Biomarker analysis of hemoglobin adducts of acrylamide and glycidamide enantiomers for mid-term internal exposure assessment by isotope dilution ultra-high performance liquid chromatography tandem mass spectrometry
Biomarker analysis of hemoglobin adducts of acrylamide and glycidamide enantiomers for mid-term internal exposure assessment by isotope dilution ultra-high performance liquid chromatography tandem mass spectrometry
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采用同位素稀释超高效液相色谱串联质谱法对丙烯酰胺和缩水甘油酰胺对映体血红蛋白加合物进行中期内暴露评估的生物标志物分析
DOI：
10.1016/j.talanta.2017.09.092
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发表时间：
2018
期刊：
影响因子：
6.1
通讯作者：
Wu Yongning
中科院分区：
文献类型：
--
作者：
Zhang Yu;Wang Qiao;Zhang Gong;Jia Wei;Ren Yiping;Wu Yongning
丙烯酰胺（AA）及其氧化代谢产物甘氨酰胺（GA）的血红蛋白（Hb）加合物是评价丙烯酰胺中期暴露毒性的重要生物标志物。以N-异硫氰酸五氟-2-甲基苯基酯为例，（2-氨甲酰乙基）缬氨酸（AAVal-PFPTH）和N-以2-氨甲酰基-2-羟乙基）缬氨酸（GAVal-PFPTH）为目标分析物，建立了同位素稀释超高效液相色谱串联质谱（UHPLC-MS/MS）同时测定AA和GA血红蛋白（Hb）加合物的方法。其中，GA-Hb加合物的对映体对首次被鉴定并在基线水平上成功分离。该方法具有较高的灵敏度，检测限和定量限分别为1.43-5.05 pmol/g Hb和4.78-16.82 pmol/g Hb。低、中和高加标水平的回收率分别计算为97.0- 105.2%、97.4-106.4%和100.3- 111.2%。可接受的实验室内重现性（RSD < 13.7%）基本上支持了当前UHPLC-MS/MS方法的耐用性，该方法已成功用于测定大鼠和人体血液中丙烯酰胺和缩水甘油酰胺对映体的血红蛋白加合物。通过将丙烯酰胺血红蛋白加合物作为变量，开发了一种线性暴露评估模型，用于估计人类每日丙烯酰胺暴露量，表明基于生物标志物的内部暴露和基于饮食的外部暴露之间存在新的联系。总体而言，本仪器分析和相关的内部暴露评估模型提供了一个实质性的方法学支持，描绘内部生物暴露和估计外部饮食暴露的丙烯酰胺。
Hemoglobin (Hb) adducts of acrylamide (AA) and its oxidative metabolite glycidamide (GA) are important biomarkers for evaluating the mid-term exposure of acrylamide toxicity in vivo. Taking pentafluoro-2-methylphenyl isothiocyanates of N-(2-carbamoylethyl)valine (AAVal-PFPTH) and N-(2-carbamoyl-2-hydroxyethy)valine (GAVal-PFPTH) as target analytes, we developed an isotope dilution ultra-high performance liquid chromatograph tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous determination of AA and GA hemoglobin (Hb) adducts under the electroscopy ionization negative (ESI‾) mode in the present work. Among them, the enantiomer pair of GA-Hb adducts was firstly identified and successfully separated at baseline level. The method achieved high sensitivity with the LOD and LOQ ranging 1.43–5.05 pmol/g Hb and 4.78–16.82 pmol/g Hb, respectively. The recovery rates with low, intermediate and high spiking levels were calculated as 97.0–105.2%, 97.4–106.4% and 100.3–111.2%, respectively. Acceptable within-laboratory reproducibility (RSD < 13.7%) substantially supported the robustness of current UHPLC-MS/MS method, which was successfully applied to measure the hemoglobin adducts of acrylamide and glycidamide enantiomers in blood of both rats and humans. A linear exposure assessment model was developed for estimating the daily exposure to acrylamide in humans via considering acrylamide hemoglobin adducts as variables, indicating a novel connect between biomarker-based internal exposure and dietary-based external exposure. Overall, the present instrumental analysis and related internal exposure assessment model provide a substantially methodological support for profiling the internal biological exposure and estimating the external dietary exposure to acrylamide.