氮芥与DNA体外加合物及其质谱裂解规律  

Mass Spectrometric Fragmentation Patterns of Nitrogen Mustard-DNA Adducts in Vitro

作　　者：王朝霞 刘玉龙 陈佳 刘艳芹 徐华 谢剑炜 WANG Zhao-xia;LIU Yu-long;CHEN Jia;LIU Yan-qin;XU Hua;XIE Jian-wei(Laboratory of Toxicant Analysis,Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences,Beijing 100850,China)

机构地区：[1]军事科学院军事医学研究院毒物药物研究所,北京100850

出　　处：《分析测试学报》2024年第5期690-702,共13页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(21974151)。

摘　　要：氮芥作为两用物质,不仅是禁止化学武器公约清单物,其衍生物至今仍是临床化疗药物。该文应用高效液相色谱-四极杆静电场轨道阱高分辨质谱法分析并鉴定了3种典型氮芥(HN1、HN2、HN3)分别与4种碱基、4种脱氧核糖核苷、4种脱氧核糖核苷酸和鲑鱼精DNA孵育后形成的加合产物。所有39个孵育体系中的HN-DNA加合产物经色谱分离,采用Full MS/dd-MS2模式进行数据采集获得其质谱特征信息。结果表明,在各孵育体系中均可鉴定到3种氮芥与鸟嘌呤、腺嘌呤和胞嘧啶特定碱基位点形成的单加合产物,并可鉴定到其与鸟嘌呤或腺嘌呤形成的双加合产物。在碱基孵育体系中虽可检测到3种氮芥与胸腺嘧啶形成的单加合产物,但在鲑鱼精DNA中并未检测到,表明其并非DNA的显著加合产物。通过探究各加合物的质谱裂解规律和特征加合离子,优化并给出了鉴定的系列DNA加合物的质谱多反应监测离子对,为氮芥暴露潜在生物标志物的鉴定和定量分析提供了重要的技术支撑,为复杂生物样品中氮芥DNA加合物的结构分析、准确溯源以及快速定量奠定了基础。As a dual-use substance,nitrogen mustard is not only on a list of the Convention on the Prohibition of Chemical Weapons,but its derivatives are traditional clinical chemotherapy drugs.In this paper,the adducts formed by incubation of three typical nitrogen mustards(HN1,HN2 and HN3)with 4 bases,4 deoxyribonucleosides,4 deoxyribonucleotides and salmon DNA respectively,were analyzed and identified by high-performance liquid chromatography-quadrupole electrostatic field orbitrap high-resolution mass spectrometry.The HN-DNA adducts in all 39 incubation systems were chromatographically separated and the data were collected by Full MS/dd-MS2 mode to obtain their mass spectrometry characteristics.The results showed that the mono-adducts formed by three kinds of nitrogen mustard with guanine,adenine and cytosine could be identified in each incubation system and the di-adducts formed by nitrogen mustard with guanine or adenine could be identified.Although the mono-adducts formed by three kinds of nitrogen mustard with thymine were detected in the base incubation system,they were not detected in the salmon DNA incubation system,indicating that adducts formed by HNs with thymine base were not primary adducts for DNA.By exploring the mass spectrometry characteristic fragments of each adduct,the specific ion pairs of series of DNA adducts for the multi-reaction monitoring mode of mass spectrometry have been summarized,which lays a foundation for the establishment of a method for structural analysis,accurate traceability and rapid quantification of nitrogen mustard-DNA adducts in complex biological samples.

关 键 词：氮芥 DNA加合物 特征加合离子 高效液相色谱-四极杆静电场轨道阱高分辨质谱 

分 类 号：O657.63[理学—分析化学] R991[理学—化学]