以反相高效液相色谱法测定生物组织及细胞中的能量代谢变化

Detection of energy metabolism in biological tissues and cells by reversed-phase high performance liquid chromatography

文章综合分析了近年来采用反相高效液相色谱检测技术测定生物组织及细胞中主要能量物质的研究进展,且从样品的预处理、流动相及色谱条件的确定等方面进行了阐述。利用反相高效液相色谱技术可同时快速、准确测定多种生物能量物质,方法简便、快捷。反相高效液相色谱检测技术要求仪器各部件性能稳定;分离色谱柱需具较高的柱效、良好的分离度和洗脱曲线;取材部位要固定,以保证结果的可比性;标本须低温下提取,并尽快将样品移入高氯酸溶液中;匀浆时要求细胞完全破碎,充分抽提细胞中的ATP,低温离心,保证测定结果的准确性。反相高效液相色谱作为一种高效分离技术,将越来越广泛地应用于复杂能量物质的分离与定量分析中。

The progress on reversed-phase high performance liquid chromatography （RP-HPLC） techniques for analyzing the main energy substances in biological tissues and cells was reviewed in this paper. From the pretreatment of samples, mobile phase and how to establish the chromatogram conditions were discussed. Many energy substances were detected rapidly and exactly by reversed-phase high performance liquid chromatography. The technique is simple and convenient, and requests for this test technique as follows： the capability of each components of the apparatus was stable; the chromatographic column was in high performance; chromatographic separation was good and elution curve was clear; the selection spots were fixed to insure the comparability; samples were extracted at a low temperature and were moved into perchloric acid solution disrupted completely when refining in order to extract ATP fully, and then centrifuged at a low temperature to make sure of the accuracy of the results. As an efficient separation technology, reversed-phase high performance liquid chromatography would be used in the area of separation and quantitative analysis of complex energy substances.