环境毒理与生态毒理
孟紫强,李瑞金,张欣.HPLC荧光检测法测定SO2吸入后小鼠脑、心和肺亚硫酸盐水平[J].环境科学学报,2005,25(3):346-350
HPLC荧光检测法测定SO2吸入后小鼠脑、心和肺亚硫酸盐水平
- The measuration of sulfite levels in brains, hearts and lungs of the mice after sulfur dioxide inhalation by HPLC with fluorescence detection
- 基金项目：国家自然科学基金资助项目(No.20477023;No.30230310);山西省自然科学基金(No.991123)
- 作者
- 单位
- 孟紫强
- 山西大学环境医学与毒理学研究所, 太原 030006
- 李瑞金
- 山西大学环境医学与毒理学研究所, 太原 030006
- 张欣
- 山西大学环境医学与毒理学研究所, 太原 030006
- 摘要：为了探讨二氧化硫(SO2)吸入后是否可进入小鼠不同组织器官,运用高效液相色谱(HPLC)荧光检测(FD)法测定了SO2动式吸入后雄性小鼠脑、心和肺组织中SO2在体内的衍生物———亚硫酸盐含量.组织匀浆液经还原、衍生和沉淀蛋白,取上清液进入色谱仪检测.亚硫酸盐测定标准曲线在0.126μg·mL-1～126μg·mL-1有良好的线性关系,检测限为0.04μg·mL-1(SN=3),测定方法的回收率在97%～101%之间,日内和日间的精密度RSD低于9%.分析结果表明,SO2吸入后小鼠3种器官组织中亚硫酸盐含量比对照组显著增加(P<0.05),且与SO2浓度呈明确的剂量效应关系(r>0.92).这说明SO2被小鼠吸入后转化为亚硫酸盐并可分布到肺和其它器官如脑和心等,从而为SO2是一种全身性毒物的观点提供支持.此外,本文对HPLC荧光检测亚硫酸盐的方法作了改进,为研究SO2及其衍生物的毒作用提供了更为有效的方法.
- Abstract：The distribution of the sulfur dioxide (SO2) permeated into the different organs in the mouse after SO2 was inhaled was studied The concentration of the sulfite in brains, hearts and lungs of the male mice were measured by high performance liquid chromatographic (HPLC) with fluorescence detection (FD). Firstly, the protein was reduced, derived and deposited from the homogenates of brains, hearts and lungs tissue from mice. And then the mixture was centrifuged, and 5μLsupernatant was directly injected into HPLC, where the flow phase was methanol-phosphoric acid(12:88,V/V),and the exciting and emitting wavelength of fluorescence detection were 392nm and 478nm, respectively. The standard curve of the sulfite was greatly linear fit in the range from 0.126μg·mL-1 to 126μg·mL-1, and the minimal detectable concentration was 0.04μg·mL-1(S/N=3) with the average recycle rates of method from 97% to 101%. The error-sensitivity within one day or between different days were less than 9%.The results showed that sulfite concentration in all the organs of mice in SO2-exposed groups was significantly increased (P<0.05) compared with the controlled groups. And it was increased in a specific dose-dependent manner(r>0.92) which indicated SO2 could transform into sulfite after it was inhaled, and distributed into lung and other organs such as brain and heart. These results supported the viewpoint that SO2 was a systemic toxic agent. Moreover, the method of the measuration of sulfite by HPLCwith fluorescence detection was improved which provided a more effective method to study the toxicological action of SO2 and its derivatives.
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