派罗宁GS双峰双波长光度法测定核酸

Study on the double-peak-double-wavelength spectrophotometry for the determination of nucleic acid with pyronine GS

采用光度法研究了派罗宁GS与核酸的结合反应,结果表明在pH6.0左右的介质中,派罗宁GS与核酸在室温下能迅速结合形成紫红色复合物,该复合物在570 nm处有正吸收峰,在540nm处有负吸收峰.据此,建立了1个测定核酸的双峰双波长光度分析新方法.用该方法测定小牛胸腺DNA（ctDNA）、鱼精子DNA（fsDNA）、酵母RNA（yt RNA）及热变性ctDNA（DctDNA）的检出限均低于0.011μg/mL.方法简单、快速,试剂及反应体系稳定、选择性好、准确度高,用于4个合成试样的分析,回收率为96.1%～103.0%.

The binding reaction of nucleic acid with.pyronine GS has been studied by the spectrophotometry. A claret complex, which exhibited a positive absorption peak located at 570 nm and a negative peak at 540 nm, was formed from pyronine GS and nucleic acid in the NaH2PO4 - K2HPO4 buffer with pH of 6.0 at room temperature. A new double-peak-double-wavelength spectrophotometry for the determination of nucleic acid was developed based on the binding reaction. The detection limit of this method for the determinatin of calf-thymus DNA （ct DNA）, fish sperm DNA （fs DNA）, yeast RNA （yt RNA） and denatured calf-thymus DNA was less than 0. 011μg/mL. The proposed method, which is simple, rapid, stable, highly selective and precise, has been applied for the determination of ct DNA in the four synthetic samples with recovery ranged from 96.1% to 103.0%.