Determination of individual long-chain fatty acyl-CoA esters in heart and skeletal muscle.
Determination of individual long-chain fatty acyl-CoA esters in heart and skeletal muscle.
复制标题
心脏和骨骼肌中单个长链脂肪酰辅酶 A 酯的测定。
作者：
Molaparast-Saless,F;Shrago,E;Spennetta,TL;Donatello,S;Kneeland,LM;Nellis,SH;Liedtke,AJ
建立了测定心肌和骨骼肌中长链脂肪酰辅酶A酯的高效液相色谱分析方法。从冻存的猪和大鼠心脏和大鼠骨骼肌组织中提取酯，用于径向压缩的C185μ反相柱上分析。在萃取物中鉴定了9个碳链长度从C12到C20的峰，这些峰随后在碱解过程中消失。乙酰辅酶A的主要峰为14：1、18：2、16：0和18：1，另外在大鼠心脏中为18：0。各分子物种的总和得到长链酰基-辅酶A酯：猪心11.34±1.48nmol/g湿重，大鼠心脏14.51±2.11nmol/g湿重，大鼠骨骼肌4.35±0.71nmol/g湿重。这些值大约是用另一种方法获得的值的132%，该方法在酯碱解后通过高效液相色谱法测定总CoA。所描述的方法证明了肌肉组织中单个酰基辅酶A物种的定量。因此，它有许多优点，因为它允许在各种营养和代谢条件下获得关于单个分子物种的信息。
A method has been developed for determination of individual long‐chain fatty acyl‐CoA esters from heart and skeletal muscle using high performance liquid chromatography (HPLC). The esters were extracted from freezeclamped tissue of pig and rat hearts and rat skeletal muscle for analysis on a radially compressed C185μ reversephase column. Nine peaks in the extract with carbon chain lengths from C12to C20that subsequently disappeared on alkaline hydrolysis were identified. The major acyl‐CoA peaks were 14∶1, 18∶2, 16∶0 and 18∶1 and additionally in rat heart 18∶0. Total long‐chain acyl‐CoA esters obtained by summation of the individual molecular species was 11.34±1.48 nmol/g wet wt. pig heart; 14.51±2.11 nmol/g wet wt. in rat heart, and 4.35±0.71 nmol/g wet wt. in rat skeletal muscle. These values were approximately 132% of those obtained using a separate procedure that measured total CoA by HPLC after alkaline hydrolysis of the esters. The described method demonstrates the quantitation of individual acyl‐CoA species in muscle tissue. Therefore, it has a number of advantages in that it permits information to be obtained on the individual molecular species under various nutritional and metabolic conditions.