BODIPY-Based Fluorometric Sensor for the Simultaneous Determination of Cys, Hcy, and GSH in Human Serum

Abstract

Cysteine (Cys), homocysteine (Hcy), and glutathione (GSH) are interconnected and play essential roles for regulating the redox balance of biological processes. However, finding a simple and effective method for the simultaneous determination for these three biothiols in biological systems is always a challenge. In this work, we report a method for the simultaneous quantitative determination of three biothiols in a mixture using a monochlorinated boron dipyrromethene (BODIPY)-based fluorometric sensor. At a specified period of time, after reacting with excess sensor, Hcy and GSH form predominantly sulfur-substituted BODIPY, while Cys generates sulfur-amino-diBODIPY due to a fast substitution-rearrangement-substitution reaction. A significant difference in polarities of these respective major products simplifies their separation by TLC, thus leading to the simultaneous determination of Cys, Hcy, and GSH readily. The sensor was successfully applied for the simultaneous quantitative detection of three biothiols in human serum, and the results were in good agreement with those obtained via high performance liquid chromatography (HPLC).

Iridium(III) Complex‐Based Activatable Probe for Phosphorescent/Time‐Gated Luminescent Sensing and Imaging of Cysteine in Mitochondria of Live Cells and Animals

“…Further enhancements in both phosphorescence and time‐gated luminescence were observed in the presence of Cys. By using the standard curve shown in Figure B, the concentration of Cys in human sera was determined to be 254±16 μ m , which is consistent with the reported value . By adding exogenous Cys into the diluted human sera, the recovery of added Cys was determined to be 89.8–90.5 % (Table S6, Supporting Information), indicating that the time‐gated luminescence analysis of Cys by using [Ir(ppy) 2 (NTY‐bpy)] + as a probe is highly accurate.…”

An inner filter effect fluorescent sensor based on g-C3N4 nanosheets/chromogenic probe for simple detection of glutathione

“…Blood GSH may serve as an accurate indicator of GSH status in human subjects, which has been considered as an important role in some human diseases [39]. The GSH level results from the strategy we proposed in the healthy human serum is demonstrated in Table 1, which is in close agreement with the normal serum GSH level by HPLC results [40]. Therefore, the prospective results by our method possessed the feasibility and reliability for real clinical sample analysis.…”

BODIPY‐Based Fluorescent Probes for Biothiols

“…The high reactivity of 3‐Cl of the BODIPY dye and the distinct spectroscopic difference between the 3‐amino and 3‐mercapto BODIPY derivatives discovered by the Yang group are the basis for the differentiation of GSH from Cys/Hcy utilized by other designs. Similar sensing mechanisms were used in several following publications using 3‐chloro‐BODIPY derivatives ( 62 – 68 ), 3‐phenylethynyl BODIPY derivative ( 69 ), and 1‐ p ‐chlorphenyl/2‐ p ‐chlorphenyl BODIPY derivatives ( 70 and 71 ; Figure ) . Even with the Michael acceptor present in probes 62 and 63 , the reacting site was found to be the 3‐position through replacement of the chlorine atom .…”