MOLECULAR DOSIMETRY OF 7-ALKYLGUANINE AND O-6-ALKYLGUANINE IN DNA BY ELECTROCHEMICAL DETECTION

被引：20

作者：

DEGROOT, AJL

JANSEN, JG

VANVALKENBURG, CFM

VANZEELAND, AA

机构：

[1] LEIDEN UNIV,MGC,DEPT RADIAT GENET & CHEM MUTAGENESIS,2333 AL LEIDEN,NETHERLANDS

[2] INTERUNIV RES INST RADIOPATHOL & RADIAT,JA COHEN INST,LEIDEN,NETHERLANDS

[3] ANTEC,LEIDEN,NETHERLANDS

来源：

MUTATION RESEARCH | 1994年 / 307卷 / 01期

关键词：

ELECTROCHEMICAL DETECTION; ALKYLGUANINE ADDUCTS; MOLECULAR DOSIMETRY;

D O I：

10.1016/0027-5107(94)90278-X

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

A methodology is described for the quantitation of 7-arkyl- and O-6-alkylguanine in DNA isolated from experimental animals exposed to alkylating agents. Following purification, the DNA is hydrolysed under acid conditions after which 7-alkyl- and O-6-alkylguanine are separated from unmodified bases by HPLC using a strong cation exchange column. The fractions containing the methylated purines are subsequently analyzed by HPLC using a reverse phase column coupled to an electrochemical detector (amperometric). This method allows the detection of 10-20 fmoles 7-alkyl- and O-6-alkylguanine, when pure markers are analyzed. In practice, the detection limit is 0.5 adducts per 10(6) nucleotides for the methylated and 1 adduct per 10(6) nucleotides for the ethylated form of 7-alkyl- and O-6-alkylguanine using 25 mu g DNA.

引用

页码：61 / 66

页数：6

共 9 条

[1] MOLECULAR DOSIMETRY OF CHEMICAL MUTAGENS - SELECTION OF APPROPRIATE TARGET MOLECULES FOR DETERMINING MOLECULAR DOSE TO GERM LINE [J].