Pietrzak Mariusz, Meyerhoff Mark E
Department of Chemistry, The University of Michigan, Ann Arbor, Michigan 48109, USA.
Anal Chem. 2009 Jul 15;81(14):5961-5. doi: 10.1021/ac900776d.
Recent studies suggest that the measurement of intracellular potassium concentrations in red blood cells (RBC-K) can be a marker for assessing the risk, development, and treatment of hypertension. In this work, the combined use of miniature potassium- and sodium-selective membrane electrodes is evaluated as a simple means to determine RBC-K. The proposed method requires two separate sets of electrode measurements: (i) potassium and sodium concentrations in the plasma phase of an unmeasured volume of a whole blood sample, and (ii) determination of potassium and sodium concentrations in the same sample of blood after complete hemolysis by ultrasonic disruption of the RBC membranes. The dilution of sodium concentration after hemolysis can be used to determine hematocrit (Hct) (volume of red cells per unit volume of blood) of the blood. The concentration of potassium within the red blood cells (RBCs) is then calculated using the measured change in potassium levels before and after RBCs lysis and the hematocrit level determined from the sodium electrode measurements and/or a conventional centrifugation method. Good correlation for RBC-K between the proposed method and traditional flame photometry is observed for animal blood samples that possess the range of potassium levels found within human RBCs (80-120 mM). However, when potassium is much lower than that found in human RBCs (known to occur for certain animal species), the Hct measured by the sodium electrode method is falsely low, compared to traditional spun hematocrit values, because of an increased level of sodium within the RBCs, necessitating use of spun Hct levels to assess RBC-K accurately. It is envisioned that this new approach could be further miniaturized into a single-use disposable cartridge type electrode system that would enable rapid point-of-care screening of RBC-K levels in human subjects.
最近的研究表明,测量红细胞内钾浓度(RBC-K)可作为评估高血压风险、病情发展及治疗效果的一项指标。在本研究中,评估了微型钾离子和钠离子选择性膜电极的联合使用,作为测定RBC-K的一种简便方法。所提出的方法需要进行两组独立的电极测量:(i)对未测定体积的全血样本血浆相中钾和钠的浓度进行测量,以及(ii)通过超声破坏红细胞膜使同一样本完全溶血后,测定其中钾和钠的浓度。溶血后钠浓度的稀释可用于测定血液的血细胞比容(Hct,即每单位体积血液中红细胞的体积)。然后,利用红细胞裂解前后测得的钾水平变化以及通过钠电极测量和/或传统离心法确定的血细胞比容水平,计算红细胞内钾的浓度。对于钾水平在人类红细胞内所发现范围(80 - 120 mM)的动物血液样本,所提出的方法与传统火焰光度法测得的RBC-K具有良好的相关性。然而,当钾含量远低于人类红细胞中的含量时(已知某些动物物种会出现这种情况),与传统离心法测得的血细胞比容值相比,通过钠电极法测得的Hct会被错误地低估,这是因为红细胞内钠水平升高,因此需要使用离心法测得的Hct水平来准确评估RBC-K。预计这种新方法可进一步小型化为一次性使用的盒式电极系统,从而能够对人类受试者进行快速的即时护理RBC-K水平筛查。