An anthracene-based fluorescent dye and a method for synthesizing the same
The anthracene fluorescent dye synthesized through specific steps solves the problems of short excitation wavelength, large spontaneous background and poor penetration in the existing technology, and realizes the application of longer excitation wavelength and lower cost excitation light source, which is suitable for high-throughput sequencing.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- DINA TECH (BEIJING) CO LTD
- Filing Date
- 2024-02-07
- Publication Date
- 2026-08-04
AI Technical Summary
Existing anthracene fluorescent dyes suffer from problems such as short excitation wavelengths, large spontaneous background in biological tissues, significant photodamage, and shallow tissue penetration during high-throughput sequencing.
Using anthracene fluorescent dyes with specific structures and their synthesis methods, anthracene fluorescent dyes with relatively long excitation wavelengths were synthesized through steps such as reduction of o-halobenzophenone, conversion of halogen to carboxyl group, dehydration and ring closure, oxidation and protection of phenolic hydroxyl groups.
The excitation wavelength of anthracene fluorescent dyes has been improved, reducing spontaneous background and photodamage in biological tissues, enhancing tissue permeability, and making them suitable for lower-cost excitation sources, thus applicable to high-throughput sequencing.
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Figure CN118005498B_ABST
Abstract
Description
Technical Field
[0001] This disclosure relates to the field of biochemical fluorescent dye technology, and in particular to an anthracene fluorescent dye and its synthesis method. Background Technology
[0002] Anthracene fluorescent dyes belong to a class of fluorescent compounds containing anthracene-like rings (carbon-bridged substitutions). These compounds exhibit characteristics such as long wavelengths, fluorescence switching, and high molar absorption coefficients. The applicant's previous patent CN201510155218.9 also disclosed such compounds and their synthesis methods.
[0003] In high-throughput sequencing technology, the fluorescent properties of anthracene fluorescent dyes can be used to label deoxynucleotides. The labeling process involves attaching different fluorescent labeling groups to specific positions on the deoxynucleotides via cleavable phosphate chains. After the DNA has recognized and recorded the fluorescence signal, the fluorescent labeling groups are removed using additional chemical methods, thus preparing for the next round of deoxynucleotide introduction.
[0004] The inventors discovered that existing anthracene fluorescent dyes have the problem of short excitation wavelengths, which leads to a large spontaneous background in biological tissues, significant photodamage, and shallow tissue penetration during high-throughput sequencing. Therefore, there is an urgent need to develop anthracene fluorescent dye with a longer excitation wavelength. Summary of the Invention
[0005] In view of this, the present disclosure provides an anthracene fluorescent dye and a method for synthesizing the same, which has a longer excitation wavelength.
[0006] In a first aspect, embodiments of this disclosure provide an anthracene-based fluorescent dye, employing the following technical solution:
[0007] The anthracene fluorescent dye has the structure shown in general formula (a):
[0008]
[0009] Wherein, R is selected from aromatic group, substituted aromatic group, C1-C6 alkyl group, substituted C1-C6 alkyl group, C1-C6 ether group, and substituted C1-C6 ether group.
[0010] Alternatively, it has the structure shown in general formula (b):
[0011]
[0012] Secondly, embodiments of this disclosure provide a method for synthesizing anthracene fluorescent dyes, employing the following technical solution:
[0013] The method for synthesizing the anthracene fluorescent dye includes:
[0014] Step S1: Reduce o-halobenzophenone to obtain the first intermediate product;
[0015] Step S2: Convert the halogen in the first intermediate product into a carboxyl group to obtain the second intermediate product;
[0016] Step S3: Dehydrate and cyclize the second intermediate to obtain the third intermediate;
[0017] Step S4: Oxidize the third intermediate product to obtain the fourth intermediate product;
[0018] Step S5: Remove the phenolic hydroxyl protecting group from the fourth intermediate to obtain the fifth intermediate;
[0019] Step S6: Protect the phenolic hydroxyl group of the fifth intermediate to obtain the sixth intermediate;
[0020] Step S7: Perform anthrone addition reaction on the sixth intermediate to obtain anthracene fluorescent dye;
[0021] The first intermediate product is a diphenylmethane derivative; the second intermediate product is a carboxyl derivative of diphenylmethane; and the fourth intermediate product is an anthrone compound.
[0022] Optionally, in step S1, the o-halogenated benzophenone is reduced by a reducing agent, wherein the reducing agent is one or more of activated iron powder, triethylsilane, and hypophosphorous acid.
[0023] Optionally, at least one of the 3 and 4 positions of the o-halobenzophenone has a methoxy group.
[0024] Optionally, in step S2, the halogen in the first intermediate product is converted into a carboxyl group by using a Grignard reagent or alkyllithium.
[0025] Optionally, in step S3, the second intermediate is subjected to a dehydration and ring-closing reaction with a ring-closing reagent to obtain a third intermediate. The ring-closing reagent is one or more of oxalyl chloride / boron tribromide, concentrated sulfuric acid, and polyphosphoric acid.
[0026] Optionally, in step S3, the second intermediate product undergoes a dehydration and ring-closing reaction at 20–50°C for no more than 1 hour.
[0027] Optionally, in step S4, the methylene group in the third intermediate product is oxidized to a carbonyl group by an oxidant at 60-70°C. The oxidant is one or more of chromium trioxide, potassium dichromate, and potassium permanganate.
[0028] Optionally, in step S7, the sixth intermediate product is subjected to an anthrone addition reaction by an addition reagent, wherein the addition reagent is a halogenated aromatic compound or a halogenated alkyl compound.
[0029] This disclosure provides an anthracene-based fluorescent dye and its synthesis method. The anthracene-based fluorescent dye has the structure shown in general formula (a). The synthesis method includes: reducing o-halobenzophenone to obtain a first intermediate; converting the halogen in the first intermediate to a carboxyl group to obtain a second intermediate; dehydrating and cyclizing the second intermediate to obtain a third intermediate; oxidizing the third intermediate to obtain a fourth intermediate; removing the phenolic hydroxyl protecting group from the fourth intermediate to obtain a fifth intermediate; protecting the two phenolic hydroxyl groups of the fifth intermediate to obtain a sixth intermediate; and performing an anthrone addition reaction on the sixth intermediate to obtain the anthracene-based fluorescent dye. This anthracene-based fluorescent dye has a hydroxyl group, and the lone pair electrons on the hydroxyl oxygen enhance the conjugation properties of the anthracene-based fluorescent dye, resulting in a longer excitation wavelength. This solves the problems of large spontaneous background radiation, significant photodamage, and shallow tissue penetration in existing technologies.
[0030] The above description is merely an overview of the technical solution disclosed herein. In order to better understand the technical means of this disclosure and to implement it in accordance with the contents of the specification, and to make the above and other objects, features and advantages of this disclosure more apparent and understandable, preferred embodiments are described below in detail with reference to the accompanying drawings. Attached Figure Description
[0031] To more clearly illustrate the technical solutions of the embodiments of this disclosure, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this disclosure. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0032] Figure 1 A flowchart illustrating the synthesis method of anthracene fluorescent dyes provided in this embodiment of the disclosure;
[0033] Figure 2 A statistical diagram of the fluorescence absorption height of anthracene fluorescent dyes at different pH values provided in the embodiments of this disclosure;
[0034] Figure 3 The excitation and emission spectra of anthracene fluorescent dyes provided in embodiments of this disclosure are shown. Detailed Implementation
[0035] The present disclosure will now be described in further detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the scope of the disclosure. Furthermore, it should be noted that, for ease of description, only the parts relevant to the present disclosure are shown in the accompanying drawings.
[0036] It should be noted that, where there is no conflict, the embodiments and features described in this disclosure can be combined with each other. The technical solutions of this disclosure will now be described in detail with reference to the accompanying drawings and embodiments.
[0037] Unless otherwise stated, the exemplary implementations / embodiments shown are to be understood as providing exemplary features of various details that provide ways in which the technical concepts of this disclosure can be implemented in practice. Therefore, unless otherwise stated, the features of various implementations / embodiments may be additionally combined, separated, interchanged and / or rearranged without departing from the technical concepts of this disclosure.
[0038] The terminology used herein is for the purpose of describing particular embodiments and is not intended to be limiting. As used herein, unless the context clearly indicates otherwise, the singular forms “a” and “the” are intended to include the plural forms as well. Furthermore, when the terms “comprising” and / or “including” and variations thereof are used in this specification, it indicates the presence of the stated features, integrals, steps, operations, parts, components, and / or groups thereof, but does not exclude the presence or addition of one or more other features, integrals, steps, operations, parts, components, and / or groups thereof. It should also be noted that, as used herein, the terms “substantially,” “about,” and other similar terms are used as approximate terms rather than as terms of degree, thus explaining the inherent biases in measurements, calculated values, and / or provided values that would be recognized by one of ordinary skill in the art.
[0039] This disclosure provides an anthracene fluorescent dye, specifically, the anthracene fluorescent dye having the structure shown in general formula (a):
[0040]
[0041] Wherein, R is selected from aromatic group, substituted aromatic group, C1-C6 alkyl group, substituted C1-C6 alkyl group, C1-C6 ether group, and substituted C1-C6 ether group.
[0042] It should be noted that the anthracene fluorescent dyes involved in the embodiments of this disclosure refer to fluorescent dyes with a structure similar to triphenylcycloanthracene. C1-C6 refers to dyes containing 1-6 carbon atoms, and so on.
[0043] In one example, the anthracene fluorescent dye in this disclosure embodiment has the structure shown in general formula (b):
[0044]
[0045] The anthracene fluorescent dyes provided in this disclosure have hydroxyl groups. The lone pair electrons on the oxygen atom of the hydroxyl group enhance the conjugated properties of the anthracene fluorescent dyes, resulting in a longer excitation wavelength. Specifically, the anthracene fluorescent dyes provided in this disclosure exhibit a red-shifted excitation wavelength of 570–580 nm and an emission wavelength of 590–610 nm, with a quantum yield of approximately 93%. Compared to existing fluorescent dyes (such as FAM, TG, and PO), the longer excitation wavelength solves the problems of large spontaneous background radiation, significant photodamage, and shallow tissue penetration in existing technologies. Furthermore, it is more suitable for lower-cost excitation light sources, which is of great significance for the sequencing field. Existing fluorescent dyes such as FAM, TG, and PO typically have excitation wavelengths concentrated in the range of 490–550 nm, while the hydrophilic anthracene fluorescent dye disclosed in patent CN112300599B has an excitation wavelength of 460–540 nm.
[0046] Furthermore, embodiments of this disclosure provide a method for synthesizing anthracene fluorescent dyes, such as... Figure 1 As shown, the synthesis method of this anthracene fluorescent dye includes:
[0047] Step S1: Reduce o-halobenzophenone to obtain the first intermediate product.
[0048] Optionally, in step S1, o-halobenzophenone is reduced using a reducing agent, which can be one or more of activated iron powder, triethylsilane, and hypophosphite. The first intermediate product is a diphenylmethane derivative. Taking triethylsilane as an example, when step S1 is carried out at room temperature, 1.5 to 3 equivalents of triethylsilane are required. Taking hypophosphite as an example, when step S1 is carried out at 60°C, 3 to 5 equivalents of hypophosphite are required.
[0049] Optionally, at least one of the 3 and 4 positions of the o-halobenzophenone has a methoxy group, which can act as a power-donating group to increase the electron cloud density of the conjugated structure, thereby increasing the excitation wavelength of the anthracene fluorescent dye.
[0050] Step S2: Convert the halogen in the first intermediate product into a carboxyl group to obtain the second intermediate product.
[0051] The second intermediate product is a carboxyl derivative of diphenylmethane. Optionally, in step S2, the halogen in the first intermediate product is converted to a carboxyl group using a Grignard reagent. For example, the Grignard reagent is used to convert the halogen in the first intermediate product to a carboxyl group at 0°C, requiring 1.5 to 3 equivalents of the Grignard reagent. The Grignard reagent can be methyl magnesium bromide, isopropyl magnesium bromide, etc. Alternatively, the halogen in the first intermediate product is converted to a carboxyl group using an alkyllithium. For example, n-butyllithium is used to convert the halogen in the first intermediate product to a carboxyl group at -78°C, requiring 1.0 to 1.5 equivalents of n-butyllithium.
[0052] Step S3: Dehydrate and cyclize the second intermediate to obtain the third intermediate.
[0053] Optionally, in step S3, the second intermediate is subjected to a dehydration cyclization reaction using a cyclization reagent to obtain the third intermediate. The cyclization reagent can be one or more of oxalyl chloride / boron tribromide, concentrated sulfuric acid, and polyphosphoric acid.
[0054] Optionally, in step S3, the second intermediate product undergoes a dehydration and ring-closing reaction at 20–50°C for no more than 1 hour to avoid excessively high reaction temperature and / or excessively long reaction time leading to an increase in byproducts.
[0055] Taking polyphosphoric acid as the cyclization reagent as an example, step S3 can specifically involve adding 10 times the mass equivalent of polyphosphoric acid to the second intermediate, stirring at room temperature for 1 hour, and then obtaining the third intermediate.
[0056] Step S4: Oxidize the third intermediate to obtain the fourth intermediate.
[0057] The fourth intermediate product is anthrone compounds.
[0058] Optionally, in step S4, the methylene group in the third intermediate is oxidized to a carbonyl group by an oxidizing agent, which may be one or more of chromium trioxide, potassium dichromate, and potassium permanganate. Optionally, the methylene group in the third intermediate is oxidized to a carbonyl group at 60–70°C by an oxidizing agent. Taking chromium trioxide as an example, step S4 may specifically involve adding three equivalents of chromium trioxide and ten times the volume of acetic acid to the third intermediate, and heating at 60°C for 2 hours.
[0059] Step S5: Remove the phenolic hydroxyl protecting group from the fourth intermediate to obtain the fifth intermediate.
[0060] For example, three equivalents of boron tribromide are added to the fourth intermediate, and a demethylation reaction is carried out at room temperature for 2 hours to remove the phenolic hydroxyl protecting group from the fourth intermediate to obtain the fifth intermediate.
[0061] Step S6: Protect the phenolic hydroxyl group of the fifth intermediate to obtain the sixth intermediate.
[0062] For example, 2.5 to 3.0 equivalents of TBSCl were added to the fifth intermediate to protect the phenolic hydroxyl group in the fifth intermediate, thereby obtaining the sixth intermediate.
[0063] Step S7: Perform anthrone addition reaction on the sixth intermediate to obtain anthracene fluorescent dye;
[0064] Optionally, in step S7, the sixth intermediate is subjected to an anthrone addition reaction by an addition reagent, which is a halogenated aromatic compound or a halogenated alkyl compound.
[0065] In step S7, 1.5 equivalents of n-butyllithium are added to the sixth intermediate product, and the reaction is carried out at -78°C for 1 hour. While maintaining the temperature at -78°C, 1.0 equivalent of tetrahydrofuran solution is added dropwise.
[0066] It should be noted that some steps mentioned in the embodiments of this disclosure may have other products. Based on the embodiments of this disclosure, those skilled in the art can obtain the target product. The specific structure of other products is not part of the core content of the embodiments of this disclosure and will not be described in detail here.
[0067] The above methods for synthesizing anthracene fluorescent dyes not only enable the synthesis of fluorescent dyes with longer excitation wavelengths, but also offer advantages such as readily available and inexpensive raw materials, simple intermediate preparation, high yield, and easy and convenient operation. Furthermore, they can effectively improve yield, increase the ease of derivatization, and reduce the cost of large-scale synthesis, which is conducive to promoting the application of anthracene fluorescent dyes in molecular labeling.
[0068] Example
[0069] The synthetic route for anthracene fluorescent dyes with the structure shown in general formula (b) is as follows:
[0070]
[0071] Its specific synthesis process includes:
[0072] (1) Synthesis of compound 1-bromo-4-methoxy-2-(3-methoxybenzyl)benzene 2:
[0073] 20 g of compound 1 was added to 200 ml of dichloromethane solution. 37 ml of triethylsilane was added under ice bath conditions, followed by 25 ml of trifluoroacetic acid. After the addition was complete, 2 ml of trifluoromethanesulfonic acid was added dropwise to the mixture. The mixture was stirred for 10 min under ice bath conditions, then heated to room temperature and stirred for another two hours. After complete conversion of the starting material as monitored by TCL, the reaction system was added to 200 ml of pure water. The mixture was separated, and the organic phase was washed once more with 200 ml of pure water. After purification by a concentration column, compound 2 (15 g, Yield: 78%) was obtained. Yield represents the yield.
[0074] The results of its one-dimensional nuclear magnetic resonance hydrogen spectrum and mass spectrometry are as follows:
[0075] 1 H NMR(500MHz, CDCl3)7.39(d,J=7.5Hz,1H),7.21(t,J=7.5Hz,1H),7.01–6.95(m,1H),6.82(ddd,J=7.6,6. 2,1.5Hz,2H),6.70(q,J=1.1Hz,1H),6.66(p,J=1.2Hz,1H),3.95(q,J=0.9Hz,2H),3.82(d,J=3.1Hz,6H).
[0076] LCMS: C15H15BrO2, (M+H): 307.0255 / 309.0235.
[0077] (2) Synthesis of compound 4-methoxy-2-(3-methoxybenzyl)benzoic acid 3:
[0078] 15 g of compound 2 was dissolved in 150 ml of ultra-dry tetrahydrofuran and stirred until dissolved. The solution was cooled to -78°C, and 20 ml of n-butyllithium in n-hexane solution (2.4 M) was added dropwise to the reaction system. After the addition was complete, the system was kept at -78°C and stirred for 30 min. Ultra-dry carbon dioxide gas was then introduced into the reaction system, and the system was kept at -78°C and stirred for 10 min. The system was then slowly heated to room temperature and the reaction was continued for 30 min. After the reaction was confirmed to be complete by TLC, the reaction system was added to 150 ml of 1 M hydrochloric acid aqueous solution, and then 100 ml of ethyl acetate was added. The mixture was separated, and the organic phase was washed once with 100 ml of saturated brine. The organic phase was concentrated to dryness to obtain crude compound 3 (14 g, yield: 100%), which was directly used in the next reaction.
[0079] The results of its one-dimensional nuclear magnetic resonance hydrogen spectrum and mass spectrometry are as follows:
[0080] 1H NMR(500MHz,Chloroform-d)δ7.79(d,J=7.4Hz,1H),7.21(t,J=7.5Hz,1H),6.97(dq,J=7.5,1.2Hz,1H),6.81(d t,J=7.5,1.5Hz,1H),6.75–6.68(m,2H),6.63(p,J=1.2Hz,1H),4.25(q,J=0.9Hz,2H),3.87(s,3H),3.80(s,3H).
[0081] LCMS: C16H16O4, (M+H): 273.1082.
[0082] (3) Synthesis of compound 3,6-dimethoxyanthracen-9(10H)-one:
[0083] 14 g of compound 3 was dissolved in 30 ml of dichloromethane solution. The dichloromethane solution of compound 3 was then added to 140 g of PPA. The reaction mixture was stirred with a glass rod at room temperature until complete reaction was observed under TLC. Ice water was then added to the mixture, and the mixture was stirred with a glass rod until the PPA was completely hydrolyzed. The mixture was then extracted once with 150 ml of dichloromethane, and the liquid phase was separated. The aqueous phase was extracted once more with 150 ml of dichloromethane, and the liquid phase was separated. The organic phases were combined and concentrated to dryness. The concentrated system was then slurried with 50 ml of MTBE and filtered to obtain compound 4 (8 g, yield: 61%), which was used directly in the next reaction step.
[0084] The results of its one-dimensional nuclear magnetic resonance hydrogen spectrum and mass spectrometry are as follows:
[0085] 1 H NMR(500MHz,Chloroform-d)δ7.88(d,J=7.4Hz,2H),6.92–6.84(m,4H),4.36(t,J=1.0Hz,2H),3.85(s,5H).
[0086] LCMS: C16H14O3, (M+H): 255.0976.
[0087] (4) Synthesis of compound 2,7-dimethoxyanthracene-9,10-dione 5:
[0088] Compound 4 (8 g) was dissolved in 40 ml of glacial acetic acid solution, and 9.4 g of chromium trioxide was added. The system was heated to 50°C and stirred for 1 hour. After the reaction was completed by TLC monitoring, the system was cooled to room temperature, and 150 ml of pure water was added. The mixture was stirred for another 30 minutes and then filtered to obtain compound 5 (8 g, yield: 95%).
[0089] The results of its one-dimensional nuclear magnetic resonance hydrogen spectrum and mass spectrometry are as follows:
[0090] 1 H NMR (500MHz, Chloroform-d) δ8.15(d,J=7.4Hz,2H),7.58(d,J=1.5Hz,2H),7.12(dd,J=7.5,1.5Hz,2H),3.82(s,5H).
[0091] LCMS: C16H12O4, (M+H): 269.0769.
[0092] (5) Synthesis of compound 2,7-dihydroxyanthracene-9,10-dione 6:
[0093] Compound 5 (8 g) was dissolved in 80 mL of dichloromethane solution, cooled to 0°C, and 6.6 mL of boron tribromide was added dropwise. After the addition was complete, the temperature was slowly raised to room temperature, and the mixture was stirred for 2 hours. After the reaction was monitored by TLC until it was complete, 150 mL of pure water was added dropwise to the reaction system, followed by 150 mL of ethyl acetate. The mixture was separated, and the aqueous phase was extracted once with 50 mL of ethyl acetate. The organic phases were combined, washed once with 50 mL of saturated brine, separated, and the organic phase was concentrated to dryness to obtain compound 6 (6 g, yield: 84%).
[0094] The results of its one-dimensional nuclear magnetic resonance hydrogen spectrum and mass spectrometry are as follows:
[0095] 1 H NMR (500MHz, Chloroform-d) δ9.04 (s, 2H), 8.08 (d, J = 7.5Hz, 2H), 7.54 (d, J = 1.4Hz, 2H), 7.29 (dd, J = 7.5, 1.5Hz, 2H).
[0096] LCMS: C14H8O4, (M+H): 241.0456.
[0097] (6) Synthesis of compound 2,7-bis((tert-butyldimethylsilyl)oxy)anthracene-9,10-dione 7:
[0098] Compound 6 (6 g) was added to 30 ml of DMF, followed by 5.1 g of imidazole and 9.4 g of TBSCl. The mixture was stirred at room temperature for 2 hours. After the reaction was completed by TLC monitoring, the reaction mixture was added to 90 ml of pure water, followed by 90 ml of MTBE. The mixture was separated into liquid and liquid phases. The organic phase was washed once with 60 ml of saturated saline solution, separated again, and concentrated to dryness. The concentrated dry matter was slurried with 30 ml of methanol and filtered to obtain compound 7 (10 g, yield: 85%).
[0099] The results of its one-dimensional nuclear magnetic resonance hydrogen spectrum and mass spectrometry are as follows:
[0100] 1 H NMR(500MHz,Chloroform-d)δ8.03(d,J=7.5Hz,2H),7.44(d,J=1.6Hz,3H),7.26(dd,J=7.5,1.5Hz,2H),1.01(s,17H),0.21(s,11H).
[0101] LCMS:C26H36O4Si2, (M+H):469.2186
[0102] (7) Synthesis of compound 10-bis(2,4-dimethylphenyl)-7,9-dihydroxyanthracen-2(9H)-onetarget 1:
[0103] 2 g of compound 7 was dissolved in 10 ml of ultra-dry tetrahydrofuran and stirred until dissolved. The solution was sealed and kept for later use. 2.4 g of compound 1-bromo-2,4-dimethylbenzene was dissolved in 10 ml of ultra-dry tetrahydrofuran and cooled to -78°C. Then, 5.4 ml of n-butyllithium in n-hexane was added dropwise, and the temperature was maintained at -78°C. Stirring was continued for 30 min. The tetrahydrofuran solution of compound 7 prepared above was added dropwise to the low-temperature system, and the temperature was maintained at -78°C. Stirring was continued for 10 min, and then the temperature was slowly raised to room temperature. Stirring was continued for 30 min. After the reaction was monitored by TLC until it was complete, 20 ml of 5M hydrochloric acid aqueous solution was added to the above reaction system. The mixture was stirred at room temperature for 1 hour and filtered to obtain compound target 1 (1.3 g, yield: 70%).
[0104] The results of its one-dimensional nuclear magnetic resonance hydrogen spectrum and mass spectrometry are as follows:
[0105] 1H NMR(500MHz,Chloroform-d)δ8.87(s,1H),7.54–7.46(m,2H),7.22(dd,J=7.5,4.4 Hz,2H),7.17(ddd,J=7.5,1.5,0.8Hz,1H),7.11–7.04(m,2H),6.93(d,J=1.5Hz,1H ),6.86(t,J=1.0Hz,1H),6.76(dd,J=7.5,1.7Hz,1H),6.62(d,J=2.3Hz,1H),6.58( dd,J=11.0,2.2Hz,1H),4.79(s,1H),2.35(d,J=4.0Hz,6H),2.30(d,J=1.5Hz,6H).
[0106] LCMS: C30H26O3, (M+H): 435.1915.
[0107] like Figure 2 As shown, anthracene fluorescent dyes with the structure shown in general formula (b) have low fluorescence absorption height at pH less than 8, significantly increased fluorescence absorption height at 8 < pH < 10, and remain basically stable after pH > 10.
[0108] The spectral properties of compound target1 were measured using a fluorescence spectrometer in a pH 8 TEAA buffer solution as follows: Figure 3 As shown, its excitation wavelength is 576nm and its emission wavelength is 594nm.
[0109] In the description of this specification, the references to terms such as "one embodiment / mode," "some embodiments / modes," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment / mode or example is included in at least one embodiment / mode or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment / mode or example. Moreover, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments / modes or examples. Furthermore, without contradiction, those skilled in the art can combine and integrate the different embodiments / modes or examples described in this specification, as well as the features of different embodiments / modes or examples.
[0110] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this application, "multiple" means at least two, such as two, three, etc., unless otherwise explicitly specified.
[0111] Those skilled in the art should understand that the above embodiments are merely for illustrating the present disclosure and are not intended to limit the scope of the disclosure. Those skilled in the art can make other changes or modifications based on the above disclosure, and these changes or modifications still fall within the scope of the present disclosure.
Claims
1. An anthracene fluorescent dye, characterized in that, It has the structure shown in general formula (b): （b）。 2. A method for synthesizing anthracene fluorescent dye, used to prepare the anthracene fluorescent dye according to claim 1, characterized in that, The synthesis route is as follows: The PPA in question is polyphosphoric acid.
3. The method for synthesizing anthracene fluorescent dyes according to claim 2, characterized in that, The dehydration and ring-closing reaction was carried out at 20~50℃ for no more than 1 hour.
4. The method for synthesizing anthracene fluorescent dyes according to claim 2, characterized in that, The oxidizing agent in the oxidation process is one or more of chromium trioxide, potassium dichromate, and potassium permanganate.