Calcium ion and zinc ion double-channel rhodamine fluorescent probe as well as preparation method and application thereof

Abstract

The invention relates to a calcium ion and zinc ion double-channel rhodamine fluorescent probe as well as a preparation method and application thereof. A structural formula of the calcium ion and zincion double-channel rhodamine fluorescent probe is as follows: the formula is shown in the description; the preparation method comprises the following steps: taking rhodamine B and hydrazine hydrate to react, so as to obtain rhodamine B hydrazide; carrying out reaction on p-aminobenzoate and the hydrazine hydrate; dissolving an obtained product into a solvent and dropwise adding salicylic aldehyde; reacting to obtain salicylic aldehyde-4-aminobenzoyl hydrazide hydrazone; dissolving cyanuric trichloride into the solvent and adding an acid binding agent; dropwise adding a THF (Tetrahydrofuran) solution of the rhodamine B hydrazide into an ice water bath under the protection of nitrogen gas; stirring to obtain RSH; dissolving the RSH and adding the acid binding agent; under the protection ofnitrogen gas, dropwise adding the salicylic aldehyde-4-aminobenzoyl hydrazide hydrazone and reacting to obtain RSHT; dissolving the RSHT and adding the acid binding agent; dropwise adding the hydrazine hydrate under the protection of N2 and reacting to obtain the probe. The fluorescent probe compound provided by the invention has very good selectivity on zinc ions and calcium ions, is convenientlyapplied to environmental applications and has a relatively good utilization effect.

Description

technical field

[0001] The invention belongs to the field of fluorescent probe materials and preparation thereof, in particular to a calcium ion and zinc ion dual-channel rhodamine-based fluorescent probe and its preparation method and application. Background technique

[0002] As an important information transmission substance in cells, calcium ions play an important role in many physiological functions, such as muscle excitation and contraction, neurotransmitter release, substance metabolism, nerve impulse conduction, etc. As the second messenger in cells, calcium ions have important biological functions, and the messenger function of calcium ions is realized through the change of the distribution of free calcium ions in cells. Accurate determination of intracellular free calcium ion concentration is an important content of current medical and biological research. The calcium ion fluorescent probe developed in recent years has the advantages of high sensitivity, fast resp...

Examples

Embodiment 1

[0049] The calcium ion and zinc ion dual-channel rhodamine-based fluorescent probes of the present embodiment have a structural formula of:

[0050]

[0051] Its preparation method, the steps are:

[0052] Step 1: Dissolve rhodamine B in absolute ethanol, add excess hydrazine hydrate (98wt%) dropwise, heat and reflux at 78°C for 5 hours, remove the solvent and unreacted hydrazine hydrate by rotary evaporation, and obtain crude rhodamine B hydrazide . Dissolve the crude product in dilute hydrochloric acid solution (4.0-5.0 wt%), adjust the pH of the system (9-10) until the product is completely precipitated, filter it with suction, and dry it to obtain the rhodamine B hydrazide product;

[0053] Step 2: Dissolve methyl p-aminobenzoate in absolute ethanol, add excess hydrazine hydrate (98wt%) dropwise, heat and reflux at 78°C for 3 hours, remove the solvent and unreacted hydrazine hydrate by rotary evaporation, and obtain the crude product p-aminobenzoate Benzohydrazide, fo...

Embodiment 2

[0058] Utilize the calcium ion and zinc ion dual-channel rhodamine fluorescent probe in embodiment 1 to test:

[0059] like figure 1 As shown, the probe molecule RSHT-NH in DMF solution 2 There is no ultraviolet absorption peak at 360-400nm itself, after the probe is complexed with zinc ions, a new ultraviolet absorption peak appears at 370nm ( figure 1 (a) shown); Probe molecule RSHT-NH in DMF solution 2 There is no ultraviolet absorption peak at 360-400nm, but a new ultraviolet absorption peak appears at 380nm after the probe is complexed with calcium ions.

[0060] exist figure 2 It can be found in the DMF solution that after the fluorescent probe is complexed with zinc ions and calcium ions, the fluorescence excitation wavelengths are 370nm and 380nm respectively, and the fluorescence emission wavelengths are 470nm and 450nm respectively, and the colors before and after the solution have changed greatly ( RSHT-NH 2 -Ca 2+ Complexation occurs light green fluorescen...

technical field

[0001] The invention belongs to the field of fluorescent probe materials and preparation thereof, in particular to a calcium ion and zinc ion dual-channel rhodamine-based fluorescent probe and its preparation method and application. Background technique

[0002] As an important information transmission substance in cells, calcium ions play an important role in many physiological functions, such as muscle excitation and contraction, neurotransmitter release, substance metabolism, nerve impulse conduction, etc. As the second messenger in cells, calcium ions have important biological functions, and the messenger function of calcium ions is realized through the change of the distribution of free calcium ions in cells. Accurate determination of intracellular free calcium ion concentration is an important content of current medical and biological research. The calcium ion fluorescent probe developed in recent years has the advantages of high sensitivity, fast resp...

Examples

Embodiment 1

[0049] The calcium ion and zinc ion dual-channel rhodamine-based fluorescent probes of the present embodiment have a structural formula of:

[0050]

[0051] Its preparation method, the steps are:

[0052] Step 1: Dissolve rhodamine B in absolute ethanol, add excess hydrazine hydrate (98wt%) dropwise, heat and reflux at 78°C for 5 hours, remove the solvent and unreacted hydrazine hydrate by rotary evaporation, and obtain crude rhodamine B hydrazide . Dissolve the crude product in dilute hydrochloric acid solution (4.0-5.0 wt%), adjust the pH of the system (9-10) until the product is completely precipitated, filter it with suction, and dry it to obtain the rhodamine B hydrazide product;

[0053] Step 2: Dissolve methyl p-aminobenzoate in absolute ethanol, add excess hydrazine hydrate (98wt%) dropwise, heat and reflux at 78°C for 3 hours, remove the solvent and unreacted hydrazine hydrate by rotary evaporation, and obtain the crude product p-aminobenzoate Benzohydrazide, fo...

Embodiment 2

[0058] Utilize the calcium ion and zinc ion dual-channel rhodamine fluorescent probe in embodiment 1 to test:

[0059] like figure 1 As shown, the probe molecule RSHT-NH in DMF solution 2 There is no ultraviolet absorption peak at 360-400nm itself, after the probe is complexed with zinc ions, a new ultraviolet absorption peak appears at 370nm ( figure 1 (a) shown); Probe molecule RSHT-NH in DMF solution 2 There is no ultraviolet absorption peak at 360-400nm, but a new ultraviolet absorption peak appears at 380nm after the probe is complexed with calcium ions.

[0060] exist figure 2 It can be found in the DMF solution that after the fluorescent probe is complexed with zinc ions and calcium ions, the fluorescence excitation wavelengths are 370nm and 380nm respectively, and the fluorescence emission wavelengths are 470nm and 450nm respectively, and the colors before and after the solution have changed greatly ( RSHT-NH 2 -Ca 2+ Complexation occurs light green fluorescen...

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