紫外分光光度法测定核酸含量的影响因素分析  

Influencing Factors of Determination of Nucleic Acid Content by Ultraviolet Spectrophotometry

作　　者：李春 刘建涛 高运华[1] 王治栋 段学欣 LI Chun;LIU Jiantao;GAO Yun-hua;WANG Zhi-dong;DUAN Xue-xin(Center for Advanced Measurement Science,College of Precision Instrument and Optoelectronics Engineering,National Institute of Metrology,Beijing 100029,China;College of Precision Instrument and Optoelectronics Engineering,Tianjin University,Tianjin 300072,China)

机构地区：[1]中国计量科学研究院前沿计量科学中心,北京100029 [2]天津大学精密仪器与光电子工程学院,天津300072

出　　处：《化学试剂》2020年第1期53-57,共5页Chemical Reagents

基　　金：国家质量研发重点专项项目(2017YFF0204604);国家质检公益行业科研专项(201510026)

摘　　要：选择寡聚核苷酸、鲑鱼精单链DNA、小牛胸腺双链DNA标准物质为样本,系统研究了溶液pH、苯酚、丙酮、蛋白质、多糖等对紫外分光光度法测量结果的影响情况,另外考察了核糖核酸(RNA)和脱氧核糖核酸(DNA)的相互影响情况。研究发现基于分光光度法的核酸定量分析,易受溶液pH、苯酚、丙酮、蛋白质、多糖等的影响,另外,DNA和RNA之间也会相互干扰。因此,紫外分光光度法测定核酸含量时,特别是该方法用于核酸标准物质特性量值确定时,需要样品高度纯化,pH确定的条件下才能保证测量结果的准确可靠。The oligonucleotide,salmon single-stranded DNA,and calf thymus double-stranded DNA were measured by UV spectrophotometry,and the effect of solution pH,phenol,acetone,proteins and polysaccharides on the measurement were investigated.In addition,the interaction between ribonucleic acid(RNA)and deoxyribonucleic acid(DNA)was examined.The studies had found that quantitative analysis of nucleic acids based on spectrophotometry was susceptible to solution pH,phenol,acetone,protein,polysaccharides,and so on.DNA and RNA also interfered with each other.Therefore,UV spectrophotometry for the determination of nucleic acid content required high purity of the sample,and the pH was determined to ensure accurate and reliable measurement results.Especially,the method was used to determine the characteristic value of nucleic acid reference material.It needed highly purified candidate of reference material,and the determination of pH could ensure the accuracy and reliability of the characteristic value of reference material.

关 键 词：紫外分光光度法 核酸 含量 影响因素 PH 有机试剂 

分 类 号：O657.32[理学—分析化学]