Analysis of inorganic polyphosphates by capillary gel electrophoresis.
Analysis of inorganic polyphosphates by capillary gel electrophoresis.
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DOI：
10.1021/ac1008018
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发表时间：
2010-08-15
影响因子：
7.4
通讯作者：
Whitesides, George M.
中科院分区：
文献类型：
--
作者：
Lee, Andrew;Whitesides, George M.
本文介绍了一种毛细管凝胶电泳（CGE）分析无机多聚磷酸盐（（Pi）n）混合物的方法。基于n（脱水磷酸盐的残基数量）的（Pi）n的分辨率通过CGE使用填充有聚（N，N-二甲基丙烯酰胺）（PDMA）溶液的毛细管和通过添加到运行缓冲液中的发色团对苯二甲酸酯的UV吸光度间接检测来实现。该方法能够分离代表（Pi）n（n高达~70）的峰;真实标准品的制备和使用能够鉴别（Pi）n（n = 1 - 10）的峰。与之前报道的分析（Pi）n混合物的方法相比，该方法的主要优点是（例如，凝胶电泳、使用聚丙烯酰胺填充的毛细管的CGE）的优点在于其分辨率、方便性和再现性;凝胶填充的毛细管易于通过泵入新鲜的低粘度PDMA溶液来再生。的分辨率是可比的离子交换色谱和检测（Pi）n抑制电导率。该方法可用于分析在低温（125 - 140 °C）下用尿素使Pi脱水产生的（Pi）n，该反应在益生元化学中可能是重要的。所述方法还可用于表征没有固有发色团的其它阴离子、低聚物或聚合物物质的混合物（例如，硫酸化多糖、低聚磷酸二酯）。
This paper describes the development of a method that uses Capillary Gel Electrophoresis (CGE) to analyze mixtures of inorganic polyphosphate ((Pi)n). Resolution of (Pi)n on the basis of n, the number of residues of dehydrated phosphate, is accomplished by CGE using capillaries filled with solutions of poly(N,N-dimethylacrylamide) (PDMA) and indirect detection by the UV-absorbance of a chromophore, terephthalate, added to the running buffer. The method is capable of resolving peaks representing (Pi)n with n up to ~70; preparation and use of authentic standards enables the identification of peaks for (Pi)n with n = 1 - 10. The main advantages of this method over previously reported methods for analyzing mixtures of (Pi)n (e.g., gel electrophoresis, CGE using polyacrylamide-filled capillaries) are its resolution, convenience, and reproducibility; gel-filled capillaries are easily regenerated by pumping in fresh, low-viscosity solutions of PDMA. The resolution is comparable to that of ion-exchange chromatography and detection of (Pi)n by suppressed conductivity. The method is useful for analyzing (Pi)n generated by the dehydration of Pi at low temperature (125 - 140 °C) with urea, in a reaction that may have been important in prebiotic chemistry. The method should also be useful for characterizing mixtures of other anionic, oligomeric or polymeric species without an intrinsic chromophore (e.g., sulfated polysaccharides, oligomeric phospho-diesters).
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