采用串联质谱法评估生理性氨基酸谱。
Evaluation of physiological amino acids profiling by tandem mass spectrometry.
作者信息
Filee Romain, Schoos Roland, Boemer François
机构信息
Biochemical Genetics Laboratory, Human Genetics, CHU Liege, University of Liege, Liege, Belgium.
出版信息
JIMD Rep. 2014;13:119-28. doi: 10.1007/8904_2013_265. Epub 2013 Nov 5.
BACKGROUND
Nowadays, the most conventional method to quantify physiological amino acids consists in ion exchange chromatography (IEC) followed by post-column ninhydrin derivatization and UV detection at two wavelengths. Unfortunately, the technique presents some drawbacks such as long run time, large sample volume, and specific costs associated to the maintenance of a dedicated instrument. Therefore, we aimed to switch towards a mass spectrometry approach.
METHODS
We have tested the aTRAQ kit for Amino Acid Analysis of Physiological Fluids (AB Sciex), affording the selective quantification of about 40 amino acids, and present here the results of our assessments.
RESULTS
Outlined accuracy profiles for each amino acid demonstrated very reliable data. A good linearity was observed from 1 to 1,000 μmol/L. RESULTS comparison with IEC showed a right concordance. Reference intervals established were very similar to those obtained by IEC and patients suffering from inborn error of metabolism have been readily identified.
CONCLUSIONS
The present approach offers a valid alternative to the IEC method, with several advantages: lower sample volume, run time reduction, and improved specificity. However, the aTRAQ method requires minute data reviewing, expending the overall time of procedure. Finally, financial and practical considerations of both techniques have to be counterbalanced before engaging any transition.
背景
如今，定量分析生理性氨基酸最常用的方法是离子交换色谱法（IEC），随后进行柱后茚三酮衍生化反应，并在两个波长下进行紫外检测。遗憾的是，该技术存在一些缺点，如运行时间长、进样量大以及维护专用仪器的特定成本。因此，我们旨在转向质谱分析法。
方法
我们测试了用于生理体液氨基酸分析的aTRAQ试剂盒（AB Sciex公司），该试剂盒能够选择性定量约40种氨基酸，在此展示我们的评估结果。
结果
每种氨基酸的精度概况表明数据非常可靠。在1至1000μmol/L范围内观察到良好的线性关系。与IEC法的结果比较显示出良好的一致性。所建立的参考区间与IEC法获得的参考区间非常相似，并且能够轻松识别患有先天性代谢缺陷的患者。
结论
本方法为IEC法提供了一种有效的替代方案，具有几个优点：进样量少、运行时间缩短以及特异性提高。然而，aTRAQ方法需要仔细审查数据，这会延长整个检测过程的时间。最后，在进行任何转变之前，必须权衡两种技术在财务和实际操作方面的因素。
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