Combination of Electrochemistry and Mass Spectrometry to Study Nitric Oxide Metabolism and Its Modulation by Compound K in Breast Cancer Cells

Abstract

The levels of L-arginine and asymmetric dimethylarginine (ADMA) and the amount of the nitric oxide (NO) production have recently been linked to breast cancer and pharmaceutical effect evaluation. Herein, a method combining electrochemistry and high-resolution mass spectrometry (HRMS) was established and used to study NO metabolism and its modulation by ginsenoside compound K (CK) in breast cancer cells. Platinum nanoparticles-decorated fluorine tin oxide was employed as an electrochemical sensor for in situ detection of NO release, while HRMS was used for the analysis of the NO-related metabolites. Through the combination of the electrochemical and HRMS results, decreases in arginine and NO and increases in ADMA and ornithine were observed after modulation by CK, and two highly correlated metabolic pathways including arginine and proline metabolism and vascular smooth muscle contraction were found. This method offers a new strategy for fast evaluation of pharmaceutical efficacy based on NO metabolism.

Bio-Inspired Electrochemical Detection of Nitric Oxide Promoted by Coordinating the Histamine-Iron Phthalocyanine Catalytic Center on Microelectrode

“…Meanwhile, the formation of irreversible adducts and their derivatives during the detection process interfered with the detection signal of NO to some extent . In comparison, electrochemical sensors are more suitable for the detection of NO, which can use the oxidation process to be oxidized to generate output signals with high sensitivity and high selectivity in a label-free manner. , Moreover, electrochemical techniques can provide broader spatiotemporal resolution and a more facile operational monitoring platform. − …”

“Iridium Signature” Mass Spectrometric Probes: New Tools Integrated in a Liquid Chromatography–Mass Spectrometry Workflow for Routine Profiling of Nitric Oxide and Metabolic Fingerprints in Cells

“…Nowadays, mass spectrometry (MS) is increasingly emerging as a powerful tool for identifying and quantifying various classes of metabolites from complex biological samples at high coverage, sensitivity, and specificity. − Despite its powerful capabilities, direct MS detection of NO is impossible since NO is highly diffusible, unstable, and a gaseous free radical. In the past decades, widespread interest in NO and its biological roles has prompted the developments of various analytical techniques capable of its measurement and quantification, such as colorimetry, electrochemical sensors, and fluorescence techniques. − Generally, these methods only measure one specific analyte and indeed show high specificity and sensitivity.…”

“…28,29 To address this problem, a straightforward approach is hyphenated to high-throughput methods such as MS. For instance, Zhao et al have reported a novel approach by combining electrochemistry and highresolution MS (HRMS), in which electrochemistry is applied for in situ detection of NO release, while HRMS can be used for metabolite profiling in breast cancer cells. 9 It should be noted that electrochemistry and fluorescence methods are susceptible to environmental changes, e.g., electrode materials and solvent conditions, which can lead to huge variability in reported NO concentrations between labs. 30,31 More importantly, data acquired through different methods or even from samples prepared by distinct protocols may lead to inconsistent outcomes.…”

Single-Particle Plasmonic Sensing of Nitric Oxide in Living Cells

“…5 Therefore, analysis of NO in biological systems is essential for the study of its multiple functions. A variety of methods have been developed to detect NO, including electrochemistry, 6,7 fluorescence, 8−13 chemiluminescence, 14 chromatography, and so on. 15 Nevertheless, monitoring NO in living cells suffers from a complex physiological microenvironment, small amount, and fluctuation of the target.…”

“…Therefore, analysis of NO in biological systems is essential for the study of its multiple functions. A variety of methods have been developed to detect NO, including electrochemistry, , fluorescence, − chemiluminescence, chromatography, and so on . Nevertheless, monitoring NO in living cells suffers from a complex physiological microenvironment, small amount, and fluctuation of the target. , For example, in the commonly used fluorescence methods, the use of fluorescent probes may be limited by the pH of the biological environment, the high autofluorescence background of complex biological fluids, and so on.…”