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使用聚醚醚酮毛细管和非接触式电导检测对各种生物源分析物进行毛细管电泳分离。

CE separation of various analytes of biological origin using polyether ether ketone capillaries and contactless conductivity detection.

作者信息

Knjazeva Tatjana, Kulp Maria, Kaljurand Mihkel

机构信息

Department of Chemistry, Tallinn Technical University, Estonia.

出版信息

Electrophoresis. 2009 Jan;30(2):424-30. doi: 10.1002/elps.200800252.

Abstract

The development of efficient and sensitive analytical methods for the separation, identification and quantification of complex biological samples is continuously a topic of high interest in biological science. In the present study, the possibility of using a polyether ether ketone (PEEK) capillary for the CE separation of peptides, proteins and other biological samples was examined. The performance of the tubing was compared with that of traditional silica capillaries. The CE analysis was performed using contactless conductivity detection (C(4)D), which eliminated any need for the detection window and was suitable for the detection of optically inactive compounds. In the PEEK capillary the cathodic EOF was low and of excellent stability even at extremes pH. In view of this fast biological anions were analyzed using an opposite end injection technique without compromising separation. A comparison of the performances of fused-silica and polymer capillaries during the separation of model sample mixtures demonstrated the efficiency and separation resolution of the latter to be higher and the reproducibility of the migration times and peak areas is better. Furthermore, PEEK capillaries allowed using simple experimental conditions without any complicated modification of the capillary surface or use of an intricate buffer composition. The PEEK capillaries are considered as an attractive alternative to the traditional fused-silica capillaries and may be used for the analysis of complex biological mixtures as well as for developing portable devices.

摘要

开发用于复杂生物样品的分离、鉴定和定量的高效灵敏分析方法一直是生物科学领域备受关注的话题。在本研究中,考察了使用聚醚醚酮(PEEK)毛细管进行肽、蛋白质及其他生物样品的毛细管电泳分离的可能性。将该毛细管的性能与传统硅胶毛细管的性能进行了比较。毛细管电泳分析采用非接触式电导检测(C(4)D),无需检测窗口,适用于检测无光学活性的化合物。在PEEK毛细管中,阴极电渗流较低,即使在极端pH条件下也具有出色的稳定性。鉴于此,使用反向端进样技术对快速生物阴离子进行分析,且不影响分离效果。对模型样品混合物分离过程中熔融石英毛细管和聚合物毛细管的性能进行比较,结果表明后者的效率和分离分辨率更高,迁移时间和峰面积的重现性更好。此外,PEEK毛细管可采用简单的实验条件,无需对毛细管表面进行任何复杂修饰,也无需使用复杂的缓冲液组成。PEEK毛细管被认为是传统熔融石英毛细管的一种有吸引力的替代物,可用于分析复杂生物混合物以及开发便携式设备。

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