Kulp Maria, Kaljurand Mihkel
Faculty of Science, Tallinn Technical University, Ehitajate Tee 5, 19086 Tallinn, Estonia.
J Chromatogr A. 2004 Apr 2;1032(1-2):305-12. doi: 10.1016/j.chroma.2003.09.031.
Capillary electrophoresis can be a valuable tool for the on-line monitoring of bioprocesses. The enzymatic conversion of nucleotide adenosine triphosphate (ATP) to adenosine diphosphate (ADP) by hexokinase (HK) was monitored in the bioreactor interfaced by a laboratory-built microsampler to a capillary electrophoresis unit. The use of this specially designed sampling device enabled rapid consecutive injections to be performed without high-voltage (HV) interruptions. No additional sample preparation was required. The method of micellar electrokinetic chromatography, employing reversed electroosmotic flow (EOF) by cationic surfactant and reversed polarity mode provided a good resolution and short analysis time of less than 5 min. The samples were injected electrokinetically, using -25 kV voltage for 3 s and detected by their UV absorbance at 254 nm. The analytes were detected at a microg/ml level with a reproducibility of about 7%. To demonstrate the potential of CE in understanding the processes of biological interest, such as nucleotide degradation and metabolism, the investigation of the efficiency and the time course of the enzymatic transformation was carried out.
毛细管电泳可以成为生物过程在线监测的一种有价值的工具。通过实验室构建的微量采样器与毛细管电泳单元相连的生物反应器中，监测了己糖激酶（HK）将三磷酸腺苷（ATP）酶促转化为二磷酸腺苷（ADP）的过程。使用这种专门设计的采样装置能够在不中断高压（HV）的情况下进行快速连续进样。无需额外的样品制备。采用阳离子表面活性剂产生反向电渗流（EOF）并采用反相模式的胶束电动色谱方法提供了良好的分离度，分析时间短于5分钟。样品通过电动进样，使用-25 kV电压进样3秒，并通过在254 nm处的紫外吸光度进行检测。分析物的检测限为微克/毫升水平，重现性约为7%。为了证明毛细管电泳在理解诸如核苷酸降解和代谢等生物学相关过程方面的潜力，对酶促转化的效率和时间进程进行了研究。
