色谱

色谱 ›› 2011, Vol. 29 ›› Issue (04): 325-329.DOI: 10.3724/SP.J.1123.2011.00325

• 特别策划 • 上一篇    下一篇

高效液相色谱-电喷雾离子阱串联质谱法检测芥子气经皮肤染毒SD大鼠肺中的DNA加合物

岳丽君, 魏玉霞, 陈佳, 林妮妮, 王瑞, 杨茜, 张雅姣, 谢剑炜*   

  1. 军事医学科学院毒物药物研究所, 北京 100850
  • 收稿日期:2010-12-22 修回日期:2011-01-21 出版日期:2011-04-28 发布日期:2011-04-28
  • 通讯作者: 谢剑炜,博士,研究员,博士生导师. E-mail: xiejw@bmi.ac.cn.
  • 基金资助:
    国家“重大新药创制”科技重大专项平台项目(No. 2009ZX09301-002)和第45批博士后科学基金(No. 20090450198).

Biomonitoring method of dermal exposure to sulfur mustard based on DNA adduct in lung of SD rats by high performance liquid chromatography-electrospray ionization tandem mass spectrometry

YUE Lijun, WEI Yuxia, CHEN Jia, LIN Nini, WANG Rui, YANG Xi, ZHANG Yajiao, XIE Jianwei*   

  1. Beijing Institute of Pharmacology and Toxicology, Beijing 100850, China
  • Received:2010-12-22 Revised:2011-01-21 Online:2011-04-28 Published:2011-04-28

PDF

513

被引次数

8

摘要： 运用高效液相色谱-电喷雾离子阱串联质谱(HPLC-ESI-MS/MS)技术,建立了快速、简单、灵敏的SD大鼠肺中N7-(2-羟乙基硫代乙基)鸟嘌呤(N7-HETEG)的检测方法。以N7-苯甲基鸟嘌呤为内标,用甲醇和水为流动相进行梯度洗脱,正离子模式检测,方法的检出限(信噪比(S/N)≥10)为300 pg/mL,定量限(S/N≥20)为850 pg/mL。在300 pg/mL～1.28 μg/mL的质量浓度范围内,N7-HETEG浓度与N7-HETEG和内标的峰面积比呈良好的线性关系(线性相关系数为0.9929)。高、中、低3个添加水平的日内测定精密度(以相对标准偏差(RSD)计)和日间测定精密度均小于10%(n=7),回收率为100%～132%。对SD大鼠背部皮肤染芥子气,剂量分别为5.5、11、22和45 mg/kg,染毒4 d后检测大鼠肺脏中N7-HETEG的含量。各个不同染毒剂量下,每克组织中分别检测到（0.56±0.16）、（0.67±0.12）、（1.36±0.68）和(5.14±0.92) ng N7-HETEG, N7-HETEG的含量随着染毒剂量的增大而增大,表明N7-HETEG可用作芥子气暴露的体内生物标志物。

关键词: N7-(2-羟乙基硫代乙基)鸟嘌呤, SD大鼠, 肺, 高效液相色谱-电喷雾离子阱串联质谱, 芥子气, 皮肤染毒

Abstract: The analysis of biological samples can provide qualitative and quantitative evidence of exposure to sulfur mustard (SM). N7-(2-hydroxyethylthioethyl)guanine (N7-HETEG) is the most abundant DNA adduct and one of the biomarkers of SM exposure. For the detection and validation of SM exposure, a sensitive high performance liquid chromatography-tandem mass spectrometry (HPLC-ESI-MS/MS) method for the determination of N7-HETEG in a biomedical sample was developed, using N7-benzylguanine as the internal standard. The method was validated and showed the limit of detection of 300 pg/mL (S/N≥10) and limit of quantitation of 850 pg/mL (S/N≥20). The linear range was 300 pg/mL~1.28 μg/mL, with both the values of intraday and interday relative standard deviations of less than 10% (n=7) over the calibration range. The recoveries varied from 100% to 132%. The method was applied for the determination of N7-HETEG in the lung of SD rats caused by dermal exposure to SM in vivo. Under the exposure to SM of 5.5, 11, 22 and 45 mg/kg, respectively, （0.56±0.16）, （0.67±0.12）, （1.36±0.68） and (5.14±0.92) ng of N7-HETEG per gram of SD rat lung tissue were found after four days of exposure. The amount of N7-HETEG increased with the exposed doses, and it can act as a biomarker of sulfur mustard exposure.

Key words: dermal exposure, lung, N7-(2-hydroxyethylthioethyl)guanine (N7-HETEG), SD rat, high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS), sulfur mustard (SM)