【作者】 江国荣；

【导师】 朱荃；

【作者基本信息】 南京中医药大学 ， 中药学， 2003， 博士

【摘要】 第一部分 FIA检测AGEs方法的建立和应用 一 FIA检测AGEs方法的建立 目的 建立糖基化终未产物(advanced glycation end products，AGEs)的一种检测方法。方法 利用高效液相色谱仪(high performance liquid chromatography,HPLC)构建流动注射分析系统，检测体外制备的AGEs，并与普通荧光法等进行比较。荧光检测的激发波长和发射波长(Ex/Em)为370/440nm和335/385nm。结果 Ex/Em为370/440nm时，在原反应液的0.1-0.0001浓度范围内仍有良好的线性，Y=67979.98x+4.38，r=0.99999；重复性CV为1.11％；日内、日间CV分别为0.94％和2.51％；平均回收率98.5％；与其它检测AGEs的方法相比，其相互之间测定的数据均有较好的相关性，结果趋势基本一致。Ex/Em 335/385nm与370/440nm结果一致。结论 流动注射分析(Flow injection assay,FIA)是较可靠的检测AGEs水平的新方法。 二 体外蛋白质非酶糖基化的建立 目的 建立可用于药物筛选的体外蛋白质非酶糖基化模型。方法 1、葡萄糖与牛血清白蛋白组成反应系统。37℃孵育1、2周后与NBT反应，用流动注射法于530nm处测定早期糖化蛋白(果糖胺)；2、甘油醛与牛血清白蛋白组成反应系统。37℃孵育2周后用流动注射法于Ex/Em：370/440，335/385nm处测定糖基化终产物；3、3-脱氧葡萄糖醛酮与牛血清白蛋白组成反应系统。37℃孵育1周后用流动注射法于Ex/Em：370/440，335/385nm处测定糖基化终产物。结果 1、反应1和2周后，反应系统内葡萄糖浓度与果糖胺的吸收度值呈显著正相关，分别为：更多还原

【Abstract】 Part 1Establishment of FIA employed as an analyzing approach for AGEs1 Establishment of an analyzing approach for advanced glycation end products formed in vitro by flow injection assayABSTRACT Objective To establish an analyzing approach for advanced glycation end products formed in vitro. Methods Flow injection system was assembled using HPLC to detect AGEs formed in vitro. Other analyzing approaches were employed as controls. Fluorescence of each sample was determined at excitation/emission wavelengths of 370/440 nm as well as 335/385 nm. Results There was a good liner between AGEs concentration (diluted to 0.1-0.0001) and peak area (obtained at 440nm with excitation at 370nm). The equation was y=67978.98x+4.38, 1=0.99999. The coefficient of variance of reproducibility, within-day and between-day assayed by flow injection assay for AGEs was 1.11%, 0.94% and 2.51% respectively. The average recovery rate was 98.5%. There were correlations between flow injection assay and other measures both in data and the results. The data, as well as the results, detected by flow injection was correlated with those detected by other measures. The same results were observed at 335/385nm. Conclusion AEGs detected by flow injection assay shows excellent accuracy and reproducibility. It is a novel method for monitoring AGEs formed in vitro.更多还原