Rabinowitz Joshua D, Kimball Elizabeth
Department of Chemistry and Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey 08544, USA.
Anal Chem. 2007 Aug 15;79(16):6167-73. doi: 10.1021/ac070470c. Epub 2007 Jul 14.
Cellular metabolome analysis by chromatography-mass spectrometry (MS) requires prior metabolite extraction. We examined a diversity of solvent systems for extraction of water-soluble metabolites from Escherichia coli. Quantitative yields of approximately 100 different metabolites were measured by liquid chromatography-tandem MS and displayed in clustered heat map format. Many metabolites, including most amino acids and components of central carbon metabolism, were adequately extracted by a broad spectrum of solvent mixtures. For nucleotide triphosphates, however, mixtures of acidic (0.1 M formic acid-containing) acetonitrile/water (80:20) or acetonitrile/methanol/water (40:40:20) gave superior triphosphate yields. Experiments involving isotopic tracers revealed that the improved triphosphate yields in the acidic acetonitrile were in part due to reduced triphosphate decomposition, which is a major problem when extracting with other solvent systems such as methanol/water. We recommend acidic solvent mixtures containing acetonitrile for extraction of the E. coli metabolome.
通过色谱 - 质谱联用（MS）进行细胞代谢组分析需要事先进行代谢物提取。我们研究了多种溶剂系统用于从大肠杆菌中提取水溶性代谢物。通过液相色谱 - 串联质谱法测量了约100种不同代谢物的定量产率，并以聚类热图形式展示。许多代谢物，包括大多数氨基酸和中心碳代谢的成分，都能被多种溶剂混合物充分提取。然而，对于三磷酸核苷酸，酸性（含0.1 M甲酸）乙腈/水（80:20）或乙腈/甲醇/水（40:40:20）的混合物能产生更高的三磷酸产率。涉及同位素示踪剂的实验表明，酸性乙腈中三磷酸产率的提高部分归因于三磷酸分解的减少，而在用其他溶剂系统（如甲醇/水）提取时，三磷酸分解是一个主要问题。我们推荐使用含乙腈的酸性溶剂混合物来提取大肠杆菌代谢组。
