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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 the capillary electrophoresis method, the determination of the contents of DNase, RNase and cytochrome C in the flower of Prunus triloba and the dynamic change of the cytochrome C content were studied. With the disodium hydrogen phosphate solution as the extract, The protein in the leaves of Plum Blossom was used as the test solution to investigate the effects of the buffer solution type, concentration, pH, separation voltage, separation temperature and injection time on the protein separation by HPCE. The best determination conditions of HPCE were as follows: the running buffer was pH = 2.5, the concentration of 30mmol / L disodium hydrogen phosphate solution, 0.5MPa pressure injection for 5s, the separation voltage was 8kV, the electrophoresis temperature was 25 ℃, the detection wavelength was 254nm. Under the best conditions, the sample can be completely separated in 20min, the linear relationship is good. The method is accurate, rapid and easy to detect DNase, RNase and cytochrome C in the sample simultaneously.