IDENTIFICATION OF PHOSPHORAMIDE MUSTARD DNA ADDUCTS USING TANDEM MASS-SPECTROMETRY
IDENTIFICATION OF PHOSPHORAMIDE MUSTARD DNA ADDUCTS USING TANDEM MASS-SPECTROMETRY
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DOI：
10.1002/rcm.1290041014
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发表时间：
1990-10-01
影响因子：
2
通讯作者：
MIRKES, PE
中科院分区：
文献类型：
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作者：
CUSHNIR, JR;NAYLOR, S;MIRKES, PE
在双功能抗癌药物的设计中，经常利用烷化剂的反应途径。这些药物形成单DNA加合物以及链间和链内交联加合物，特别是通过在DNA碱基上反应，包括鸟嘌呤（G）的N-7-位。磷酰胺芥烷基化DNA体外制备的正离子快原子轰击（FAB）质谱（抗癌药物环磷酰胺的活性代谢产物）表明存在两个单DNA加合物N-（2-氯乙基）-N-[2-（7-鸟嘌呤基）乙基]胺，命名为NOR G，和N-（2-羟乙基）-N-[2-（7-鸟嘌呤基）乙基]胺，命名为NOR G OH（MH+分别为257/259和239），但不存在交联加合物N，N-双-[2-（7-鸟嘌呤基）乙基]胺，命名为NOR G（MH+372）。使用合成标准品，通过正离子FAB串联质谱法（FAB-MS/MS）获得NOR G、NOR G OH和G NOR G的子光谱（基质0.2 Mp-甲苯磺酸甘油溶液）。单DNA加合物NOR G和NOR G OH的子离子光谱都含有一个碎片离子atm/z152 [G + H]+，而交联加合物G NOR G则显示一个离子atm/z221 [MH − G]+。通过对m/z 152和221进行双母离子扫描，获得了体外制备中存在NOR G、NOR G OH和G NOR G的证据。通过对m/z 221进行单个母离子扫描，进一步支持了G-NOR-G的存在。这种MS/MS技术的使用应该消除了在生物提取物中鉴定G β NOR β G的复杂样品纯化的需要。
The reaction pathway of alkylating agents is often exploited in the design of bifunctional anti‐cancer drugs. These drugs form mono‐DNA adducts as well as inter‐ and intra‐strand cross‐linked adducts, notably by reaction at DNA bases, including theN‐7‐position of guanine (G). A positive‐ion fast‐atom bombardment (FAB) mass spectrum of anin vitropreparation of DNA alkylated with phosphoramide mustard (the active metabolite of the anti‐cancer drug cyclophosphamide) indicated the presence of the two mono‐DNA adductsN‐(2‐chloroethyl)‐N‐[2‐(7‐guaninyl)ethyl] amine, designated NORG, andN‐(2‐hydroxyethyl)‐N‐[2‐(7‐guaninyl)ethyl] amine, designated NORGOH, (MH+257/259 and 239, respectively) but not the presence of the cross‐linked adductN,N‐bis‐[2‐(7‐guaninyl)ethyl] amine, designated GNORG (MH+372). Using synthetic standards, daughterion spectra of NORG, NORGOH and GNORG were obtained (matrix 0.2 Mp‐toluene sulphonic acid in glycerol) by positive‐ion FAB tandem mass spectrometry (FAB‐MS/MS). The daughter‐ion spectra of both mono‐DNA adducts NORG and NORGOH contained a fragment ion atm/z152 [G + H]+, whereas the cross‐linked adduct, GNORG, showed an ion atm/z221, [MH − G]+. Evidence for the presence of NORG, NORGOH and GNORG in thein vitropreparation was obtained by performing a double parent‐ion scan onm/z152 and 221. The presence of GNORG was further supported by performing a single parent‐ion scan onm/z221. The use of this MS/MS technique should eliminate the need for intricate sample purification in the identification of GNORG in biological extracts.