A phospholuciferin fluorescent probe for rapid and efficient detection of H2S and a synthesis method and application thereof

Abstract

The application belongs to the technical field of fluorescent probes, and particularly relates to a phosphofluorene fluorescent probe for rapidly and efficiently detecting H2S as well as a synthesis method and application thereof. The phosphofluorene fluorescent probe has the structure of formula (1): the probe can selectively recognize H2S molecules, the fluorescent probe takes a phosphofluorene molecule as a fluorescent group, and then links a 2,4-dinitrophenyl ether group at the position of a hydroxyl group to form a phosphofluorene fluorescent compound. The phosphofluorene fluorescent probe is a fluorescent probe based on a sulfurolysis reaction, can rapidly occur the sulfurolysis reaction in the presence of H2S, and can emit strong fluorescence by removing the 2,4-dinitrophenyl ether group. The phosphofluorene fluorescent probe can be widely applied to detection of H2S in cells, soil and water bodies.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of fluorescent probes, and particularly relates to a phosphofluorene fluorescent probe for rapidly and efficiently detecting H2S as well as a synthesis method and application thereof. BACKGROUND

[0002] Hydrogen sulfide (H2S) is ubiquitous, and it not only exists in nature but also plays an important role in our life activities. In vivo, endogenous H2S is an important signal transduction molecule, and is involved in various pathophysiological processes. H2S is the third important signal transduction molecule in vivo after CO and NO. It plays an important role in the nervous system, cardiovascular system, endocrine system, gastrointestinal system and immune system, and H2S concentration disorder is closely related to various diseases such as neurodegenerative diseases, cirrhosis, Parkinson's disease, pulmonary hypertension, diabetes, etc. With the progress of science and technology, H2S is found in more and more fields. In order to specifically understand the specific role of H2S in physiological processes, it is very valuable to track the specific changes of H2S in complex environments and physiological processes.

[0003] At present, the commonly used methods for detecting hydrogen sulfide (H2S) mainly include an electrochemical method based on a sulfide selective electrode, a methylene blue determination method and a gas chromatography method. These methods cannot simply and efficiently qualitatively and quantitatively analyze H2S in cells due to time, labor and detection cost. We need a simple and fast way to detect hydrogen sulfide in cell tissues. The fluorescent probe detection method is widely concerned due to high sensitivity.

[0004] The common H2S fluorescent probes on the market are mainly based on the detection mechanism of reduction reaction and nucleophilic reaction. However, these reactions usually have a problem that their reaction rates are relatively slow, and the detection process needs dozens of minutes or even several hours, and cannot efficiently detect H2S. Now we use phosphofluorene as a fluorescent group to synthesize an H2S fluorescent probe with good selectivity and fast detection effect, which has certain challenge and research significance. SUMMARY

[0005] The application aims at the deficiencies in the prior art, and provides a phosphofluorene fluorescent probe for rapidly and efficiently detecting H2S and a synthesis method thereof. The probe can selectively recognize H2S molecules. The fluorescent probe takes phosphofluorene molecules as fluorescent groups, and then links a 2,4-dinitrophenyl ether group at the position of a hydroxyl group to form a phosphofluorene fluorescent compound. The phosphofluorene fluorescent probe is a fluorescent probe based on a sulfurolysis reaction. When H2S exists, the sulfurolysis reaction can quickly occur, the 2,4-dinitrophenyl ether group is removed, and strong fluorescence is emitted. The phosphofluorene fluorescent probe can be widely applied to the detection of H2S in cells, soil and water bodies.

[0006] To achieve the above object, the specific technical scheme of the present application is as follows:

[0007] A phosphole fluorescent probe compound for rapid and efficient detection of H2S, which is 3-(2,4-dinitrophenoxy)-5-phenylbenzo[b]phosphole-5-oxide, has the structure of formula (1):

[0008]

[0009] (1); the compound is used for rapid and efficient detection of H2S.

[0010] A preparation method of a phosphole fluorescent probe compound for rapid and efficient detection of H2S, and the synthetic route is shown in formula (2):

[0011]

[0012] A preparation method of a phosphole fluorescent probe compound for rapid and efficient detection of H2S, comprising the following steps:

[0013] (1) K2CO3 and Pd(PPh)3Cl2 are added to the mixture of compound A and compound B, and then the foregoing substances are dissolved in a DME-containing aqueous solution; and the dissolved substances are refluxed under argon, detected by TLC, extracted with ethyl acetate and columned to obtain transparent oily compound C;

[0014] (2) Mg and I2 are added under argon, THF is used as a solvent, and the foregoing substances are stirred at a certain temperature until the iodine fades; compound C is dissolved in THF and slowly added dropwise into a three-necked flask, heated and refluxed, cooled to room temperature, and the supernatant is taken into a clean round-bottom flask and slowly added dropwise with PPhCl2 dissolved in THF under argon; the foregoing substances are heated and refluxed, detected by TLC, cooled to 0°C, oxidized with H2O, stirred, and then Et3N is added, detected by TLC, extracted with ethyl acetate and columned to obtain compound E;

[0015] (3) Compound E is dissolved in THF under argon, Pd(OAC)2 is added as a catalyst, stirred at a certain temperature, detected by TLC, and columned to obtain yellow oily compound F;

[0016] (4) Compound F is dissolved in dichloromethane under argon, BBr3 is added as a catalyst, stirred at a certain temperature, detected by TLC, and the dichloromethane is dried and extracted to obtain yellow oily compound G;

[0017] (5) Compound G and 2,4-dinitrophenyl chloride are dissolved in DMF, potassium carbonate is added, stirred at a certain temperature, extracted with ethyl acetate and columned to obtain yellow solid compound H.

[0018] As a preferred embodiment in the present application, in step (1) of the preparation method of the phosphofluorene fluorescent probe compound for rapid and efficient detection of H2S, the molar ratio of DME to H2O in the aqueous solution containing DME is 8-9:1-2 (more preferably 8.5:1.5); the molar ratio of compound A, compound B, K2CO3 and Pd(PPh)3Cl2 is 1-1.5:1-1.5:2-3:0.01-0.02 (more preferably 1:1.1:2.5:0.015); the reflux condition is 85℃±5℃, 8±0.5h (more preferably 85℃, 8h).

[0019] As a preferred embodiment in the present application, in step (2) of the preparation method of the phosphofluorene fluorescent probe compound for rapid and efficient detection of H2S, stirring is carried out at 40℃ until the iodine fades; TLC detection must be cooled to 0℃, H2O is added for oxidation, and the oxidation time is 15-30min.

[0020] The molar ratio of compound C, Mg, I2, PPhCl2 and Et3N is 1-1.5:1-1.5:0.02-0.04:1.0-1.4:1.5-2.5 (more preferably 1:1.1:0.03:1.2:2); 2-3mL of THF is added per millimole of Mg, 0.5-1 times (volume) of THF is used to dissolve per millimole of compound C, the reflux condition is 80±5℃, 2±0.5h (more preferably 80℃, 2h); 0.5-1 times (volume) of THF is used to dissolve per millimole of PPhCl2, the reflux condition is 80±5℃, 3±0.5h (more preferably 80℃, 3h).

[0021] As a preferred embodiment in the present application, in step (3) of the preparation method of the phosphofluorene fluorescent probe compound for rapid and efficient detection of H2S, the molar ratio of compound E and Pd(OAC)2 is 1:0.03-0.2; 5-10 times the volume of THF is used to dissolve per millimole of compound E; the reflux condition is 65±5℃, 5±0.5h (more preferably 65℃, 5h).

[0022] As a preferred embodiment in the present application, in step (4) of the preparation method of the phosphofluorene fluorescent probe compound for rapid and efficient detection of H2S, the molar ratio of compound F and BBr3 is 1:1.2-3.0; 5-10 times the volume of dichloromethane is used to dissolve per millimole of compound F, and the reaction condition is 0℃, 4h.

[0023] As a preferred embodiment in the application, in step (5) of the preparation method of the fast and efficient H2S phosphofluorene fluorescent probe compound, the molar ratio of compound G, 2,4-dinitrochlorobenzene and potassium carbonate is 1-1.5:1-1.5:1.0-2.0 (more preferably 1:1.2:1.5), 5-15 volumes of DMF are used per millimole of compound G, the reaction condition is 100±5℃, 6±0.5h (more preferably 100℃, 6h).

[0024] The application also protects the use of the fast and efficient H2S phosphofluorene fluorescent probe compound in the fast and efficient detection of H2S in cells, soil or water bodies.

[0025] Further, the reaction mechanism of the application is shown in formula (3);

[0026]

[0027] Compared with the prior art, the application has the following beneficial effects:

[0028] (I) The phosphofluorene molecule is used as a fluorescent group in the application, and the final structure is obtained after the introduction of the 2,4-dinitrophenyl ether group into the fluorescent group. The two nitro groups on the introduced 2,4-dinitrophenyl ether group have a strong electron-withdrawing effect, so that the phosphofluorene fluorescent group emits fluorescence is quenched. The fluorescence quenching effect disappears after the sulfidation reaction of the fluorescent probe with H2S and the 2,4-dinitrophenyl ether group is removed, and the fluorescence effect is obviously enhanced at 528nm.

[0029] (II) The phosphofluorene fluorescent probe in the application has the following characteristics: the maximum ultraviolet absorption peak before the reaction is 298nm, the excitation wavelength is 298nm, and the fluorescence is quenched by the 2,4-dinitrophenyl ether group. The maximum ultraviolet absorption after the reaction is 406nm, and the fluorescence maximum emission is 528nm. In a 10% (mass percent) DMF PBS buffer solution at room temperature, after the addition of H2S, the fluorescence at 528nm is enhanced by 12.8 times;

[0030] (III) The phosphofluorene fluorescent probe in the application is not disturbed by various analytes such as inorganic salts, reducing agents, amino acids, oxidizing agents, metal ions, etc., and can realize high-selectivity detection of H2S, and can exclude the interference of the external environment in the detection process. BRIEF DESCRIPTION OF DRAWINGS

[0031] Figure 1 NMR spectrum of the phosphofluorene fluorescent probe 1 H-NMR spectrum;

[0032] Figure 2 NMR spectrum of the phosphofluorene fluorescent probe 1 C-NMR spectrum;

[0033] Figure 3 UV absorption spectra of phospholuenin fluorescent probe before and after adding H2S for 15 min;

[0034] Figure 4 Fluorescence emission spectra of phospholuenin fluorescent probe before and after adding H2S for 15 min;

[0035] Figure 5 Kinetic experiment spectrum of phospholuenin fluorescent probe and H2S reaction;

[0036] Figure 6 Experimental results of phospholuenin fluorescent probe and H2S reaction with different concentration gradients;

[0037] Figure 7 Fluorescence emission spectra of phospholuenin fluorescent probe and H2S reaction under different pH values;

[0038] Figure 8 Selectivity results of phospholuenin fluorescent probe and H2S reaction under the condition of different interferents; DETAILED DESCRIPTION

[0039] All features disclosed in this specification, and / or all steps of any methods or processes disclosed in this specification, may be combined in any combination, except combinations where at least some of such features and / or steps are mutually exclusive.

[0040] Any feature disclosed in this specification, unless stated otherwise, can be replaced by any equivalent or similar feature, or combination thereof. That is, unless stated otherwise, each feature disclosed is one example only of a generic series of equivalent or similar features.

[0041] The features and performance of the present application will be further described in detail below with reference to the examples. In the following examples, % means volume percent unless otherwise specified; the steps not described in detail are conventional techniques.

[0042] Example 1:

[0043] A phospholuenin fluorescent probe compound for rapid and efficient detection of H2S, which is 3-(2,4-dinitrophenoxy)-5-phenylbenzo[b]phospholuenin-5-oxide, and its structural formula is:

[0044]

[0045] Example 2:

[0046] A phospholuenin fluorescent probe compound for rapid and efficient detection of H2S, and its preparation route is as follows: A phospholuenin fluorescent probe compound for rapid and efficient detection of H2S, which is 3-(2,4-dinitrophenoxy)-5-phenylbenzo[b]phospholuenin-5-oxide, and its structural formula is:

[0047] Example 3:

[0048] A method for preparing a fast and efficient H2S phospholene fluorescent probe compound, the preparation route is the same as example 2, and the specific method comprises the following steps:

[0049] Preparation of compound C:

[0050]

[0051] Compound A (5.66 g, 20 mmol) and compound B (3.344 g, 22 mmol) were mixed to obtain a mixture, then K2CO3 (6.9 g, 50 mmol), Pd(PPh)3Cl2 (0.221 g, 0.3 mmol) were added, the above-mentioned substances were dissolved in 60 mL of ethylene glycol dimethyl ether and 8 mL of water, and then added to a 250 mL round-bottom flask, and refluxed at 85°C under argon for 8 h, TLC detection, ethyl acetate extraction and column chromatography to obtain transparent oily compound C (4.8 g, 91.2%); 1 H NMR (600 MHz, CDCl3) δ 7.58 (dd, J = 8.0, 0.8 Hz, 1H), 7.30-7.22 (m, 4H), 7.09 (ddd, J = 8.0, 6.9, 2.2 Hz, 1H), 6.90-6.87 (m, 2H), 3.78 (s, 3H).

[0052] Preparation of compound E:

[0053]

[0054] Mg (515 mg, 21.5 mmol) was added under argon, and I2 (163.83 mg, 0.645 mmol) was dissolved in 20 mL of THF as a solvent, and stirred at 40°C until the iodine faded. Compound C (4.71 g, 17.9 mmol) was dissolved in 10 mL of THF and slowly added to a three-necked flask (at least 5 min), heated to 80°C and refluxed for 2 h, cooled to room temperature, and the supernatant was slowly added to a clean round-bottom flask under argon THF solution of PPhCl2 (4.8 g, 26.85 mmol) (at least 5 min), heated to 80°C and refluxed for 3 h, TLC detection, cooled to 0°C, added with H2O oxidation, stirred for 15 min, then added with Et3N (10 mL), stirred for 20 min, TLC detection, extracted with ethyl acetate and column chromatography to obtain compound E (2.6 g, 47.1%); 1H NMR (600 MHz, CDC13) δ 8.27 (s, 1H), 7.89 (dd, J = 14.1, 7.6 Hz, 1H), 7.56 (t, 1H), 7.47 (d, J = 7.6 Hz, 1H), 7.45 (s, 1H), 7.41 (t, 1H), 7.36 - 7.27 (m, 5H), 7.16 (d, J = 8.6 Hz, 2H), 6.82 (d, J = 8.7 Hz, 2H), 3.81 (s, 3H).

[0055] Preparation of compound F:

[0056]

[0057] Compound E (1.019 g, 3.3 mmol) was dissolved in 20 mL THF under argon, Pd(OAC)2 (37 mg, 0.165 mmol) was added as catalyst, stirred at 65 °C for 5 h, TLC detection, rotary evaporation and column chromatography to obtain compound F (980 mg, 96%) as yellow oil; 1 H NMR (600 MHz, CDC13) δ 7.73 - 7.68 (m, 2H), 7.68 - 7.62 (m, 3H), 7.53 (t, 1H), 7.48 (td, 1H), 7.38 (td, J = 9.2 Hz, 2H), 7.29 (td, 1H), 7.20 (dd, J = 10.9 Hz, 1H), 7.08 (dd, J = 8.5 Hz, 1H), 3.79 (s, 3H).

[0058] Preparation of compound G:

[0059]

[0060] Compound F (1.12 g, 3.6 mmol) was dissolved in 20 mL dichloromethane under argon, BBr3 (1.09 g, 4.35 mmol) was added slowly dropwise at 0 °C as catalyst, stirred at 0 °C for 4 h, TLC detection, rotary evaporation and dichloromethane extraction to obtain compound G (746 mg, 71%) as yellow oil; 1 H NMR (600 MHz, CDC13) δ 10.18 (s, 1H), 7.97 - 7.89 (m, 2H), 7.63 (p, J = 26.5 Hz, 2H), 7.58 - 7.49 (m, 3H), 7.47 (td, 2H), 7.34 (td, J = 9.1 Hz, 1H), 7.04 (ddd, J = 24.2 Hz, 2H).

[0061] Preparation of compound H:

[0062]

[0063] Compound G (200 mg, 0.68 mmol) and 2,4-dinitrochlorobenzene (166.4 mg, 0.82 mmol) were dissolved in 7 mL of DMF and added to a round-bottom flask, potassium carbonate (141 mg, 1.03 mmol) was added, stirred at 100°C for 6 h, extracted with ethyl acetate and columned to obtain yellow solid compound H (267 mg, 85.6%), H is 3-(2,4-dinitrophenoxy)-5-phenylbenzo[b]phosphaphenanthrene-5-oxide, phosphole fluorescent probe; 1 H NMR (600 MHz, CDC13) δ 8.78 (d, J = 2.7 Hz, 1H), 8.28 (dd, J = 9.2, 2.7 Hz, 1H), 7.86 (dd, J = 8.4, 3.2 Hz, 1H), 7.78 (dd, J = 7.7, 2.8 Hz, 1H), 7.68 (dd, J = 9.8, 7.6 Hz, 1H), 7.57 (dt, J = 8.2, 4.4 Hz, 4H), 7.47 (td, J = 7.4, 1.2 Hz, 1H), 7.41 - 7.34 (m, 5H), 7.29 (dd, J = 8.4, 2.2 Hz, 1H), 7.06 (d, J = 9.2 Hz, 1H).

[0064] Example 4:

[0065] Compound H (phosphole fluorescent probe) obtained in Example 3 was detected, and it was detected that the water solubility of the probe was good. In a PBS solution (pH = 7.40) containing 10% (mass percentage) DMSO, the probe concentration was 20.0 μΜ in DMF:PBS buffer solution = 1:9 (mass percentage) (2 x 10 -5 , pH = 7.4), the detection and recognition of H2S could be realized at room temperature, the response speed to H2S was fast, the reaction time was 10 min, and the response to H2S was fluorescence enhancement type; the recognition mechanism is as follows;

[0066]

[0067] Example 5

[0068] Characteristics and performance of phosphole H2S fluorescent probe

[0069] 1. Fluorescence emission in aqueous solution

[0070] The fluorescent probe (compound H, 20.0 μM, PBS solution of 10% (w / w) DMSO) prepared in Example 3 was used to detect different concentrations of hydrogen sulfide (0-200 μM sodium hydrosulfide, water as solvent). The excitation wavelength was 298 nm and the emission wavelength was 528 nm. The results are as follows: Figure 6 As shown.

[0071] Depend on Figure 6 It can be seen that the fluorescent probe has a very weak emission peak at 528 nm. After adding H2S, the fluorescence at 528 nm is significantly enhanced, reaching 15 times the original value, indicating that the fluorescent probe can detect H2S in aqueous solution.

[0072] 2. Probe recognition of H2S under the influence of pH

[0073] The fluorescence intensity was measured at different pH values ​​using the fluorescent probe (compound H, 20.0 μM, PBS solution of 10% (w / w) DMSO) prepared in Example 3, with a buffer solution pH range of 2.0-13.0 (pH adjusted with sodium hydroxide and hydrochloric acid, water as solvent). The sodium hydrosulfide concentration was 200 μM. The results are as follows: Figure 7 As shown.

[0074] Depend on Figure 7 It can be seen that when the pH range is 4.0-10.0, the probe is basically unaffected by pH, and the fluorescence at 528 nm is significantly enhanced after the addition of H2S. However, when the pH range is 1.0-4.0, the fluorescence intensity of both the probe and probe + H2S decreases significantly with increasing acidity. This is presumably because the phosphoryl fluorene structure of the probe's fluorophore is disrupted. When the pH range is 10.0-13.0, the fluorescence intensity of both the probe and probe + H2S increases significantly with increasing alkalinity. This is presumably because the structure of 2,4-dinitrophenyl ether is disrupted, the PET process is blocked, and the fluorescence of the fluorophore is restored.

[0075] 3. Selectivity

[0076] The fluorescent probe (compound H, 20.0 μM, PBS solution of 10% (w / w) DMSO) prepared in Example 3 was mixed with 200 μM K. + Cu + Ca 2+ Na + I - F - Cl - CO3 2- HCO 3- S2O8 2- HSO 4- HSO3- , SCN - , S2O3 2- , S2O5 2- , SO3 2- , NO2 - , L-Cys, D-Cys, GSH, Zinc Citrate), respectively, and the fluorescence spectra at 528 nm after 200 μM NaHS was added for 10 min, as shown in Fig. 4. Figure 8

[0077] As can be seen from Figure 8 , the addition of organic and inorganic salts has no significant effect on the fluorescence intensity of the fluorescent probe prepared in Example 3; the addition of L-Cys and D-Cys can enhance the fluorescence intensity of the probe system, but far less than the fluorescence intensity enhancement caused by the addition of NaHS. After the addition of interfering ions in the probe, the coexistence of NaHS will greatly enhance the fluorescence intensity, and the increase in intensity is approximately equal to the fluorescence intensity without the addition of interfering ions. Therefore, the probe has good selectivity for the detection of H2S. The probe can accurately recognize H2S under most adverse conditions, and can exclude the interference of the external environment.

[0078] The above examples only express the specific embodiments of the present application, which are described in detail, but should not be construed as limiting the scope of protection of the present application. It should be noted that for ordinary skilled persons in the art, without departing from the technical concept of the present application, several modifications and improvements can be made, which are all within the scope of protection of the present application.

[0079] This background section is provided to generally present the context of the application, the work of the current named inventors, the work described in this background section to the extent it is described, and nothing in this section is to be construed as an admission that the present application is not entitled to antedate such work by virtue of prior application.​

Claims

1. A phospholucene fluorescent probe compound for rapid and efficient detection of H2S, characterized in that, The compound is 3-(2,4-dinitro-phenoxy)-5-phenyl-benzo[b]phosphinindole-5-oxide, having the structure of formula (1): (1), and the compound is used for detecting H2S.

2. A preparation method of a fast and efficient H2S phospholane fluorescent probe compound, characterized in that, The synthesis route is shown in (2):

3. The application of the fast and efficient H2S phosphole fluorescent probe compound in claim 1 in fast and efficient detection of H2S in soil or water.

4. The preparation method of the phospholene fluorescent probe compound for rapid and efficient detection of H2S according to claim 2, characterized in that The steps include: (1) K2CO3 and Pd(PPh)3Cl2 are added to the mixture of compound A and compound B, and then the foregoing substances are dissolved in a DME-containing aqueous solution; and the dissolved substances are refluxed under argon, detected by TLC, extracted with ethyl acetate and over-columned to obtain transparent oily compound C; (2) Mg and I2 are added under argon, THF is used as a solvent, and the foregoing substances are stirred at a certain temperature until the iodine fades; compound C is dissolved in THF and slowly added dropwise into a three-necked flask, heated and refluxed, cooled to room temperature, and the supernatant is taken into a clean round-bottom flask and slowly added dropwise with PPhCl2 dissolved in THF under argon; the foregoing substances are heated and refluxed, detected by TLC, cooled to 0 DEG C, oxidized with H2O, stirred, added with Et3N, detected by TLC, extracted with ethyl acetate and over-columned to obtain compound E; (3) under argon, compound E is dissolved in THF, Pd(OAC)2 is added as a catalyst, stirred at a certain temperature, detected by TLC, and over-columned to obtain yellow oily compound F; (4) under argon, compound F is dissolved in dichloromethane, BBr3 is added as a catalyst, stirred at a certain temperature, detected by TLC, and the dichloromethane is extracted and dried to obtain yellow oily compound G; (5) compound G and 2,4-dinitrophenyl chloride are dissolved in DMF, added with K2CO3, stirred at a certain temperature, extracted with ethyl acetate and over-columned to obtain yellow solid compound H. 5.The application of the phosphorus-containing fluorene fluorescent probe compound for rapid and efficient detection of H2S in soil or water according to claim 3, characterized in that, The reaction mechanism of the application is described in formula (3); 6. The preparation method of the phospholene fluorescent probe compound for rapid and efficient detection of H2S according to claim 4, characterized in that: In step (1), the molar ratio of DME to H2O in the DME-containing aqueous solution is 8-9:1-2; the molar ratio of compound A, compound B, K2CO3 and Pd(PPh)3Cl2 is 1-1.5:1-1.5:2-3:0.01-0.02; and the refluxing condition is 85 DEG C+ / -5 DEG C for 8+ / -0.5 h.

7. The preparation method of the phospholene fluorescent probe compound for rapid and efficient detection of H2S according to claim 4, characterized in that: In step (2), the stirring temperature is 40 DEG C until the iodine fades; compound C is dissolved in THF and slowly added dropwise into a flask, the heating and refluxing temperature is 80 DEG C for 2 h; after the PPhCl2 dissolved in THF is added dropwise, the heating and refluxing temperature is 80 DEG C for 3 h; the TLC detection is performed after the foregoing substances are cooled to 0 DEG C, oxidized with H2O, and oxidized for 15-30 min; The molar ratio of compound C, Mg, I2, PPhCl2 and Et3N is 1-1.5:1-1.5:0.02-0.04:1.0-1.4:1.5-2.5; 2-3 mL THF is added per millimole of Mg, 0.5-1 times THF is used to dissolve per millimole of compound C, the reflux condition is 80℃, 2h; 0.5-1 times THF is used to dissolve per millimole of PPhCl2, the reflux condition is 80℃, 3h.

8. The preparation method of the phospholene fluorescent probe compound for rapid and efficient detection of H2S according to claim 4, characterized in that: In step (3), the molar ratio of compound E and Pd(OAC)2 is 1:0.03-0.2; 5-10 times THF is used to dissolve per millimole of compound E; the reflux condition is 65±5℃, 5±0.5h.

9. The preparation method of the phospholene fluorescent probe compound for rapid and efficient detection of H2S according to claim 4, characterized in that: In step (4), the molar ratio of compound F and BBr3 is 1:1.2-3.0; 5-10 times volume of dichloromethane is used to dissolve per millimole of compound F, the reaction condition is 0℃, 4h.

10. The preparation method of the phospholene fluorescent probe compound for rapid and efficient detection of H2S according to claim 4, characterized in that: In step (5), the molar ratio of compound G, 2,4-dinitrobenzene chloride and potassium carbonate is 1-1.5:1-1.5:1.0-2.0, 5-15 times volume of DMF is used to dissolve per millimole of compound G, the reaction condition is 100±5℃, 6±0.5h.

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