Department of Nutrition, Case Western Reserve University, Cleveland, OH 44106.
J Lipid Res. 2014 Mar;55(3):592-602. doi: 10.1194/jlr.D045112. Epub 2013 Dec 23.
A metabolomic approach to selectively profile all acyl-CoAs was developed using a programmed multiple reaction monitoring (MRM) method in LC-MS/MS and was employed in the analysis of various rat organs. The programmed MRM method possessed 300 mass ion transitions with the mass difference of 507 between precursor ion (Q1) and product ion (Q3), and the precursor ion started from m/z 768 and progressively increased one mass unit at each step. Acyl-dephospho-CoAs resulting from the dephosphorylation of acyl-CoAs were identified by accurate MS and fragmentation. Acyl-dephospho-CoAs were also quantitatively scanned by the MRM method with the mass difference of 427 between Q1 and Q3 mass ions. Acyl-CoAs and dephospho-CoAs were assayed with limits of detection ranging from 2 to 133 nM. The accuracy of the method was demonstrated by assaying a range of concentrations of spiked acyl-CoAs with the results of 80-114%. The distribution of acyl-CoAs reflects the metabolic status of each organ. The physiological role of dephosphorylation of acyl-CoAs remains to be further characterized. The methodology described herein provides a novel strategy in metabolomic studies to quantitatively and qualitatively profile all potential acyl-CoAs and acyl-dephospho-CoAs.
采用程序多反应监测(MRM)方法在 LC-MS/MS 中开发了一种代谢组学方法,用于选择性地对所有酰基辅酶 A 进行分析,并将其应用于各种大鼠器官的分析。该程序 MRM 方法具有 300 个质量离子跃迁,其前体离子(Q1)和产物离子(Q3)之间的质量差为 507,前体离子从 m/z 768 开始,每个步骤递增一个质量单位。酰基辅酶 A 去磷酸化产物通过准确的 MS 和碎片分析进行鉴定。酰基辅酶 A 去磷酸化产物也可以通过 MRM 方法进行定量扫描,Q1 和 Q3 质量离子之间的质量差为 427。酰基辅酶 A 和去磷酸化的辅酶 A 的检测限范围为 2 至 133 nM。通过测定一系列不同浓度的加标酰基辅酶 A 的结果,证明了该方法的准确性为 80-114%。酰基辅酶 A 的分布反映了每个器官的代谢状态。酰基辅酶 A 去磷酸化的生理作用仍有待进一步研究。本文所述的方法为代谢组学研究提供了一种新的策略,可用于定量和定性地分析所有潜在的酰基辅酶 A 和酰基辅酶 A 去磷酸化产物。