CN102533249A - Preparation of water soluble saccharide fluorescent molecular probe and rapid detection for iron ions in water and biological tissue - Google Patents

Abstract

其中糖基配体不单是氨基葡萄糖，还可以是其他所有的糖类化学物及其衍生物。The invention provides the preparation of a class of water-soluble carbohydrate fluorescent molecular probes and the direct and rapid detection of iron ions in water and biological tissues. This kind of fluorescent molecular probe has high selectivity and sensitivity to iron ions in water and serum. After selective combination with iron ions, the color of the solution can be observed from yellow to reddish brown with the naked eye, and the detection limit can reach up to 10 ^-6 m. On-site real-time, direct and rapid detection can be performed without the aid of instruments. The fluorescent molecular probe can be prepared into an iron ion detection test paper and a detection kit, and can also be made into a portable solid detector by using the viscosity of the sugar compound itself and coating it. The specific structure of the carbohydrate fluorescent molecular probe is as follows (taking the glucosamine ligand as an example):

Wherein the glycosyl ligand is not only glucosamine, but also all other saccharide chemicals and their derivatives.

Description

Preparation of a class of water-soluble carbohydrate fluorescent molecular probes and its rapid detection of iron ions in water and biological tissues

technical field

The invention relates to the preparation of a class of water-soluble carbohydrate fluorescent molecular probes and the rapid detection of iron ions in water and biological tissues.

Background technique

Iron ion is one of the ions necessary for living organisms. It is a necessary substance for the manufacture of heme and myoglobin and the promotion of vitamin B metabolism. Many biological and chemical processes at the cellular level require the participation of iron ions, so the research can be used for iron ions The detected fluorescent molecular probes are of great significance to the environment and life sciences. It is reported in the literature that the synthesis method of fluorescent molecular probes for iron ions is complex and costly. The detection of iron ions by the synthesized products is outside the visible spectrum range, and the color change cannot be observed with the naked eye, and expensive detection instruments must be used. In addition, there are still disadvantages such as sample pretreatment, detection is not fast enough, water solubility is poor, and detection is expensive. Therefore, in many important applications, there is an urgent need for rapid, accurate, low-cost and selective methods for the analysis and detection of iron ions. Carbohydrates are abundant in nature. Because of containing multiple hydroxyl groups, these compounds have high solubility in water, and the sugar compounds are non-toxic and harmless, and will not cause secondary pollution to the environment. To this end, we combine the non-toxicity and high solubility of sugar compounds with fluorescent ligands to synthesize highly selective and sensitive fluorescent molecular probes for the rapid qualitative detection of iron ions in water and biological tissues. The combination of these fluorescent ligands and metal cations will cause a huge change in the absorption spectrum band, so that the obvious color change can be observed with the naked eye, without the need for an instrument, and is especially suitable for on-site real-time detection.

Contents of the invention

The invention provides the preparation of a class of water-soluble carbohydrate fluorescent molecular probes and the direct and rapid detection of iron ions in water and biological tissues. This kind of fluorescent molecular probe has good water solubility, and has high selectivity and sensitivity to iron ions in water and serum. After selective combination with iron ions, the color of the solution can be observed from yellow to red by naked eyes. Brown, the detection limit can reach 10 ^-6 M. On-site real-time, direct and rapid detection can be performed without the aid of instruments. The molecular probe can be prepared into an iron ion detection test paper and a detection kit, and can also be made into a portable detector by using the viscosity of the sugar compound itself and coating it. The specific structure of the carbohydrate fluorescent molecular probe is as follows (taking the glucosamine ligand as an example):

Wherein the glycosyl ligand is not only glucosamine, but also all other saccharide chemicals and their derivatives.

The method of the invention is simple and easy to operate, has low cost and low toxicity, further reduces environmental pollution, and is especially suitable for on-site real-time emergency detection.

For the selectivity experiment of the synthesized carbohydrate fluorescent molecular probes to metal ions, equimolar amounts of carbohydrate fluorescent molecular probes and various metal ions (Na ⁺ , Ag ⁺ , Ca ²⁺ , Hg ²⁺ , Mg ²⁺ , Mn ²⁺ , Cu ²⁺ , Pb ²⁺ , Cd ²⁺ , Cr ²⁺ , Fe ²⁺ , Fe ³⁺ , Zn ²⁺ , Co ²⁺ , Ni ²⁺ ) to form 10 ^-4 M solution, then mix and observe the color change of the solution. The color of the fluorescent molecular probe solution is yellow. After the fluorescent molecular probes were added to various ionic solutions, the color of the Fe ³⁺ solution changed from yellow to reddish-brown quickly, and the colors of other solutions hardly changed or did not change significantly. Through this experiment, it can be seen that the synthesized sugar fluorescent molecular probe has good selectivity to iron ions, and other metal ions will not cause interference. In addition, the concentration of iron ions was adjusted to 10 ^-8 M, 10 ^-7 M, 10 ^-6 M, 10 ^-5 M, 10 ^-4 M, 10 ^-3 M, and then an equimolar amount of sugar fluorescent molecules were added probe, and observe the color change of the solution. According to this experiment, it can be easily concluded that the lower detection limit of iron ions can reach 10 ^-7 M, and the color becomes darker and darker as the concentration increases, and finally turns purple.

Detailed ways

Optical rotation was measured at 25°C with a Perkin-Elmer 241MC automatic polarimeter. ¹ H NMR was measured by Bruker ARX 400 in CDCl ₃ with tetramethylsilane as internal standard. The mass spectrometer adopts VG PLATFORM mass spectrometer, and the sample is injected by ESI technique. Thin-layer chromatography (TLC) was performed on HF ₂₅₄ silica gel plate with 30% (v/v) methanolic sulfuric acid solution or ultraviolet (UV) detector. Column chromatography adopts 100-200 mesh silica gel, uses ethyl acetate-petroleum ether (60-90°C) as the eluent, and the solution is distilled under reduced pressure when the solution is less than 60°C. The compounds not specified below are all commercially available or available Materials prepared with reference to literature.

Example 1:

Compound 1 (1.5 g) was dissolved in trifluoroacetic acid, and hexamethylenetetramine (1.9 g) was added at room temperature to react for about 10 hours. The progress of the reaction was monitored by thin layer chromatography. After the reaction was complete, it was concentrated and purified on a silica gel column. Ethyl acetate and petroleum ether were used as eluents to obtain compound 2 (800 mg, 57%).

Example 2:

Dissolve compound 2 (350 mg) in water, add D-glucosamine hydrochloride (796 mg) and sodium hydroxide (67 mg) at room temperature, and react at a certain temperature for about 5 hours. Then, it was filtered, washed, and the solid was collected to obtain compound 3 (592 mg, 97%), which is a fluorescent molecular probe for carbohydrates.

Example 3:

Equimolar amounts of carbohydrate fluorescent molecular probes and various metal ions (Na ⁺ , Ag ⁺ , Cat ²⁺ , Hg ²⁺ , Mg ²⁺ , Mn ²⁺ , Cu ²⁺ , Pb ²⁺ , Cd ²⁺ , Cr ²⁺ , Fe ²⁺ , Fe ³⁺ , Zn ²⁺ , Ni ²⁺ ) were made into 10 ^-4 M solutions respectively, then mixed, and the color change of the solution was observed. The color of the fluorescent molecular probe solution is yellow. After the fluorescent molecular probes were added to various ionic solutions, the color of the iron ion solution changed from yellow to reddish-brown quickly, and the colors of other solutions hardly changed or did not change significantly. Through this experiment, it can be seen that the synthesized sugar fluorescent molecular probe has good selectivity to iron ions, and other metal ions will not cause interference.

Example 4:

Set the concentration of iron ions to 10 ^-8 M, 10 ^-7 M, 10 ^-6 M, 10 ^-5 M, 10 ^-4 M, 10 ^-3 M, and then add an equimolar amount of carbohydrate fluorescent molecular probes , and then observe the color change of the solution. According to this experiment, it can be easily concluded that the detection limit of iron ions can reach 10 ^-6 M, and the color gradually deepens with the increase of the concentration, and finally turns purple.

Claims (5)

1. the invention provides the preparation of one type of water soluble saccharide fluorescent molecular probe and be used for water and biological tissue in the direct rapid detection of iron ion.This type of fluorescent molecular probe has very high selectivity and susceptibility to iron ion in water He in the serum, after the iron ion selective binding, promptly can be observed solution colour through naked eyes and become sorrel by yellow, detects lower limit and can reach 10 ^-6M.Need just can not carry out the direct rapid detection of on-site real-time by means of instrument.Can this fluorescent molecular probe be prepared into iron ion and detect test paper and detection kit, also can utilize the viscosity of saccharide compound itself, film, process the portable solid detector.The concrete structure of carbohydrate fluorescent molecular probe is (is example with the GS part) as follows:

Wherein the glycosyl part singly is not a GS, can also be other all carbohydrate chemicals and verivates thereof.

2. water soluble saccharide fluorescent molecular probe according to claim 1 is characterized in that, the glycosyl part singly is not a GS, can also be other all carbohydrate chemicals and verivates thereof.

3. water soluble saccharide fluorescent molecular probe according to claim 1 is characterized in that, uses it for the detection of the iron ion in the sharp biological tissue of environment.

4. water soluble saccharide fluorescent molecular probe according to claim 1 is characterized in that, compounds forms reagent and the method that iron ion detects test paper and detection kit and portable solid fluorescent molecular probe therewith.

5. water soluble saccharide fluorescent molecular probe according to claim 1 is characterized in that, is specially adapted to the emergent detection of the on-the-spot iron ion of environment.