Zhang Jing, Wei Xuan, Wang Yuanhang, Ren Bo, Zhao Litao, Wang Zhaoyan, Pu Qiaosheng
School of Pharmacy, Lanzhou University Lanzhou 730000 China
Department of Chemistry, Tonghua Normal University Tonghua 134002 China.
RSC Adv. 2018 May 18;8(33):18266-18271. doi: 10.1039/c8ra00492g. eCollection 2018 May 17.
Change in cation concentration, including that of potassium and sodium, is characteristic of apoptosis, therefore it is significant to detect cation concentration changes. In this work a rapid, sensitive, and practical method was developed for the determination of Na and K concentration in HeLa cells during emodin induced apoptosis by a low-cost capillary electrophoresis device with capacitively coupled contactless conductivity detection (CE-CD). Under the optimized conditions, both ions were baseline separated in 4 min with 40 mM MES/40 mM His containing 1 mM 18-crown-6 as the separation buffer at pH 6.0. The limit of detections (LODs) and limit of quantifications (LOQs) were 0.47-1.15 μM and 1.58-3.86 μM, respectively. The precision for migration times and peak areas was below 0.56% and 3.74%, respectively. The data proved that the concentration of cations in cells can be accurately quantified. It was found that the K concentration decreased from 82.2 μM to 52.7 μM, and the Na concentration increased from 62.4 μM to 127.2 μM during the process of apoptosis when the cell density was 1 × 10 cells per mL. The low-cost CE-CD provides a convenient way to decipher the interaction of Na and K in the regulation of cell apoptosis.
阳离子浓度的变化,包括钾和钠的浓度变化,是细胞凋亡的特征,因此检测阳离子浓度变化具有重要意义。在这项工作中,开发了一种快速、灵敏且实用的方法,用于通过具有电容耦合非接触电导检测(CE-CD)的低成本毛细管电泳装置测定大黄素诱导HeLa细胞凋亡过程中的钠和钾浓度。在优化条件下,两种离子在4分钟内实现基线分离,分离缓冲液为含1 mM 18-冠-6的40 mM MES/40 mM His,pH值为6.0。检测限(LOD)和定量限(LOQ)分别为0.47 - 1.15 μM和1.58 - 3.86 μM。迁移时间和峰面积的精密度分别低于0.56%和3.74%。数据证明细胞中的阳离子浓度可以被准确量化。结果发现,当细胞密度为每毫升1×10个细胞时,在细胞凋亡过程中,钾浓度从82.2 μM降至52.7 μM,钠浓度从62.4 μM升至127.2 μM。低成本的CE-CD为解读钠和钾在细胞凋亡调控中的相互作用提供了一种便捷的方法。