摘要目的 建立一种通过高效液相色谱法(HPLC)检测人视网膜中辅酶Q10(CoQ10)含量的方法 .方法 10例新鲜健康人尸眼(年龄20～28岁)视网膜经分离、匀浆、冻干后,以甲醇脱蛋白,再经己烷萃取,萃取液进样进行HPLC分析.采用Onyx Monolithic C18柱,流动相采用含0.42% 醋酸钠的甲醇-己烷-乙酸-异丙醇溶液(体积比为55:9:1:1),紫外线检测波长为275nm.结果 采用HPLC从人视网膜中成功地提取出CoQ10,此方法 CoQ10的检测限为0.14mg/L.在0.20～395.00mg/L质量浓度范围内,CoQ10峰面积与相应质量浓度呈良好的线性关系.重复性检测获得CoQ10质量浓度在0.86、2.59、3.45mg/L时的相对标准差分别为2.7 %、0.1%和3.3%.日内和日间相对标准差分别为1.6%和3.7%.回收率为101%～113%.用此方法 检测10例人尸眼视网膜CoQ10 平均含量为(0.51±0.20)μg/眼.结论 人视网膜中CoQ10的含量可以通过反相HPLC法进行定量检测.

abstractsBackground Researches have demonstrated that age-related macular degeneration (AMD) is associated with the oxidative stress injury of retina.Coenzyme Q10 (CoQ10) is an important antioxidant agent.CoQ10 level in blood plasma is a primary index of reflecting the oxidative stress ability of human.However,the study on CoQ10 content in retina has not been seen yet.ObjectiveThe aim of this study is to establish a method of detecting CoQ10 content in retina by the high-performance liquid chromatography (HPLC). Methods The retinas were isolated from 10 healthy eyes of donors aged 20-28 years.The donor eyes were obtained from National Development and Research Institute,Inc.USA.Isolated retina tissue was prepared into homogenate then lyophilized and deproteinized with methanol.Samples were extracted with heptane prior to the HPLC analysis with the chromatographic conditions as follows:RP-18 column,a mobile phase consisted of methanol-hexane-acetic acid-isopropanol (V/V=55:9:1:1) and 0.42% sodium acetate,ultraviolet rays (UV) detector at 275 nm.Results CoQ10 was effectively isolated from human retina.The limit of detection of CoQ10 was 0.14mg/L.The peak area and concentration of CoQ10 showed a good linear correlation within the concentration range of 0.2-395.00mg/L (R~2＝0.9943).Repeatability study showed that the relative standard deviations for CoQ10 at the concentration of 0.86mg/L,2.59mg/L and 3.45mg/L were 2.7%,0.1% and 3.3%,respectively.The within- and inter-day standard deviations for the analysis of CoQ10 were 1.6% and 3.7%,respectively.The recovery was 101%-113% for the human retina samples.The concentration of CoQ10 in 10 retinas from human donors was 0.51±0.20μg/eye in average.Conclusion A HPLC method for the quantified analysis of CoQ10 in human retina is developed.