LC-MS/MS法测定大鼠血浆中偏二甲肼浓度及毒代动力学研究  

LC-MS/MS determination of unsymmetrical dimethylhydrazine concentrations in rat plasma and toxicokinetic studies

作　　者：石沅叆 石富江 宋妃灵 李亚娟 廖沙 杜克贺 尹纪业 SHI Yuanai;SHI Fujiang;SONG Feiling;LI Yajuan;LIAO Sha;DU Kehe;YIN Jiye(Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China;Beijing Huizhi Taikang Pharmaceutical Technology Co.,Ltd,Beijing 100176,China)

机构地区：[1]军事科学院军事医学研究院,北京100850 [2]北京汇智泰康医药技术有限公司,北京100176

出　　处：《军事医学》2024年第11期826-831,共6页Military Medical Sciences

摘　　要：目的建立快速灵敏的测定大鼠血浆中偏二甲肼(UDMH)含量的液相色谱‐串联质谱(LC‐MS/MS)分析方法,并探究UDMH在大鼠体内的毒代动力学特征。方法22只SD大鼠分为静脉给药10 mg/kg剂量组(4只雌性)和灌胃给药组(低、中、高剂量组,每组6只,雌雄各半)。以10 mg/kg剂量单次静脉注射及以10、30、90 mg/kg剂量灌胃给药UDMH,在给药前0 h以及给药后不同时间点进行眼眶静脉丛取血。用蛋白沉淀和衍生化法处理血浆样品并进行LC‐MS/MS分析。色谱柱:ZORBAX柱(4.6 mm×75 mm,3.5μm);流动相:0.3%甲酸水‐乙腈;流速:1 mL/min;内标:盐酸普萘洛尔;离子源:ESI源,正离子MRM模式检测;定量分析UDMH和盐酸普萘洛尔的检测离子分别为m/z:192.0→148.1,m/z:260.2→116.1。采用DAS 2.1软件计算毒动学参数。结果UDMH在50~50000 ng/mL浓度范围内线性良好,线性相关系数大于0.9900,定量下限为50 ng/mL。UDMH平均回收率为98.1%,内标盐酸普萘洛尔的回收率为100.5%,标准曲线样品的批间精密度为0.7%~6.3%,相对误差为-7.1%~6.2%,质控样品的批间、批内精密度为1.8%~19.8%,相对误差为-9.8%~0.2%。大鼠10、30、90 mg/kg灌胃染毒UDMH的主要毒代动力学参数分别为UC(0‐t):(7624.99±2569.31)、(34284.04±6657.15)、(84720.88±22354.80)μg/L·h,t_(1/2):(0.07±0.15)、(2.24±1.45)、(3.04±0.90)h,T_(max):(0.75±0.27)、(0.51±0.29)、(0.29±0.10)h,C_(max):(4454.14±1329.45)、(19442.45±9121.07)、(32334.35±9882.41)μg/L,生物利用度(F):(77.34±26.06)%、(115.92±22.51)%、(95.48±25.19)%。结论LC‐MS/MS法准确度高、专属性强,适用于UDMH在大鼠体内的毒代动力学研究。单次灌胃给药UDMH在高剂量下存在吸收和消除饱和。本研究为UDMH的相关毒理学研究提供了数据支持。Objective To establish a rapid and sensitive liquid chromatography-tandem mass spectrometry(LC-MS/MS)analysis method for determination of unsymmetrical dimethylhydrazine(UDMH)contents in rat plasma and investigate the toxicokinetic characteristics of UDMH in rats.Methods Twenty-two SD rats were divided into the intravenous injection of 10 mg/kg dose group(4 females)and intragastric administration groups(low,medium and high dose,with 6 rats in each group,half males and half females).The rats were given 10mg/kg by intravenous administration and 10 mg/kg,30 mg/kg,and 90 mg/kg single dose of UDMH by gavage.Blood samples were collected from the orbital venous plexus at 0 hour before administration and at different time points after administration.The plasma samples were extracted with protein precipitation and derivatization before being analyzed using the LC-MS/MS method.Separation was carried out on a ZORBAX column(4.6mm×75mm,3.5µm),with a mobile phase composed of 0.3%acetonitrile/formic acid at a flow rate of 1 mL/min.Propranolol was used as the internal standard.An electrospray ionization(Turbo Ionspray)source was applied and the mass spectrometer was operated in a positive MRM mode.Quantitative analysis showed that the ionization source unsymmetrical dimethylhydrazine and propranolol was at m/z:192.0→148.1,m/z:260.2→116.1,respectively.The toxicokinetic parameters were analyzed with the DAS 2.1 software.Results Quantification of UDMH exhibited a good linearity within the concentration range of 50-50000 ng/mL,with a linear correlation coefficient greater than 0.9900 and a lower limit of quantification of 50 ng/mL.The average recovery rate of UDMH was 98.1%,compared with 100.5%for the internal standard propranolol hydrochloride.The inter batch precision of standard curve samples ranged from 0.7%to 6.3%,and the relative error was between-7.1%and 6.2%.The inter batch and intra batch precision of quality control samples ranged from1.8%to 19.8%,and the relative error from-9.8%to 0.2%.The main pharmacokinetic param

关 键 词：偏二甲肼 血药浓度 毒代动力学 液相色谱-串联质谱 

分 类 号：R917[医药卫生—药物分析学]