血中N-[4-(4-氨基苄基)-苯基]乙酰胺血红蛋白加合物的超高效液相色谱-串联质谱测定法  

Determination of N-[4-(4-aminobenzyl)-phenyl]acetamide hemoglobin adduct in blood by ultra performance liquid chromatography-tandem mass spectrometry

作　　者：王小康 李海斌[1] 闫慧芳[1] Wang Xiaokang;Li Haibin;Yan Huifang(National Institute of Occupational Health and Poison Control,Chinese Center for Disease Control and Prevention,Beijing 100050,China)

机构地区：[1]中国疾病预防控制中心职业卫生与中毒控制所,北京100050

出　　处：《中华劳动卫生职业病杂志》2020年第10期772-776,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家科技支撑计划项目(2014BAI12B00)。

摘　　要：目的建立血中二苯基甲烷二异氰酸酯(MDI)代谢产物N-[4-(4-氨基苄基)-苯基]乙酰胺(AcMDA)血红蛋白加合物的超高效液相色谱-串联质谱测定方法。方法准确称取20 mg血红蛋白样品置于15 ml离心管中,加入10μl 20μg/L AcMDA-D8内标溶液,再加入1 ml 0.1 mol/L氢氧化钠溶液在37℃下水解0.5 h,经二氯甲烷液液萃取后,真空浓缩至近干,乙腈复溶后超高效液相色谱-串联质谱测定,内标标准曲线法定量。结果该方法线性范围为0.01~25.00μg/L,相关系数为r=0.9996。方法的检出限和定量下限分别为0.07和0.2 ng/g Hb;方法的加标回收率为91.0%~95.4%;批内精密度为4.5%~6.3%,批间精密度为3.7%~4.4%。结论该方法的各项技术指标符合GBZ/T 210.5-2008《职业卫生标准制定指南第5部分:生物材料中化学物质测定方法》的要求。Objective To establish the method for the determination of N-Acetyl-4,4'-diaminodiphenylmethane(AcMDA)adduct in the hemoglobin by ultra-performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS).Methods The 20 mg hemoglobin sample was weighed into a 15 ml centrifuge tube,adding 20μg/L internal standard solution AcMDA-D810μl,then hydrolyzed with 1 ml 0.1 mol/L sodium hydroxide solution at 37℃ for 0.5 hours,extracted with dichloromethane and concentrated by vacuum concentrator.The residue was dissolved in acetonitrile and detected by UPLC-MS/MS,then quantitative by internal standard method.Results The linearity of the method was good at the range of 0.05-25.00μg/L with a correlation coefficient of 0.9996,the detection limit(LOD)and limit of quantitation(LOQ)were 0.07 and 0.2 ng/g Hb,respectively.The recovery rate was ranged from 91.0%-95.4%;the relative standard deviation(RSD)of intra-and inter-batch precision were 4.5%-6.3% and 3.7%-4.4%,respectively.Conclusion The determination method meet the requirement of GBZ/T 210.5-2008.

关 键 词：血红蛋白 加合物 二苯基甲烷二异氰酸酯 N-[4-(4-氨基苄基)-苯基]乙酰胺 超高效液相色谱-串联质谱 

分 类 号：O657.63[理学—分析化学]