High-selectivity nitrite colorimetric fluorescent probe with good water solubility as well as preparation method and application of high-selectivity nitrite colorimetric fluorescent probe

Abstract

The invention relates to a colorimetric fluorescent probe with good water solubility and high selectivity for recognizing nitrite as well as a preparation method and application of the colorimetric fluorescent probe. Specifically, the colorimetric fluorescent probe provided by the invention can be used for measuring, detecting or screening nitrite, can realize high-selectivity recognition of nitrite, and has relatively strong anti-interference capability; high-sensitivity and naked-eye analysis on nitrite can be realized; the water solubility is good, and testing can be carried out in a pure water system; meanwhile, the method has the advantages of simple synthesis, stable property and facilitation of commercial application.

Description

Technical Field

[0001] The present invention belongs to the field of fluorescent probes, and particularly relates to a highly selective colorimetric fluorescent probe with good water solubility for nitrite and its application in measuring or detecting nitrite; the present invention also provides a method for preparing the colorimetric fluorescent probe. Background Art

[0002] Nitrite (NO2) is an important environmental pollutant widely present in water and soil environments, mainly derived from agricultural fertilization, sewage treatment, industrial emissions, etc. As a common water pollutant, although nitrite has certain uses as a fertilizer in agriculture, excessive concentration in water will have a serious impact on aquatic organisms and the ecological environment. In addition, nitrite also poses a threat to human health. Excessive intake may lead to nitrite poisoning, affecting the oxygen-carrying capacity of the blood, thus causing various health problems.

[0003] Currently, the methods for detecting nitrite mainly include spectrophotometry, ion chromatography, fluorescence method, electrochemical detection method, etc. Among them, the colorimetric fluorescent probe method has become a research hotspot for detecting nitrite due to its high sensitivity, rapid response, and good selectivity. However, the existing nitrite colorimetric fluorescent probes still have some deficiencies, such as poor water solubility, poor detection selectivity, complex synthesis, and long response time. Therefore, developing a colorimetric fluorescent probe for nitrite with good water solubility, high sensitivity, high selectivity, and simple synthesis has become an important topic in the current research field. Summary of the Invention

[0004] In view of the above background, the present invention aims to provide a colorimetric fluorescent probe with good water solubility for highly selective recognition of nitrite, its preparation method and uses. The probe has the characteristics of good water solubility, high sensitivity, strong selectivity, strong anti-interference ability, and can be analyzed with the naked eye, and is particularly suitable for the effective measurement and detection of nitrite in water environments. This new type of probe can play an important role in water body monitoring and provide reliable technical support for water quality safety.

[0005] Specifically, the present invention provides a colorimetric fluorescent probe for measuring, detecting or screening nitrite, having the structure shown in formula (I):

[0006]

[0007] In formula (I), R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 , R 11 and R 12 are hydrogen atoms, straight-chain or branched-chain alkyl groups, straight-chain or branched-chain alkoxy groups, sulfonic acid groups, ester groups, carboxyl groups; R1, R2, R3, R4, R5, R6, R7, R8, R9, R10 ,R 11 and R 12 may be the same or different.

[0008] In some specific embodiments of the present invention, the compound of the present invention is a compound of formula (II) in which R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 ,R 11 and R 12 are all hydrogen atoms, and its structural formula is as follows:

[0009]

[0010] The present invention also provides a method for preparing the colorimetric fluorescent probe of formula (I), which includes the following steps: reacting the compound of formula (III) with the compound of formula (IV) to prepare the compound of formula (I), and its reaction formula is as follows:

[0011]

[0012] Wherein, R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 ,R 11 ,R 12 and R 13 are hydrogen atoms, straight-chain or branched-chain alkyl groups, straight-chain or branched-chain alkoxy groups, sulfonic acid groups, ester groups, carboxyl groups; R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 ,R 11 ,R 12 and R 13 may be the same or different.

[0013] In some specific embodiments of the present invention, the method for preparing the colorimetric fluorescent probe of formula (I) includes the following steps:

[0014] Put a certain molar ratio of the compound of formula (III) and the compound of formula (IV) into a reaction vessel, then add methanesulfonic acid, react under heating conditions, and after the reaction is completed, rotary evaporate the organic solvent to obtain a crude product, and perform column chromatography separation on the crude product to obtain a pure compound of formula (I).

[0015] In some specific embodiments of the present invention, the molar ratio of the compound of formula (III) to the compound of formula (IV) is 1:1 - 1:3.

[0016] In some specific embodiments of the present invention, the reaction time is 24 - 28 hours.

[0017] In some specific embodiments of the present invention, the reaction temperature is high temperature (100 °C).

[0018] In some specific embodiments of the present invention, the reaction environmental condition is nitrogen.

[0019] The present invention also provides a colorimetric fluorescent probe composition for measuring, screening or detecting nitrite, which comprises a colorimetric fluorescent probe of formula (I) or formula (II).

[0020] In some specific embodiments of the present invention, the colorimetric fluorescent probe composition further comprises a solvent, an acid, a base, a buffer solution or a combination thereof.

[0021] The present invention also provides the use of the colorimetric fluorescent probe of formula (I) or formula (II) in the preparation of a reagent for measuring, screening or detecting nitrite.

[0022] The present invention also provides a kit for detecting the presence of nitrite in a sample or determining the content of nitrite in a sample, which comprises the colorimetric fluorescent probe of formula (I) or formula (II).

[0023] The present invention also provides a method for detecting the presence of nitrite in a sample or determining the content of nitrite in a sample, which comprises:

[0024] a) contacting the colorimetric fluorescent probe of formula (I) or formula (II) with the sample to form a fluorescence quenching compound;

[0025] b) measuring the fluorescence property of the fluorescence quenching compound.

[0026] In some specific embodiments of the present invention, the sample is a chemical sample.

[0027] In some specific embodiments of the present invention, the sample is a water environmental sample.

[0028] In some specific embodiments of the present invention, the water environmental sample is a river, lake or groundwater water environmental sample.

[0029] The present invention has the following remarkable advantages and effects compared with the prior art:

[0030] (1) Good water solubility

[0031] The nitrite colorimetric fluorescent probe of the present invention has good water solubility and can detect or measure nitrite in a pure water system, which is very suitable for detecting or measuring nitrite in water environmental samples, especially river, lake or groundwater water samples.

[0032] (2) High sensitivity

[0033] The nitrite colorimetric fluorescent probe of the present invention reacts very sensitively with nitrite, which is beneficial to the detection of nitrite, and is especially suitable for detecting, screening or measuring nitrite in water environments with a concentration of 0-10 μM.

[0034] (3) High selectivity and strong anti-interference ability

[0035] The nitrite colorimetric fluorescence probe of the present invention can selectively react specifically with nitrite to generate a product with fluorescence change. Compared with other common substances in the water environment, including but not limited to silver ions, copper ions, ferrous ions, ferric ions, iodide ions, magnesium ions, manganese ions, sodium ions, zinc ions, chromium ions, lead ions, potassium ions, calcium ions, sulfate ions, carbonate ions, chloride ions, the colorimetric fluorescence probe of the present invention shows high selectivity and strong anti-interference ability.

[0036] (4) Good stability

[0037] The nitrite colorimetric fluorescence probe of the present invention has good stability, and thus can be stored and used for a long time.

[0038] (5) Simple synthesis

[0039] The nitrite colorimetric fluorescence probe of the present invention has simple synthesis, which is beneficial to commercial promotion and application.

[0040] (6) Can be analyzed with the naked eye

[0041] After the colorimetric fluorescence probe of the present invention reacts with nitrite, the solution produces an obvious color change, and nitrite can be identified with the naked eye. Description of the drawings

[0042] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0043] Figure 1 shows the change of the fluorescence spectrum before and after adding nitrite (20 μM) to the probe (5 μM);

[0044] Figure 2 shows the change of the absorption spectrum before and after adding nitrite (20 μM) to the probe (5 μM); The inset shows the change of the solution color before and after adding nitrite (20 μM) to the probe (5 μM);

[0045] Figure 3 is the time kinetic spectrum of the probe (5 μM) for nitrite (20 μM) at 625 nm;

[0046] Figure 4 is the fluorescence spectrum after adding nitrite (0 - 25 μM) to the probe (5 μM);

[0047] Figure 5 It is the working curve of the probe (5 μM) for quantitative analysis of nitrite at different concentrations (0 - 10 μM).

[0048] Figure 6 (B) is the test of the ability of the probe (5 μM) to selectively recognize nitrite. Among them, numbers 1 - 18 are respectively: 1. Blank; 2. Silver ion Ag + ; 3. Calcium ion Ca 2+ ; 4. Chloride ion Cl - ; 5. Carbonate ion CO3 2- ; 6. Chromium ion Cr 3+ ; 7. Copper ion Cu 2+ ; 8. Ferrous ion Fe 2+ ; 9. Ferric ion Fe 3+ ; 10. Iodide ion I - ; 11. Potassium ion K + ; 12. Magnesium ion Mg 2+ ; 13. Manganese ion Mn 2+ 14. Sodium ion Na +- ; 15. Lead ion Pb 2+ ; 16. Sulfate ion SO4 2- ; 17. Zinc ion Zn 2+ ; 18. Nitrite NO2 (the analyte concentration is 20 μM for all). The bar graph represents the fluorescence intensity values of the probe at 625 nm in the presence of different analytes;

[0049] Figure 6 (A) is the anti - interference ability test of the probe (5 μM) to recognize nitrite (20 μM) in the presence of substances common in the aqueous environment. 1 - 18 are respectively: 1. Blank; 2. Silver ion Ag+; 3. Calcium ion Ca 2+ ; 4. Chloride ion Cl - ; 5. Carbonate ion CO3 2- ; 6. Chromium ion Cr 3+ ; 7. Copper ion Cu 2+ ; 8. Ferrous ion Fe 2+ ; 9. Ferric ion Fe 3+ ; 10. Iodide ion I - ; 11. Potassium ion K + ; 12. Magnesium ion Mg 2+ ; 13. Manganese ion Mn 2+ 14. Sodium ion Na +- ; 15. Lead ion Pb 2+ ; 16. Sulfate ion SO4 2- ; 17. Zinc ion Zn 2+; 18. Nitrite NO2 (analyte concentration is 20 μM). The bar graph represents the fluorescence intensity values of the probe at 625 nm in the co - existence of different analytes and nitrite. Detailed implementation mode

[0050] The following will combine the accompanying drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention and should not be used to limit the protection scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the protection scope of the present invention.

[0051] Example 1: Synthesis of the compound of formula (II)

[0052] The synthetic design route is as follows:

[0053]

[0054] Embodiment 1: Add 407 mg (1.3 mmol) of 4 - diethylaminoketone acid and 206 mg (1.3 mmol) of 6 - amino - 1 - naphthol to the mixed reactants, and add 7 ml of methanesulfonic acid (MeSO3H).

[0055] Pass nitrogen to expel all the internal gases, reflux at 100 °C for 24 h, rotary evaporate to dry the organic solvent to obtain the crude product, and finally separate the solid by liquid chromatography column with a mixed system of dichloromethane and anhydrous methanol to obtain the pure product. 180 mg of purple pure compound of formula (II) is obtained, and the conversion rate is 32%.

[0056] Embodiment 2: Add 407 mg (1.3 mmol) of 4 - diethylaminoketone acid and 412 mg (2.6 mmol) of 6 - amino - 1 - phenol to the mixed reactants, add 7 ml of methanesulfonic acid (MeSO3H), pass nitrogen to expel all the internal gases, reflux at 100 °C for 24 h, rotary evaporate to dry the organic solvent to obtain the crude product, and finally separate the solid by liquid chromatography column with a mixed system of dichloromethane and anhydrous methanol to obtain the pure product. 240 mg of purple pure compound of formula (II) is obtained, and the yield is 42%.

[0057] Embodiment 3: Add 407 mg (1.3 mmol) of 4 - diethylaminoketone acid and 618 mg (3.9 mmol) of 6 - amino - 1 - phenol to the mixed reactants, add 7 ml of methanesulfonic acid (MeSO3H), pass nitrogen to expel all the internal gases, reflux at 100 °C for 24 h, rotary evaporate to dry the organic solvent to obtain the crude product, and finally separate the solid by liquid chromatography column with a mixed system of dichloromethane and anhydrous methanol to obtain the pure product. 225 mg of purple pure compound of formula (II) is obtained, and the yield is 40%.

[0058] Example 4: 407 mg (1.3 mmol) of 4 - diethylaminoketone acid and 412 mg (2.6 mmol) of 6 - amino - 1 - phenol were taken. 7 ml of methanesulfonic acid (MeSO3H) was added to the mixed reactants. Nitrogen was introduced to expel all the internal gases. The mixture was refluxed at 100 °C for 28 h. The organic solvent was evaporated to dryness by rotary evaporation to obtain the crude product. Finally, the solid was separated by liquid chromatography column using a mixed system of dichloromethane and anhydrous methanol to obtain the pure product. 250 mg of the purple pure compound (II) was obtained, and the yield was 44%.

[0059] Example 5: 407 mg (1.3 mmol) of 4 - diethylaminoketone acid and 206 mg (1.3 mmol) of 6 - amino - 1 - phenol were taken. 7 ml of methanesulfonic acid MeSO3H was added to the mixed reactants. Nitrogen was introduced to expel all the internal gases. The mixture was refluxed at 100 °C for 28 h. The organic solvent was evaporated to dryness by rotary evaporation to obtain the crude product. Finally, the solid was separated by liquid chromatography column using a mixed system of dichloromethane and anhydrous methanol to obtain the pure product. 195 mg of the purple pure compound (II) was obtained, and the yield was 34%.

[0060] Example 2: To test the changes in the fluorescence spectrum and absorption spectrum of the probe before and after the recognition of nitrite

[0061] 25 μL was taken from the probe mother liquor (1 mM) and placed in a 5 mL pure water test system. Then, 10 μL of the nitrite stock solution (10 mM) was pipetted into the test system. After shaking well, it was left standing for 40 min. The changes in its fluorescence spectrum and absorption spectrum were measured using a fluorescence spectrometer and an ultraviolet absorption spectrometer respectively. The above measurements were carried out in a pure water system (pH = 1). The probe used was the probe prepared in Example 1, and all spectral measurements were carried out at room temperature (25 °C). The test results are shown respectively as Figure 1 and Figure 2 shown.

[0062] It can be clearly seen from Figure 1 that when nitrite was added, the change in fluorescence intensity at 625 nm was very obvious; it can be seen from Figure 2 that the absorption intensity value at 568 nm decreased significantly, and through the change in color, naked - eye recognition can be achieved. The color change after adding nitrite was: red before adding nitrite, and light red after adding nitrite.

[0063] Example 3: To test the time - dynamics of the colorimetric fluorescence probe

[0064] First, 25 μL was taken out from the probe mother liquor (1 mM) and placed into a 5 mL test system. Then, 10 μL of the nitrite stock solution (10 mM) was pipetted into the test system. After shaking well, the fluorescence intensity change was immediately measured using a fluorescence spectrometer. The above measurement was carried out in a pure water system (pH = 1). The probe used was the probe prepared in Example 1, and all spectral tests were measured at room temperature (25 °C). The results are as Figure 3 shown.

[0065] It can be Figure 3 clearly seen that when nitrite was added, the fluorescence intensity basically reached the lowest value after about 40 min of detection.

[0066] Example 4: Testing the concentration gradient of the colorimetric fluorescent probe for nitrite

[0067] First, 25 μL was taken out from the probe mother liquor (1 mM) and placed into a 5 mL test system. Then, according to the different concentrations of nitrite to be prepared (concentrations were: 0 μM, 0.5 μM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 4 μM, 5 μM, 6 μM, 8 μM, 10 μM, 15 μM, 20 μM, 25 μM), different volumes of the nitrite stock solution were added. After shaking well, it was left standing for 40 min. Then, the fluorescence intensity change was measured using a fluorescence spectrometer. The above measurement was carried out in a pure water system (pH = 1). The probe used was the probe prepared in Example 1, and all spectral tests were measured at room temperature (25 °C). The test results are as Figure 4 shown.

[0068] It can be Figure 4 clearly seen that as the concentration of added nitrite increased, the fluorescence intensity at 625 nm gradually decreased; and, it can be Figure 5 seen that at 625 nm, after the colorimetric fluorescent probe (5 μM) was added with nitrite (0 - 10 μM), the fluorescence intensity showed a good linear relationship, which proved that the colorimetric fluorescent probe could be used for quantitative analysis of nitrite.

[0069] Example 5: Testing the selectivity of the colorimetric fluorescent probe for nitrite

[0070] The analytes were respectively: 1. blank; 2. silver ion Ag + ; 3. calcium ion Ca 2+ ; 4. chloride ion Cl - ; 5. carbonate ion CO3 2- ; 6. chromium ion Cr 3+ ; 7. copper ion Cu 2+ ; 8. ferrous ion Fe 2+ ; 9. ferric ion Fe3+ ; 10. Iodide ion I - ; 11. Potassium ion K + ; 12. Magnesium ion Mg 2+ ; 13. Manganese ion Mn 2+ 14. Sodium ion Na +- ; 15. Lead ion Pb 2+ ; 16. Sulfate ion SO4 2- ; 17. Zinc ion Zn 2+ ; 18. Nitrite NO2 (analyte concentrations are all 20 μM). The bar graph represents the fluorescence intensity values of the probe at 625 nm in the presence of different analytes. The above measurements were carried out in a pure water system (pH = 1), the probe used was the probe prepared in Example 1, and all spectral tests were measured at room temperature (25 °C). Specifically, multiple parallel samples with a probe concentration of 5 μM were prepared in 10 mL colorimetric tubes, then a certain amount of analyte was added, shaken well, and after standing for 40 min, the fluorescence intensity value was measured. The test results are as Figure 6 (B) shown.

[0071] From Figure 6 it can be seen that the probe of the present invention has high selectivity for nitrite, can specifically react with nitrite, and there are obvious changes in the fluorescence spectrum before and after the reaction, while the fluorescence intensity does not change significantly after the interaction between other common analytes in the water environment and the probe.

[0072] Example 6: Testing the anti-interference ability of the colorimetric fluorescence probe

[0073] The interfering analytes are respectively: 1. Blank; 2. Silver ion Ag + ; 3. Calcium ion Ca 2+ ; 4. Chloride ion Cl - ; 5. Carbonate ion CO3 2- ; 6. Chromium ion Cr 3+ ; 7. Copper ion Cu 2+ ; 8. Ferrous ion Fe 2+ ; 9. Ferric ion Fe 3+ ; 10. Iodide ion I - ; 11. Potassium ion K + ; 12. Magnesium ion Mg 2+ ; 13. Manganese ion Mn 2+ 14. Sodium ion Na +- ; 15. Lead ion Pb 2+ ; 16. Sulfate ion SO4 2- ; 17. Zinc ion Zn 2+; 18. Nitrite NO2 (analyte concentration is 20 μM for all). The bar graph represents the fluorescence intensity values of the probe at 625 nm in the co - existence of different analytes and nitrite. The above - mentioned determination was carried out in a pure water system (pH = 1). The probe used was the one prepared in Example 1, and all spectral tests were measured at room temperature (25 °C). Specifically, multiple parallel samples with a probe concentration of 5 μM were prepared in 10 - mL colorimetric tubes, then the above - mentioned analytes (except nitrite) were added. Immediately afterwards, 20 μL of nitrite stock solution (1 mM) was pipetted into the test system (except for the first group of blank groups), shaken well, and left standing for 40 min to measure the fluorescence intensity values. The results are as Figure 6 (A) shown.

[0074] From Figure 6 (A), it can be seen that the probe of the present invention has excellent anti - interference ability. Ions and other substances commonly found in the water environment do not interfere with the quantitative and qualitative detection of nitrite by the probe of the present invention.

[0075] Example 7: Detection limit test and calculation of the probe

[0076] The fluorescence titration method was used to calculate the detection limit. The detection limit calculation formula is as follows:

[0077] Detection limit = 3σ / k

[0078]

[0079] σ is the standard deviation of the fluorescence intensity of the blank probe, and k is Figure 5 the slope of the linear relationship graph.

[0080] From this calculation formula (Ⅱ), the detection limit of the probe for nitrite is 0.16 μM.

[0081] Example 8: Analysis and test of nitrite in three real water samples by the probe

[0082] Three real water samples were collected respectively. Water sample A was collected from the Jinxiu River in Jinan City, water sample B was collected from the Zihe River in Jinan City, and water sample C was collected from the Pearl Spring in Jinan City. These water samples were all detected, and no nitrite was found. Then, three test systems were configured for each water sample. Each test system was added with 5 μM of the probe. The probe used was the one prepared in Example 1. Then, 2 μM, 5 μM, and 8 μM of nitrite were added to each test system of each water sample respectively, shaken well, left standing for 40 minutes, and then the change in fluorescence intensity was measured to calculate the nitrite content of each test system.

[0083] The above steps were repeated three times.

[0084] The test results are shown in the following table. The test results indicate that the recovery rates of the three water samples are 88.32%-111%, further confirming that the colorimetric fluorescent probe of the present invention can effectively detect nitrite in real water samples.

[0085]

[0086]

[0087] Although the present invention has been described by the above embodiments, it should be understood that the present invention can be further modified and changed without departing from the spirit of the present invention, and these modifications and changes are within the protection scope of the present invention.

Claims

1. A colorimetric fluorescent probe for measuring, detecting or screening nitrite, characterized in that: The chemical structure is shown in formula (I): Among them, R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 , R 11 and R 12 is a hydrogen atom, a straight-chain or branched alkyl group, a straight-chain or branched alkoxy group, a sulfonic acid group, an ester group, or a carboxyl group; R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 , R 11 and R 12 Can be the same or different.

2. The colorimetric fluorescent probe according to claim 1, characterized in that R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 , R 11 and R 12 All are hydrogen atoms.

3. A method for preparing the colorimetric fluorescent probe according to claim 1 or 2, characterized in that: The preparation is carried out according to the following reaction formula: characterized in that it comprises the following steps: reacting a compound of formula (III) with a compound of formula (IV) to prepare a compound of formula (I), and the reaction formula is as follows: Among them, R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 , R 11 , R 12 and R 13 is a hydrogen atom, a straight-chain or branched alkyl group, a straight-chain or branched alkoxy group, a sulfonic acid group, an ester group, or a carboxyl group; R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 , R 11 , R 12 and R 13 Can be the same or different.

4. The preparation method according to claim 3, characterized in that: The steps include: A certain molar ratio of the compound of formula (III) and the compound of formula (IV) are placed in a reaction container, and then methanesulfonic acid is added to react under heating conditions. After the reaction is completed, the organic solvent is evaporated to dryness to obtain a crude product, which is then separated by column chromatography to obtain a pure compound of formula (I).

5. A colorimetric fluorescent probe composition for measuring, screening or detecting nitrite, comprising the colorimetric fluorescent probe according to any one of claims 1 or 2.

6. The colorimetric fluorescent probe composition according to claim 5, characterized in that: The colorimetric fluorescent probe composition further comprises a solvent, an acid, a base, a buffer or a combination thereof.

7. Use of the colorimetric fluorescent probe according to any one of claim 1 or claim 2 in the preparation of a reagent for measuring, screening or detecting nitrite.

8. A kit for detecting the presence of nitrite in a sample or determining the nitrite content in a sample, characterized in that: It comprises the colorimetric fluorescent probe of formula (I) or formula (II).

9. A method for detecting the presence of nitrite in a sample or determining the nitrite content in a sample, comprising: a) contacting the colorimetric fluorescent probe according to any one of claims 1 to 2 with a sample to form a fluorescence quenching compound; b) determining the fluorescence property of the fluorescence quenching compound.

10. The method according to claim 9, wherein the sample is a chemical sample or an aqueous environment sample.

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