journal: J Inherit Metab Dis Date: 2022-04-06 Impact factor: 4.750