眼科研究
眼科研究
안과연구
CHINESE OPHTHALMIC RESEARCH
2010年
3期
253-256
,共4页
曲进锋%黎晓新%Ilyas Washington
麯進鋒%黎曉新%Ilyas Washington
곡진봉%려효신%Ilyas Washington
高效液相色谱法%辅酶Q10%视网膜%氧化应激
高效液相色譜法%輔酶Q10%視網膜%氧化應激
고효액상색보법%보매Q10%시망막%양화응격
high-performance liquid chromatography%coenzyme Q10%retina%oxidative stress
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.
目的 建立一種通過高效液相色譜法(HPLC)檢測人視網膜中輔酶Q10(CoQ10)含量的方法 .方法 10例新鮮健康人尸眼(年齡20～28歲)視網膜經分離、勻漿、凍榦後,以甲醇脫蛋白,再經己烷萃取,萃取液進樣進行HPLC分析.採用Onyx Monolithic C18柱,流動相採用含0.42% 醋痠鈉的甲醇-己烷-乙痠-異丙醇溶液(體積比為55:9:1:1),紫外線檢測波長為275nm.結果 採用HPLC從人視網膜中成功地提取齣CoQ10,此方法 CoQ10的檢測限為0.14mg/L.在0.20～395.00mg/L質量濃度範圍內,CoQ10峰麵積與相應質量濃度呈良好的線性關繫.重複性檢測穫得CoQ10質量濃度在0.86、2.59、3.45mg/L時的相對標準差分彆為2.7 %、0.1%和3.3%.日內和日間相對標準差分彆為1.6%和3.7%.迴收率為101%～113%.用此方法 檢測10例人尸眼視網膜CoQ10 平均含量為(0.51±0.20)μg/眼.結論 人視網膜中CoQ10的含量可以通過反相HPLC法進行定量檢測.
목적 건립일충통과고효액상색보법(HPLC)검측인시망막중보매Q10(CoQ10)함량적방법 .방법 10례신선건강인시안(년령20～28세)시망막경분리、균장、동간후,이갑순탈단백,재경기완췌취,췌취액진양진행HPLC분석.채용Onyx Monolithic C18주,류동상채용함0.42% 작산납적갑순-기완-을산-이병순용액(체적비위55:9:1:1),자외선검측파장위275nm.결과 채용HPLC종인시망막중성공지제취출CoQ10,차방법 CoQ10적검측한위0.14mg/L.재0.20～395.00mg/L질량농도범위내,CoQ10봉면적여상응질량농도정량호적선성관계.중복성검측획득CoQ10질량농도재0.86、2.59、3.45mg/L시적상대표준차분별위2.7 %、0.1%화3.3%.일내화일간상대표준차분별위1.6%화3.7%.회수솔위101%～113%.용차방법 검측10례인시안시망막CoQ10 평균함량위(0.51±0.20)μg/안.결론 인시망막중CoQ10적함량가이통과반상HPLC법진행정량검측.
Background 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.
