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作 者:杨婉琪 彭利娟 王亚南[2] 李青 陈季旺 吴波 YANG Wanqi;PENG Lijuan;WANG Ya’nan;LI Qing;CHEN Jiwang;WU Bo(College of Food Science and Engineering,Wuhan Polytechnic University,Hubei Key Laboratory for Processing and Transformation of Agricultural Products,Wuhan 430023,China;Department of Clinical Laboratory,The Affiliated Suzhou Hospital of Nanjing Medical University,Suzhou Municipal Hospital,Gusu School,Nanjing Medical University,Suzhou 215002,China)
机构地区:[1]武汉轻工大学食品科学与工程学院,农产品加工与转化湖北省重点实验室,湖北武汉430023 [2]南京医科大学姑苏学院,苏州市立医院,南京医科大学附属苏州医院,江苏苏州215002
出 处:《现代食品科技》2023年第1期320-325,共6页Modern Food Science and Technology
基 金:国家自然科学基金项目(21405118)。
摘 要:采用超高效液相色谱-串联质谱(UPLC-MS/MS)技术,建立了人体摄入吸收后的食物致癌物杂环胺(HAAs)与血清白蛋白(HSA)形成的亚磺酰胺加合物的准确、高灵敏度检测方法,用于评估HAAs的摄取,研究HAAs与相关癌症风险的关系。选取2-氨基-1-甲基-6-苯基咪唑并[4,5-b]吡啶(PhIP)为研究对象,依据PhIP在人体内的代谢途径,体外合成PhIP-HSA亚磺酰胺加合物。使用UPLC-MS/MS检测PhIP-HSA亚磺酰胺加合物的酸水解产物PhIP,对PhIP-HSA亚磺酰胺加合物进行定量。结果表明,HiTrap Blue亲和柱对HSA的回收率高于90%;优化的条件下,PhIP-HSA亚磺酰胺加合物的水解效率高达96%;PhIP-HSA亚磺酰胺加合物的含量与PhIP的信号强度呈良好线性关系;线性回归方程是y=1.0117x+3.2563,R^(2)=0.9987。该方法的检测限和定量限分别低至5×10^(-3) fmol/mg HAS和1.50×10^(-3) fmol/mg HSA。该方法对人血浆中PhIP-HSA亚磺酰胺加合物的测定具有较高的灵敏度,在HAAs摄入与相关癌症风险关系的研究中,是用于评估常食用熟肉个体的HAAs摄入的可行方法。An accurate and highly sensitive assay was developed for the determination of sulfinamide adducts formed by heterocyclic aromatic amines(HAAs)and human serum albumin(HSA)after human ingestion and absorption,using ultra-performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS).The aim of this study was to assess exposure to HAAs,and further investigate the related cancer risks.Using 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine(PhIP)as a research subject,the PhIP-HSA sulfinamide adduct was synthesized in vitro according to the metabolic pathway of the human body and was quantified using UPLC-MS/MS based on the detection of PhIP derived from its acid hydrolysis.The results showed that the recovery of HSA by the HiTrap Blue affinity columns was greater than 90%,and the hydrolysis efficiency of the PhIP-HSA sulfinamide adduct reached 96%under optimized conditions.There was a positive linear relationship between the PhIP-HSA sulfinamide adduct content and the signal intensity of PhIP,and the linear regression equation was y=1.0117x+3.2563,R^(2)=0.9987.The respective detection and quantitation limits for the PhIP-HSA sulfinamide adduct in plasma were as low as 5×10^(-3) fmol/mg HSA and 1.50×10^(-3) fmol/mg HSA.This method showed a high degree of sensitivity when determining PhIP-HSA sulfinamide adducts in human plasma,and proved to be a feasible method for the evaluation of exposure to HAAs in individuals who frequently eat cooked meat,as well as the associated cancer risks.
关 键 词:杂环胺 加合物 人血清白蛋白 超高效液相质谱联用法
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