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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†
Analytical Methods ( IF 2.7 ) Pub Date : 2018-02-01 00:00:00 , DOI: 10.1039/c7ay02434g Min Zhang 1, 2, 3, 4 , Lei Wang 1, 2, 3, 4 , Pei Pei 1, 2, 3, 4 , YiHua Bao 1, 2, 3, 4 , Jin Guo 1, 2, 3, 4 , Li Wang 1, 2, 3, 4 , ShaoYan Chang 1, 2, 3, 4 , XiaoLu Xie 1, 2, 3, 4 , HaiQin Cheng 1, 2, 3, 4 , Li Quan 1, 2, 3, 4 , Ting Zhang 1, 2, 3, 4
Analytical Methods ( IF 2.7 ) Pub Date : 2018-02-01 00:00:00 , DOI: 10.1039/c7ay02434g Min Zhang 1, 2, 3, 4 , Lei Wang 1, 2, 3, 4 , Pei Pei 1, 2, 3, 4 , YiHua Bao 1, 2, 3, 4 , Jin Guo 1, 2, 3, 4 , Li Wang 1, 2, 3, 4 , ShaoYan Chang 1, 2, 3, 4 , XiaoLu Xie 1, 2, 3, 4 , HaiQin Cheng 1, 2, 3, 4 , Li Quan 1, 2, 3, 4 , Ting Zhang 1, 2, 3, 4
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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.
中文翻译：
同时测定NTD组织中一碳相关氨基酸代谢物的LC-MS / MS方法的开发和临床应用†
背景：神经管缺陷是一组严重的神经系统先天性缺陷，伴有一碳代谢紊乱。缺乏有关病变组织中一碳代谢物分布的可用信息，无法进一步阐明NTD的发病机理。我们的目标是开发和验证一种简单而可靠的方法，用于在细胞内和细胞外生物样品中进行一碳相关氨基酸的测定，并将该方法应用于NTD组织进行风险代谢物筛查。方法：同时测试九种氨基酸（同型半胱氨酸，S-细胞内和细胞外基质中的α-腺苷同型半胱氨酸，丝氨酸，组氨酸，蛋氨酸，胱硫醚，半胱氨酸，甘氨酸和色氨酸）是通过液相色谱串联质谱法在多反应监测模式下开发的。使用此方法分析了叶酸缺乏和受NTD感染的胚胎以及NTD高风险区域中的对照的三种组织。结果：方法的回收率在87.10％至118.85％之间，准确度（相对误差，RE％）在-14.96％至13.96％之间。校准曲线是线性的（R 2> 0.99）。批内和批间精度均低于15％。在低，中和高浓度标准加标血浆，HEK-293细胞以及脑，肺，心脏组织中，重现性CV％范围从0.29％到13.37％。NTD脑组织中高半胱氨酸和胱硫醚的浓度很高，而组氨酸，甘氨酸，蛋氨酸和色氨酸的浓度却很低。结论：该高通量方法可同时测定细胞内和细胞外生物样品中的一碳氨基酸代谢产物，可靠，准确。
更新日期：2018-02-01
中文翻译：
同时测定NTD组织中一碳相关氨基酸代谢物的LC-MS / MS方法的开发和临床应用†
背景：神经管缺陷是一组严重的神经系统先天性缺陷，伴有一碳代谢紊乱。缺乏有关病变组织中一碳代谢物分布的可用信息，无法进一步阐明NTD的发病机理。我们的目标是开发和验证一种简单而可靠的方法，用于在细胞内和细胞外生物样品中进行一碳相关氨基酸的测定，并将该方法应用于NTD组织进行风险代谢物筛查。方法：同时测试九种氨基酸（同型半胱氨酸，S-细胞内和细胞外基质中的α-腺苷同型半胱氨酸，丝氨酸，组氨酸，蛋氨酸，胱硫醚，半胱氨酸，甘氨酸和色氨酸）是通过液相色谱串联质谱法在多反应监测模式下开发的。使用此方法分析了叶酸缺乏和受NTD感染的胚胎以及NTD高风险区域中的对照的三种组织。结果：方法的回收率在87.10％至118.85％之间，准确度（相对误差，RE％）在-14.96％至13.96％之间。校准曲线是线性的（R 2> 0.99）。批内和批间精度均低于15％。在低，中和高浓度标准加标血浆，HEK-293细胞以及脑，肺，心脏组织中，重现性CV％范围从0.29％到13.37％。NTD脑组织中高半胱氨酸和胱硫醚的浓度很高，而组氨酸，甘氨酸，蛋氨酸和色氨酸的浓度却很低。结论：该高通量方法可同时测定细胞内和细胞外生物样品中的一碳氨基酸代谢产物，可靠，准确。
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