反相高效液相色谱法测定大鼠肝组织中DNA的碱基含量
Determination of the Base Contents of Liver DNA of Rats by Reversed-Phase High Performance liquid Chromatography

王超云胡凤祖师治贤

文摘	利用反相高效液相色谱法，采用Supelcosil LC-18色谱柱（250 mm * 4.6 mm i.d.，5μm ），以甲醇-0.05 mol/L KH_2PO_4缓冲液（体积比为20：80）为流动相，流速0.8 mL/min，在254 nm波长处检测，对生活在高原（海拔2.3 km）的习服大鼠肝组织中脱氧核糖核酸（DNA）的碱基含量进行了检测，发现各碱基在DNA中所占的比例是相对稳定的：腺嘌呤（A）28.8%、鸟嘌呤（G）23.3%、胞嘧啶（C）17.4%、胸腺嘧啶（T）25.3%，并利用内标法对DNA甲基化水平进行了测定。
其他语种文摘	The base contents of liver deoxyribonucleic acid (DNA) of rats living at an altitude of 2.3 km were determined by reversed-phase high performance liquid chromatography. At first, 0.05 mol/L KH_2PO_4 (pH 4.0) was used to dissolve the DNA acid hydrolysis products with 8-bromoguanosine (Br~8G) as an internal standard. Then the DNA hydrolysis products with Br~8G were chromatographed on a Supelcosil LC-18 column with UV detection at 254 nm and eluted by the mobile phase of MeOH 0.05 mol/L KH_2PO_4 (pH 4.0) (20:80, V/V) at the flow rate of 0.8 mL/min. Under these conditions, several bases were separated effectively. From the results, the relatively constant proportions of the bases in DNA were found. The contents were 17.4% of cytosine (C), 28.8% of adenine (A), 23.3% of guanine (G) and 25.3% of thymine (T). RSDs of the deteremination of these bases were 1.7%, 1.5%, 1.3% and 2.1%, respectively. At the same time the methylation level of liver DNA of the rats determined by the internal standard method was 6.2%.
来源	色谱 ,2002,20(4):348-349 【核心库】
关键词	反相高效液相色谱法 ; 脱氧核糖核酸 ; 碱基 ; 甲基化水平 ; 肝组织 ; 大鼠
地址	中国科学院西北高原生物研究所, 青海, 西宁, 810001
语种	中文
文献类型	研究性论文
ISSN	1000-8713
学科	化学
文献收藏号	CSCD:1043520