Synthesis and application of multifunctional fluorescent molecular probe for simultaneously distinguishing and detecting Cys, Hcy and GSH

Abstract

The invention discloses a multifunctional fluorescent molecular probe for simultaneously distinguishing and detecting cysteine (Cys), homocysteine (Hcy) and glutathione (GSH) in cells by using three different fluorescent emission signals. The structural formula of the molecular probe is shown as follows: (the formula is shown in the description). Under the same detection conditions, the probe cangenerate a product with different fluorescence properties by carrying out different chemical reactions between the probe and Cys, Hcy and GSH, and emit different fluorescence signals under the excitation of different excitation wavelengths, so that the aim of simultaneously distinguishing and detecting the three biological thiols is achieved. After a mixture of the three thiols is added for reacting at room temperature for 15 minutes, blue fluorescence with the wavelength of 457nm is emitted for detecting the Cys at 360nm excitation wavelength; under the condition of 480nm excitation wavelength, 559nm yellow-green fluorescence is emitted for detecting the Hcy; under the condition of 400nm excitation wavelength, 529nm green fluorescence is emitted for detecting the GSH; the molecular probehas extremely-high sensitivity and selectivity to the Cys, the Hcy and the GSH and is successfully used for fluorescence imaging analysis of the Cys, the HCY and the GSH in different living cells.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry, and in particular relates to the synthesis of a multifunctional fluorescent molecular probe for the simultaneous differential detection of Cys, Hcy and GSH, and the simultaneous differential detection of Cys by different reaction phenomena and fluorescent signals of the probe in serum and cells , Hcy and GSH applications. Background technique [0002] Small molecule biothiols such as cysteine ​​(Cys), homocysteine ​​(Hcy) and glutathione (GSH) play complex and important roles in biological systems. The concentration of Cys in normal cells is 30-200 μM, abnormal content can lead to slow growth, edema, lethargy, cardiovascular and liver damage and other diseases (Chemical Communication, 2013, 49, 9176-9178). The normal concentration of Hcy in serum is about 5-12 μM, and the content in normal cells is the lowest of the three biothiols, but excessive Hcy can cause Alzheimer's, folic aci...

AI Technical Summary

Problems solved by technology

Since the structures and chemical properties of cysteine ​​(Cys), homocysteine ​​(Hcy) and glutathione (GSH) are similar, although some specific fluorescent probes based on different mechanisms have been proposed to detect Cys / Hcy and GSH, but there are few reports in the published patents / literatures that can simultaneously detect the three biothiols of Cys, Hcy and GSH, and most of the reported fluorescent probes can only selectively detect one of them or Two types (such as Chinese patents CN105524055A, CN105588823A, CN105601658A, CN105712964A, CN105820810A and CN105906534A), although it has been reported that fluorescent probes can detect Cys, Hcy and GSH at the same time, under the same detection conditions, Cys and Hcy with very similar chemical structures can be distinguished The effect is not ideal, or it is necessary to add admixtures to assist detection (Journal of the American Chemical Society, 2014, 136, 574-577; Chemical Science, 2016, 7, 256-260), but it is impossible to use three fluorescent signals to distinguish detection at the same time Three biothiols (Analytical Chemistry, 2015, 87, 3460–3466), there are few patents / literatures that can simultaneously distinguish and detect the three biothiols in cells Cys, Hcy and GSH

Images