摘要
目的:建立微量 DNA 含量测定方法,用于载基因纳米粒中 DNA 的含量和包封率测定。方法:利用 SYBR Green Ⅰ与 pD-NA 结合后能激发出强烈荧光,荧光分光光度计检测荧光强度建立标准曲线,测定载基因纳米粒提取液中的 pDNA 含量,计算纳米粒的包封率。结果:SYBR Green Ⅰ荧光分光光度法的检测灵敏度为10.188 ng·mL^(-1),线性范围20.375～1062.5 ng·mL^(-1)(r=0.9998),高、中、低3个浓度的回收率分别为101.0%,97.5%,99.6%;RSD 分别为1.26%,1.09%,1.19%(n=3);日内精密度的 RSD 分别为2.13%,1.88%,2.25%(n=5);日间精密度的 RSD 分别为3.86%,4.97%,3.41%(n=5)。结论:本方法准确可靠、灵敏度高、简便快速,可用于载基因纳米粒中 pDNA 的含量和包封率测定。
Objective：To develop a fluorescence method for determining DNA content and the entrapment efficency of DNA loaded nanoparticles. Method：The fluorescence intensity of the mixture of pDNA and SYBR Green Ⅰ was detected by a spectrofluoromrter, a standard curve was established between the fluorescence intensities and the concentrations of the pDNA. The pDNA content in the DNA loaded nanoparticles and the entrapment efficency of the nanoparticles were determined. Results： The SYBR Green Ⅰ - Fluorospectrophotometry method had high sensitivity up to 10. 188 ng·mL^-1 of pDNA with a good linear relation in a range at 20. 375 - 1062. 5 ng·mL^-1（r = 0. 9998 ）. The recovery at the high, middle and low concentrations were 101.0% ,97. 5 % and 99.6%, respectively, RSD were 1.26%, 1.09% and 1.19%, respectively （ n = 3）. The intra - day RSD were 2. 13%, 1.88% and 2.25 %, respectively （ n = 5 ） and the inter - day RSD were 3.86 % ,4. 97 % and 3.41%, respectively （ n = 5 ）. Conclusions： SYBR Green I - Fluorospectrophotometry is an accurate, sensitive, convenient method for determination the DNA content and the entrapment efficiency of DNA loaded nanoparticles.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2007年第11期1769-1772,共4页
Chinese Journal of Pharmaceutical Analysis
基金
国家自然科学基金(30572267)
