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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 NTDs tissues

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 one carbon metabolites distribution in lesion tissues precludes further elucidation of pathogenesis of NTDs. Our aims were to develop and validate a simple and robust method for one carbon related amino acids measurement in both intracellular and extracellular bio-samples and apply this method in NTDs tissues for risk metabolites screening. Method: A simultaneous test for nine amino acids (homocysteine, S-adenosylhomocysteine, serine, histidine, methionine, cystathionine, cysteine, glycine, tryptophan) in both intracellular and extracellular matrix was developed using liquid chromatography tandem mass spectrometry in a multiple reaction monitoring mode. Three kinds of tissues from folate deficiency and NTDs-affected embryos and controls in a NTDs 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, brain, lung, heart and HEK-293 cells. NTDs brain showed signif-icantly high concentration of homocysteine and cystathionine, and low concentration of histidine, glycine, methionine and tryptophan. Conclusion: This high-throughput method is reliable and accuracy for simultaneous determination of one carbon amino acid metabolites in both intracellular and extracellular bio-samples.

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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, Accepted Manuscript
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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 NTDs 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, Accepted Manuscript , DOI: 10.1039/C7AY02434G

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