检验医学与临床2017，Vol.14Issue(7)：903-905,908,4.DOI:10.3969/j.issn.1672-9455.2017.07.001
高效液相色谱法测定A549细胞中的胆固醇水平研究
Study on HPLC method for determining cholesterol level in A549 cells
摘要
Abstract
Objective To establish a HPLC method for detecting intracellular cholesterol level in A549 cells so as to provide the technical support for the tissue injury by identifying the foaming degree of tissues cells with intracellular cholesterol relative level change as the indicator and specific tissue foaming.Methods Protein and triglycerides in the cell lysis solution were removed by using trichloroacetic acid and KOH ethanolic solution,the mixed solvent of normal hexane and isopropanol was used to extract intracellular cholesterol.The cholesterol level was detected by adopting the HPLC method.The chromatographic column was ZORBAX SB-C18(4.6 mm×150.0 mm,5 μm),the mobile phase was acetonitrile-isopropanol(67∶33,V/V),the flow rate was1 mL/min and the detection wavelength was at 206 nm.Results The linear range of intracellular cholesterol level showed good linearity in the range of 5-100 ng/μL(r=0.999 9).RSD was less than 3.0%.The average recovery rate was 94.89% and RSD was 2.86%(n=6).Conclusion The used method is convenient,simple and accurate with good repeatability,can be used to identify foam cell,and lays the foundation for exploring the lipid metabolism abnormality and organ injury and future studying the cholesterol metabolism disturbance and pulmonary injury.关键词
高效液相色谱/A549细胞/胆固醇Key words
high performance liquid chromatography/A549 cells/cholesterol引用本文复制引用
魏思宇,卢蕾,蔺晓菁,何芸芸,丁世家,阮雄中,练雪梅..高效液相色谱法测定A549细胞中的胆固醇水平研究[J].检验医学与临床,2017,14(7):903-905,908,4.基金项目
国家自然科学基金资助项目(81071907) （81071907）
教育部春晖计划合作科研启动项目(Z2012118). （Z2012118）
