尿液游离DNA加合物定量测定揭示3种氮芥暴露损伤及其特点OA

The identification of HN-DNA adducts formed in animals after exposure to nitrogen mus‑tards(NMs,codenamed HN)enables the traceability and confirmatory analysis of HN exposure.Six-week-old Sprague-Dawley(SD)rats,with a mean weight of approximately 200 g,were given intra‑peritoneal injections of HN1,HN2,and HN3,respectively,to establish an exposure animal mod‑el.The nitrogen mustard exposure groups(0.3 LD50,0.5 LD50,or LD50 exposure dose)as well as a blank solvent control group were set up.A high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method employing multiple reaction monitoring(MRM)mode was es‑tablished to quantitatively analyze the types and concentrations of HN-DNA adducts in rat urine sam‑ples.The results demonstrated a good linearity for the 12 HN-DNA adduct standards within the con‑centration range of 0.02-500 ng/mL,with limits of detection(LODs)of 0.01-0.02 ng/mL and limits of quantification(LOQs)of 0.02-0.05 ng/mL.The relative standard deviations(RSDs)of intra-assay and inter-assay precision were less than 15%.Urine analysis of HN-exposed SD rats at 12 hours post-exposure revealed the presence of HN-AN3,HN-GN7,HN-GO6,and HN-Bis-GN7 adducts,with concentrations decreasing in the order of HN-GN7>HN-Bis-GN7>HN-AN3>HN-GO6.The concentrations of the four DNA adducts demonstrated a dose-dependent positive correlation and a gradual decrease over time.The application of HPLC-MS/MS technique to detect these urinary DNA adducts allows for the non-invasive early detection,accurate traceability,and damage assessment of HN exposure.These findings provide an experimental basis for further study of HN-induced DNA damage repair processes and mechanisms.