Simultaneous extraction of cellular lipids and water-soluble metabolites: Evaluation by NMR spectroscopy
Simultaneous extraction of cellular lipids and water-soluble metabolites: Evaluation by NMR spectroscopy
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DOI：
10.1002/mrm.1910350210
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发表时间：
1996-02-01
影响因子：
3.3
通讯作者：
Salomon, Y
中科院分区：
文献类型：
--
作者：
Tyagi, RK;Azrad, A;Salomon, Y
建立了一种从单细胞样品中同时提取脂类和水溶性代谢物的方法，并对其进行了优化，以用于核磁共振波谱。用P-31和C-13核磁共振仪研究了培养的小鼠黑色素瘤细胞M2R的中间代谢产物及其在黑素刺激后的变化。细胞用甲醇、氯仿和水(1：1：1，v/v/v)提取。氯仿和甲醇-水相的含量被分离和定量回收。上、下相的含量与高氯酸法和Folch‘s脂抽提法得到的同系物相吻合。萃取物的pH值保持在生理范围内，消除了对细胞代谢物的潜在有害影响，水相含有最少量的盐，使这些萃取物适合后续分析程序。从同一样品中获得脂类和水溶性代谢物，可以定量描述连接这两种细胞成分的代谢途径。
A method for simultaneous extraction of lipids and water-soluble metabolites from a single cell sample was developed and optimized for NMR spectroscopy. Intermediary metabolites in cultured M2R mouse melanoma cells and changes therein in response to challenge with melanotropin were studied by P-31 and C-13 NMR. Cells were extracted with methanol, chloroform, and water (1:1:1, v/v/v). The contents of the chloroform and methanol-water phases were separated and quantitatively recovered. The contents of the upper and lower phases compared well with the homologous fractions obtained by perchloric acid and Folch's lipid extraction methods. The pH of the extracts remained within the physiologic range, eliminating potential deleterious effect on cellular metabolites, The water phase contained minimal amounts of salts, making these extracts amenable to subsequent analytical procedures. Obtaining lipid- and water-soluble metabolites from the same sample enables characterization of metabolic pathways that bridge the two cellular components in a quantitative manner.