Highly Sensitive GC/MS/MS Method for Quantitation of Amino and Nonamino Organic Acids
Highly Sensitive GC/MS/MS Method for Quantitation of Amino and Nonamino Organic Acids
复制标题
DOI：
10.1021/ac103245b
复制
发表时间：
2011-04-01
影响因子：
7.4
通讯作者：
Bruheim, Per
中科院分区：
文献类型：
--
作者：
Kvitvang, Hans F. N.;Andreassen, Trygve;Bruheim, Per
代谢物图谱方法是测量生物系统中代谢物库的重要工具。虽然大多数代谢物分析方法报告的是相对强度或依赖于代表所有代谢物的少数内部标准，但对生物细胞和液体中代谢物池的定量描述的最终要求是绝对浓度测定。我们在这里报告了一种高通量和灵敏的气相色谱/串联质谱仪(GC/MS/MS)目标代谢物分析方法，能够对所有检测到的代谢物进行绝对定量。该方法以氯甲酸甲酯衍生化为基础，用代谢物标准品加标定量，分别用氢化衍生化试剂衍生化。传统的电子碰撞电离被正化学电离所取代，因为后者在更大程度上保留了分子离子和其他高分子量碎片。这使得在许多洗脱代谢物中选择独特的MS/MS转变变得更容易。目前，新的GC/MS/MS方法包括67种常见的初级代谢物，其中大部分属于氨基酸和非氨基有机酸类。通过对尿液和血清样品的检测，验证了该方法的适用性。该方法是对目前氨基酸和非氨基酸的GC/MS代谢物定量分析方法的重大改进。
Metabolite profiling methods are important tools for measurement of metabolite pools in biological systems. While most metabolite profiling methods report relative intensities or depend on a few internal standards representing all metabolites, the ultimate requirement for a quantitative description of the metabolite pool in biological cells and fluids is absolute concentration determination. We report here a high-throughput and sensitive gas chromatography/tandem mass spectrometry (GC/MS/MS) targeted metabolite profiling method enabling absolute quantification of all detected metabolites. The method is based on methyl chloroformate derivatization and quantification by spiking samples with metabolite standards separately derivatized with deuterated derivatization reagents. The traditional electron impact ionization is replaced with positive chemical ionization since the latter to a much larger extent preserve the molecular ion and other high molecular weight fragments. This made it easier to select unique MS/MS transitions among the many coeluting metabolites. Currently, the novel GC/MS/MS method comprises 67 common primary metabolites of which most belong to the groups of amino and nonamino organic acids. We show the applicability of the method on urine and serum samples. The method is a significant improvement of present methodology for quantitative GC/MS metabolite profiling of amino acids and nonamino organic acids.