Preparation method and application of Cu-BNNs chemiluminescent probe for detecting nicotine

Abstract

The invention discloses a preparation method and application of a Cu-BNNs chemiluminescent probe for detecting nicotine. Boron nitride and copper chloride dihydrate are used as raw materials, oil bath heating is carried out in N-methyl pyrrolidone (NMP) to synthesize a Cu-BNNs probe, the Cu-BNNs probe can generate strong chemiluminescence signals with nicotine and sodium hypochlorite, a chemiluminescence detection platform of nicotine is constructed based on the strong chemiluminescence signals, and the chemiluminescence detection platform is used for detecting the nicotine. The method can be used for detecting the content of a nicotine solution and the content of nicotine in cigarette smoke. The method has the advantages of high sensitivity, strong anti-interference capability, low detection cost, simplicity and convenience in operation and the like.

Description

Technical Field

[0001] The present invention belongs to the technical field of chemiluminescent probes, and particularly relates to a preparation method and application of a Cu-BNNs chemiluminescent probe for detecting nicotine. Background Art

[0002] Nicotine (NIC), a major component of cigarette smoke, has numerous adverse effects on human health, causing respiratory and cardiovascular diseases and various types of cancer, including lung, liver, kidney, breast, and cervical cancer. Nicotine also severely affects the central nervous system. Therefore, the determination of nicotine is necessary and important. To date, numerous analytical methods have been reported for measuring nicotine, including spectrophotometry, chromatography, radioimmunoassay, and electrochemiluminescence. Each of these methods has certain advantages. However, none of these methods has been reported to be suitable for the rapid, on-site detection of nicotine in cigarette smoke. Therefore, the development of a novel, rapid, and simple nicotine sensor is of significant importance.

[0003] Chemiluminescence (CL) is the emission of light caused by a chemical reaction. Its advantages include ultra-high sensitivity, ease of use, rapid operation, the absence of complex equipment or radioactive labeling, and strong anti-interference capabilities, making it an ideal choice for high-precision trace analysis. It is currently widely used in pharmacology, genetics, biotechnology, clinical chemistry, food, and environmental chemistry.

[0004] Currently, there are no reports in existing patent literature of chemiluminescent probes synthesized using copper chloride dihydrate and boron nitride as raw materials for monitoring nicotine content in cigarette smoke. Therefore, the present invention synthesized a copper-modified boron nitride chemiluminescent probe (Cu-BNNs probe) using boron nitride and copper chloride dihydrate as raw materials via a solvothermal method. This method offers advantages such as high selectivity, low detection cost, and ease of operation. Summary of the Invention

[0005] The present invention aims to develop a chemiluminescent probe for nicotine detection and to use it for nicotine detection in cigarette smoke. Detection is performed using a peristaltic pump static injection method. NIC reacts with reactive oxygen species released by NaClO solutions, and this probe amplifies the CL intensity of the reaction. This successfully constructed a chemiluminescent probe for NIC detection.

[0006] The technical solution of the present invention is a method for preparing a Cu-BNNs chemiluminescent probe for detecting nicotine. The preparation method of the chemiluminescent probe is to use boron nitride and copper chloride dihydrate as raw materials, grind them in an N-methylpyrrolidone solvent, reflux heat, and then centrifuge, rotary evaporate and dry to obtain a dry solid, which is dissolved in deionized water to obtain the Cu-BNNs probe.

[0007] The aforementioned preparation method is carried out according to the following steps:

[0008] (1) Weigh 0.5-1.5 g of boron nitride and 0.017-0.106 g of copper chloride dihydrate in a mortar, add 1-3 mL of N-methylpyrrolidone, and grind for 50-70 min to obtain product A;

[0009] (2) Add 24-72 mL of N-methylpyrrolidone to product A and heat under reflux at 180-220°C for 10-14 h to obtain product B;

[0010] (3) Take product B and centrifuge it at 4000-6000 rpm for 25-35 min. Take the supernatant and perform rotary evaporation at 95-100 °C for 25-40 min. Finally, dry it at 55-65 °C for 12-15 h to remove NMP. Take 4-6 mg of the dried solid and dissolve it in 8-12 mL of deionized water to obtain the Cu-BNNs probe.

[0011] Specifically, the above preparation method is carried out according to the following steps:

[0012] (1) Weigh 0.5-1.5 g of boron nitride and 0.035-0.088 g of copper chloride dihydrate in a mortar, add 1-3 mL of N-methylpyrrolidone, and grind for 50-70 min to obtain product A;

[0013] (2) Add 24-72 mL of N-methylpyrrolidone to product A and heat under reflux at 180-220°C for 10-14 h to obtain product B;

[0014] (3) Take product B and centrifuge it at 4000-6000 rpm for 25-35 min. Take the supernatant and perform rotary evaporation at 95-100 °C for 25-40 min. Finally, dry it at 55-65 °C for 12-15 h to remove NMP. Take 4-6 mg of the dried solid and dissolve it in 8-12 mL of deionized water to obtain the Cu-BNNs probe.

[0015] More specifically, the above preparation method is carried out according to the following steps:

[0016] (1) Weigh 0.1 g of boron nitride and 0.071 g of copper chloride dihydrate in a mortar, add 2 mL of N-methylpyrrolidone, and grind for 60 min to obtain product A.

[0017] (2) Add 48 mL of N-methylpyrrolidone to product A and heat under reflux at 200°C for 12 h to obtain product B;

[0018] (3) Take product B and centrifuge it at 5000 rpm for 30 min, then perform rotary evaporation at 97 °C for 30 min, and finally dry it at 60 °C for 14 h to remove NMP. Take 5 mg of the dried solid and dissolve it in 10 mL of deionized water to obtain the Cu-BNNs probe.

[0019] The aforementioned Cu-BNNs chemiluminescent probe is used to detect nicotine content. The aforementioned chemiluminescent probe can detect nicotine content and nicotine content in cigarette smoke.

[0020] The aforementioned method for detecting nicotine content comprises the following steps:

[0021] (1) The chemical reaction was carried out using a peristaltic pump static injection method. The luminescence signal generated by the reaction was monitored by a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube. The operating voltage of the photomultiplier tube was set to -1000 V, and the data integration time of the BPCL ultra-weak luminescence analyzer was set to 0.1 s.

[0022] (2) Dissolve 0.5-1.5 mg of nicotine in 5-15 mL of water to obtain a nicotine solution. Mix 100-300 μL of the nicotine solution and 100-300 μL of the Cu-BNNs probe in a dedicated luminescent dish to obtain the sample to be tested.

[0023] (3) Dissolve 30-40 μL of 1.3-1.5 mol / L sodium hypochlorite solution in 10 mL of water to obtain a sodium hypochlorite solution. Take 100-300 μL of the sodium hypochlorite solution and place it in a plastic tube. Start the peristaltic pump and quickly inject it into the C sample of the luminescent dish to collect the NIC chemiluminescence signal.

[0024] Specifically, the aforementioned method for detecting nicotine content includes the following steps:

[0025] (1) The chemical reaction was carried out using a peristaltic pump static injection method. The luminescence signal generated by the reaction was monitored by a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube. The operating voltage of the photomultiplier tube was set to -1000 V, and the data integration time of the BPCL ultra-weak luminescence analyzer was set to 0.1 s.

[0026] (2) Dissolve 1.0 mg of nicotine in 10 mL of water to obtain a nicotine solution. Mix 200 μL of the nicotine solution and 200 μL of the Cu-BNNs probe in a dedicated luminescent dish to obtain the sample to be tested.

[0027] (3) Dissolve 36 μL of 1.39 mol / L sodium hypochlorite solution in 10 mL of water to obtain a sodium hypochlorite solution. Take 200 μL of the sodium hypochlorite solution and place it in a plastic tube. Start the peristaltic pump and quickly inject it into the C sample of the luminescent dish to collect the NIC chemiluminescence signal.

[0028] The specific detection method for detecting the nicotine content in cigarette smoke comprises the following steps:

[0029] (1) Construction of chemiluminescence platform:

[0030] S1: Prepare 100-300 μL of nicotine standard solutions with concentrations of 0.0, 0.1, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 μg / mL, respectively, and then mix them with 100-300 μL of Cu-BNNs probes, respectively, to obtain 9 groups of Cu-BNNs / NIC hybrid materials with different concentrations;

[0031] S2: 9 groups of Cu-BNNs / NIC mixed materials with different concentrations were placed in the luminescent dishes of the reaction system, and then 4×10 -3 -6×10 -3 100-300 μL of mol / L sodium hypochlorite solution was reacted with nine groups of Cu-BNNs / NIC mixed materials with different concentrations. The chemiluminescence signal was monitored using a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube. The operating voltage of the photomultiplier tube was set to -1000 V, and the data integration time of the BPCL ultra-weak luminescence analyzer was set to 0.1 s.

[0032] S3: Finally, a standard curve with a range of 0.0 μg-6.0 μg / mL was drawn, and the detection limit was determined to be 0.016 μg / mL, resulting in the regression equation I = 2050.21 [NIC] + 2453.10;

[0033] (2) Smoke detection:

[0034] SS1: Burn 0.5-1.5g of cigarette smoke in a natural environment and use a pump to pump the smoke into a closed round-bottom flask containing 200-300mL of deionized water for 4-6 minutes to simulate the human smoking process. After the smoke is completely dissolved in deionized water, the cigarette smoke sample is obtained;

[0035] SS2: Mix the cigarette smoke sample with the Cu-BNNs probe, place it in a luminescent dish of the reaction system, and then introduce 4×10 -3 -6×10 -3 100-300 μL of 1.5 mol / L sodium hypochlorite solution was added, and the light signal was collected using a chemiluminescence signal detection instrument. The obtained signal was substituted into the regression equation I=2050.21[NIC]+2453.10 to calculate the nicotine content in the cigarette smoke.

[0036] Specifically, the aforementioned specific detection method for detecting the nicotine content in cigarette smoke includes the following steps:

[0037] (1) Construction of chemiluminescence platform:

[0038] S1: 200 μL of nicotine standard solutions with concentrations of 0.0, 0.1, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 μg / mL were prepared, and then mixed with 200 μL of Cu-BNNs probe to obtain 9 groups of Cu-BNNs / NIC hybrid materials with different concentrations;

[0039] S2: 400 μL of 9 groups of Cu-BNNs / NIC mixed materials with different concentrations were placed in the luminescent dishes of the reaction system, and then 5×10 -3 200 μL of mol / L sodium hypochlorite was reacted with nine groups of Cu-BNNs / NIC mixed materials with different concentrations. The chemiluminescence signal was monitored using a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube. The operating voltage of the photomultiplier tube was set to -1000 V, and the data integration time of the BPCL ultra-weak luminescence analyzer was set to 0.1 s.

[0040] S3: Finally, a standard curve with a range of 0.0 μg-6.0 μg / mL was drawn, and the detection limit was determined to be 0.016 μg / mL, resulting in the regression equation I = 2050.21 [NIC] + 2453.10;

[0041] (2) Smoke detection:

[0042] SS1: 0.9 g of cigarette smoke was burned in a natural environment and the smoke was pumped into a closed round-bottom flask containing 250 mL of deionized water for 5 minutes to simulate the human smoking process. After the smoke was completely dissolved in the deionized water, the cigarette smoke sample was obtained;

[0043] SS2: Mix the cigarette smoke sample with the Cu-BNNs probe, place it in a luminescent dish of the reaction system, and then introduce 5×10 -3 200 μL of 1.5 mol / L sodium hypochlorite solution was added, and the light signal was collected using a chemiluminescence signal detection instrument. The obtained signal was substituted into the regression equation I=2050.21[NIC]+2453.10 to calculate the nicotine content in the cigarette smoke.

[0044] Compared with the prior art, the present invention has the following beneficial effects:

[0045] 1. For the first time, it was proposed to use boron nitride and copper chloride dihydrate as raw materials to react in N-methylpyrrolidone solvent to generate Cu-BNNs probes, which can accurately detect nicotine content and nicotine content in cigarette smoke. The nicotine detection is specific and is not affected by other substances in cigarette smoke, and has strong anti-interference ability.

[0046] 2. The Cu-BNNs probe method for detecting nicotine involves the reaction of nicotine with sodium hypochlorite to produce superoxide anions and hydrogen peroxide. The cuprous ions in the Cu-BNNs probe catalyze the decomposition of hydrogen peroxide to produce hydroxyl radicals and superoxide anions. The hydroxyl radicals and superoxide anions then react to generate singlet oxygen, which then polymerizes into a singlet oxygen dimer. Ultimately, the singlet oxygen dimer transfers energy to the BNNs. When the excited BNNs return to the ground state, the system emits light at 475 nm. Thus, a chemiluminescent detection platform for nicotine in cigarette smoke was established. This method has the advantages of good selectivity, low detection cost, and simple operation.

[0047] 3. Nicotine exhibited good linearity over the concentration range of 0.0 mg / mL to 6.0 μg / mL, with a detection limit of 0.016 μg / mL, demonstrating the high sensitivity of this method for nicotine detection. Furthermore, this method was used to detect and recover nicotine in cigarette smoke, with recoveries ranging from 99.1% to 104.0%, demonstrating its reliability. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 : TEM image of Cu-CNNSs probe;

[0049] Figure 2: Chemiluminescence curves of sodium hypochlorite and Cu-BNNs / nicotine mixture;

[0050] Figure 3: Anti-interference ability spectrum of Cu-BNNs probe to other substances in smoke except NIC (NNN is N-nitrosonornicotine; NNK is 4-(methylnitrosamino)-3-pyridine-1-butanone; Sp is scopolamine; CA is caffeic acid; CGA is chlorogenic acid; Kaempferrol is kaempferol; Solanesol is solanesol; rutin is rutin; quercetin is quercetin);

[0051] Figure 4: NIC and CL intensity spectra at different concentrations;

[0052] Figure 5 : Linear relationship between different concentrations of NIC and CL intensity; DETAILED DESCRIPTION

[0053] The present invention will be further described below with reference to the examples, but they are not intended to limit the present invention.

[0054] Example 1:

[0055] Preparation of Cu-BNNs probe:

[0056] Weigh 100 mg of bulk boron nitride and 71 mg of copper chloride dihydrate and place them in a mortar. Take 2 mL of N-methylpyrrolidone (NMP) and place them in a mortar. After mixing and grinding, transfer them to a round-bottom flask containing 48 mL of N-methylpyrrolidone (NMP) solvent and reflux at 200 ° C for 12 hours. The resulting mixture is centrifuged at 5000 rpm for 30 minutes, then rotary evaporated at 97 ° C for 30 minutes, and finally dried at 60 ° C for 14 hours. Finally, 5 mg of solid is dissolved in 10 mL of deionized water to obtain the Cu-BNNs probe.

[0057] Example 2:

[0058] Preparation of Cu-BNNs probe:

[0059] Weigh 200 mg of bulk boron nitride and 142 mg of copper chloride dihydrate and place them in a mortar. Take 2 mL of N-methylpyrrolidone (NMP) and place them in a mortar. After mixing and grinding, transfer them to a round-bottom flask containing 98 mL of N-methylpyrrolidone (NMP) solvent and reflux at 200 ° C for 12 hours. The resulting mixture is centrifuged at 5000 rpm for 30 minutes, then rotary evaporated at 97 ° C for 30 minutes, and finally dried at 60 ° C for 14 hours. Finally, 5 mg of solid is dissolved in 10 mL of deionized water to obtain the Cu-BNNs probe.

[0060] Example 4:

[0061] Preparation of Cu-BNNs probe:

[0062] Weigh 100 mg of bulk boron nitride and 17 mg of copper chloride dihydrate and place them in a mortar. Take 2 mL of N-methylpyrrolidone (NMP) and place them in a mortar. After mixing and grinding, transfer them to a round-bottom flask containing 48 mL of N-methylpyrrolidone (NMP) solvent and reflux at 200 ° C for 12 hours. The resulting mixture is centrifuged at 5000 rpm for 30 minutes, then rotary evaporated at 97 ° C for 30 minutes, and finally dried at 60 ° C for 14 hours. Finally, 5 mg of solid is dissolved in 10 mL of deionized water to obtain the Cu-BNNs probe.

[0063] Example 5:

[0064] Preparation of Cu-BNNs probe:

[0065] Weigh 50 mg of bulk boron nitride and 106 mg of copper chloride dihydrate and place them in a mortar. Take 3 mL of N-methylpyrrolidone (NMP) and place them in a mortar. After mixing and grinding, transfer them to a round-bottom flask containing 72 mL of N-methylpyrrolidone (NMP) solvent and reflux at 220 ° C for 10 hours. The resulting mixture is centrifuged at 4000 rpm for 35 minutes, then rotary evaporated at 95 ° C for 40 minutes, and finally dried at 65 ° C for 12 hours. Finally, 4 mg of solid is dissolved in 12 mL of deionized water to obtain the Cu-BNNs probe.

[0066] Example 6:

[0067] Preparation of Cu-BNNs probe:

[0068] Weigh 150 mg of bulk boron nitride and 35 mg of copper chloride dihydrate and place them in a mortar. Take 1 mL of N-methylpyrrolidone (NMP) and place it in a mortar. After mixing and grinding, transfer them to a round-bottom flask containing 24 mL of N-methylpyrrolidone (NMP) solvent and reflux at 180 ° C for 14 hours. The resulting mixture is centrifuged at 6000 rpm for 25 minutes, then rotary evaporated at 100 ° C for 25 minutes, and finally dried at 55 ° C for 15 hours. Finally, 6 mg of solid is dissolved in 8 mL of deionized water to obtain the Cu-BNNs probe.

[0069] Example 7:

[0070] Preparation of Cu-BNNs probe:

[0071] Weigh 100 mg of bulk boron nitride and 88 mg of copper chloride dihydrate and place them in a mortar. Take 3 mL of N-methylpyrrolidone (NMP) and place them in a mortar. After mixing and grinding, transfer them to a round-bottom flask containing 60 mL of N-methylpyrrolidone (NMP) solvent and reflux at 190 ° C for 13 hours. The resulting mixture is centrifuged at 4500 rpm for 25 minutes, then rotary evaporated at 95 ° C for 35 minutes, and finally dried at 60 ° C for 13 hours. Finally, 5 mg of solid is dissolved in 10 mL of deionized water to obtain the Cu-BNNs probe.

[0072] Example 8:

[0073] Preparation of Cu-BNNs probe:

[0074] Weigh 100 mg of bulk boron nitride and 52 mg of copper chloride dihydrate and place them in a mortar. Take 2 mL of N-methylpyrrolidone (NMP) and place them in a mortar. After mixing and grinding, transfer them to a round-bottom flask containing 48 mL of N-methylpyrrolidone (NMP) solvent and reflux at 200 ° C for 12 hours. The resulting mixture is centrifuged at 5000 rpm for 30 minutes, then rotary evaporated at 97 ° C for 30 minutes, and finally dried at 60 ° C for 14 hours. Finally, 5 mg of solid is dissolved in 10 mL of deionized water to obtain the Cu-BNNs probe.

[0075] Example 9:

[0076] Nicotine detection method:

[0077] (1) The chemical reaction was carried out using a peristaltic pump static injection method. The luminescence signal generated by the reaction was monitored by a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube. The operating voltage of the photomultiplier tube was set to -1000 V, and the data integration time of the BPCL ultra-weak luminescence analyzer was set to 0.1 s.

[0078] (2) Dissolve 1.0 mg of nicotine (NIC) in 10 mL of water to obtain a concentration of 6.17 × 10 -4 mol / L NIC solution; 200 μL of Cu-BNNs probe (prepared in Example 1) and a solution with a concentration of 6.17×10 -4 mol / L NIC (200 μL) solution was mixed and placed in a special luminescent dish for later use;

[0079] (3) Dissolve 36 μL of 1.39 mol / L sodium hypochlorite solution in 10 mL of water to obtain a concentration of 5×10 -3 mol / L sodium hypochlorite solution, take 5×10 -3 200 μL of 1.39 mol / L sodium hypochlorite solution was placed in a plastic tube. 200 μL of 1.39 mol / L sodium hypochlorite solution was placed in a plastic tube. Then the peristaltic pump was started to quickly inject the sodium hypochlorite solution in the disposable plastic tube into the luminescent dish. At the same time, the chemiluminescence signal detection instrument was turned on to collect the light signal, and 6.17×10 -4 mol / L NIC chemiluminescent signal.

[0080] Example 10:

[0081] Nicotine detection method:

[0082] (1) The chemical reaction was carried out using a peristaltic pump static injection method. The luminescence signal generated by the reaction was monitored by a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube. The operating voltage of the photomultiplier tube was set to -1000 V, and the data integration time of the BPCL ultra-weak luminescence analyzer was set to 0.1 s.

[0083] (2) Dissolve 1.0 mg of nicotine (NIC) in 10 mL of water to obtain a concentration of 6.17 × 10 -4 mol / L NIC solution; 200 μL of Cu-BNNs probe (prepared in Example 1) and a solution with a concentration of 6.17×10 -4 mol / L NIC (200 μL) solution was mixed and placed in a special luminescent dish for later use;

[0084] (3) Dissolve 36 μL of 1.39 mol / L sodium hypochlorite solution in 10 mL of water to obtain a concentration of 5×10 -3 mol / L sodium hypochlorite solution, take 5×10 -3 mol / L

[0085] 200 μL of sodium hypochlorite solution was placed in a plastic tube. 200 μL of 1.39 mol / L sodium hypochlorite solution was placed in a plastic tube. Then the peristaltic pump was started to quickly inject the sodium hypochlorite solution in the disposable plastic tube into the luminescent dish. At the same time, the chemiluminescence signal detection instrument was turned on to collect the light signal, and 6.17×10 -4 mol / L NIC chemiluminescent signal.

[0086] Example 11: Detection of cigarette smoke

[0087] 1. Construction of chemiluminescence platform

[0088] (1) Nicotine standard solutions with concentrations of 0.0, 0.1, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 μg / mL were prepared respectively. 200 μL of each of the nine nicotine standard solutions with different concentrations was mixed with 200 μL of the Cu-BNNs probe (prepared in Example 1) to obtain nine groups of Cu-BNNs / NIC mixed materials (400 μL) with different concentrations.

[0089] (2) Nine groups of Cu-BNNs / NIC mixed materials with different concentrations (400 μL) were placed in the luminescent dishes of the reaction system, and then 5×10 -3 200 μL of mol / L sodium hypochlorite was reacted with 9 groups of Cu-BNNs / NIC mixed materials with different concentrations, and the chemiluminescence signal was monitored by a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube;

[0090] (3) Finally, a standard curve was drawn with a range of 0.0 μg-6.0 μg / mL, and the detection limit was determined to be 0.016 μg / mL, resulting in the regression equation I = 2050.21 [NIC] + 2453.10.

[0091] 2. Testing process

[0092] (1) 0.9 g of cigarette smoke was burned in a natural environment and the smoke was pumped into a closed round-bottom flask containing 250 mL of deionized water for 5 minutes to simulate the human smoking process. After the smoke was completely dissolved, a cigarette smoke sample was obtained;

[0093] (2) The cigarette smoke sample was mixed with 200 μL of the Cu-BNNs probe of Example 1, placed in a luminescent dish of the reaction system, and then introduced into 5×10 -3 200 μL of 1.5 mol / L sodium hypochlorite solution was added, and the chemiluminescence signal was monitored using a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube. Substituting the obtained signal into the regression equation I = 2050.21 [NIC] + 2453.10, the nicotine content in the cigarette smoke was calculated to be 2.43 μg / mL.

[0094] The technical solution of the present invention is further described in detail below in conjunction with specific embodiments to better understand how the present invention achieves its objectives and solves related technical problems:

[0095] 1. Reagents

[0096] Sodium hypochlorite and boron nitride were purchased from Shanghai Titan Technology Co., Ltd., China; nicotine (NIC) was purchased from Chengdu Desite Biotechnology Co., Ltd., China.

[0097] 2 Instruments

[0098] The chemiluminescence signals were detected using an ultra-weak chemiluminescence instrument (BPCL-2-TGG, Guangzhou Weiguang Technology Co., Ltd., China); the TEM images of Cu-BNNs were taken by Tecnai G2 F20 S-Twin (Thermo Fisher, USA) with the accelerating voltage set to 200 kV.

[0099] 3Cu-BNNs probe preparation method

[0100] Weigh 100 mg of bulk boron nitride and 71 mg of copper chloride dihydrate and place them in a mortar. Take 2 mL of N-methylpyrrolidone (NMP) and place them in a mortar. After mixing and grinding, transfer them to a round-bottom flask containing 48 mL of N-methylpyrrolidone (NMP) solvent and reflux at 200 ° C for 12 hours. The resulting mixture is centrifuged at 5000 rpm for 30 minutes, then rotary evaporated at 97 ° C, and finally dried at 60 ° C. Finally, 5 mg of solid is dissolved in 10 mL of deionized water to obtain the Cu-BNNs probe.

[0101] TEM image characterization of 4 pairs of Cu-BNN probes

[0102] The TEM image experiment of Cu-BNNs probe is shown in the attached figure. Figure 1The nanosheet structure is shown in Figure 2, which indicates that the copper-modified boron nitride nanosheet material (Cu-BNNs probe) was successfully synthesized.

[0103] 5Cu-BNNs probe detection method and results of NIC

[0104] 5.1 Luminescent probes of Cu-BNNs modified with different copper contents

[0105] The present invention uses a traditional chemiluminescence device and a peristaltic pump static injection method for detection, including an injection system, a reaction system and a detection system. The main function of the injection system is to inject 200 μL of 5×10 -3 A 1.5 mol / L NaClO solution was introduced into the reaction system. A mixed solution of 200 μL of Cu-BNNs luminescent probes and 200 μL of NIC was placed in a luminescent dish in the reaction system and reacted with the solution introduced by a peristaltic pump, forming the reaction system. The luminescence signal generated by the reaction was monitored by a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube (PMT). The PMT operating voltage was set to -1000 V, and the data integration time of the BPCL ultra-weak luminescence analyzer was set to 0.1 s.

[0106] Preparation of Cu-BNNs luminescent probe:

[0107] 17 mg, 35 mg, 52 mg, 71 mg and 88 mg of copper chloride dihydrate were weighed respectively, and placed in a mortar with 100 mg of bulk boron nitride. 2 mL of N-methylpyrrolidone (NMP) was placed in a mortar. After mixing and grinding, the mixture was transferred to a round-bottom flask containing 48 mL of N-methylpyrrolidone (NMP) solvent and refluxed at 200 ° C for 12 h. The resulting mixture was centrifuged at 5000 rpm for 30 min, then rotary evaporated at 97 ° C for 30 min, and finally dried at 60 ° C for 14 h. Finally, 5 mg of the solid was dissolved in 10 mL of deionized water to obtain 5 groups of Cu-BNNs probes modified with different copper chloride dihydrate contents.

[0108] The CL response signals of Cu-BNNs luminescent probes modified with different contents of copper chloride dihydrate to NIC are shown in Appendix 1.

[0109] Table 1: CL intensity and cupric chloride dihydrate content parameters of Cu-BNNs luminescent probe

[0110] Sample name Copper chloride dihydrate content Chemiluminescence intensity Cu-BNNs-1 17.0mg 712 Cu-BNNs-2 35.0mg 6129 Cu-BNNs-3 52.0mg 3337 Cu-BNNs-4 71.0mg 14157 Cu-BNNs-5 88.0mg 6094

[0111] 5.2 Detection of NIC in the Cu-BNNs-NIC-NaClO System

[0112] First, 200 μL of Cu-BNNs probe (prepared in Example 1) and 1 μg / ml nicotine (200 μL) were mixed and placed in a special luminescent dish.-3 200 μL of mol / L sodium hypochlorite solution was placed in a plastic tube, and then the peristaltic pump was started to quickly inject the sodium hypochlorite solution in the disposable plastic tube into the luminescent dish. At the same time, the chemiluminescence signal detector was turned on to collect the light signal. The chemiluminescence curve of the mixture of sodium hypochlorite and Cu-BNNs / nicotine (such as Figure 2 (shown), sodium hypochlorite reacts with the Cu-BNNs / nicotine mixture to produce a strong chemiluminescent signal. In addition, the chemiluminescent signal response of substances other than nicotine in the smoke mixed with nicotine was measured. Other substances showed no significant effect on the chemiluminescent signal of Cu-BNNs-NIC-NaClO, proving that the Cu-BNNs chemiluminescent probe has good anti-interference ability for nicotine (Solanesol, rutin, and quercetin are not easily soluble in water, eliminating their interference). ( Figure 3 Show)

[0113] 5.3 Detection of Nicotine in Cigarette Smoke

[0114] Finally, the applicability of the chemiluminescence method was verified by measuring nicotine in cigarette smoke. Cigarettes were purchased from supermarkets and the smoke was dissolved in pure water solvent by burning. The chemiluminescence sensor constructed as follows ( Figure 4 Figure 5 ) Nicotine monitoring and recovery experiments were performed on the actual sample solution obtained.

[0115] 5.3.1 Construction of chemiluminescence platform

[0116] (1) Nicotine standard solutions with concentrations of 0.0, 0.1, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 μg / mL were prepared, and then mixed with 200 μL of Cu-BNNs probe (prepared in Example 1) to obtain 9 Cu-BNNs / NIC hybrid materials with different concentrations.

[0117] (2) The 9 prepared Cu-BNNs / NIC mixed materials with different concentrations were placed in the luminescent dish of the reaction system, and then 5×10 -3 200 μL of mol / L sodium hypochlorite was reacted with it, and the chemiluminescence signal was monitored by a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube;

[0118] (3) Finally, a standard curve was drawn with a range of 0.0-6.0 μg / mL, the detection limit was determined to be 0.016 μg / mL, and the regression equation was obtained.

[0119] 5.3.2 Specific testing process

[0120] (1) First, the cigarette smoke sample (prepared in Example 11) and the Cu-BNNs chemiluminescent probe (prepared in Example 1) were mixed and placed in a luminescent dish of the reaction system, and then 5×10 -3 200 μL of 1.5 mol / L sodium hypochlorite was added, and the chemiluminescent signal generated by the reaction was recorded using an ultra-weak luminescence analyzer. The obtained chemiluminescent signal intensity was substituted into the regression equation I = 2050.21 [NIC] + 2453.10, and the nicotine concentration detected in the cigarette smoke was calculated to be 2.43 μg / mL.

[0121] (2) Nicotine solutions of known concentrations (0.2, 0.3, and 1.5 μg / mL) were added to cigarette smoke samples using the standard addition method. The same experimental steps as in (1) were used to detect Cu-BNNs and samples with different spiked concentrations, and then 5×10 -3 The chemiluminescence intensity generated by the reaction of 10 mol / L sodium hypochlorite was calculated according to the standard curve to obtain the recovery rate and relative standard deviation (RSD) of the chemiluminescence method in actual samples, as shown in Table 2. The recovery rate of nicotine was between 99.1% and 104.0%.

[0122] Table 2: Recovery and determination results of nicotine in cigarette smoke samples

[0123]

[0124] 6 Conclusion

[0125] This invention develops a novel chemiluminescence technology to detect nicotine content in cigarette smoke. A Cu-BNNs chemiluminescent probe with catalytic and luminescent capabilities was designed. This probe catalyzes the reaction of nicotine with sodium hypochlorite to produce hydrogen peroxide, which then transfers energy to the BNNs, effectively amplifying the chemiluminescent signal. Consequently, a highly selective chemiluminescent platform was constructed for detecting nicotine in cigarette smoke, with recoveries ranging from 99.1% to 104.0%, demonstrating the reliability of this method. This invention provides a new method for nicotine detection using chemiluminescence.

Claims

1. A method for preparing a Cu-BNNs chemiluminescent probe for detecting nicotine, characterized by: The preparation method of the chemiluminescent probe is to use boron nitride and copper chloride dihydrate as raw materials, grind them in N-methylpyrrolidone solvent, reflux heat, and then centrifuge, rotary evaporate and dry to obtain a dry solid that is dissolved in deionized water to obtain the Cu-BNNs probe.

2. The method for preparing the Cu-BNNs chemiluminescent probe for detecting nicotine according to claim 1, wherein: The preparation method is carried out according to the following steps: (1) Weigh 0.5-1.5 g of boron nitride and 0.017-0.106 g of copper chloride dihydrate in a mortar, add 1-3 mL of N-methylpyrrolidone, and grind for 50-70 min to obtain product A; (2) Add 24-72 mL of N-methylpyrrolidone to product A and heat under reflux at 180-220°C for 10-14 h to obtain product B; (3) Take product B and centrifuge it at 4000-6000 rpm for 25-35 min. Take the supernatant and perform rotary evaporation at 95-100 °C for 25-40 min. Finally, dry it at 55-65 °C for 12-15 h to remove NMP. Take 4-6 mg of the dried solid and dissolve it in 8-12 mL of deionized water to obtain the Cu-BNNs probe.

3. The method for preparing the Cu-BNNs chemiluminescent probe for detecting nicotine according to claim 2, wherein: The preparation method is carried out according to the following steps: (4) Weigh 0.5-1.5 g of boron nitride and 0.035-0.088 g of copper chloride dihydrate in a mortar, add 1-3 mL of N-methylpyrrolidone, and grind for 50-70 min to obtain product A; (5) Add 24-72 mL of N-methylpyrrolidone to product A and heat under reflux at 180-220°C for 10-14 h to obtain product B; (3) Take product B and centrifuge it at 4000-6000 rpm for 25-35 min. Take the supernatant and perform rotary evaporation at 95-100 °C for 25-40 min. Finally, dry it at 55-65 °C for 12-15 h to remove NMP. Take 4-6 mg of the dried solid and dissolve it in 8-12 mL of deionized water to obtain the Cu-BNNs probe.

4. The method for preparing a Cu-BNNs chemiluminescent probe for detecting nicotine according to claim 3, wherein: The preparation method is carried out according to the following steps: (1) Weigh 0.1 g of boron nitride and 0.071 g of copper chloride dihydrate in a mortar, add 2 mL of N-methylpyrrolidone, and grind for 60 min to obtain product A. (2) Add 48 mL of N-methylpyrrolidone to product A and heat under reflux at 200°C for 12 h to obtain product B; (3) Take product B and centrifuge it at 5000 rpm for 30 min, then perform rotary evaporation at 97 °C for 30 min, and finally dry it at 60 °C for 14 h to remove NMP. Take 5 mg of the dried solid and dissolve it in 10 mL of deionized water to obtain the Cu-BNNs probe.

5. Use of the Cu-BNNs chemiluminescent probe according to any one of claims 1 to 4 for detecting nicotine content, characterized in that: The chemiluminescent probe can detect the nicotine content and the nicotine content in cigarette smoke.

6. The use according to claim 5, characterized in that: The method for detecting nicotine content comprises the following steps: (1) The chemical reaction was carried out using a peristaltic pump static injection method. The luminescence signal generated by the reaction was monitored by a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube. The operating voltage of the photomultiplier tube was set to -1000 V, and the data integration time of the BPCL ultra-weak luminescence analyzer was set to 0.1 s. (2) Dissolve 0.5-1.5 mg of nicotine in 5-15 mL of water to obtain a nicotine solution. Mix 100-300 μL of the nicotine solution and 100-300 μL of the Cu-BNNs probe in a dedicated luminescent dish to obtain the sample to be tested. (3) Dissolve 30-40 μL of 1.3-1.5 mol / L sodium hypochlorite solution in 10 mL of water to obtain a sodium hypochlorite solution. Take 100-300 μL of the sodium hypochlorite solution and place it in a plastic tube. Start the peristaltic pump and quickly inject it into the C sample of the luminescent dish to collect the NIC chemiluminescence signal.

7. The use according to claim 6, characterized in that: The method for detecting nicotine content comprises the following steps: (1) The chemical reaction was carried out using a peristaltic pump static injection method. The luminescence signal generated by the reaction was monitored by a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube. The operating voltage of the photomultiplier tube was set to -1000 V, and the data integration time of the BPCL ultra-weak luminescence analyzer was set to 0.1 s. (2) Dissolve 1.0 mg of nicotine in 10 mL of water to obtain a nicotine solution. Mix 200 μL of the nicotine solution and 200 μL of the Cu-BNNs probe in a dedicated luminescent dish to obtain the sample to be tested. (3) Dissolve 36 μL of 1.39 mol / L sodium hypochlorite solution in 10 mL of water to obtain a sodium hypochlorite solution. Take 200 μL of the sodium hypochlorite solution and place it in a plastic tube. Start the peristaltic pump and quickly inject it into the C sample of the luminescent dish to collect the NIC chemiluminescence signal.

8. The use according to claim 5, characterized in that: The specific detection method for detecting the content of nicotine in cigarette smoke comprises the following steps: (1) Construction of chemiluminescence platform: S1: Prepare 100-300 μL of nicotine standard solutions with concentrations of 0.0, 0.1, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 μg / mL, respectively, and then mix them with 100-300 μL of Cu-BNNs probes, respectively, to obtain 9 groups of Cu-BNNs / NIC hybrid materials with different concentrations; S2: 9 groups of Cu-BNNs / NIC mixed materials with different concentrations were placed in the luminescent dishes of the reaction system, and then 4×10 -3 -6×10 -3 100-300 μL of mol / L sodium hypochlorite solution was reacted with nine groups of Cu-BNNs / NIC mixed materials with different concentrations. The chemiluminescence signal was monitored using a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube. The operating voltage of the photomultiplier tube was set to -1000 V, and the data integration time of the BPCL ultra-weak luminescence analyzer was set to 0.1 s. S3: Finally, a standard curve with a range of 0.0 μg-6.0 μg / mL was drawn, and the detection limit was determined to be 0.016 μg / mL, resulting in the regression equation I = 2050.21 [NIC] + 2453.10; (2) Smoke detection: SS1: Burn 0.5-1.5g of cigarette smoke in a natural environment and use a pump to pump the smoke into a closed round-bottom flask containing 200-300mL of deionized water for 4-6 minutes to simulate the human smoking process. After the smoke is completely dissolved in deionized water, the cigarette smoke sample is obtained; SS2: Mix the cigarette smoke sample with the Cu-BNNs probe, place it in a luminescent dish of the reaction system, and then introduce 4×10 -3 -6×10 -3 100-300 μL of 1.5 mol / L sodium hypochlorite solution was added, and the light signal was collected using a chemiluminescence signal detection instrument. The obtained signal was substituted into the regression equation I=2050.21[NIC]+2453.10 to calculate the nicotine content in the cigarette smoke.

9. The use according to claim 8, characterized in that: The specific detection method for detecting the content of nicotine in cigarette smoke comprises the following steps: (1) Construction of chemiluminescence platform: S1: 200 μL of nicotine standard solutions with concentrations of 0.0, 0.1, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 μg / mL were prepared, and then mixed with 200 μL of Cu-BNNs probe to obtain 9 groups of Cu-BNNs / NIC hybrid materials with different concentrations; S2: 400 μL of 9 groups of Cu-BNNs / NIC mixed materials with different concentrations were placed in the luminescent dishes of the reaction system, and then 5×10 -3 200 μL of mol / L sodium hypochlorite was reacted with nine groups of Cu-BNNs / NIC mixed materials with different concentrations. The chemiluminescence signal was monitored using a BPCL ultra-weak luminescence analyzer equipped with a photomultiplier tube. The operating voltage of the photomultiplier tube was set to -1000 V, and the data integration time of the BPCL ultra-weak luminescence analyzer was set to 0.1 s. S3: Finally, a standard curve with a range of 0.0 μg-6.0 μg / mL was drawn, and the detection limit was determined to be 0.016 μg / mL, resulting in the regression equation I = 2050.21 [NIC] + 2453.10; (2) Smoke detection: SS1: 0.9 g of cigarette smoke was burned in a natural environment and the smoke was pumped into a closed round-bottom flask containing 250 mL of deionized water for 5 minutes to simulate the human smoking process. After the smoke was completely dissolved in the deionized water, the cigarette smoke sample was obtained; SS2: Mix the cigarette smoke sample with the Cu-BNNs probe, place it in a luminescent dish of the reaction system, and then introduce 5×10 -3 200 μL of 1.5 mol / L sodium hypochlorite solution was added, and the light signal was collected using a chemiluminescence signal detection instrument. The obtained signal was substituted into the regression equation I=2050.21[NIC]+2453.10 to calculate the nicotine content in the cigarette smoke.

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