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Liquid chromatography-mass spectrometry with triazole-bonded stationary phase for N-methyl-d-aspartate receptor-related amino acids: development and application in microdialysis studies

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Abstract

We aimed to monitor changes in the levels of amino acid neurotransmitters or neuromodulators simultaneously at the synaptic clefts of experimental animals. We developed a method for the simultaneous determination of the levels of amino acids, such as d-Ser, Gly, and l-Glu, which were involved in neurotransmission via the N-methyl-d-aspartate (NMDA) receptor, and other protein-constituted amino acids in a rat brain microdialysis (MD) sample. We used a liquid chromatography-mass spectrometry (LC-MS)/MS device equipped with a triazole-bonded column. The determination was achieved without using stable isotope-labeled compounds. We instead used suitable amino acid analogues as internal standards (ISs). We examined various analyte-IS combinations to improve reproducibility. We found a positive correlation (r = 0.720, **p < 0.0001) between relative standard deviation (%) of the area ratio and the analyte-IS retention time differences. Using the proposed method, we were able to accurately analyze trace amounts of amino acids in MD samples using ISs that were structurally similar to the analytes. Furthermore, we observed that the peripheral administration of S-methyl-l-cysteine, which was an inhibitor of the amino acid transporter Asc-1, caused some amino acid level changes in the rat brain. The proposed LC-MS/MS method can be applied in vivo to study the effects of novel therapeutic agents with monitoring the levels of amino acid neuromodulators, such as Glu, Gly, GABA, and d-Ser, in the brain.

LC-MS/MS analysis of amino acid enantiomers in microdialysis samples from rat striatum using triazole-bonded stationary phase

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References

  1. Wroblewski JT, Fadda E, Mazzetta J, Lazarewicz JW, Costa E. Glycine and D-serine act as positive modulators of signal transduction at N-methyl-D-aspartate sensitive glutamate receptors in cultured cerebellar granule cells. Neuropharmacology. 1989;28:447–52.

    Article  CAS  Google Scholar 

  2. Van Horn MR, Sild M, Ruthazer ES. D-serine as a gliotransmitter and its roles in brain development and disease. Front Cell Neurosci. 2013;7:39.

    Article  Google Scholar 

  3. Schell M, Brady R, Molliver M, Snyder S. D-Serine as a neuromodulator: regional and developmental localizations in rat brain glia resemble NMDA receptors. J Neurosci. 1997;17:1604–15.

    CAS  Google Scholar 

  4. Schell MJ, Molliver ME, Snyder SH. D-serine, an endogenous synaptic modulator—localization to astrocytes and glutamate-stimulated release. Proc Natl Acad Sci U S A. 1995;92:3948–52.

    Article  CAS  Google Scholar 

  5. Pålsson E, Jakobsson J, Södersten K, Fujita Y, Sellgren C, Ekman CJ, et al. Markers of glutamate signaling in cerebrospinal fluid and serum from patients with bipolar disorder and healthy controls. Eur Neuropsychopharmacol. 2015;25:133–40.

    Article  Google Scholar 

  6. Chouinard ML, Gaitan D, Wood PL. Presence of the N-methyl-D-aspartate-associated glycine receptor agonist, D-serine, in human temporal cortex: comparison of normal, Parkinson, and Alzheimer tissues. J Neurochem. 1993;61:1561–4.

    Article  CAS  Google Scholar 

  7. Bardaweel SK, Alzweiri M, Ishaqat AA. D-Serine in neurobiology: CNS neurotransmission and neuromodulation. Can J Neurol Sci. 2014;41:164–76.

    Article  Google Scholar 

  8. Kegel ME, Bhat M, Skogh E, Samuelsson M, Lundberg K, Dahl ML, et al. Imbalanced kynurenine pathway in schizophrenia. Int J Tryptophan Res. 2014;7:15–22.

    Article  CAS  Google Scholar 

  9. Nilsson LK, Linderholm KR, Engberg G, Paulson L, Blennow K, Lindström LH, et al. Elevated levels of kynurenic acid in the cerebrospinal fluid of male patients with schizophrenia. Schizophr Res. 2005;80:315–22.

    Article  CAS  Google Scholar 

  10. Hashimoto K, Fukushima T, Shimizu E, Komatsu N, Watanabe H, Shinoda N, et al. Decreased serum levels of D-serine in patients with schizophrenia—evidence in support of the N-methyl-D-aspartate receptor hypofunction hypothesis of schizophrenia. Arch Gen Psychiatry. 2003;60:572–6.

    Article  CAS  Google Scholar 

  11. Fukushima T, Iizuka H, Yokota A, Suzuki T, Ohno C, Kono Y, et al. Quantitative analyses of schizophrenia-associated metabolites in serum: serum D-lactate levels are negatively correlated with gamma-glutamylcysteine in medicated schizophrenia patients. PLoS One. 2014;9:e101652.

    Article  Google Scholar 

  12. Calcia Marilia A, Madeira C, Alheira Flavio V, Silva Thuany CS, Tannos Filippe M, Vargas-Lopes C, et al. Plasma levels of D-serine in Brazilian individuals with schizophrenia. Schizophr Res. 2012;142:83–7.

    Article  CAS  Google Scholar 

  13. Fukushima T, Kawai J, Imai K, Toyo'oka T. Simultaneous determination of D- and L-serine in rat brain microdialysis sample using a column-switching HPLC with fluorimetric detection. Biomed Chromatogr. 2004;18:813–9.

    Article  CAS  Google Scholar 

  14. Heemskerk AA, Busnel JM, Schoenmaker B, Derks RJ, Klychnikov O, Hensbergen PJ, et al. Ultra-low flow electrospray ionization-mass spectrometry for improved ionization efficiency in phosphoproteomics. Anal Chem. 2012;84:4552–9.

    Article  CAS  Google Scholar 

  15. Li Z, Han J, Sun SA, Chen K, Tang DQ. Hydrophilic interaction liquid chromatography/mass spectrometry: an attractive and prospective method for the quantitative bioanalysis in drug metabolism. Curr Drug Metab. 2016;17:386–400.

    Article  CAS  Google Scholar 

  16. Simon R, Enjalbert Q, Biarc J, Lemoine J, Salvador A. Evaluation of hydrophilic interaction chromatography (HILIC) versus C18 reversed-phase chromatography for targeted quantification of peptides by mass spectrometry. J Chromatogr A. 2012;1264:31–9.

    Article  CAS  Google Scholar 

  17. Huang ZY, Francis R, Zha Y, Ruan J. Development of a simple method for quantitation of methanesulfonic acid at low ppm level using hydrophilic interaction chromatography coupled with ESI-MS. J Pharm Biomed Anal. 2015;102:17–24.

    Article  CAS  Google Scholar 

  18. Iwasaki M, Kashiwaguma Y, Nagashima C, Izumi M, Uekusa A, Iwasa S, et al. A high-performance liquid chromatography assay with a triazole-bonded column for evaluation of d-amino acid oxidase activity. Biomed Chromatogr. 2015;30:384–9.

    Article  Google Scholar 

  19. Sakamoto T, Kuwabara R, Takahashi S, Onozato M, Ichiba H, Iizuka H, et al. Determination of D-serine in human serum by LC-MS/MS using a triazole-bonded column after pre-column derivatization with (S)-4-(3-isothiocyanatopyrrolidin-1-yl)-7- (N, N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole. Anal Bioanal Chem. 2016;408:517–26.

    Article  CAS  Google Scholar 

  20. Godfrey AR, Jones L, Davies M, Townsend R. Miltefosine: a novel internal standard approach to lysophospholipid quantitation using LC-MS/MS. Anal Bioanal Chem. 2017;409:2791–800.

    Article  CAS  Google Scholar 

  21. Krautbauer S, Büchler C, Liebisch G. Relevance in the use of appropriate internal standards for accurate quantification using LC-MS/MS: tauro-conjugated bile acids as an example. Anal Chem. 2016;88:10957–61.

    Article  CAS  Google Scholar 

  22. Ishii K, Iizuka H, Ogaya T, Song Z, Fukushima T. Comparative study on kynurenic acid production in the rat striatu by tryptophan enantiomers: an in vivo microdialysis study. Chirality. 2011;23:E12–5.

    Article  CAS  Google Scholar 

  23. Chamkasem N, Harmon T. Direct determination of glyphosate, glufosinate, and AMPA in soybean and corn by liquid chromatography/tandem mass spectrometry. Anal Bioanal Chem. 2016;408:4995–5004.

    Article  CAS  Google Scholar 

  24. Jin D, Nagakura K, Murofushi S, Miyahara T, Toyo'oka T. Total resolution of 17 DL-amino acids labelled with a fluorescent chiral reagent, R(-)-4-(3-isothiocyanatopyrrolidin-1-y1)-7-(N,N-dimethylaminosulfonyl)- 2,1,3-benzoxadiazole, by high-performance liquid chromatography. J Chromatogr A. 1998;822:215–24.

    Article  CAS  Google Scholar 

  25. Zhang M, Fang C, Smagin G. Derivatization for the simultaneous LC/MS quantification of multiple neurotransmitters in extracellular fluid from rat brain microdialysis. J Pharm Biomed Anal. 2014;100:357–64.

    Article  CAS  Google Scholar 

  26. Berna MJ, Ackermann BL. Quantification of serine enantiomers in rat brain microdialysate using Marfey’s reagent and LC/MS/MS. J Chromatogr B Anal Technol Biomed Life Sci. 2007;846:359–63.

    Article  CAS  Google Scholar 

  27. Song P, Mabrouk OS, Hershey ND, Kennedy RT. In vivo neurochemical monitoring using benzoyl chloride derivatization and liquid chromatography-mass spectrometry. Anal Chem. 2012;84:412–9.

    Article  CAS  Google Scholar 

  28. Utsunomiya-Tate N, Endou H, Kanai Y. Cloning and functional characterization of a system ASC-like Na+-dependent neutral amino acid transporter. J Biol Chem. 1996;271:14883–90.

    Article  CAS  Google Scholar 

  29. Fukasawa Y, Segawa H, Kim JY, Chairoungdua A, Kim DK, Matsuo H, et al. Identification and characterization of a Na(+)-independent neutral amino acid transporter that associates with the 4F2 heavy chain and exhibits substrate selectivity for small neutral D- and L-amino acids. J Biol Chem. 2000;275:9690–8.

    Article  CAS  Google Scholar 

  30. Ishiwata S, Ogata S, Umino A, Shiraku H, Ohashi Y, Kajii Y, et al. Increasing effects of S-methyl-L-cysteine on the extracellular D-serine concentrations in the rat medial frontal cortex. Amino Acids. 2013;44:1391–5.

    Article  CAS  Google Scholar 

  31. Onozato M, Ishimaru K, Nagashima C, Fukumoto M, Nakazawa H, Shishikura M, et al. S-Methyl-l-cysteine levels in plasma and striatum following its intraperitoneal administration and the effects on striatal D-serine levels in rats: an in vivo microdialysis study. RRJPPS. 2016;5:6.

    Google Scholar 

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Acknowledgements

The authors thank Mr. Tago for his technical assistance. The authors would also like to thank Shimadzu Access Corporation for the equipment conservation and technical advice. We also thank Editage (www.editage.jp) for the English language editing.

Sources of funding

This work was supported by the Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C) (Grant Number 25460224), and the Faculty of Pharmaceutical Science, Toho University.

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Correspondence to Takeshi Fukushima.

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The authors declare that they have no conflicts of interest.

Animal experiments

All of the animal experiments were performed in accordance with the Guide for the Care and Use of Laboratory Animals at Faculty of Pharmaceutical Sciences, Toho University. The project of animal experiments was approved by the Animal Care and Ethics Committee of Toho University (approval number: 16-55-165).

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Sakamoto, T., Onuma, R., Furukawa, S. et al. Liquid chromatography-mass spectrometry with triazole-bonded stationary phase for N-methyl-d-aspartate receptor-related amino acids: development and application in microdialysis studies. Anal Bioanal Chem 409, 7201–7210 (2017). https://doi.org/10.1007/s00216-017-0682-2

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  • DOI: https://doi.org/10.1007/s00216-017-0682-2

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