Rosendal J, Knudsen J
Institute of Biochemistry, University of Odense, Denmark.
Anal Biochem. 1992 Nov 15;207(1):63-7. doi: 10.1016/0003-2697(92)90500-7.
A method for the extraction of acyl-CoA esters from tissue, and their subsequent analysis by HPLC is described. The lipids are removed by a two-phase extraction in a chloroform/methanol/water system. The long-chain acyl-CoA esters are extracted using methanol and a high salt concentration (2 M ammonium acetate). Reextraction of the dry residue after evaporation of extraction solvent results in low overall recoveries (20%). By adding 1 mg/ml acyl-CoA-binding protein to the extraction solvent the overall recovery was increased to 55%. The method is easy and fast to perform and is thereby suitable for analysis of a large number of samples. The advantages of the method over previously published methods are discussed.
本文描述了一种从组织中提取酰基辅酶A酯并随后通过高效液相色谱法进行分析的方法。脂质通过在氯仿/甲醇/水系统中的两相萃取去除。长链酰基辅酶A酯使用甲醇和高盐浓度(2M醋酸铵)进行提取。萃取溶剂蒸发后对干燥残渣进行再萃取,整体回收率较低(20%)。通过向萃取溶剂中添加1mg/ml酰基辅酶A结合蛋白,整体回收率提高到了55%。该方法操作简便、快速,因此适用于大量样品的分析。文中还讨论了该方法相对于先前发表的方法的优点。