National Engineering Laboratory of Intelligent Food Technology and Equipment, Zhejiang Key Laboratory for Agro-Food Processing; Fuli Institute of Food Science, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China.
Key Laboratory of Food Safety Risk Assessment, Ministry of Health; China National Center for Food Safety Risk Assessment, Beijing 100022, China.
Talanta. 2018 Feb 1;178:825-833. doi: 10.1016/j.talanta.2017.09.092. Epub 2017 Oct 1.
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 1.43-5.05pmol/g Hb and 4.78-16.82pmol/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.
血红蛋白（Hb）与丙烯酰胺（AA）及其氧化代谢物丙烯醛（GA）的加合物是评估体内丙烯酰胺毒性中期暴露的重要生物标志物。本工作以 N-（2-氨乙基）-缬氨酸戊基异硫氰酸酯（AAVal-PFPTH）和 N-（2-氨乙基-2-羟乙基）-缬氨酸（GAVal-PFPTH）为目标分析物，建立了电喷雾负离子（ESI‾）模式下同时测定血红蛋白（Hb）中 AA 和 GA 加合物的同位素稀释超高效液相色谱串联质谱（UHPLC-MS/MS）方法。其中，首次在基线水平上成功分离并鉴定了 GA-Hb 加合物的对映体。该方法具有较高的灵敏度，LOD 和 LOQ 分别为 1.43-5.05pmol/g Hb 和 4.78-16.82pmol/g Hb。低、中、高添加水平下的回收率分别为 97.0-105.2%、97.4-106.4%和 100.3-111.2%。实验室内部重复性（RSD<13.7%）可接受，充分证明了当前 UHPLC-MS/MS 方法的稳健性，该方法成功应用于大鼠和人血液中丙烯酰胺和丙烯醛加合物的测定。建立了线性暴露评估模型，通过将丙烯酰胺血红蛋白加合物作为变量来估计人体对丙烯酰胺的每日暴露量，这表明生物标志物内暴露与饮食外暴露之间存在新的联系。总的来说，本研究中的仪器分析和相关的内暴露评估模型为分析体内生物暴露和估计丙烯酰胺的饮食外暴露提供了重要的方法学支持。
