Acidic acetonitrile for cellular metabolome extraction from Escherichia coli
Acidic acetonitrile for cellular metabolome extraction from Escherichia coli
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DOI：
10.1021/ac070470c
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发表时间：
2007-08-15
影响因子：
7.4
通讯作者：
Kimball, Elizabeth
中科院分区：
文献类型：
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作者：
Rabinowitz, Joshua D.;Kimball, Elizabeth
通过色谱-质谱法（MS）进行的细胞代谢组分析需要预先提取代谢物。我们研究了从大肠杆菌中提取水溶性代谢物的溶剂系统的多样性。通过液相色谱-串联MS测量类似于100种不同代谢物的定量产率，并以聚类热图格式显示。许多代谢产物，包括大多数氨基酸和中心碳代谢组分，被广泛的溶剂混合物充分提取。然而，对于核苷酸三磷酸，酸性（含0.1M甲酸）乙腈/水（80：20）或乙腈/甲醇/水（40：40：20）的混合物给出了上级的三磷酸产率。涉及同位素示踪剂的实验表明，在酸性乙腈中三磷酸盐产率的提高部分是由于三磷酸盐分解减少，这是用其他溶剂系统（如甲醇/水）提取时的主要问题。我们建议用含有乙腈的酸性溶剂混合物提取大肠杆菌。大肠杆菌代谢组。
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 similar to 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.