大体积样品堆积-毛细管电泳-紫外检测分析鼠肝中不同亚型金属硫蛋白

制备了均匀稳定的聚丙烯酰胺涂层毛细管,有效抑制了毛细管内壁对金属硫蛋白(MT)的吸附,大大提高了MT的分离度和分离重现性。基于此,建立了大体积样品堆积-毛细管电泳-紫外检测法(LVSS-CE-UV)分析鼠肝中MT的新方法。在最优化的条件下,该方法对两种MT亚型(MT-1/2)的富集倍数分别为13、11倍,检出限分别为0.80、1.01μg·m L~(-1)。将所建立的LVSS-CE-UV方法用于经Cd~(2+)、Zn~(2+)诱导的Sprague Dawley大鼠肝脏中MT-1/2的定量分析,结果表明经Cd~(2+)、Zn~(2+)诱导的鼠肝内均检测出MT-1/2。其中,经Zn~(2+)诱导的鼠肝内MT-1与MT-2的含量分别为31.9、24.3μg·g~(-1);Cd~(2+)诱导组的MT-1与MT-2分别为15.9、31.2μg·g~(-1)。

Large Volume Sample Stacking-Capillary Electrophoresis-Ultraviolet Detection for Analysis of Metallothionein Isoforms in Rats' Liver

A stable and homogeneous polyacrylamide(PAA) coated capillary was prepared for separating metallothionein isoforms (MT-1 and MT-2),effectively suppressing the adsorption of metallothionein onto the inner wall of capillary and highly improving the separation resolution and reproducibility.Large volume sample stacking(LVSS) was combined with the prepared PAA coated capillary for on-column enrichment of MT-1/2 in rat livers followed by CE-UV detection.The EFs for MT-1/2 was 13-and 11-fold,with LODs of 0.80 and 1.01 μg·mL 1,respectively.For real sample analysis,different dose of Zn2+ and Cd2+ were respectively intraperitoneally injected to Sprague Dawley rats to induce metallothionein,and the livers were then analyzed by the proposed LVSS-CE-UV method after suitable treatment.The two metallothionein isoforms were detected after inducement.For Zn2+-inducing group,MT-1 and MT-2 in wet liver were found to be 31.9 and 24.3 μg· g-1,respectively;and for Cd2+-inducing group,MT-1 and MT-2 in wet liver were found to be 15.9 and 31.2 μg·g-1,respectively.The proposed method has been demonstrated to be effective and efficient for direct analysis of metallithionien in real complex samples.