Chemiluminescent system based on H2O2-luminol and detection method of triphenyl phosphate

Abstract

The present invention discloses a method based on H 2 O 2 The chemiluminescent system of ‑luminol and the detection method of triphenyl phosphate solve the technical problem that there is still no effective detection method for TPHP in the prior art. 2 O 2 The chemiluminescent system of luminol comprises a first component and a second component, wherein the first component comprises a first mixed solution after ultraviolet light irradiation, and the first mixed solution comprises H 2 O 2 and a solution of a substance to be detected; the second component comprises a second mixed solution, the second mixed solution comprises luminol and a buffer solution; wherein the substance to be detected can weaken the chemiluminescent signal. 2 O 2 Oxidation effectively promotes the generation of ·OH, accelerates the reaction of the analyte (such as TPHP) and ·OH, and quickly consumes the active oxygen free radicals in the system, thereby reducing the active oxygen free radicals participating in the luminol chemiluminescence, thereby reducing the 3‑APA* generated by the oxidative decomposition of the luminol anion, and ultimately causing the chemiluminescence system to exhibit a greater luminescence inhibition effect.

Description

Technical Field

[0001] The present invention relates to the technical field of chemiluminescence, and in particular to a method for producing a chemiluminescent light-emitting diode based on H 2 O 2 -The chemiluminescent system of luminol and the detection method of triphenyl phosphate. Background Art

[0002] Organophosphorus flame retardants (OPFRs) have excellent thermal insulation and plasticizing effects. They are mainly used as additives in industrial and household consumer products such as plastics, fabrics, electronic products and building materials, and have been widely used worldwide. Triphenyl phosphate (TPHP) is an aromatic organophosphate containing three benzene rings. It is widely used as an external plasticizer in furniture, printed circuit boards, hydraulic oils, paints, coatings and nail polish. In addition, since the structure of TPHP does not contain covalent bonds, TPHP is easily broken and enters the environment. TPHP residues have been detected in the air, soil, water environment and organisms. Airborne TPHP is adsorbed on soil, making it one of the main OPFRs detected in soil. TPHP is biodegradable and does not accumulate permanently in the environment, but its continued release into the environment may pose a risk to organisms due to long-term human use.

[0003] Luminol, also known as luminol, has a chemical formula of C 8 H 7 N 3 O 2 , can be oxidized by hydrogen peroxide under alkaline conditions to produce excited 3-aminophthalate ions (3-APA*). When 3-APA* releases energy and returns to the ground state, it produces chemiluminescence with a wavelength of 425nm. Therefore, luminol-H 2 O 2 The system is one of the classic chemiluminescence systems. Luminol-H 2 O 2 The current applications of the system mainly include the following two types:

[0004] First, under normal circumstances, the chemiluminescent reaction between luminol and hydrogen peroxide is very rapid in the presence of certain catalysts. The most commonly used catalyst is metal ions. Within a wide concentration range, the metal ion concentration is proportional to the luminescence intensity, so chemiluminescent analysis of certain metal ions can be performed. Therefore, using this reaction, organic compounds containing metal ions can be analyzed with high sensitivity.

[0005] Secondly, the inhibitory effect of organic compounds on the chemiluminescent reaction of luminol is used to determine the organic compounds that have a quenching effect on the chemiluminescent reaction. 2 O 2The chemiluminescence reaction of the system has a quenching effect, so in theory, quantitative detection of TPHP can be achieved.

[0006] However, the applicant of the present application has found in practice that although TPHP has no significant effect on luminol-H 2 O 2 The chemiluminescence reaction of the system has a quenching effect, but when luminol-H 2 O 2 When the system detects TPHP, the detection limit and linear range cannot meet the ideal requirements, resulting in the lack of effective means of detecting TPHP. Summary of the invention

[0007] The main purpose of the present invention is to provide a 2 O 2 -The chemiluminescent system of luminol and the detection method of triphenyl phosphate are used to solve the technical problem that there is still no effective detection method for TPHP in the prior art.

[0008] In order to achieve the above object, the present invention first provides a method based on H 2 O 2 -The chemiluminescent system of luminol, the technical scheme is as follows:

[0009] Based on H 2 O 2 -luminol chemiluminescence system, comprising a first component and a second component, wherein the first component comprises a first mixed solution after ultraviolet light irradiation, and the first mixed solution comprises H 2 O 2 and a solution of an analyte; the second component comprises a second mixed solution, the second mixed solution comprises luminol and a buffer solution; wherein the analyte can weaken a chemiluminescent signal.

[0010] As the above-mentioned H 2 O 2 - Further improvement of the chemiluminescent system of luminol, the ultraviolet lamp is an ultraviolet high-pressure mercury lamp with a power of 300-500W and a main wavelength of 365nm; the ultraviolet light irradiation time is 5-10min; the distance between the ultraviolet light and the liquid surface of the first mixed liquid is 15-20cm.

[0011] As the above-mentioned H 2 O 2 -A further improvement of the chemiluminescent system of luminol, wherein the first mixed solution contains H 2 O 2 The volume ratio of the sample to the test solution is 1:(7-12).

[0012] As the above-mentioned H 2 O 2-A further improvement of the chemiluminescent system of luminol, wherein the first mixed solution contains H 2 O 2 The concentration is 100-400mmol / L.

[0013] As the above-mentioned H 2 O 2 - Further improvement of the chemiluminescent system of luminol, the concentration of luminol in the second mixed solution is 40-50 μmol / L; the buffer solution is Na 2 CO 3 -NaHCO 3 Buffer solution; the pH of the second mixed solution is 11-12.

[0014] As the above-mentioned H 2 O 2 -A further improvement of the chemiluminescent system of luminol, wherein the volume ratio of the first component to the second component is (1-3):1.

[0015] In order to achieve the above object, the present invention secondly provides a detection method for triphenyl phosphate, and the technical scheme is as follows:

[0016] Using the above-mentioned H-based 2 O 2 The method for detecting triphenyl phosphate by a chemiluminescent system of -luminol comprises the following steps: triphenyl phosphate, a solvent and H 2 O 2 Prepare a first mixed solution; irradiate the first mixed solution with an ultraviolet lamp to obtain a first component; prepare a second mixed solution with luminol and a buffer solution to obtain a second component; add the second component to a luminescent container; add the first component to the luminescent container; and then detect the chemiluminescent signal.

[0017] As a further improvement of the above detection method, the first component is added by a peristaltic pump with a rotation speed of 60-80 rpm.

[0018] As a further improvement of the above detection method, the solvent is acetonitrile or acetonitrile aqueous solution.

[0019] As a further improvement of the above detection method, two lifting platforms are symmetrically arranged below the ultraviolet lamp, and the first mixed liquid is arranged between the two lifting platforms.

[0020] It has been verified that the present invention uses ultraviolet light to 2 O 2 The oxidation process effectively promotes the generation of ·OH with a higher redox potential, accelerates the reaction of the analyte (such as TPHP) with ·OH, and quickly consumes H 2 O 2and ·OH and other active oxygen free radicals, reducing the 1 O 2 and O 2 ·- The conversion reduces the active oxygen free radicals involved in the luminol chemiluminescence, thereby reducing the 3-APA* generated by the oxidative decomposition of the luminol anion, and finally makes the chemiluminescence system show a greater luminescence inhibition effect, and the generated chemiluminescence signal is significantly weakened, thereby improving the sensitivity of the test object. It can be seen that the present invention is simple and easy to implement, effectively solves the technical obstacles of TPHP difficult monitoring, and has the value of promotion and use.

[0021] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments. Additional aspects and advantages of the present invention will be partially given in the following description, partially become apparent from the following description, or be learned through the practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The drawings constituting a part of the present invention are used to assist in understanding the present invention. The contents provided in the drawings and their related descriptions in the present invention can be used to explain the present invention, but do not constitute improper limitations on the present invention. In the drawings:

[0023] Figure 1 It is a structural schematic diagram of the ultraviolet lamp irradiation device.

[0024] Figure 2 For UV, luminol, H 2 O 2 The chemiluminescence signal test results of the related system when and TPHP are used as variables.

[0025] Figure 3 H 2 O 2 UV-TPHP-H when concentration is variable 2 O 2 -Chemiluminescent signal test results of the luminol system.

[0026] Figure 4 UV-TPHP-H when luminol concentration is variable 2 O 2 -Chemiluminescent signal test results of the luminol system.

[0027] Figure 5 When pH is a variable, UV-TPHP-H 2 O 2 -Chemiluminescent signal test results of the luminol system.

[0028] Figure 6 When the irradiation time is variable, UV-TPHP-H 2 O2 -Chemiluminescent signal test results of the luminol system.

[0029] Figure 7 UV-TPHP-H when the peristaltic pump speed is variable 2 O 2 -Chemiluminescent signal test results of the luminol system.

[0030] Figure 8 H before and after UV irradiation 2 O 2 -EPR characterization results of ·OH in the luminol system.

[0031] Fig. 9 H before and after UV irradiation 2 O 2 - Luminol system 1 O 2 EPR characterization results.

[0032] Fig.10 H before and after UV irradiation 2 O 2 -O of the luminol system 2 ·- EPR characterization results.

[0033] Fig.11 UV-H 2 O 2 -Luminol system (a) and UV-TPHP-H 2 O 2 -Luminol system (b) Fluorescence spectrum.

[0034] Fig.12 UV-TPHP-H at different TPHP concentrations 2 O 2 -Chemiluminescent signal test results of the luminol system.

[0035] Fig.13 TPHP concentration and UV-TPHP-H 2 O 2 -Linear fitting curve of the chemiluminescence intensity of the luminol system.

[0036] Fig.14 UV-H 2 O 2 -Selectivity test results of the luminol system for the analytes.

[0037] Fig.15 UV-H 2 O 2 -The test results of the anti-interference of the luminol system on the analytes. DETAILED DESCRIPTION

[0038] The present invention is described clearly and completely below in conjunction with the accompanying drawings. A person skilled in the art will be able to implement the present invention based on these descriptions. Before describing the present invention in conjunction with the accompanying drawings, it should be particularly noted that:

[0039] The technical solutions and technical features provided in each part of the present invention, including the following description, may be combined with each other if there is no conflict.

[0040] In addition, the embodiments of the present invention involved in the following description are generally only a part of the embodiments of the present invention, rather than all the embodiments. Therefore, based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0041] About the terms and units in the present invention: The terms "include", "have" and any variations thereof in the description and claims of the present invention and related parts are intended to cover non-exclusive inclusions.

[0042] The present invention is based on H 2 O 2 The specific embodiment of the chemiluminescent system of -luminol comprises a first component and a second component, wherein the first component comprises a first mixed solution irradiated with ultraviolet light (UV), and the first mixed solution comprises H 2 O 2 and a solution of an analyte; the second component comprises a second mixed solution, the second mixed solution comprises luminol and a buffer solution; wherein the analyte can weaken a chemiluminescent signal.

[0043] The ultraviolet lamp is an ultraviolet high-pressure mercury lamp with a power of 300-500W and a main wavelength of 365nm. In specific implementation, the power of the ultraviolet lamp can be but is not limited to any one of 300W, 320W, 350W, 380W, 400W, 430W, 450W, 470W, and 500W.

[0044] The ultraviolet light irradiation time is 5-10 minutes. In specific implementation, the irradiation time can be, but is not limited to, any one of 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, and 10 minutes.

[0045] The distance between the ultraviolet light and the liquid surface of the first mixed liquid is 15-20 cm. In specific implementation, the distance can be, but is not limited to, any one of 15 cm, 16 cm, 17 cm, 18 cm, 19 cm, and 20 cm.

[0046] The first mixed solution contains H2 O 2 The volume ratio of the solution to be tested is 1:(7-12). In specific implementation, the volume ratio can be, but is not limited to, any one of 1:7, 1:8, 1:9, 1:10, 1:11, and 1:12.

[0047] The first mixed solution contains H 2 O 2 The concentration is 100-400mmol / L. In specific implementation, the concentration can be but is not limited to any one of 100mmol / L, 150mmol / L, 200mmol / L, 250mmol / L, 300mmol / L, 350mmol / L, and 450mmol / L.

[0048] The concentration of luminol in the second mixed solution is 40-50 μmol / L. In specific implementation, the concentration can be, but is not limited to, any one of 40 μmol / L, 42 μmol / L, 44 μmol / L, 46 μmol / L, 48 μmol / L, and 50 μmol / L.

[0049] The buffer is Na 2 CO 3 -NaHCO 3 Buffer.

[0050] The pH value of the second mixed liquid is 11-12. In specific implementation, the pH value can be, but is not limited to, any one of 11, 11.2, 11.4, 11.6, 11.8, and 12.

[0051] The volume ratio of the first component to the second component is (1-3):1. In a specific implementation, the volume ratio can be, but is not limited to, any one of 1:1, 1.5:1, 2:1, 2.5:1, and 3:1.

[0052] The specific implementation of the detection method of triphenyl phosphate (TPHP) of the present invention is to use the above-mentioned H-based 2 O 2 -The chemiluminescence system of luminol is used to detect triphenyl phosphate. The chemiluminescence signal is collected by the ultra-weak chemiluminescence instrument of BPCL. The detection process specifically includes the following steps:

[0053] (1) TPHP, solvent and H 2 O 2 Prepare a first mixed solution;

[0054] (2) irradiating the first mixed solution with an ultraviolet lamp to obtain a first component;

[0055] Figure 1FIG. 1 is a schematic diagram of the structure of the ultraviolet lamp irradiation device. Figure 1 As shown, two lifting platforms are symmetrically arranged below the ultraviolet lamp, and a container containing the first mixed liquid is placed between the two lifting platforms. The cross-sectional diameter of the container is 24 mm. The model of the ultraviolet lamp is GGZ (Shanghai Ji Guang Special Lighting Appliance Factory), and the distance H between the ultraviolet light and the liquid surface of the first mixed liquid is 18 cm.

[0056] (3) preparing a second mixed solution with luminol and a buffer solution to obtain a second component;

[0057] (4) adding the second component into the luminescent dish of the ultra-weak chemiluminescence instrument and then closing the light window;

[0058] (4) using a peristaltic pump to add the first component to the luminescent dish containing the second component;

[0059] (5) Then the BPCL software of the ultra-weak chemiluminescence instrument starts to detect the chemiluminescence signal. The chemiluminescence signal acquisition interval is 0.1 s, and the negative high voltage is set to 800 V.

[0060] The peristaltic pump has a rotation speed of 60-80 rpm. In specific implementation, the rotation speed of the peristaltic pump can be, but is not limited to, any one of 60 rpm, 63 rpm, 65 rpm, 67 rpm, 70 rpm, 72 rpm, 75 rpm, 78 rpm, and 80 rpm.

[0061] The solvent is acetonitrile or acetonitrile aqueous solution. One use method is: firstly, acetonitrile is used to dissolve TPHP to obtain a stock solution, and then according to the required concentration, the stock solution is taken at any time and adjusted to the required concentration with acetonitrile aqueous solution.

[0062] The beneficial effects of the present invention are described below through specific experimental data.

[0063] In the following experiment: the power of the UV lamp is 400W, the distance between the UV light and the surface of the first mixed liquid is 18cm; 2 O 2 The volume ratio of the first component to the test solution is 1:9; the volume ratio of the first component to the second component is 2:1 (specifically 100 μL luminol solution, 100 μL buffer solution and 400 μL first mixed solution); the concentration of TPHP in the stock solution is 20000 ng / L, and each dilution uses a 10% acetonitrile aqueous solution.

[0064] Figure 2 For UV, luminol, H 2 O 2 The chemiluminescence signal test results of the relevant system when and TPHP are used as variables. Among them, curve (a) belongs to H 2 O 2-Luminol system, i.e. without UV and TPHP; curve (b) belongs to TPHP-H 2 O 2 -Luminol system, that is, no UV is used; curve (c) belongs to UV-H 2 O 2 -Luminol system, i.e. without TPHP; curve (d) belongs to UV-TPHP-H 2 O 2 -Luminol system; curve (e) belongs to UV-TPHP-H 2 O 2 system, i.e., luminol is not used; curve (f) belongs to the UV-TPHP-luminol system, i.e., H 2 O 2 .

[0065] from Figure 2 It can be seen that by comparing curve (a) and curve (b), it can be found that in the absence of UV, the introduction of TPHP will make H 2 O 2 -The chemiluminescent signal of the luminol system is weakened by about 40%. Comparing curve (c) and curve (d), it can be found that when the UV-H 2 O 2 The chemiluminescent signal of the luminol system was reduced by more than 70% after the addition of TPHP, which indicates that the introduction of UV has an important effect on the H 2 O 2 -Luminol system plays an important role in detecting TPHP. Comparison of curve (d) and curve (e) and curve (d) and curve (f) also shows that luminol and H 2 O 2 The presence of is also important for the detection of TPHP.

[0066] Figure 3 H 2 O 2 UV-TPHP-H when concentration is variable 2 O 2 -Chemiluminescent signal test results of the luminol system. Figure 4 UV-TPHP-H when luminol concentration is variable 2 O 2 -Chemiluminescent signal test results of the luminol system. Figure 5 When pH is a variable, UV-TPHP-H 2 O 2 -Chemiluminescent signal test results of the luminol system. Figure 6 When the irradiation time is variable, UV-TPHP-H 2 O 2 -Chemiluminescent signal test results of the luminol system. Figure 7UV-TPHP-H when the peristaltic pump speed is variable 2 O 2 -Chemiluminescent signal test results of the luminol system.

[0067] like Figure 3-7 As shown, the optimal conditions for detecting TPHP are: 2 O 2 The concentration was 200mmol / L, the luminol concentration was 40μmol / L, the pH value was 11, the peristaltic pump speed was 70rpm and the irradiation time was 5min.

[0068] Figure 8 H before and after UV irradiation 2 O 2 -EPR characterization results of ·OH in the luminol system. Fig. 9 H before and after UV irradiation 2 O 2 - Luminol system 1 O 2 EPR characterization results. Fig.10 H before and after UV irradiation 2 O 2 -O of the luminol system 2 ·- EPR characterization results. DMPO is the scavenger of ·OH; TEMP is 1 O 2 The capture agent is NBT. 2 ·- EPR is electron paramagnetic resonance, and the equipment model is Bruker EMXPLUS (Bruker, Germany).

[0069] from Figure 8-10 It can be seen that without UV (0min) assistance, the characteristic peak of ·OH in the chemiluminescence system is not obvious, but after UV irradiation for 5min, the characteristic peak of ·OH, 1 O 2 and O 2 ·- The EPR characteristic peaks indicate that the UV-assisted method effectively promotes the generation of active oxygen free radicals in the chemical system.

[0070] Fig.11 UV-H 2 O 2 -Luminol system (a) and UV-TPHP-H 2 O 2 -Luminol system (b) Fluorescence spectrum, equipment model is F-7000 (Hitachi Scientific Instruments Co., Ltd.).

[0071] from Fig.11It can be seen that the peak positions of curve (a) and curve (b) are the same, indicating that the introduction of TPHP does not change the luminophore, that is, the chemiluminescence signal of the system is caused by luminol.

[0072] Fig.12 UV-TPHP-H at different TPHP concentrations 2 O 2 -Chemiluminescent signal test results of the luminol system. Fig.13 TPHP concentration and UV-TPHP-H 2 O 2 -Linear fitting curve of the chemiluminescence intensity of the luminol system.

[0073] like Figure 12-13 As shown, UV-TPHP-H 2 O 2 The chemiluminescence intensity of the -luminol system gradually weakened with the increase of TPHP concentration, and there was a good linear relationship in the range of 50-1000 ng / L: I = -81.8178C + 164037.4936 (R 2 =0.988), where I is the chemiluminescent signal intensity and C is the TPHP concentration. The calculated detection limit is 17.2 ng / L.

[0074] Fig.14 UV-H 2 O 2 -Selectivity test results of the luminol system for the analytes. The concentration of the analytes was 1000ng / L.

[0075] like Fig.14 As shown, TPHP added to UV-H 2 O 2 -luminol system has a significant inhibitory effect on the chemiluminescence, while other analytes have a significant inhibitory effect on UV-H 2 O 2 -The chemiluminescent signal of the luminol system was less affected.

[0076] Fig.15 UV-H 2 O 2 -The test results of the anti-interference of the luminol system on the analytes.

[0077] like Fig.15 As shown, when 1000 ng / L TPHP coexists with anti-interference ions, it exhibits good anti-interference ability against 10 times of bicarbonate, 20 times of sulfate, magnesium, sodium and calcium ions, and 40 times of potassium, chloride, nitrate and phosphate.

[0078] For the analysis of environmental samples, water samples were extracted from the lake water of Southwest Jiaotong University and filtered through a 0.45 μm filter membrane to remove impurities in the water. Soil samples were taken from the soil of the lawn on campus and dried, ground, and sieved (using a sieve with a pore size of 2 mm) before use.

[0079] For the spiked test of soil samples, specifically: add the soil sample and TPHP solution of different concentrations into a centrifuge tube, vortex for 10 minutes to fully mix it, and then take the supernatant for detection after solid-liquid separation (centrifugation at 4000r / min for 15 minutes).

[0080] UV-H 2 O 2 The test results of the spiked experiment of the -luminol system are shown in Table 1. As can be seen from Table 1, no TPHP was detected in the original water samples and soil samples without spikes; when spiked with TPHP solutions of 100ng / L, 700ng / L and 900ng / L, the recovery rates of the detection were 91.55-108.71%, and the relative standard deviations (RSDs) were all less than 1.12%. It can be seen that the UV-H 2 O 2 -Luminol system can be used for the analysis of TPHP in environmental samples to a certain extent.

[0081] Table 1

[0082]

[0083] The above is a description of the relevant contents of the present invention. A person skilled in the art will be able to implement the present invention based on these descriptions. Based on the above contents of the present invention, all other embodiments obtained by a person skilled in the art without creative work shall fall within the scope of protection of the present invention.

Claims

1. The detection method of triphenyl phosphate is characterized in that: The chemiluminescent system using H2O2-luminol includes a first component and a second component, wherein: The first component includes a first mixed solution after ultraviolet light irradiation, and the first mixed solution includes H2O2 and triphenyl phosphate solution; The second component includes a second mixed solution, and the second mixed solution includes luminol and a buffer solution; The detection method includes the following steps: triphenyl phosphate, a solvent and H2O2 are prepared into a first mixed solution; The first mixed solution is irradiated with an ultraviolet lamp to obtain a first component; The luminol and the buffer solution are prepared into a second mixed solution, so as to obtain a second component; adding the second component to the light-emitting container; adding a first component to a light-emitting container; The chemiluminescent signal is then detected.

2. The detection method according to claim 1, characterized in that: The ultraviolet lamp is an ultraviolet high-pressure mercury lamp with a power of 300-500W and a main wavelength of 365nm; the ultraviolet light irradiation time is 5-10min; and the distance between the ultraviolet light and the liquid surface of the first mixed liquid is 15-20cm.

3. The detection method according to claim 1, characterized in that: The volume ratio of H2O2 to the test substance solution in the first mixed solution is 1:(7-12).

4. The detection method according to claim 1, characterized in that: The concentration of H2O2 in the first mixed solution is 100-400mmol / L.

5. The detection method according to claim 1, characterized in that: The concentration of luminol in the second mixed solution is 40-50 μmol / L; the buffer solution is Na2CO3-NaHCO3 buffer solution; and the pH of the second mixed solution is 11-12.

6. The detection method according to claim 1, characterized in that: The volume ratio of the first component to the second component is (1-3):

1.

7. The detection method according to claim 1, characterized in that: The first component is added by a peristaltic pump, and the speed of the peristaltic pump is 60-80 rpm.

8. The detection method according to claim 1, characterized in that: The solvent is acetonitrile or acetonitrile aqueous solution.

9. The detection method according to claim 1, characterized in that: Two lifting platforms are symmetrically arranged below the ultraviolet lamp, and the first mixed liquid is arranged between the two lifting platforms.

Images