摘要
建立了用反相高效液相色谱法 (RP HPLC)测定大肠杆菌 (E .coli)脱氧核糖核酸 (DNA)和核糖核酸 (RNA)水解产物中核苷的方法 .色谱柱为SupelcoInc.C18反相柱 ,2 5 4nm和 2 80nm紫外吸收波长同时检测 ,分析条件经选择在流速为 1mL/min ,流动相A(2 .5 %甲醇 +0 .0 1mol/LKH2 PO4,pH 4 .6 ) 2 8min ,流动相B(8.0 %甲醇 +0 .0 1mol/LKH2 PO4,pH 4 .6 ) 37min ,柱温为 2 0℃条件下 6 5min可实现核苷的完全分离 .回收率为 80 .19%～ 99.31% ,相对标准偏差RSD为 0 .86 %～ 2 .6 2 % .该方法具有高灵敏度、高选择性的特点 。
A highly sensitive and selective reversed phase high performance liquid chromatography (RP HPLC) method for the analysis of nucleosides in DNA and RNA of E.coli is described. The analytical column was Supelco Inc.C\- 18 (250 mm×4.5 mm) column(at 20 ℃). The separation of the nucleosides was achieved in 65 min using a two buffer step gradient system. Buffer A (2.5 % v/v methanol, 0.01 mol/L KH\-2PO\-4, pH 4.6) was pumped for the first 28 min of the run, followed by 37 min of Buffer B (8.0 % v/v methanol, 0.01 mol/L KH\-2PO\-4, pH 4.6). Absorption was measured simultaneously at 254 nm and 280 nm with sensitivity of 0.01 aufs and the flow rate was 1.0 mL/min. The relative standard deviation (RSD) were from 0.86 % to 2.62 % and the recoveries ranged between 80.20 %～99.31 %.
出处
《分析测试技术与仪器》
CAS
2002年第2期107-110,共4页
Analysis and Testing Technology and Instruments
