HPLC法研究DNA及其甲基修饰物的酸性介质降解  

Study on the Degradation of DNA in an Acidic Medium by HPLC Using Gradient Elution

作　　者：龙耀庭[1] 赏珩 杨广红 沈彤 陆妙琴[1] 

机构地区：[1]中国科学院生态环境研究中心

出　　处：《环境科学》1995年第4期35-38,共4页Environmental Science

基　　金：国家自然科学基金

摘　　要：用ＨＰＬＣ梯度洗脱法研究了ＤＮＡ在盐酸、高氨酸、三氯乙酸、三氟乙酸以及甲酸等酸性介质中的降解碱基的分离和确定。给出了ＭＮＵ与ＤＮＡ的加合产物的降解产率。对流动相组成、流率、柱温、梯度速率、ｐＨ值、盐浓度以及缓冲液浓度对容量因子（Ｋ＇）的影响分别进行了讨论。结果表明，ＤＮＡ在盐酸介质中可以得到最好的碱基回收率，ＭＮＵ与ＤＮＡ的加合产物在１ｍｏｌ／Ｌ盐酸介质中、２０ｍｉｎ条件下，可获得较好的碱基回收效果。he determination and separation of bases from the degradation products of DNA in hydrochloric acid, perchloric acid, trichloroacetic acid, trifluoroacetic acid and formic acid were described. The hydrolysis yields of DNA adduct products of MNU also were presented. The effects of mobile phase composition, flow rate, column temperature, gradient ramp, PH value, salt concentration, and buffer concentration on capacity factors(K') were respectively discussed. The experimental results showed that hydrochloric acid is a .favorable hydrolytical medium for DNA degradation and high recovery of DNA bases can be obtained. The optimum hydrolytical conditions for adduct products of DNA of MNU were 1 mol/L hydrochloric acid, 20 min, and 80-100℃.

关 键 词：DNA 降解 高速液体色谱 HPLC法 

分 类 号：Q523[生物学—生物化学]