Inorganic polyphosphates (polyPs) have been identified in eukaryotic and prokaryotic cells alike. Various extraction methods have been optimized as a necessary step before identification and measurement of these polymers. Three commercially available sodium polyP glasses were either dissolved or dissolved and extracted by two commonly used polyP extraction techniques – perchloric acid or buffered phenol–chloroform. The products were separated by polyacrylamide gel electrophoresis (PAGE), stained with toluidine blue O, and the migration results quantitatively compared. Both extraction processes reduced the relative migration distances of the peak and leading edges, and the stained band lengths, suggesting reduced polyP migration and dispersion. ³¹P diffusion-ordered spectroscopy nuclear magnetic resonance confirmed that polyP extraction by perchloric acid or phenol–chloroform processes reduced polyP diffusion coefficients and suggested hydrolytic degradation with stronger end-chain signals. Reduced polyP diffusivity after extraction makes possible an overestimation of synthetic polyP chain length assignment when compared to unextracted polyP ladders with PAGE. The mechanism(s) for reduced synthetic polyP diffusion after extraction and intracellular chemical environment effects on migration are not known.

中文翻译：

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凝胶电泳和 31P DOSY NMR 检测到萃取过程减少了聚磷酸盐离子迁移、分散和扩散

无机多磷酸盐 (polyPs) 已在真核和原核细胞中被鉴定出来。在识别和测量这些聚合物之前，已经优化了各种提取方法作为必要步骤。通过两种常用的聚磷提取技术——高氯酸或缓冲苯酚-氯仿——溶解或溶解并提取三种市售钠聚磷玻璃。产物经聚丙烯酰胺凝胶电泳（PAGE）分离，甲苯胺蓝O染色，定量比较迁移结果。两种提取过程都减少了峰和前缘的相对迁移距离，以及染色带长度，表明减少了 polyP 迁移和分散。³¹P 扩散有序光谱核磁共振证实，通过高氯酸或苯酚 - 氯仿过程提取 polyP 降低了 polyP 扩散系数，并表明水解降解具有更强的端链信号。与使用 PAGE 的未提取的 polyP 梯子相比，提取后 polyP 扩散率降低使得合成 polyP 链长度分配的高估成为可能。提取后减少合成 polyP 扩散的机制和细胞内化学环境对迁移的影响尚不清楚。