高效毛细管电泳法研究重瓣榆叶梅花中蛋白质的动态变化

Study on dynamic changes of protein contents in the flower of Amygdalus triloba f.multiplex by HPCE

为验证毛细管电泳方法,研究测定重瓣榆叶梅花朵生长过程中的DNA酶、RNA酶和细胞色素C含量及动态变化的有效性,以磷酸氢二钠溶液为提取液,超声提取重瓣榆叶梅花朵中的蛋白质作为待测液,考察缓冲溶液种类、浓度、pH、分离电压、分离温度及进样时间等因素对HPCE分离检测蛋白质的影响。确定了HPCE最佳测定条件为:运行缓冲液为pH为2.5、浓度为30mmol/L的磷酸氢二钠溶液,0.5MPa压力进样5s,分离电压为8kV,电泳温度为25℃,检测波长为254nm。在最佳条件下,样品可在20min内完全分离,线性关系良好。该方法准确、快速、简便,可同时检测样品中的DNA酶、RNA酶和细胞色素C。

In order to verify HPCE method and study the effectiveness of measuring,RNase and Cytochrome C and study the effectiveness of their dynamic changes in flower of Amygdalus triloba f. multiplex, samples extracted with Disodium hydrogen phosphate solution by ultrasonic treatment were used as the test solutions after separation and purification. The effects of the concentration and pH of buffer, separation voltage, separation temperature and injection time on the separation efficiency were confirmed. The optimal conditions were., the running buffer, 30 mmol/L disodium hydrogen phosphate with pH 2.5 ; injection time, 5 s with the pressure of 0.5 MPa; Separation voltage, 8 kV ; Electrophoresis temperature,25 ℃ ;Detection wave length,254 nm. The samples could be completely separated in 20 min with a good linear relationship under the above conditions. The proposed determination of DNase, RNase and Cytochrome C in flowers of accuracy, fast separation and simple extraction. method could be simultaneously applied for the A. triloba f. multiplex with the advantages of high accuracy,fast separation and simple extraction.