CAJ | 학술논문

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/L 3-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。
건립료이갑기갑선알( N,N-dimethyl formamide,DMF)직업접촉자혈홍단백중N-갑기안갑선가합물초고효액상색보-질보련용(UPLC/MS/MS)쾌속검측방법。준학칭취0.1 g 혈홍단백양품치우15 mL리심관중,가입40μmol/L 3-갑기-이정기해인(3-methy-5-isobutylhydantion,MIH)내표용액250μL,재가입HCl-작산(2∶1, V/V)용액4.75 mL,진탕혼균3 min。장시관치우비수욕중가열강해1 h,취출냉각。혼균후취200μL도96공판,가입600μL1%갑산을정용액,혼균。공제진공빙압력,사양품재2~4 min류출。장정화후양품혼균전이도양품병,용 UPLC/MS/MS 검측 N-갑기안갑선가합물적강해산물3-갑기-이병기을내산뇨(3-Methyl-5-isopropylhydantoin, MVH)。질보채집방식위다반응감측모식,내표법정량。결과표명,MVH농도재0.01~1.0μmol/L범위내선성관계량호,상관계수위0.999。공백인군혈홍단백 MVH 가표회수솔위97.3%,상대표준편차위1.7%,혈홍단백중MVH검출한위0.01μmol/g。본방법능흔호지응용우이갑기갑선알직업접촉자혈홍단백중신형생물표지물N-갑기안갑선가합물적쾌속、준학측정。
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 1h. 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 was transferred into the sample vial, and 3-methyl-5-isopropylhydantoin ( MVH ) as degration product of 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 μmol/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.