摘要
多年来,电泳技术在分离和鉴定荷电粒子中已经得到了广泛的应用。但其应用范围主要在于微量荷电粒子的分析与鉴定,而对于连续制备过程应用较少,因为在此条件时,电场中离子的迁移规律直接影响分离、鉴定的结果,因此,研究电泳过程中荷电粒子的迁移规律对优化电泳条件具有重要意义。
本文首先利用容积为30L的大垂直电泳装置对苦咸水中氯离子的迁移规律进行了研究。考查了电压、进水流速和填料对氯离子浓度的影响,得出了氯离子浓度与电导率的拟合曲线。根据电导-浓度曲线,测得了电泳室内不同位置的氯离子浓度,研究氯离子浓度在垂直电泳中的迁移规律。接着设计新的微型(容积为60mL)电泳装置,与原来的装置相比,在结构上增加了两冷却室以减小电泳时焦耳热的影响。以硫酸钠和氨基酸混合物为研究对象,考查了硫酸根离子和甘氨酸、赖氨酸混合物的迁移规律。
大电泳装置的实验结果表明:保持进水流速不变,电压越大,氯离子的迁移程度越大;对于金刚砂填料,随着进水流速增大,氯离子的迁移程度先增大后减小,存在一个较佳的进水流速,在该流速条件下,氯离子迁移的程度最大,相应的脱盐效果也较好;填料的粒径越小,其作为承载填料对进分离液的分散效果越好,从而在垂直电泳过程中荷电粒子的迁移程度越明显;在一定pH值条件下,电导与氯离子的浓度成良好的线性关系,氯离子在电泳箱内横向和纵向分布规律与理论上基本相符,但石英砂填料下氯离子浓度与理论的偏离比金刚砂填料下的偏离大。
微型电泳装置的实验结果表明:支持介质——滤纸在反应初期对实验结果的影响较大,当稳定运行时,可不考虑它的影响;随着电压的增大,硫酸根离子在电泳中的迁移程度较大,出样比较集中,硫酸根离子与钠离子的迁移速率之比与理论分析存在一定的差异;两性电解质氨基酸在电泳过程中的迁移规律较无机离子复杂,在本实验过程中没有发现明显的规律性;电场中赖氨酸在比甘氨酸容易带电,电泳时的迁移程度大;电压对氨基酸的迁移有很大影响;新设计的垂直电泳装置适合用于无机离子的分离,如用于分离两性电解质氨基酸还需进一步研究电泳条件或改进装置。
In the last few years, electrophoresis technology has been used widely in the separation and determination of charged ions. However, it is applied mainly in the microanalysis and there are few reports in the continuous operation. Since the migration of charged ions has the direct influence on the efficiency of separation and determination, therefore, the research on the migration of charged ions in the electrophoresis can help to optimize the electrophoresis conditions to obtain good results.Firstly, the migration of Cl* in seawater is studied using the large electrophoresis equipment with a volume of 30L. The influence of voltage, flow rate, padding properties on concentration of Cl" is researched. The fitted curve of relationship between conductivity and Cl" concentration is obtained. According to the curve, the concentration of Cl" at different location of electrophoresis chamber is measured. The migration of Cl" in the process of electrophoresis is studied. Secondly, a new subminiature (the volume is 60mL) electrophoresis equipment (SEE) has been designed. The distinguishing configuration feature of SEE compared with the former is the addition of two cooling chambers to reduce the effect of forming heat on electrophoresis. Finally, selecting Na2SO4 and mixture of amino acid as the research object, the migration of charged ions in Na2SO4 and mixture of lysine and glycin is investigated.The conclusions obtained by the large electrophoresis equipment are as follows: the bigger the voltage, the easier the migration of Cl" while keeping the fixed flow rate; the migration extent of Cl" is strengthened at first and then weakened with the increase of flow rate when the carborundum is used; the optimal flow rate is found at which the maximum migration quantity of Cl' is obtained, and the desalting efficiency is well; the comparison of two kinds of padding show that the smaller the diameter of padding, the better effect of disperse for influx, therefore the bigger the migration extent of charged ion in vertical electrophoresis; at certain pH, the relationship of conductivity and Cl" concentration is linear; the distribution of Cl" in transverse and longitudinal direction of electrophoresis chamber is nearly the same with the theoretical values, but the departure from theoretical values using quartz sand is bigger than using carborundum.The conclusions obtained by SEE are as follows: the supporting medium--filter paperhas obvious influence on the experiment results at the beginning of reaction, but when the equipment run steadily, the influence can be neglected; the migration extent of SO42- is strengthened and the distribution of SO42- is concentrated with the increase of voltage; the ratio of migration rate of SO42- to Na+ is different from that of the theory analysis; the migration
regularity of ampholyte amino acid is more complex than that of inorganic ion and the obvious regularity is not found in the experiment; in electrical field, the lysine is charged easier than glycin, so the migration quantity of lysine is larger than glycin; the voltage has obviously influence on the migration of amino acid; the new equipment can be applied to separate inorganic ions and some improvements for equipment and further research on conditions of electrophoresis should be undertaken to separate ampholyte amino acid.
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