Abstract A method to determine the content of nucleic bases Cytosine (Cyt) and Adenine (Ade) by electrode potential titration in non-aqueous soolvent is established.The factors affecting the experiments, such as non-aqueous solvent, titrant, the concentration of titrant, and end point of the titration are determined.The optimal experiment conditions determined are glacial acetic acid (HAc) as the non-aqueous solvent,perchloric acid as titrant (HClO4),titrant concentration 0.05 mol/L,and equivalence point of the electrode potential titration curve as endpoint.Through 15 times measurements and uncertainty evaluation, the purity of Ade is 99.40% ±0.38% (k=2), Cyt 99.58%±0.42% (k=2).
ZHANG Ling,LIU Fang-fang,CHEN Da-zhou, et al. Determine the Content of Nucleic Bases Cytosine and Adenine by Electrode Potential Titration in Non-aqueous Solvent[J]. Acta Metrologica Sinica, 2013, 34(5): 508-512.
［1］ Holden M J, Rabb S A, Tewari Y B, et al. Traceable phosphorus measurements by ICP-OES and HPLC for the quantitation of DNA［J］. Analytical Chemistry, 2007, 79(4):1536-1541.
［2］ Yang I, Han M S, Yim Y H, et al. A strategy for establishing accurate quantitation standards of oligonucleotides: quantitation of phosphorus of DNA phosphodiester bonds using inductiVely coupled plasma-optical emission spectroscopy［J］. Anlytical Biochemistry, 2004,335(1):150-161.
［3］ Kinumi T, Narukawa T, Fujii S, et al. Quantification of oligonucleotide containing a sequence failure product: comparison of isotope dilution mass spectrometry with other quantification methods［J］. European Journal of Mass Spectrometry,2009, 15(3): 399-407.
［4］ O′Connor G, Dawson C, Woolford A, et al. Quantitation of oligonucleotides by phosphodiesterase digestion followed by isotope dilution mass spectrometry: Proof of concept［J］.Analytical Chemistry,2002,74(15):3670-3676.
［5］ Sigel H. Acid-base properties of purine residues and the effect of metal ions: Quantification of rare nucleobase tautomers［J］.Pure Appl Chem, 2004, 76(10):1869–1886.
［6］ 梁忆,张皋,陈智群,等.非水溶剂电位滴定法测定标准物质RDX和HMX的含量［J］.火炸药学报,2008, 31(2):105-108.
［7］ 梁忆,胡玲,张皋,等.标准物质RDX、HMX定值的不确定度评估［J］.化学分析计量,2007,16(5): 4-6.
［8］ JJG1006—1994,一级标准物质［s］.
［9］ 全浩,韩永志.标准物质及其应用［M］.(第2版).北京:中国标准出版社,2003.
2013, Vol. 34 
