基于HPLC-AFS技术研究南极磷虾油对大鼠脏器中砷形态分布的影响

目的:建立动物组织脏器中砷形态的分析方法,探讨摄食南极磷虾油尤其是砷甜菜碱对大鼠脏器中砷形态分布的影响。方法:雄性Wistar大鼠随机分为大豆油对照组(7 d组、30 d组)、南极磷虾油组(7 d组、30 d组)、添加100 mg/kg砷甜菜碱的南极磷虾油组(7 d组、30 d组),每组5只,灌胃剂量2.625 g油/kg·bw,喂养7 d或30 d后,剥离肝脏、肾脏。采用建立的高效液相色谱-原子荧光光谱法,经Hamilton PRPx-100色谱柱分离,磷酸二氢铵缓冲液洗脱,对脏器中砷甜菜碱(AsB)、二甲基砷酸(DMA)、一甲基砷酸(MMA)、亚砷酸盐(AsⅢ)和砷酸盐(AsⅤ)的含量进行测定。结果:大鼠脏器中五种砷形态的检出限为0.35~0.50 μg/L,定量限为1.0~2.0 μg/L,加标回收率为80.2%~113.0%,RSD为1.34%~3.97%,方法线性良好,R²>0.999。采用该方法测得,大鼠肝脏和肾脏中主要砷形态为AsB和DMA。与对照组相比,大鼠摄食磷虾油和砷甜菜碱磷虾油7或30 d后,脏器中总砷、AsB、AsⅢ、DMA、MMA和AsⅤ含量均无显著性变化。结论:摄食南极磷虾油后,大鼠肝脏和肾脏中不会出现砷蓄积现象。南极磷虾油及其砷甜菜碱不会对大鼠脏器中砷形态含量以及各砷形态之间的转化产生显著影响。本研究可为南极磷虾油食用安全性的科学认识及其应用领域拓展提供理论支撑。

Effects of the Antarctic Krill Oil on the Arsenic Speciation Composition in the Rat Viscera Evaluated by HPLC-AFS Technique

Objective: The effects of dietary intake with the Antarctic krill oil and its arsenobetaine( AsB) on the arsenic speciation composition in the rat viscera were studied using a new established high performance liquid chromatography-atomic fluorescence spectrometry( HPLC-AFS) method. Methods: Male Wistar rats were randomly divided into six groups( 5 each):soybean oil control groups( 7,30 d),krill oil groups( 7,30 d),krill oil added 100 mg/kg arsenobetaine groups( 7,30 d).After7 and 30 d of feeding at the same dose of 2.625 g/kg·bw,the rats were sacrificed and the livers and kidneys were excised.The contents of AsB,dimethyl arsenic acid( DMA),methyl arsenic acid( MMA),arsenite( AsⅢ) and arsenate( AsⅤ) in the viscera were analyzed by the established HPLC-AFS method.The separation column was Hamilton PRPx-100,while the mobile phase was ammonium dihydrogen phosphate buffer. Results: Under the established analysis conditions,the LOD of the five arsenic compounds in the rat viscera was 0.35 ~ 0.50 μg/L and the LOQ was 1.0 ~ 2.0 μg/L. The spike recoveries of the five arsenic compounds at three fortified concentrations ranged from 80.2% to 113.0%.The RSD was 1.34% to 3.97%,the method showed a good linear relationship,R^2> 0.999.The main arsenic speciation in the rat livers and kidneys were AsB and DMA.After 7 and30 d of feeding with the krill oil and its AsB,the contents of total arsenic,AsB,AsⅢ,DMA,MMA and AsⅤin the viscera showed no significant differences compared to the control groups.Conclusion: No arsenic accumulation effects in the rat viscera would appear after dietary intake with the krill oil. Furthermore,the krill oil and its AsB had no significant effects on the changes of the arsenic speciation contents and the transformation between the different arsenic compounds in the rat viscera.This research provided theoretical supports for the scientific recognition of the edible safety of the krill oil as well as the expansion of its application fields.