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Issue 11, 2018
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Development and clinical application of a LC-MS/MS method for simultaneous determination of one-carbon related amino acid metabolites in NTD tissues

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Abstract

Background: Neural tube defects are a group of severe neurological birth defects with one-carbon metabolism disturbance. The lack of information available on the distribution of one-carbon metabolites in lesion tissues precludes further elucidation of the pathogenesis of NTDs. Our aims were to develop and validate a simple and robust method for one-carbon related amino acid measurements in both intracellular and extracellular bio-samples and apply this method to NTD tissues for risk metabolite screening. Method: A simultaneous test for nine amino acids (homocysteine, S-adenosylhomocysteine, serine, histidine, methionine, cystathionine, cysteine, glycine, and tryptophan) in both intracellular and extracellular matrices was developed using liquid chromatography tandem mass spectrometry in a multiple reaction monitoring mode. Three kinds of tissues from folate deficient and NTD-affected embryos and controls in a NTD high-risk area were analyzed using this method. Results: Method recoveries ranged from 87.10% to 118.85% and accuracy (relative error, RE%) ranged from −14.96% to 13.96%. Calibration curves were linear (R2 > 0.99). The within-run and between-run precision were both lower than 15%. Reproducibility CV% ranged from 0.29% to 13.37% in low, medium and high concentration standard spiked plasma, HEK-293 cells, and brain, lung, heart tissues. NTD brain tissue showed significantly high concentration of homocysteine and cystathionine, and low concentration of histidine, glycine, methionine and tryptophan. Conclusion: This high-throughput method is reliable and accurate for simultaneous determination of one-carbon amino acid metabolites in both intracellular and extracellular bio-samples.

Graphical abstract: Development and clinical application of a LC-MS/MS method for simultaneous determination of one-carbon related amino acid metabolites in NTD tissues

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Publication details

The article was received on 16 Oct 2017, accepted on 01 Feb 2018 and first published on 01 Feb 2018


Article type: Paper
DOI: 10.1039/C7AY02434G
Citation: Anal. Methods, 2018,10, 1315-1324
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    Development and clinical application of a LC-MS/MS method for simultaneous determination of one-carbon related amino acid metabolites in NTD tissues

    M. Zhang, L. Wang, P. Pei, Y. Bao, J. Guo, L. Wang, S. Chang, X. Xie, H. Cheng, L. Quan and T. Zhang, Anal. Methods, 2018, 10, 1315
    DOI: 10.1039/C7AY02434G

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