Abundance of Four Sulfur Mustard-DNA Adducts ex Vivo and in Vivo Revealed by Simultaneous Quantification in Stable Isotope Dilution鈥揢ltrahigh Performance Liquid Chromatography鈥揟andem Mas

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• 作者：Lijun Yue ; Yuxia Wei ; Jia Chen ; Huiqin Shi ; Qin Liu ; Yajiao Zhang ; Jun He ; Lei Guo ; Tingfen Zhang ; Jianwei Xie ; Shuangqing Peng
• 刊名：Chemical Research in Toxicology
• 出版年：2014
• 出版时间：April 21, 2014
• 年：2014
• 卷：27
• 期：4
• 页码：490-500
• 全文大小：641K
• 年卷期：v.27,no.4(April 21, 2014)
• ISSN：1520-5010

文摘

Sulfur mustard (SM) is a highly reactive alkylating vesicant and causes blisters upon contact with skin, eyes, and respiratory organs. It covalently links with DNAs by forming four mono- or cross-link adducts. In this article, the reference standards of SM-DNA adducts and deuterated analogues were first synthesized with simplified procedures containing only one or two steps and using less toxic chemical 2-(2-chloroethylthio)ethanol or nontoxic chemical thiodiglycol as starting materials. A sensitive and high-throughput simultaneous quantification method of N⁷-[2-[(2-hydroxyethyl)thio]-ethyl]guanine (N⁷-HETEG), O⁶-[2-[(2-hydroxyethyl)thio]-ethyl]guanine (O⁶-HETEG), N³-[2-[(2-hydroxyethyl)thio]-ethyl]adenine (N³-HETEA), and bis[2-(guanin-7-yl)ethyl]sulfide (Bis-G) in the Sprague鈥揇awley rat derma samples was developed by stable isotope dilution鈥搖ltrahigh performance liquid chromatography鈥搕andem mass spectrometry (ID-UPLC-MS/MS) with the aim of revealing the real metabolic behaviors of four adducts. The method was validated, the limit of detection (S/N ratio greater than 10) was 0.01, 0.002, 0.04, and 0.11 fmol on column for N⁷-HETEG, O⁶-HETEG, Bis-G, and N³-HETEA, respectively, and the lower limit of quantification (S/N ratio greater than 20) was 0.04, 0.01, 0.12, and 0.33 fmol on column for N⁷-HETEG, O⁶-HETEG, Bis-G, and N³-HETEA, respectively. The accuracy of this method was determined to be 76% to 129% (n = 3), and both the interday (n = 6) and intraday (n = 7) precisions were less than 10%. The method was further applied for the quantifications of four adducts in the derma of adult male Sprague鈥揇awley rats exposed to SM ex vivo and in vivo, and all adducts had time鈥?and dose鈥揺ffect relationships. To the best of our knowledge, this is the first time that the real presented status of four DNA adducts was simultaneously revealed by the MS-based method, in which Bis-G showed much higher abundance than the result previously reported and N³-HETEA showed much less. It should be noted that since the interstrand cross-linked adduct is believed to stall DNA replication and finally induce a double-strand break, the higher abundance of Bis-G is a great indication of a more serious DNA lesion by SM alkylation.