Determination of amino acid neurotransmitters in mouse brain tissue using high-performance liquid chromatography with fluorescence detection

doi:10.5246/jcps.2013.03.034

Abstract

Abstract:

Amino acid neurotransmitters facilitate the transmission of nerve messages across the synapses and play essential roles in the control and regulation of a variety of functions in the central and peripheral nervous system. In this study, we developed a sensitive and efficient method using high-performance liquid chromatography (HPLC) with fluorescence detection for the assay of five important amino acid neurotransmitters. After derivatization with ο-phthaldialdehyde (OPA), aspartate (Asp), glutamic acid (Glu), glycine (Gly), taurine (Tau) and γ-aminobutyric acid (GABA) were simultaneously detected in the presence of the internal standard homoserine (Hse). Precise separation of these five amino acids was achieved using isocratic elution within 24 min. Good linearity was found over the concentration range with correlation coefficients (r²) not less than 0.9998. The limit of detection (LOD) values were no more than 10 nmol/L. The intra- and inter-day reproducibility was adequate with the relative standard deviation (RSD) of 10.5% or below. This method has also been applied to the analysis of amino acids in the substantia nigra and striatum samples obtained from C57BL/6 mice.

Key words: HPLC, Fluorescence detection, ο-Phthaldialdehyde, Amino acids

CLC Number:

R969

Supporting:

Mengmeng Wang, Wangchun Du, Jie Shen^*, Yi Dong, Wenshi Wei, Zhongjuan Song . Determination of amino acid neurotransmitters in mouse brain tissue using high-performance liquid chromatography with fluorescence detection[J]. , DOI: 10.5246/jcps.2013.03.034.

References

[1] Perry, M.; Li, Q.; Kennedy, R.T. Anal. Chim. Acta. 2009, 653, 1-22.
[2] Buck, K.; Voehringer, P.; Ferger, B. J. Neurosci. Methods. 2009, 182, 78-84.
[3] Grant, S.L.; Shulman, Y.; Tibbo, P.; Hampson, D.R.; Baker, G.B. J. Chromatogr. B. 2006, 844, 278-282.
[4] de Freitas, S.D.; Ferraz, V.P.; Ribeiro, A.M. J. Neurosci. Methods. 2009, 177, 289-293.
[5] Jimenez-Martin, E.; Ruiz, J.; Perez-Palacios, T.; Silva, A.; Antequera, T. J. Agric. Food Chem. 2012, 60, 2456-2463.
[6] Fatemi, M.H.; Elyasi, M. J. Sep. Sci. 2011, 34, 3216-3220.
[7] Hemmati, P.; Shilliam, C.S.; Hughes, Z.A.; Shah, A.J.; Roberts, J.C.; Atkins, A.R.; Hunter, A.J.; Heidbreder, C.A. Neurochem. Int. 2001, 39, 199-208.
[8] Kang, X.; Xiao, J.; Huang, X.; Gu, Z. Clin. Chim. Acta. 2006, 366, 352-356.
[9] Welty, N.; Shoblock, J.R. Psychopharmacology (Berl). 2009, 207, 433-438.
[10] Choi, J.; Choi, K.; Kim, J.; Ahmed, A.Y.; Al-Othman, Z.A.; Chung, D.S. J. Chromatogr. A. 2011, 1218, 7227-7233.
[11] Li, H.; Yan, Z.Y. Biomed. Chromatogr. 2010, 24, 1185-1192.
[12] Huang, Y.; Shi, M.; Zhao, S. J. Sep. Sci. 2009, 32, 3001-3006.
[13] Monge-Acuna, A.A.; Fornaguera-Trias, J. J. Neurosci. Methods. 2009, 183, 176-181.
[14] Tcherkas, Y.V.; Denisenko, A.D. J. Chromatogr. A. 2001, 913, 309-313.
[15] Deng, Y.H.; Wang, H.; Zhang, H.S. J. Sep. Sci. 2008, 31, 3088-3097.
[16] Hapuarachchi, S.; Aspinwall, C.A. Electrophoresis. 2007, 28, 1100-1106.
[17] Sun, Z.; You, J.; Li, G.; Zhao, X.; Suo, Y.; Wang, X. J. Chromatogr. B. 2011, 879, 1367-1374.
[18] Song, Y.; Feng, Y.; Lu, X.; Zhao, S.; Liu, C.W.; Liu, Y.M. Neurosci. Lett. 2008, 445, 53-57.
[19] Zhang, Z.X.; Gao, P.F.; Guo, X.F.; Wang, H.; Zhang, H.S. Anal. Bioanal. Chem. 2011, 401, 1905-1914.
[20] Zhao, X.E.; Suo, Y.R. Talanta. 2008, 76, 690-697.
[21] Musgrave, T.; Tenorio, G.; Rauw, G.; Baker, G.B.; Kerr, B.J. Neurochem. Int. 2011, 59, 28-38.
[22] Vallee, N.; Rostain, J.C.; Boussuges, A.; Risso, J.J. Neurochem. Res. 2009, 34, 835-844.
[23] Burton, N.C.; Schneider, J.S.; Syversen, T.; Guilarte, T.R. Toxicol. Sci. 2009, 111, 131-139.
[24] Schwarz, E.L.; Roberts, W.L.; Pasquali, M. Clin. Chim. Acta. 2005, 354, 83-90.
[25] Mercolini, L.; Mandrioli, R.; Amore, M.; Raggi, M.A. J. Pharm. Biomed. Anal. 2010, 53, 62-67.
[26] Oreiro-Garcia, M.T.; Vazquez-Illanes, M.D.; Sierra-Paredes, G.; Sierra-Marcuno, G. Biomed. Chromatogr. 2005, 19, 720-724.
[27] Sierra-Paredes, G.; Oreiro-Garcia, M.T.; Vazquez-Illanes, M.D.; Sierra-Marcuno, G. Epilepsy Res. 2007, 77, 36-43.
[28] Siri, N.; Lacroix, M.; Garrigues, J.C.; Poinsot, V.; Couderc, F. Electrophoresis. 2006, 27, 4446-4455.
[29] Jambor, A.; Molnar-Perl, I. J. Chromatogr. A. 2009, 1216, 3064-3077.
[30] Kehr, J. J. Chromatogr. B. 1998, 708, 49-54.
[31] Kehr, J. J. Chromatogr. B. 1998, 708, 27-38.
[32] Toner, C.C.; Connelly, K.; Whelpton, R.; Bains, S.; Michael-Titus, A.T.; McLaughlin, D.P.; Stamford, J.A. Br. J. Anaesth. 2001, 86, 550-554.
[33] Rauw, G.A.; Grant, S.L.; Labrie, V.; Roder, J.C.; Antflick, J.E.; Hampson, D.R.; Baker, G.B. Anal. Biochem. 2010, 405, 260-262.
[34] Devall, A.J.; Blake, R.; Langman, N.; Smith, C.G.; Richards, D.A.; Whitehead, K.J. J. Chromatogr. B. 2007, 848, 323-328.
[35] Guidance for Industry, Bioanalytical Method Validation, U.S. Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), Center for Veterinary Medicine (CVM). 2001.
[36] Brodie, R.R.; Hill, H.M. Chromatographia. 2002, 55, S91-S94.