摘要
建立了二甲基甲酰胺（N,N-dimethyl formamide,DMF）职业接触者血红蛋白中N-甲基氨甲酰加合物超高效液相色谱-质谱联用（UPLC/MS/MS）快速检测方法。准确称取0.1 g血红蛋白样品置于15 mL离心管中,加入40μmol/L3-甲基-异丁基海因（3-methy-5-isobutylhydantion,MIH）内标溶液250μL,再加入HCl-醋酸（2∶1,V/V）溶液4.75 mL,振荡混匀3 min。将试管置于沸水浴中加热降解1 h,取出冷却。混匀后取200μL到96孔板,加入600μL1%甲酸乙腈溶液,混匀。控制真空泵压力,使样品在2~4 min流出。将净化后样品混匀转移到样品瓶,用UPLC/MS/MS检测N-甲基氨甲酰加合物的降解产物3-甲基-异丙基乙内酸脲（3-Methyl-5-isopropylhydantoin,MVH）。质谱采集方式为多反应监测模式,内标法定量。结果表明,MVH浓度在0.01~1.0μmol/L范围内线性关系良好,相关系数为0.999。空白人群血红蛋白MVH加标回收率为97.3%,相对标准偏差为1.7%,血红蛋白中MVH检出限为0.01μmol/g。本方法能很好地应用于二甲基甲酰胺职业接触者血红蛋白中新型生物标志物N-甲基氨甲酰加合物的快速、准确测定。
A rapid analytical method was developed for the determination of N-methylcarbamoyl adduct in Hemoglobin of workers exposed to N, N-dimethylformamide by ultra high performance liquid chromatography- mass spectrometry （UPLC/MS/MS）. About 0.1 g of hemoglobin sample, 40 μmol/L of 3-methyl-5- isopropylhydantoin （MIH） as the internal standard and 4.75 mL of HCl-acetic acid （2： 1, V/V） were added in the centrifuge tube, and mixed for 3 min. Then the tube was heated in boiling water bath for 1 h. After cooling down, 200 μL of the mixture and 600 μL of formic acid-acetonitrile （ 1% ） were added into 96-well extract plate. The vacuum pump pressure was controlled to make the sample collection elute within 2-4 min.The purified collection degration product of was transferred into the sample vial, and 3-methyl-5-isopropylhydantoin （MVH） as N-methylcarbamoyl adduct was quantified by UPLC/MS/MS in multiple reaction monitoring （MRM） by internal standard method. A good linear relationship was obtained in the MVH concentration range of 0.01 - 1.0 p, mol/L with the correlation coefficient of 0. 999. The recovery of added MVH in the blank sample was 97.3% and the relative standard deviation was 1.7%. The limit of detection （LOD） was 0.01 μmoL/g. This method was proved to be fast and efficient.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2014年第9期1326-1331,共6页
Chinese Journal of Analytical Chemistry
基金
国家自然科学基金面上项目(No.81372963)
江苏省预防医学科研基金项目(No.Y200720)
江苏省卫生厅科研项目(No.H200866)资助~~
