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A high-throughput fluorimetric assay for histidine was developed, using a 96-well plates platform. The analyte reacts selectively with -phthalaldehyde under mild alkaline conditions to form a stable derivative. Instrumental-free detection was carried out using a smartphone after illumination under UV light (365 nm). The method was proved to be linear up to 100 μM histidine, with an LLOQ (lower limit of quantification) of 10 μM. The assay was only prone to interference from glutathione and histamine that exist in the urine samples at levels that are orders of magnitude lower compared to histidine. Human urine samples were analyzed following minimum treatment and were found to contain histidine in the range of 280 to 1540 μM. The results were in good agreement with an HPLC corroborative method.
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http://dx.doi.org/10.3390/molecules28176205 | DOI Listing |
Biochem J
June 2025
Department of Pharmaceutical Sciences, Washington State University, Spokane, WA 99202, U.S.A.
We aimed to create a high-throughput fluorimetric assay for the activity of CYP4A11, the major 20-HETE-producing enzyme. To this end, we probed 3-(6-methoxynaphthalen-2-yl)acrylic acid (MONACRA) as a potential CYP4A11 substrate. We studied its metabolism using human liver microsomes (HLM) and recombinant P450 enzymes.
View Article and Find Full Text PDFBiomedicines
March 2025
Associated Laboratory for Green Chemistry (LAQV), Network of Chemistry and Technology (REQUIMTE), Laboratory of Applied Chemistry, Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, 4050-313 Porto, Portugal.
: Recent interest in dietary components and their effects on xenobiotic metabolism has highlighted their role in modulating drug pharmacokinetics. Cytochrome P450 3A4, a key isoform of the cytochrome P450 superfamily, is involved in the metabolism of over 50% of xenobiotics. Flavonoids, present in various foods and supplements, exhibit diverse biological activities influenced by the structural modifications in their scaffold.
View Article and Find Full Text PDFAnalyst
February 2025
Huzhou Key Laboratory of Medical and Environmental Application Technologies, School of Life Sciences, Huzhou University, Huzhou 313000, China.
Water-soluble and biocompatible protein carbon dots (P-CDs) were simply prepared from egg white by a rapid one-step neutralization heat reaction. Unexpectedly, the thus-fabricated P-CDs could present excitation-dependent tunable fluorescence that could be quenched specifically by Fe and Fe ions with obvious color changes. A high-throughput fluorimetric platform was thereby developed by coating the P-CDs onto a capillary array for detection of total iron ions in fish blood samples, with a linear concentration range of 0.
View Article and Find Full Text PDFSci Rep
November 2024
DNA Replication Group, Molecular Oncology Programme, Spanish National Cancer Research Center (CNIO), 3 Melchor Fernandez Almagro, 28029, Madrid, Spain.
The COVID-19 pandemic highlighted the need for the rapid development of antiviral therapies. Viral RNA-dependent RNA polymerases (RdRp) are promising targets, and numerous virtual screenings for potential inhibitors were conducted without validation of the identified hits. Here we have tested a set of presumed RdRp inhibitors in biochemical assays based on fluorometric detection of RdRp activity or on the electrophoretic separation or RdRp products.
View Article and Find Full Text PDFEnviron Int
November 2024
Institute of Water Chemistry, Dresden University of Technology, D-01062 Dresden, Germany. Electronic address:
Growing concerns about the environmental impact of ionic liquids (ILs) have spurred research into their (eco)toxic effects, but studies on their mode of toxic action (MOA) still remain limited. However, understanding the MOA and identifying structural features responsible for enhanced toxicity is crucial for characterising the hazard and designing safer alternatives. Therefore, 45 ILs, with systematically varied chemical structures, were tested for cytotoxicity and two specific endpoints in reporter gene assays targeting the Nrf2-ARE mediated oxidative stress response (AREc32) and aryl hydrocarbon receptor activation (AhR-CALUX).
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