酸水解同位素稀释质谱法测量基因组DNA含量  

Accurate Quantitation of Genome DNA Mass Concentration by Acid Hydrolysis–Isotope Dilution Mass Spectrometry

作　　者：张玲[1] 陈大舟[1] 武利庆 高运华[1] 汤桦[1] 王晶[1] 刘新海 

机构地区：[1]中国计量科学研究院,北京100013 [2]中国医学科学院整形外科医院,北京100144

出　　处：《化学分析计量》2013年第5期9-13,共5页Chemical Analysis And Meterage

基　　金：科技部支撑项目(2008BAK41B03;2013BAK12B05);中国计量科学院基本业务费项目(AKY0915;AKY1219)

摘　　要：建立了测量噬菌体入基因组DNA含量的方法。样品添加同位素标记碱基内标之后，用体积分数为88％的甲酸溶液在170℃水解30min，解离出的核酸碱基通过反向柱分离，电喷雾四级杆质谱法测定，用多反应监测模式分别检测碱基及其同位素标记物的母离子和碎片子离子，从而建立了基因组DNA水解一同位素稀释质谱法测量长片段核酸含量的方法，并将DNA浓度溯源至碱基浓度。方法的线性范围为1—1000μg／g，检出限可低至100ng／g。测定的入DNA含量标准物质为（2．51±0．06）μg／支（k=2），该方法可用于长片段核酸含量标准物质定值。The method for determining genomic DNA mass content in phage was set up. Phage λ genome DNA samples were added by isotope labeled nucleic bases internal standard, then hydrolyzed in 88% formic acid solution for 30 min under 170℃ . The DNA hydrolysis products nucleic bases were separated in C18 reverse phase column and analyzed by electrospray quadrupore isotope dilution mass spectrometry （HPLC-IDMS） by multiple reaction monitoring mode to scan parent ion and product ion of nucleic bases and the isotope labeled nucleic bases. The method was established for traceability of the genomic DNA to 4 nucleic bases mass content. The method linear range was 1-1 000 μg / g, and the detection limit could be as low as 100 ng/g. The DNA mass content in each vial was measured （2.51±0.06）μg （k=2）. This method could be used as a primary method for determining DNA mass as a reference material.

关 键 词：噬菌体入DNA 核酸碱基 核苷酸 高效液相色谱法 同位素稀释质谱法 

分 类 号：O657.6[理学—分析化学]