Synthesis method of multi-fluorescence unit composite molecules as well as application thereof
A technology of composite molecules and synthesis methods, applied in the field of synthesis of multi-fluorescence unit composite molecules, can solve the problems of inconsistent standard curves between batches of sensors, and achieve the effect of stable performance and efficient coupling between batches
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Embodiment 1
[0047]The preparation of embodiment 1 single fluorescein-ethylenediamine
[0048] The reaction formula is as follows:
[0049]
[0050] The specific steps are:
[0051] (1) Add 1 mL of water to a 2 mL centrifuge tube, add 4.72 μL of ethylenediamine to the centrifuge tube, then add 8.92 μL of triethylamine, transfer to a flask, and mix evenly by ultrasonication.
[0052] (2) Weigh 25 mg of fluorescein isothiocyanate (FSITC), dissolve it in 4 mL of absolute ethanol, add it to the flask in (1) after fully dissolving, stir at room temperature in the dark for 24 hours, and evaporate with a rotary evaporator Dry the solvent, use 1-2mL dichloromethane:ethanol=1:1 (V / V) mixed solution for reconstitution.
[0053] (3) Fix the glass chromatography column on the iron stand, put 100-200nm silica gel into the glass chromatography column, the height of the silica gel accounts for half of the height of the glass chromatography column, and then use a circulating water type multi-purpose ...
Embodiment 2
[0054] The preparation of embodiment 2 single fluorescein-ethylenediamine-azide butyric acid
[0055] The reaction formula is as follows:
[0056]
[0057] The specific steps are:
[0058] (1) Weigh 9 mg of monofluorescein-ethylenediamine, dissolve it in 6 mL of water and 100 μL of ethanol, add 20 μL of ammonia water (25-28%), and dissolve it fully by ultrasonication.
[0059] (2) Weigh 2.5 mg of azidobutyric acid and dissolve it in 1 mL of water, add 7.16 mg of EDC to activate for 1 hour, then add 2.3 mg of NHS to activate for 1 hour, and add it to the monofluorescein-ethylenediamine solution of (1), Stir at room temperature in the dark for 24 h. After lyophilization, reconstitute with a mixed solution of 1-2 mL dichloromethane:ethanol=1:1 (V / V).
[0060] (3) Fix the glass chromatography column on the iron stand, put 100-200nm silica gel into the glass chromatography column, the height of the silica gel accounts for half of the height of the glass chromatography column,...
Embodiment 3
[0061] The preparation of embodiment 3 monorhodamine-ethylenediamine
[0062] The reaction formula is as follows:
[0063]
[0064] The specific steps are:
[0065] (1) Add 3 mL of water to a 5 mL centrifuge tube, add 12.6 μL of ethylenediamine to the centrifuge tube, then add 11.7 μL of triethylamine, transfer to a flask, and mix well by ultrasonication.
[0066] (2) Weigh 30 mg of isothiocyanatorhodamine, dissolve it in 6 mL of absolute ethanol, add it into the flask in (1) after fully dissolving, stir at room temperature in the dark for 24 hours, and evaporate the solvent to dryness with a rotary evaporator. Use 1-2 mL of dichloromethane:methanol=11:1 (V / V) mixed solution for reconstitution.
[0067] (3) Fix the glass chromatography column on the iron stand, put 100-200nm silica gel into the glass chromatography column, the height of the silica gel accounts for half of the height of the glass chromatography column, and then use a circulating water type multi-purpose va...
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