从组织中分离短链、中链和长链酰基辅酶A酯的新型方法。
Novel isolation procedure for short-, medium-, and long-chain acyl-coenzyme A esters from tissue.
作者信息
Minkler Paul E, Kerner Janos, Ingalls Stephen T, Hoppel Charles L
机构信息
Department of Pharmacology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
出版信息
Anal Biochem. 2008 May 15;376(2):275-6. doi: 10.1016/j.ab.2008.02.022. Epub 2008 Feb 29.
Abstract
A novel procedure for the quantitative isolation and purification of acyl-coenzyme A esters is presented. The procedure involves two steps: (1) tissue extraction using acetonitrile/2-propanol (3+1, v+v) followed by 0.1M potassium phosphate, pH 6.7, and (2) purification using 2-(2-pyridyl)ethyl-functionalized silica gel. Recoveries determined by adding radiolabeled acetyl-, malonyl-, octanoyl-, oleoyl-, palmitoyl-, or arachidonyl-coenzyme A to powdered rat liver varied 93-104% for tissue extraction and 83-90% for solid-phase extraction. The procedure described allows for isolation and purification, with high recoveries, of acyl-coenzyme A esters differing widely in chain length and saturation.
摘要
本文介绍了一种用于定量分离和纯化酰基辅酶A酯的新方法。该方法包括两个步骤：（1）先用乙腈/2-丙醇（3+1，v/v），然后用pH 6.7的0.1M磷酸钾进行组织提取；（2）用2-（2-吡啶基）乙基功能化硅胶进行纯化。通过向大鼠肝脏粉末中添加放射性标记的乙酰辅酶A、丙二酰辅酶A、辛酰辅酶A、油酰辅酶A、棕榈酰辅酶A或花生四烯酰辅酶A来测定回收率，组织提取的回收率在93%-104%之间，固相提取的回收率在83%-90%之间。所述方法能够以高回收率分离和纯化链长和饱和度差异很大的酰基辅酶A酯。
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