Adenine nucleotides (ANs)—adenosine 5′-triphosphate (ATP), adenosine 5′-diphosphate (ADP), and adenosine 5′-monophosphate (AMP)—are essential for energy transfer and the supply of countless processes within cellular metabolism. Their concentrations can be expressed as adenylate energy charge (AEC), a measure of cellular metabolic energy that directly correlates with the homeostasis of the organism. AEC index has broad diagnostic potential, as reduced ATP levels are associated to various conditions, such as inflammatory diseases, metabolic disorders, and cancer. We introduce a novel methodology for rapid isolation, purification, and quantification of ANs from a single drop of capillary blood. Of all the stationary phases tested, activated carbon proved to be the most efficient for the purification of adenine nucleotides, using an automated micro-solid phase extraction (µ-SPE) platform. An optimized µ-SPE method, coupled with RP-HPLC and a run time of 30 min, provides a reliable analytical framework for adenine nucleotide analysis of diverse biological samples. AN concentrations measured in capillary blood samples were 1393.1 µM, 254.8 µM, and 76.9 µM for ATP, ADP, and AMP molecules aligning with values reported in the literature. Overall, this study presents a streamlined and precise approach for analyzing ANs from microliters of blood, offering promising applications in clinical diagnostics.

We present a simple method for modification of 2D materials by drop-casting of the organic molecule in solution on the 2D material under ambient conditions. Specifically, we investigated the adsorption of 6-(4,5-Dihydro-1H-imidazol-3-ium-2-yl)-2-(naphthalene-2-yl)benzothiazole methanesulfonate (L63MS) organic molecule on 2D MoS${}_2$. To better understand the effect of the organic molecule on the 2D material, we also investigated the impact of solvents alone on the materials’ properties. The MoS${}_2$ samples were synthesized using ambient pressure chemical vapor deposition. Atomic force microscopy, Raman spectroscopy, photoluminescence spectroscopy and optical microscopy were used to characterize the samples. The measurements were performed after synthesis, after the drop-casting of solvents and after the drop-casting of organic molecule solutions. Our results indicate that the used organic molecule effectively adsorbs on and prompts discernible changes in the (opto)electronic properties of the 2D material. These changes encompass variations in the Raman spectra shape, alterations in the photoluminescence (PL) signal characteristics and modifications in excitonic properties. Such alterations can be linked to various phenomena including doping, bandgap modifications, introduction or healing of defects and that the solvent plays a crucial role in the process. Our study provides insights into the modification of 2D materials under ambient conditions and highlights the importance of solvent selection in the process.

Adenine nucleotides—adenosine monophosphate, diphosphate, and triphosphate—are of utmost importance to all living organisms, where they play a critical role in the energy metabolism and are tied to allosteric regulation in various regulatory enzymes. Adenylate energy charge represents the precise relationship between the concentrations of adenosine monophosphate, diphosphate, and triphosphate and indicates the amount of metabolic energy available to an organism. The experimental conditions of adenylate extraction in freshwater amphipod Gammarus fossarum are reported here for the first time and are crucial for the qualitative and quantitative determination of adenylate nucleotides using efficient and sensitive ion-pair reverse phase LC. It was shown that amphipod calcified exoskeleton impeded the neutralization of homogenate. The highest adenylate yield was obtained by homogenization in perchloric acid and subsequent addition of potassium hydroxide and phosphate buffer to achieve a pH around 11. This method enables separation and accurate detection of adenylates. Our study provides new insight into the complexity of adenylate extraction and quantification that is crucial for the application of adenylate energy charge as a confident physiological measure of environmental stress and as a health index of G. fossarum.

Adenine nucleotides (AN): adenosine 5’-triphosphate (ATP), adenosine 5’-diphosphate (ADP), and adenosine 5’-monophosphate (AMP) are essential for energy transfer and supply of countless processes within the cellular metabolism. Their concentrations can be expressed as adenylate energy charge (AEC), a measure of cellular metabolic energy that directly correlates with the homeostasis of the organism. AEC index can be used as a clinical health indicator with broad diagnostic potential, as a decrease in ATP levels is associated with various conditions, such as inflammatory diseases, metabolic disorders, and cancer. We introduce a novel methodology for rapid isolation, purification, and quantification of AN from just a drop of capillary blood obtained from a healthy volunteer. Of all the stationary phases tested, activated carbon proved to be the most efficient for the purification of adenine nucleotides, using an automated micro-solid phase extraction (µ-SPE) platform. By removing interfering substances, we achieved high chromatographic selectivity and resolution, facilitating the quantification of AN. The analyzed AN concentrations from capillary blood correspond to the values found in the literature, which corroborates reliability of the developed extraction method. Overall, this study presents a streamlined and precise approach for analyzing AN from microliters of blood that offers promising applications in clinical diagnostics.