HPLC Study of Product Formed in the Reaction of NBD-Derived Fluorescent Probe with Hydrogen Sulfide, Cysteine, N-acetylcysteine, and Glutathione

Abstract

Hydrogen sulfide (H2S) and its bioderivatives analogs, such as L-cysteine (L-Cys) and glutathione (GSH), are ubiquitous biological thiols in the physiological and pathological processes of living systems. Their aberrant concentration levels are associated with many diseases. Although several NBD-based fluorescence probes have been developed to detect biological thiols, the HPLC-detection of H₂S, GSH, L-Cys, and N-acetylcysteine-specific products has not been described. Herein, a novel NBD-derived pro-coumarin probe has been synthesized and used to develop a new strategy for the triple mode detection of H₂S and such thiols as GSH, L-Cys, and NAC. Hydrogen sulfide and those biothiols at physiological pH release fluorescent coumarin from the probe and cause a significant fluorescence enhancement at 473 nm. The appropriate NBD-derived product for H₂S, L-Cys, GSH, and NAC has a different color and retention time that allows distinguishing these biological thiols meaning the probe has a great possibility in the biological application. Fluorescent imaging combined with colorimetric and HPLC detection of H₂S/biothiol-specific product(s) brings a potential tool for confirming the presence of biological thiols and determining concentrations in various aqueous biological samples.

A New and Fast-Response Fluorescent Probe for Monitoring Hypochlorous Acid Derived from Myeloperoxidase

“…The limitation of known boronate-based in vivo sensors is their lack of specificity [ 48 ]. In addition, the detection of HOCl in vivo is complicated as the existence of various antioxidants, such as GSH [ 49 , 50 ] and compounds present in the cellular environment at high concentrations [ 3 , 51 ]. Several reports [ 52 , 53 , 54 , 55 , 56 , 57 ] described that sulfur- and selenium-oxides formed in reaction with HOCl are reduced via GSH to a sulfur and selenide group, respectively.…”

Association between Micronutrients and Hyperhomocysteinemia: A Case-Control Study in Northeast China

“…Hcy and other nutrients were completed in the clinical laboratory of Shengjing Hospital according to clinical testing standards. Plasma Hcy levels were determined through chemiluminescence using a commercial kit (Jiuqiang Biological Company, Beijing, China) and an automatic chemiluminassay analyzer (Abbott-i16200, Shanghai, China), following the detection methods reported by Yin [33,34]. Plasma Zn, Cu, Ca, P, Mg, and Pb levels were determined using an atomic absorption spectrometer (Jingbohui Biotechnology Company, Beijing, China), following the method by Komarova [35,36].…”

Comparison of Colorimetric, Spectroscopic and Electrochemical Techniques for Quantification of Hydrogen Sulfide

“…Thus, an effective method to accurately measure H 2 S concentration in each sample is important for this gaseous molecule that can be lost easily to the environment. Literature suggests that approaches such as gas chromatography and ion chromatography [70], fluorescent probes [ 7 1], methylene blue assay [ 7 2] and monobromobimane assay [ 7 3] with HPLC, ion-selective or polarographic electrodes [ 7 4] can be used for the detection of H 2 S [ 7 5]. However, the primary limitation posed by these approaches is their inability to measure the real-time concentration of H 2 S while achieving the necessary sensitivity and response time.…”