A comparison of cell and tissue extraction techniques using high-resolution 1H-NMR spectroscopy
A comparison of cell and tissue extraction techniques using high-resolution 1H-NMR spectroscopy
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DOI：
10.1002/nbm.740
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发表时间：
2002-02-01
影响因子：
2.9
通讯作者：
Bhakoo, KK
中科院分区：
文献类型：
--
作者：
Le Belle, JE;Harris, NG;Bhakoo, KK
通过广泛的分析技术分析大脑代谢物通常是使用生化提取方法来实现的，该方法需要两个单独的样品或两个单独的提取步骤来制备水和有机代谢物组分。然而，在许多大脑病理中，水的代谢物和脂类都会发生变化，因此同时提取这两个组分将是有价值的。甲醇-氯仿(M/C)技术可以同时提取水中的代谢物和脂类。它已经被很好地应用于细胞和组织的脂肪提取，但其提取水中代谢物的效率和重复性尚不清楚。因此，我们使用成年大鼠脑和纯化的大鼠星形胶质细胞培养提取物的H-1-核磁共振波谱，比较了M/C法和常用的高氯酸(PCA)法的水代谢物产量和重复性。结果表明，与PCA法相比，M/C法是一种从脑组织和细胞中提取水中代谢物的更好的技术。M/C提取技术能够使用较小的溶剂体积从单个样品中同时提取脂类和水代谢物，使其非常适合于组织和细胞的核磁共振研究。版权所有(C)2002 John Wiley Sons，Ltd.
Analysis of brain metabolites by a wide range of analytical techniques is typically achieved using biochemical extraction methodologies that require either two separate samples or two separate extraction steps to prepare both aqueous and organic metabolite fractions. However there are a number of brain pathologies in which both aqueous metabolite and lipid changes occur so that a simultaneous extraction of both fractions would be valuable. The methanol-chloroform (M/C) technique enables extraction of both aqueous metabolites and lipids simultaneously. It is already well established for lipid extraction of cells and tissue but its efficiency and reproducibility for extraction of aqueous metabolites is unknown. Therefore, we compared the aqueous metabolite yield and the reproducibility of the M/C method to the commonly used perchloric acid (PCA) method, using H-1-NMR spectroscopy of adult rat brain and purified rat astrocyte culture extracts. The results indicate that M/C is a superior technique for aqueous metabolite extraction from both brain tissue and cells when compared to the PCA method. The M/C extraction technique enables the simultaneous extraction of both lipids and aqueous metabolites from a single sample using small solvent-volumes, making it well suited for NMR investigations of both tissues and cells. Copyright (C) 2002 John Wiley Sons, Ltd.