Feil Family Brain and Mind Research Institute, Weill Cornell Medicine, New York, NY 10065, USA.
Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, Russia.
Molecules. 2017 Aug 23;22(9):1388. doi: 10.3390/molecules22091388.
Coenzyme A (CoA) and acetyl-coenzyme A (acetyl-CoA) play essential roles in cell energy metabolism. Dysregulation of the biosynthesis and functioning of both compounds may contribute to various pathological conditions. We describe here a simple and sensitive HPLC-UV based method for simultaneous determination of CoA and acetyl-CoA in a variety of biological samples, including cells in culture, mouse cortex, and rat plasma, liver, kidney, and brain tissues. The limits of detection for CoA and acetyl-CoA are >10-fold lower than those obtained by previously described HPLC procedures, with coefficients of variation <1% for standard solutions, and 1-3% for deproteinized biological samples. Recovery is 95-97% for liver extracts spiked with Co-A and acetyl-CoA. Many factors may influence the tissue concentrations of CoA and acetyl-CoA (e.g., age, fed, or fasted state). Nevertheless, the values obtained by the present HPLC method for the concentration of CoA and acetyl-CoA in selected rodent tissues are in reasonable agreement with literature values. The concentrations of CoA and acetyl-CoA were found to be very low in rat plasma, but easily measurable by the present HPLC method. The method should be useful for studying cellular energy metabolism under normal and pathological conditions, and during targeted drug therapy treatment.
辅酶 A（CoA）和乙酰辅酶 A（acetyl-CoA）在细胞能量代谢中发挥着重要作用。这两种化合物的生物合成和功能失调可能导致各种病理状况。我们在这里描述了一种简单而灵敏的 HPLC-UV 方法，可同时测定多种生物样品中的 CoA 和乙酰-CoA，包括培养细胞、小鼠皮层和大鼠血浆、肝脏、肾脏和脑组织。与以前描述的 HPLC 方法相比，CoA 和乙酰-CoA 的检测限高出 10 多倍，标准溶液的变异系数 <1%，去蛋白生物样品的变异系数为 1-3%。用 Co-A 和乙酰-CoA 对肝提取物进行加标回收率为 95-97%。许多因素可能影响 CoA 和乙酰-CoA 的组织浓度（例如，年龄、进食或禁食状态）。然而，本 HPLC 方法测定的选定啮齿动物组织中 CoA 和乙酰-CoA 的浓度与文献值基本一致。本 HPLC 方法可轻松测定大鼠血浆中 CoA 和乙酰-CoA 的浓度，其浓度非常低。该方法可用于研究正常和病理条件下以及靶向药物治疗期间的细胞能量代谢。
