Two complementary reversed-phase separations for comprehensive coverage of the semipolar and nonpolar metabolome.
Two complementary reversed-phase separations for comprehensive coverage of the semipolar and nonpolar metabolome.
复制标题
两种互补的反相分离，用于全面覆盖半极性和非极性代谢组。
DOI：
10.1007/s00216-017-0768-x
复制
发表时间：
2018-03
影响因子：
4.3
通讯作者：
Patti GJ
中科院分区：
文献类型：
--
作者：
Naser FJ;Mahieu NG;Wang L;Spalding JL;Johnson SL;Patti GJ
虽然它是常见的非靶向代谢组学应用反相液相色谱法（RPLC）和亲水相互作用液相色谱法（HILIC）的方法，已被系统优化的脂质和中心碳代谢物，在这里，我们表明，这些既定的协议提供不良的覆盖率半极性代谢物由于保留不足。我们的目标是开发一种RPLC方法，在不牺牲脂质覆盖率的情况下改善这些代谢物的检测。我们最初通过分析均匀跨越非极性和半极性范围的47种小分子标准品，评估了沃茨最近在CORTECS系列下发布的色谱柱。非靶向代谢组学中常用的RPLC方法被视为基准参考。我们发现，由于进样溶剂的溶解度和保留限制，无法用任何单一方法可靠地分析高度非极性和半极性代谢物。相反，我们使用CORTECS T3柱分析半极性化合物和CORTECS C8分析脂质，优化了多重方法。引人注目的是，我们确定，结合这些方法能够检测到47个标准品中的41个，而我们的参考RPLC方法仅检测到47个标准品中的10个。然后，我们应用认证来比较方法在综合尺度上的性能。串联方法显示，相对于我们的RPLC基准，认证覆盖率增加了5倍以上。我们的研究结果表明，全面覆盖的代谢产物适合反相分离需要两个重建溶剂和色谱方法。因此，我们建议用T3和C8双重RPLC方法补充HILIC方法，以增加非靶向代谢组学的半极性代谢物和脂质的覆盖范围。半极性和非极性代谢物的分析需要两种RPLC方法，其将代谢物组覆盖范围扩展>5倍，用于非靶向分析。
Although it is common in untargeted metabolomics to apply reversed-phase liquid chromatography (RPLC) and hydrophilic interaction liquid chromatography (HILIC) methods that have been systematically optimized for lipids and central carbon metabolites, here we show that these established protocols provide poor coverage of semipolar metabolites due to inadequate retention. Our objective was to develop a RPLC approach that improved detection of these metabolites without sacrificing lipid coverage. We initially evaluated columns recently released by Waters under the CORTECS line by analyzing 47 small-molecule standards that evenly span the nonpolar and semipolar ranges. A RPLC method commonly used in untargeted metabolomics was considered a benchmarking reference. We found that highly nonpolar and semipolar metabolites cannot be reliably profiled with any single method due to solubility and retention limitations of the injection solvent. Instead, we optimized a multiplexed approach using the CORTECS T3 column to analyze semipolar compounds and the CORTECS C8 to analyze lipids. Strikingly, we determined that combining these methods enabled detection of 41 of the total 47 standards whereas our reference RPLC method only detected 10 of 47. We then applied credentialing to compare method performance at the comprehensive scale. The tandem method showed over a 5-fold increase in credentialing coverage relative to our RPLC benchmark. Our results demonstrate that comprehensive coverage of metabolites amenable to reversed-phase separation necessitates two reconstitution solvents and chromatographic methods. Thus, we suggest complementing HILIC methods with a dual T3 and C8 RPLC approach to increase coverage of semipolar metabolites and lipids for untargeted metabolomics. Analysis of semipolar and nonpolar metabolites necessitates two RPLC methods, which extend metabolome coverage >5 fold for untargeted profiling.
登录
查看更多内容
影响因子：
7.4
作者：
Nikolskiy, Igor;Mahieu, Nathaniel G.;Chen, Ying-Jr;Tautenhahn, Ralf;Patti, Gary J.
通讯作者：
Patti, Gary J.
DOI：
10.1093/bioinformatics/btu370
发表时间：
2014-10
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Daly R;Rogers S;Wandy J;Jankevics A;Burgess KE;Breitling R
通讯作者：
Breitling R
影响因子：
29
作者：
Johnson CH;Dejea CM;Edler D;Hoang LT;Santidrian AF;Felding BH;Ivanisevic J;Cho K;Wick EC;Hechenbleikner EM;Uritboonthai W;Goetz L;Casero RA Jr;Pardoll DM;White JR;Patti GJ;Sears CL;Siuzdak G
通讯作者：
Siuzdak G
影响因子：
7.4
作者：
Masson, Perrine;Alves, Alexessander Couto;Want, Elizabeth J.
通讯作者：
Want, Elizabeth J.
影响因子：
7.4
作者：
Broeckling, C. D.;Afsar, F. A.;Prenni, J. E.
通讯作者：
Prenni, J. E.