Chemically Induced pH Perturbations for Analyzing Biological Barriers Using Ion-Sensitive Field-Effect Transistors

Potentiometric pH measurements have long been used for the bioanalysis of biofluids, tissues, and cells. A glass pH electrode and ion-sensitive field-effect transistor (ISFET) can measure the time course of pH changes in a microenvironment as a result of physiological and biological activities. However, the signal interpretation of passive pH sensing is difficult because many biological activities influence the spatiotemporal distribution of pH in the microenvironment. Moreover, time course measurement suffers from stability because of gradual drifts in signaling. To address these issues, an active method of pH sensing was developed for the analysis of the cell barrier in vitro. The microenvironmental pH is temporarily perturbed by introducing a low concentration of weak acid (NH4+) or base (CH3COO−) to cells cultured on the gate insulator of ISFET using a superfusion system. Considering the pH perturbation originates from the semi-permeability of lipid bilayer plasma membranes, induced proton dynamics are used for analyzing the biomembrane barriers against ions and hydrated species following interaction with exogenous reagents. The unique feature of the method is the sensitivity to the formation of transmembrane pores as small as a proton (H+), enabling the analysis of cell–nanomaterial interactions at the molecular level. The new modality of cell analysis using ISFET is expected to be applied to nanomedicine, drug screening, and tissue engineering.

Voltammetric pH Measurements in Unadulterated Foodstuffs, Urine, and Serum with 3D-Printed Graphene/poly(lactic Acid) Electrodes

The pH of a system is a critical descriptor of its chemistry – impacting reaction rates, solubility, chemical speciation, and homeostasis. As a result, pH is one of the most commonly measured parameters in food safety, clinical, and environmental laboratories. Glass pH probes are the gold standard for pH measurements, but suffer drawbacks including frequent recalibration, wet storage of the glass membrane, difficulty in miniaturization, and interferences from alkali metals. In this work, we describe a voltammetric pH sensor that uses a 3D-printed graphene/poly(lactic acid) filament electrode that is pretreated to introduce quinone functional groups to the graphene surface. After thoroughly characterizing the pretreatment parameters using outer-sphere and inner-sphere redox couples, we measured pH by reducing the surface-bound quinones, which undergo a pH-dependent 2e^–/2H⁺ reduction. The position of the redox peak was found to shift –60 ± 2 mV pH^-1 at 25 ºC, which is in excellent agreement with the theoretical value predicted by the Nernst Equation (–59.2 mV pH^-1). Importantly, the sensors did not require the removal of dissolved oxygen prior to successful pH measurements. We investigated the impact of common interfering species (Pb²⁺ and Cu²⁺) and found that there was no impact on the measured pH. We subsequently challenged the sensors to measure the pH of unadulterated complex samples including cola, vinegar, serum, and urine, and obtained excellent agreement compared to a glass pH electrode. In addition to the positive analytical characteristics, the sensors are extremely cheap and easy to fabricate, making them highly accessible to a wide range of researchers. These results pave the way for customizable pH sensors that can be fabricated in (nearly) any geometry for targeted applications using 3D-printing.

Voltammetric pH Measurements in Unadulterated Foodstuffs, Urine, and Serum with 3D-Printed Graphene/poly(lactic Acid) Electrodes

The pH of a system is a critical descriptor of its chemistry – impacting reaction rates, solubility, chemical speciation, and homeostasis. As a result, pH is one of the most commonly measured parameters in food safety, clinical, and environmental laboratories. Glass pH probes are the gold standard for pH measurements, but suffer drawbacks including frequent recalibration, wet storage of the glass membrane, difficulty in miniaturization, and interferences from alkali metals. In this work, we describe a voltammetric pH sensor that uses a 3D-printed graphene/poly(lactic acid) filament electrode that is pretreated to introduce quinone functional groups to the graphene surface. After thoroughly characterizing the pretreatment parameters using outer-sphere and inner-sphere redox couples, we measured pH by reducing the surface-bound quinones, which undergo a pH-dependent 2e^–/2H⁺ reduction. The position of the redox peak was found to shift –60 ± 2 mV pH^-1 at 25 ºC, which is in excellent agreement with the theoretical value predicted by the Nernst Equation (–59.2 mV pH^-1). Importantly, the sensors did not require the removal of dissolved oxygen prior to successful pH measurements. We investigated the impact of common interfering species (Pb²⁺ and Cu²⁺) and found that there was no impact on the measured pH. We subsequently challenged the sensors to measure the pH of unadulterated complex samples including cola, vinegar, serum, and urine, and obtained excellent agreement compared to a glass pH electrode. In addition to the positive analytical characteristics, the sensors are extremely cheap and easy to fabricate, making them highly accessible to a wide range of researchers. These results pave the way for customizable pH sensors that can be fabricated in (nearly) any geometry for targeted applications using 3D-printing.