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目的:建立柱前衍生化-HPLC法检测肝脏GSH的方法,探讨雄黄中砷对小鼠肝脏GSH含量的影响。方法:将40只ICR小鼠随机分成对照组、雄黄低、中、高剂量组(0.15 g/kg、0.45 g/kg、1.35 g/kg雄黄),连续灌胃8周;采用柱前衍生化-高效液相色谱-紫外检测法测定小鼠肝脏GSH含量,同时采用氢化物发生-火焰原子吸收法(HGFAAS)测定雄黄染毒小鼠肝脏砷含量。结果:GSH在10~500μmol/L范围内线性关系良好,日内、日间精密度的RSD均小于5%,加标回收率在81.9%~96.4%之间;随着雄黄染毒剂量的升高,GSH含量呈现出先升高后降低的趋势,且雄黄低、中剂量组较对照组有显著性差异(P<0.05),而雄黄高剂量组较对照组未表现出显著性差异(P>0.05);雄黄染毒小鼠肝脏砷含量随着染毒剂量增加而逐渐升高,且低、中、高剂量均较对照组有显著性差异(P<0.05)。结论:该方法操作简单、快速、准确、重现性好、灵敏度高,适合肝脏中GSH含量的测定;雄黄染毒8周,可引起小鼠肝脏中GSH含量变化。
Objective: To establish a method of precolumn derivatization-HPLC for detection of hepatic GSH and to explore the effect of arsenic in realgar on the hepatic GSH in mice. Methods: Forty ICR mice were randomly divided into control group, low, medium and high dose realgar (0.15 g / kg, 0.45 g / kg, 1.35 g / kg and realgar) The contents of GSH in the liver of mice were determined by high performance liquid chromatography with ultraviolet detection. The arsenic content in the liver of realgar was measured by hydride generation - flame atomic absorption spectrometry (HGFAAS). Results: The linearity of GSH in the range of 10 ~ 500μmol / L was good. The intra-day and inter-day RSDs were less than 5%, and the recoveries were between 81.9% and 96.4%. With the increase of the dose of realgar (P <0.05), while there was no significant difference between the high-dose realgar and the control group (P> 0.05) ). The arsenic content in the male and female mice increased gradually with the increase of exposure dose, and there was significant difference between the low, medium and high dose groups compared with the control group (P <0.05). Conclusion: The method is simple, rapid, accurate, reproducible and sensitive. It is suitable for the determination of GSH in the liver. When andrographolide is applied for 8 weeks, it can cause the change of GSH in the liver of mice.