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

摘要: 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pH 6.0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540 nm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(Dct DNA)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%.

Abstract: 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 570nm and a negative peak at 540nm,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%.