KR102278620B1 - Colorimetric sensor for detecting mercury ions or arsenic ions comprising surface-modified nanoparticles, method for detecting mercury ions or arsenic ions using the same and method for preparing the same - Google Patents

Abstract

Disclosed herein are a colorimetric sensor for detecting mercury ions or arsenic ions including surface-modified nanoparticles, a method for detecting mercury ions or arsenic ions using the same, and a method for manufacturing the same.

Description

Surface modification-colorimetric sensor for detecting mercury ions or arsenic ions including nanoparticles, method for detecting mercury ions or arsenic ions using the same, and method for manufacturing the same MERCURY IONS OR ARSENIC IONS USING THE SAME AND METHOD FOR PREPARING THE SAME}

The present specification relates to a colorimetric sensor for detecting mercury ions or arsenic ions including surface-modified nanoparticles, a method for detecting mercury ions or arsenic ions using the same, and a method for manufacturing the same.

More specifically, by surface-modifying the surface of gold nanoparticles with D-penicillamine to activate an amine group and a carboxyl group as a receptor for a specific metal ion, a colorimetric sensor that selectively reacts with a metal ion to be detected, a detection method and to a method for manufacturing the same.

Mercury ion (Hg ^{2 +} ) is the only metal that exists in a liquid state at room temperature and is released into the environment in the form of inorganic mercury from natural or artificial sources. Such inorganic mercury is converted into organic mercury, which is an organic form, due to the activity of microorganisms in the sediment, and is also converted back to inorganic mercury while remaining for a long time in the environment around us. Mercury is a heavy metal that causes fatal damage to the nervous system when accumulated in the human body. It is closely used in real life such as paints, pigments, fluorescent lamps, and thermometers. Mercury used as a catalyst for the production of plastic products in factories is ingested by plankton, fish and shellfish, and is harmful to the ecosystem. It can cause damage by causing the accumulation of methylmercury in the body through the food chain process.

Mercury accumulates in the body and is not easily excreted. When the total mercury content exceeds 30 mg/kg, mercury poisoning occurs. It is mainly absorbed into the digestive tract through food or drinking water or enters the respiratory tract in the form of vapor, of which 80% Abnormalities accumulate in the kidneys and liver and paralyze the function of the cerebellum. As mercury accumulates in the human body, movement disorders, etc. are expressed due to chronic nervous system diseases, and death occurs when exposed to 150-300 mg/70 kg or more.

Arsenic (As ^{3 +} ) is detected in large amounts in polluted areas through coal mines, smelting of non-ferrous metals, and combustion of fossil fuels. When pesticides containing arsenic are used for a long period of time, they become a source of contamination in farmland, and when preserving wood for wood preservation, using products containing arsenic ions is a cause of environmental pollution.

The general public is exposed to arsenic ions mainly from food and drinking water. Arsenic in drinking water is an important source of exposure to inorganic arsenic. The daily intake of total arsenic in food and drinking water is about 20-300 μg/day (WHO, 2001). Arsenic has different degrees of toxicity depending on its chemical form. Metallic arsenic and organic arsenic are relatively low in toxicity, but when water-soluble inorganic arsenic enters the human body, it causes acute toxicity and causes cardiovascular abnormalities, peripheral vascular disease, and nervous system dysfunction, and eventually leads to death.

Mercury and arsenic ions are mainly analyzed by High Performance Liquid Chromatography (HPLC), Inductively Coupled Plasma Mass Spectrometer (ICP/MS), Electrochemical assay, and atomic absorption spectrometry. (Atomic Absorption Spectrometer, AAS) has been proposed.

Nanoparticle colorimetric sensor analysis using surface plasmon resonance such as gold nanoparticles theoretically induces free electron vibration on the surface of nanoparticles by light waves absorbed by nano-sized particles. use the principle At this time, a resonance phenomenon occurs and a specific wavelength is emitted, and the particle takes on various colors depending on the size, shape, and type of the particle. However, when metal nanoparticles are combined with an external compound, the nanoparticles may aggregate and the size of the particles may change, and thus the color may change, so it can be applied to sensors that measure and monitor environmental pollutants. (Environmental Technology Technology Trend Report 2011).

Penicillamine, sold under the trade name Camprimine, is mainly used for the treatment of Wilson's disease, a genetic disorder of copper metabolism, rheumatoid arthritis, and kidney stones in patients with various heavy metal poisoning. is also used for In addition, it is on the World Health Organization's list of essential medicines and is divided into D-penicillamine and L-penicillamine, which is a stereoisomer, depending on the structure.

Although the toxicity of D-penicillamine is relatively low, in the case of L-penicillamine, it induces neurotoxin due to inhibition of a pyridoxine-dependent enzyme in animal experiments. In 1957, two cases were published on the effect of D-penicillamine on lead chelation, a small controlled study was published in the early 1960s, and in the 1950s the chelation of mercury with N-acetyl-penicillamine was also has been proven [Boulding JE, Baker RA. Lancet. 1957;2:985.] [Goldberg A, et al. Treatment of lead-poisoning with oral Penicillamine. Br. Med. J. 1963; 1:1270] [Ohlsson WTL. Penicillamine as lead-chelating substance in man. Br. Med. J. 1962; 1:1454].

D-penicillamine, commonly referred to as penicillamine, is a heavy metal chelate and is particularly effective in copper and arsenic, iron, mercury and lead chelates. D-penicillamine is a naturally occurring penicillin base, also called 3-Mercapto-D-valine. During the action of D-penicillamine, thiol binds to nanoparticles, and amines and carboxyl groups (COOH) exist on the surface of the nanoparticles to activate the function.

Due to the recent advancement of the industry, the environmental problems we are facing are increasing, and in order to live a pleasant and safe life, we need to build a system that can prepare ourselves from the environment. Skills are needed. For this, several element technologies are required, and one of them is environmental sensor technology.

^{Therefore, mercury ions (Hg 2 +} ) and arsenic ions (As ^{3 +} ), which are toxic substances, can be rapidly detected and analyzed selectively or simultaneously in environmental pollution samples, forensic science samples, drinking water, pharmaceuticals, chemical substances handling industrial sites, etc. , development of sensor technology capable of real-time measurement and recycling of unreacted sensors, high stability, and small-scale production is required.

Korean Patent Publication No. 10162117000

Sensors and Actuators B, 249 (2017) 339-347 Talanta, 151 (2016) 106-113 Sensors and Actuators B, 284 (2019) 217-280 Environmental Technology Technology Trend Report 2011 Boulding JE, Baker RA. Lancet. 1957;2:985. Goldberg A, et al. Treatment of lead-poisoning with oral Penicillamine. Br. Med. J. 1963; 1:1270 Ohlsson WTL. Penicillamine as lead-chelating substance in man. Br. Med. J. 1962;1:1454

Embodiments of the present invention provide a colorimetric sensor for detecting mercury ions or arsenic ions including surface-modified nanoparticles, a method for detecting mercury ions or arsenic ions using the same, and a method for manufacturing the same.

In one embodiment of the present invention, a colorimetric sensor for detecting mercury ions or arsenic ions, including surface-modified nanoparticles, wherein the surface-modified nanoparticles include gold nanoparticles; and a modifier represented by Formula 1 below, wherein the modifier is bound to the surface of the gold nanoparticles, and provides a colorimetric sensor for detecting mercury ions or arsenic ions.

In another embodiment of the present invention, there is provided a method for detecting mercury ions or arsenic ions, the method comprising: inputting a sample to be detected into the colorimetric sensor for detecting mercury ions or arsenic ions; a reaction and waiting step between the colorimetric sensor and the sample to be detected; and a sensing step of detecting mercury ions or arsenic ions in the detection target sample by the color change of the colorimetric sensor.

In another embodiment of the present invention, as a first solution, preparing a solution in which gold nanoparticles are dispersed; As a second solution, preparing a solution containing a modifier represented by the following formula (1); and mixing the first solution and the second solution to prepare the surface-modified nanoparticles in which the modifier is bonded to the surface of the gold nanoparticles; it provides a method for manufacturing a colorimetric sensor for detecting mercury ions or arsenic ions, comprising:

The present invention uses gold nanoparticles modified with D-Penicillamine to selectively react with mercury ions or arsenic ions to be detected, so that it can be combined with biological samples, beverages, and pharmaceuticals through color change. Specific heavy metal ion components contained or dissolved in food, etc. can be easily detected, and the sensor can be recycled.

In particular, it has excellent selectivity, sensitivity and quantitation for ^{mercury (Hg 2 +} ) ions and arsenic ions (As ^{3 + ) components.} It can enable rapid detection, real-time measurement and continuous management.

1 shows (a) gold nanoparticles (b) a nanoprobe surface-modified with DPL, (c) aggregated nanoprobes due to ^{mercury ions (Hg 2 + ) according to an embodiment of the present invention; (} d) Photographs and absorbance graphs of nanoprobes aggregated due to arsenic ions (As ^{3 + ).}
2 is (a) gold nanoparticles (b) nanoprobes, (c) mercury ions (Hg ^{2 +} ) aggregated nanoprobes according to an embodiment of the present invention, (d) arsenic ions (As ^{3 +} ) This is a particle distribution graph and TEM image of the nanoprobe aggregated due to the aggregation.
3 shows (a) color change, (b) UV-vis spectrum of gold nanoparticles when added to a nanoprobe according to an embodiment of the present invention for a selectivity test of a nanoprobe using various metal ions and anions; , and (c) the absorbance ratio (A ₇₃₀ /A ₅₂₅ ).
_{4 is a diagram showing absorbance ratios (A 730} /A ₅₂₅ ) according to various surface modifiable materials according to an embodiment of the present invention, (a) when mercury ions (Hg ^{2 +} ) are added, (b ) is a diagram showing the addition of arsenic ions (As ^{3 + ).} (Conditions: AuNPs in ^{1 ml 2 μM Hg 2+, 2} μM As 3 +, 5 mM DPL, addition of 5 mM other ingredients)
5 is a diagram showing (a) color change, and (b) absorbance ratio (A ₇₃₀ /A ₅₂₅ ) ^{of the colorimetric sensor solution according to pH change after mercury ion (Hg 2 +} ) is added in Example 4 of the present specification .
6 is a graph showing (a) a color change photograph of the colorimetric sensor solution according to pH change after addition ^{of arsenic ions (As 3 +} ) in Example 4 of the present specification _{, and (b) absorbance ratio (A 730} /A _{525 )} to be.
7 is a reaction experiment to the addition ^{of mercury ions (Hg 2 +} ) of the colorimetric sensor solution according to the concentration of the D-penicillamine solution in Example 5 of the present specification. (a) Color change of colorimetric sensor solution according to DPL concentration change, (b) UV-vis spectrum according to DPL concentration change, and (c) absorbance ratio (A ₇₃₀ /A ₅₂₅ ) according to DPL concentration change.
8 is an experiment showing the reactivity ^{of the colorimetric sensor solution to arsenic ions (As 3 +} ) according to the concentration of the D-penicillamine solution in Example 5 of the present specification. (a) Color change of colorimetric sensor solution according to DPL concentration change, (b) UV-vis spectrum according to DPL concentration change, and (c) absorbance ratio (A ₇₃₀ /A ₅₂₅ ) according to DPL concentration change.
9 shows the absorbance ratio of the colorimetric sensor solution according to the reaction temperature of the nanoprobe after addition of (a) mercury ions (Hg ^{2 +} ) and (b) arsenic ions (As ³⁺ _{) in Example 6 of the present specification (A 730} / A ₅₂₅ ) is shown.
10 shows the absorbance ratio of the colorimetric sensor solution over time according to the concentration of (a) mercury ions (Hg ^{2 +} ) and (b) arsenic ions (As ³⁺ _{) in Example 7 of the present specification (A 730} /A ₅₂₅ ) This is a picture showing the change.
Figure 11 (a) color change picture of the colorimetric sensor solution according to the change in concentration of the mercury ion (Hg ^{2 +)} in the eighth embodiment of the present disclosure, (b) UV-vis spectra, (C) the absorbance ratio (A ₇₃₀ / A ₅₂₅ ) is a quantitative curve graph.
12 is (a) a color change photograph, (b) UV-vis spectrum, (c) absorbance ratio (A ₇₃₀ /A ^{) of the colorimetric sensor solution according to the concentration of arsenic ions (As 3 +} ) in Example 9 of the present specification ₅₂₅ ) is a quantitative curve graph.
_{13 shows the absorbance ratio of the colorimetric sensor solution (A 730} / A ₅₂₅ ) when various metal ions and anions are added after the addition of 5 μM of ^{mercury ions (Hg 2 +} ) in Example 10 of the present specification (a) Graph, (b) When ^{5 μM of arsenic ions (As 3 +} ) is added and then 50 μM of various metal ions and anions are added, the absorbance ratio (A ₇₃₀ /A ₅₂₅ ) of the colorimetric sensor solution is a graph.
14 is a diagram showing gold nanoparticles and nanoprobes having a surface modified with DPL according to an embodiment of the present invention ^{aggregation due to mercury ions (Hg 2 +} ) and arsenic ions (As ^{3 +} ) through color change. It is a schematic diagram showing a series of colorimetric sensor processes.

In the present specification, when a part "includes" a certain component, this means that other components may be further included, rather than excluding other components, unless otherwise stated.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. Embodiments of the present invention have been described with reference to the accompanying drawings, which are described for purposes of illustration, and the technical spirit of the present invention and its configuration and application are not limited thereby.

mercury ions or For arsenic ion detection colorimetric sensor

In exemplary embodiments of the present invention, there is provided a colorimetric sensor for detecting mercury ions or arsenic ions, including surface-modified nanoparticles, wherein the surface-modified nanoparticles include gold nanoparticles; and a modifier represented by Formula 1 below, wherein the modifier is bound to the surface of the gold nanoparticles, and provides a colorimetric sensor for detecting mercury ions or arsenic ions.

[Formula 1]

The gold nanoparticle probe modified with DPL shows a distinct color change as the overall size increases by about 5 times due to aggregation by ^{mercury ions (Hg 2 +} ) and arsenic ions (As ^{3 + ), making it easy to detect mercury ions or arsenic ions} can do.

Specifically, referring to FIG. 14 , the thiol, amine group, and carboxyl group of D-penicillamine bound to the surface of the gold nanoparticles impart functionality to the gold nanoparticles, thereby inducing aggregation between the gold nanoparticles to induce aggregation. , and the color emission wavelength of the colorimetric sensor is changed by the surface plasmon resonance.

In one embodiment, the surface-modified nanoparticles may be uniformly dispersed in the colorimetric sensor solution, and the surface-modified nanoparticles may have a spherical shape, increasing the particle surface area, and may be relatively easy to surface-modify. In addition, it is easy to control the wavelength of absorption and scattering according to the size of the spherical particle, and biocompatibility can be improved.

In an exemplary embodiment, the size of the gold nanoparticles may be 20 nm or less, and may be about 1 nm or more, and when the size of the nanoparticles is reduced to 20 nm or less, it acts as subwavelength antennas. You can see the LSPR effect.

Specifically, localized surface plasmon resonance (LSPR) is an optical phenomenon caused by light when it interacts with conductive nanoparticles (NPs) with an incident wavelength and smaller than this. As in surface plasmon resonance (SPR), the electric field of incident light can collectively excite electrons in the conduction band to generate an increased electric field, and this phenomenon depends on the composition, size, shape, dielectric environment and It is very close to localized plasmon oscillations with a resonant frequency that is highly dependent on the interparticle distance. Therefore, it can be used as a high-sensitivity and high-efficiency technology in bioassays, biosensing, and solar cells using nanoparticles.

In an exemplary embodiment, the concentration of the modifier may be 1 to 10 mM based on the total volume of the colorimetric sensor solution, for example, 2 mM or more, 3 mM or more, 4 mM or more, 9 mM or less, 7 mM or less Hereinafter, it may be 6 mM or less, preferably 5 mM. When the concentration of the modifier is less than 1 mM, the detection color change of arsenic ions is small, the reaction time may be very long, and when the concentration of the modifier is more than 10 mM, stability due to aggregation between nanoparticles may be lowered.

In an exemplary embodiment, the color of the colorimetric sensor may be red, and when mercury ions or arsenic ions are added, the color may be changed to dark blue.

In an exemplary embodiment, the mercury ion detection pH of the colorimetric sensor may be 3 to 7, and the arsenic ion detection pH of the colorimetric sensor may be 3 to 4.5. Therefore, in the range of pH 5 to pH 7, ^{only the concentration of mercury ions (Hg 2 +} ) can be selectively detected, and at pH 3 to pH 4.5, the concentration of mercury ions (Hg ^{2 +} ) and arsenic ions (As ^{3 +} ) is sum can be measured.

In an exemplary embodiment, the absorbance ratio of the colorimetric sensor may be 0.2 to 1.2. This extinction ratio means that the detection sensor of the present invention has very good selectivity for mercury or arsenic ions compared to other metal ions.

In an exemplary embodiment, the colorimetric sensor is at 20 to 50 °C, mercury Since ions or arsenic ions can be detected, preferably mercury ions or arsenic ions can be detected at 30° C., a separate heating or the like is not required for the colorimetric sensor during detection.

In an exemplary embodiment, the color emission wavelength of the colorimetric sensor is 450 to 600 nm, the color emission wavelength when detecting mercury ions of the colorimetric sensor is 520 to 620 nm, and the color emission wavelength when detecting arsenic ions of the colorimetric sensor is It may be 650 to 800 nm, so that it exhibits a different color emission wavelength unlike other ions, so that selective detection is possible.

In an exemplary embodiment, when the colorimetric sensor reacts with mercury ions or arsenic ions, the reaction time may be within 10 minutes, within 7 minutes, within 5 minutes, or preferably within 3 minutes. Therefore, it is possible to detect ions within a relatively short time, and thus has a high utility value.

In an exemplary embodiment, the detectable concentration of mercury ions or arsenic ions of the colorimetric sensor may be 25 nM or more, and a minute amount of mercury ions or arsenic ions may be detected.

As such, in the case of the detection sensor of the present invention, mercury Since the presence or absence of an ion or arsenic ion can be known and two ions can be detected with one sensor, it can be widely used in various fields.

mercury ions or Arsenic ion detection Way

In another embodiment of the present invention, there is provided a method for detecting mercury ions or arsenic ions, the method comprising: inputting a sample to be detected into the colorimetric sensor for detecting mercury ions or arsenic ions; a reaction and waiting step between the colorimetric sensor and the sample to be detected; and a sensing step of detecting mercury ions or arsenic ions in the detection target sample by the color change of the colorimetric sensor.

In an exemplary embodiment, before the inputting step, in order to improve the stability and sensitivity of the nanoparticles, adjusting at least one of pH and temperature of the colorimetric sensor; may further include.

In this way, mercury or arsenic ion detection can be performed through a very simple method using the colorimetric sensor for detecting mercury ions or arsenic ions. The selectivity and sensitivity to ions are very high, so that mercury or arsenic ions can be easily detected with the naked eye and/or with a spectrophotometer or a colorimeter alone.

mercury ions or For arsenic ion detection Method of manufacturing colorimetric sensor

In another embodiment of the present invention, as a first solution, preparing a solution in which gold nanoparticles are dispersed; As a second solution, preparing a solution containing a modifier represented by the following formula (1); and mixing the first solution and the second solution to prepare the surface-modified nanoparticles in which the modifier is bonded to the surface of the gold nanoparticles; it provides a method for manufacturing a colorimetric sensor for detecting mercury ions or arsenic ions, comprising:

Specifically, a gold nanoparticle solution was prepared by adding trisodium citrate as a reducing agent to an aqueous chlorauric acid hydrate solution (HAuCl ₄ ), and a thiol group, an amine group, and an amine group were added to the prepared gold nanoparticle solution (first solution). The surface of the gold nanoparticles may be modified by adding a D-penicillamine mixed solution (second solution) having a carboxyl group.

Hereinafter, the present invention will be described in detail with reference to preferred embodiments so that those skilled in the art can easily carry out the present invention. However, the present invention may be embodied in various different forms, and is not limited to the embodiments described herein.

production example 1: Preparation of gold nanoparticles

A gold nanoparticle solution was prepared by adding trisodium citrate, a reducing agent, to an aqueous chloroauric acid hydrate solution (HAuCl ₄ ), and then a DPL mixed solution having a thiol group, an amine group, and a carboxyl group was added to the prepared gold nanoparticle solution. Thus, surface-modified gold nanoparticles were prepared.

_{More specifically, 40 mg of aqueous chloroauric acid hydrate (HAuCl 4} ) and 100 ml of distilled water are added to a 2-neck round bottom flask of 250 ml and heated while stirring. After about 15 minutes, when the solution boils, a solution of 10 ml of 38.8 mM trisodium citrate is added with a syringe and stirred rapidly.

And when stirred for about 5 minutes, the color of the solution changes from light yellow to black to dark red, and the aqueous chlorauric acid hydrate solution (HAuCl ₄ ) is gold (III) (gold(III)) ions reduced by trisodium citrate. Nanoparticles were synthesized.

1 mM of gold nanoparticles having an average size of about 20 nm were uniformly prepared by the above method. The prepared solution was used after cooling for 2 hours at room temperature.

production example 2: Preparation of colorimetric sensor ( surface modification -Nanoparticle manufacturing)

Dissolve D-penicillamine having a molecular weight (MW) of 149.212 g/mol in a neutral aqueous solution (pH 7, tertiary distilled water) to prepare a D-penicillamine solution, but add 1 mL of 5 mM DPL solution to 9 mL of gold nanoparticle aqueous solution A colorimetric sensor solution containing gold nanoparticles was prepared by culturing with stirring for 30 minutes. Unreacted D-penicillamine and chloroauric acid hydrate were removed by centrifugation at 4,000 rpm for 10 minutes, and then the above process was repeated three times.

TEM images of the prepared gold nano-particle colorimetric sensor and nano-probe are shown in FIG. 2 . The size of the prepared gold nanoparticles was in the form of a round circle, and the diameter was up to 22 nm, and the solution was dark red due to the surface plasmon resonance phenomenon of the gold nanoparticles. Gold nanoparticles surface-modified with DPL were also not significantly different in size and color.

Example 1: D- with penicillamine modified Size Characteristics of Gold Nanoparticle Colorimetric Sensor

1 is TEM images of gold nanoparticles and functionalized gold nanoparticles prepared in Preparation Examples 1 and 2 according to the present invention, and mercury ions (Hg ^{2 +} ) and arsenic ions (As ^{3 + ) reacted therewith.} As can be seen from the photo, the color was changed from red before the reaction to blue after the reaction. 1 shows the colors according to the reaction between the prepared gold nanoparticles and the DPL-modified nanoprobe, and thus mercury ions and arsenic ions.

The gold nanoparticles and the nanoprobes modified with DPL are each red in color, and it can be seen that the color change occurs clearly due to the reaction between the ^{mercury ion (Hg 2 +} ) and the arsenic ion (As ^{3 + ), which are the detection target ions.} .

FIG. 2A shows the gold nanoparticles obtained in Preparation Example 1, and as shown, all gold nanoparticles were uniformly prepared as gold nanoparticles having a diameter of about 22 nm, and it can be seen that these particles were well separated. FIG. 2b is a diagram showing functionalized gold nanoparticles (DPL-AuNPs) obtained in Preparation Example 2. FIG.

Figure 2c is a particle size distribution diagram and TEM photograph of gold nanoparticles (DPL-AuNPs) modified with DPL after reacting with the addition of mercury ions (Hg ^{2 + ) 2μM, Figure 2d is mercury ions (Hg 2 +} ) 1 μM It is a particle size distribution diagram and TEM photograph of gold nanoparticles (DPL-AuNPs) after reacting by adding .

Comparing FIGS. 2b and 2c to 2d, ^{the average size of nanoparticles is maintained at 22 nm before the reaction with mercury ions (Hg 2 +} ) and arsenic ions (As ^{3 +} ), but after the reaction, the size of the nanoparticles is 98 nm. It can be seen that the nanoparticles are agglomerated with each other and become very large.

Example 2: Experiments on the reactivity of gold nanoparticles to various ions

^{23 kinds of cations Li +} (1), Na ⁺ (2), K ⁺ (3), Ag ⁺ (4), Ba ^{2 +} (5) in the nanoprobe colorimetric sensor solution surface-modified with DPL of Preparation Example 2 at room temperature ^{, Ca 2 + (6),} Cd 2 + (7), Co 2 + (8), Cu 2+ (9), Hg 2 + (10), Mn 2 + (11), Mg 2 + (12), ^{Ni 2 + (13), Pb} 2 + (14), Sn 2 + (15), Zn 2 + (16), Al 3+ (17), As 3 + (18), Fe 3 + (19), Ga ^{2 +} (20), Ti ^{3 +} (21), Ge ^{4 +} (22), Cr ^{6 +} (23) and 8 anions NO ₂ ^- (24), NO ₃ ^- (25), PO ₄ ^3- (26) ), SO ₄ ^2- (27), F ^- (28), Cl ^- (29), Br ^- (30), I ^- (31) were added and reacted for 30 minutes, and the color change was observed. 3a is a photograph of each sample after the reaction, FIG. 3b is a UV-vis spectrum, and FIG. 3c is an absorbance ratio (A ₇₃₀ /A ₅₂₅ ) graph.

In FIG. 3A , the colorimetric sensor solution to which mercury ions (Hg ^{2 +} ) and arsenic ions (As ^{3 +} ) were added was different from the colorimetric sensor solution to which other anions and metal ions were added. This means that the aggregation of the nanoprobe occurs only by ^{mercury ions (Hg 2+} ) and arsenic ions (As ^{3 + ).}

In FIG. 3b, the solutions to which other ions are added show an absorption spectrum very similar to that of the colorimetric sensor solution, and a very strong absorbance at 525 nm. On the other hand, the solution to which mercury ions (Hg ²⁺ ) and arsenic ions (As ^{3 +} ) are added has no absorption peak at 525 nm, a weak peak at 550 nm, and a strong absorption peak at 730 nm.

3c shows the spectrum of each sample as an absorbance ratio (A ₇₃₀ /A ₅₂₅ ). Unlike other samples, the solution to which mercury ions (Hg ^{2 +} ) and arsenic ions (As ^{3 +} ) are added is at least 10 times higher. It shows the absorbance ratio. The high absorbance ratio means ^{that the selectivity to mercury ions (Hg 2 +} ) and arsenic ions (As ^{3 +} ) is very good.

Example 3: Reactivity experiment of various metal ions according to the surface modification material

As the surface modifying material of the gold nanoparticles used in the present invention, there are a total of 15 amino acids having a structure similar to that of D-penicillamine. More specifically, D-penicillamine, alanine, lysine, phenylalanine, glutathione, glutamic acid, glycine, valine , Methionine, Serine, Leucine, Isoleucine, Tryptophan, Proline, and Threonine were used.

In FIG. 4A , the reactivity of mercury with solutions to which other surface modifying materials are added is shown as an absorbance ratio (A ₇₃₀ /A ₅₂₅ ). When the surface was modified with D-Penicillamine ^{and reacted with mercury ions (Hg 2 +} ), the absorbance ratio was as high as 1.2, and after surface modification with other materials, mercury ions (Hg ^{2 + )} ) and the reaction was as low as 0.22 to 0.48, which resulted in the same result when reacted with ^{arsenic ions (As 3 + ) (FIG. 4b).}

Therefore, it was confirmed that it was preferable to detect ^{mercury ions (Hg 2 +} ) and arsenic ions (As ^{3 +} ) when the gold nanoparticle aqueous solution was surface-modified with D-Penicillamine.

Example 4: Mercury ions of colorimetric sensor according to pH change ( Hg ^{2 +} ) and arsenic ions ( As ^{3 +} ) for reactivity experiments

The color change and stability of the gold nanoparticles according to the pH of the colorimetric sensor solution obtained in Preparation Example 2 were examined. More specifically, a colorimetric sensor was prepared by adding 1 ml of a 5 mM DPL solution to 9 ml of an aqueous gold nanoparticle solution and stirring for 30 minutes. The pH was adjusted using 0.5M HNO ₃ and 0.5M NaOH, and samples were prepared to be pH 3, 4, 5, 6, 7, 8, 9 and 10, respectively.

A photograph of the sample according to each pH is shown in FIG. 5A, and the absorbance spectrum of each sample was measured by UV-vis, and the absorbance spectrum is shown in FIG. 5B.

Referring to FIG. 5A , at pH 3, 4, 5, 6, 7, 8, 9 to 10, the gold nanoparticle colorimetric sensor was stable without changing its intrinsic color, and the concentration was adjusted to 1.0 μM of ^{mercury ions (Hg 2 + ).} After the sample showed a blue color at pH 3, 4, 5, 6 to 7. Referring to the wavelength of 525 nm in FIG. 5B , the nanoparticles formed at a pH of 6 exhibit the strongest absorbance.

As for arsenic ions (As ^{3 +} ), a photograph of the sample according to each pH is shown in FIG. 6a, and the absorbance spectrum of each sample was measured by UV-vis, and the absorbance spectrum was shown in FIG. 6b.

6a, at pH 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9 to 10, the gold nanoparticle colorimetric sensor was stable without changing its intrinsic color, and arsenic ion (As ^{3 +} ) 2.0 μM After adjusting the concentration to , the sample showed a blue color at pH 3, 4, to 4.5. Referring to the wavelength of 525 nm in FIG. 6b , the nanoparticles formed at a pH of 4.5 exhibit the strongest absorbance.

5 and 6 above ^{, the optimum pH for detecting mercury ions (Hg 2 +} ) is 3 to 7, and the ^{optimum conditions for detecting arsenic ions (As 3+} ) are at pH 3 to 4.5, respectively, overlapping mercury ions and arsenic ions Selectivity can be increased without detection. And in the case of pH 4 and 4.5, it is an environmental condition that can detect mercury ions and arsenic ions simultaneously.

Example 5: D- penicillamine (D- Penicillamine ) Mercury ions in the colorimetric sensor solution according to the solution concentration ( Hg ^{2 +} ) and arsenic ions ( As ^{3 +} ) for reactivity experiments

After adjusting the pH of the colorimetric sensor solution obtained in Preparation Example 2 to 6, the color change and stability of the colorimetric sensor were tested to search for optimal conditions for detecting ^{mercury ions (Hg 2 + ) according to the concentration of DPL, a surface modification material.} More specifically, a colorimetric sensor solution surface-modified with 0, 1, 2, 3, 4, 5, 6, 7, 8 to 9 mM D-penicillamine was prepared. A photograph of the sample according to each D-penicillamine concentration is shown in FIG. 7A, and the absorbance spectrum of each sample was measured by UV-Vis, and the absorbance spectrum is shown in FIG. 7B.

Referring to FIG. 7a, the colorimetric sensor of gold nanoparticles at 0, 1, 2, 3, 4, 5, 6, 7, 8 to 9 mM did not change its intrinsic color and was stable, and mercury ions (Hg ^{2 +} ) at 1.0 μM After adjusting the concentration, the sample exhibited a blue color at 1, 2, 3, 4, 5, 6, 7, 8 to 9 mM.

Referring to the wavelength of 525 nm in FIG. 7b , the nanoparticles formed under the condition that the concentration of D-penicillamine is 5 mM exhibits the strongest absorbance.

In Figure 7c, the modifier concentration and reaction color change according to the addition of 2 μM ^{of Hg 2 +} ions at the modifier D-penicillamine concentration (1 ~ 9 mM) were observed. It can be seen that the stability of the particles at 1 to 9 mM was very good, and the unmodified solution (ctrl) used as a control did not react with mercury.

At pH 4.5, arsenic ions (As ^{3 +} ) are also shown in FIG. 8a for pictures of samples according to each D-penicillamine concentration, and absorbance spectra were obtained by measuring each sample with UV-Vis. 8b.

Referring to FIG. 8a, the gold nanoparticle colorimetric sensor at 0, 1, 2, 3, 4, 5, 6, 7, 8 to 9 mM was stable without changing its intrinsic color, and arsenic ion (As ^{3 +} ) 2.0 μM Samples after adjusting the concentration to 1, 2, 3, 4, 5, 6, 7, 8 to 9 mM showed a blue color.

Referring to the wavelength of 525 nm in FIG. 8B , the nanoparticles formed under the condition that the concentration of D-Penicillamine is 5 mM exhibits the strongest absorbance.

In Figure 8c, the modifier concentration and reaction color change according to the addition of 2 μM ^{of As 2 +} ions at the modifier D-penicillamine (D-Penicillamine) concentration (1 ~ 9 mM) were observed. It can be seen that the stability of the particles at 1 to 9 mM was very good, and the unmodified solution (ctrl) used as a control did not react with mercury.

Example 6: Gold nanoparticle reactivity according to reaction temperature

After adjusting the pH of the colorimetric sensor solution obtained in Preparation Example 2 to 6, 5mM DPL, 8 samples were collected, the mercury ion (Hg ^{2 +} ) concentration was also 1 μM, and the reaction temperature was adjusted to 10, 20, 30, 40, 50, 60, 70, and 80 ° C., respectively, were reacted for 30 minutes, and the absorbance ratio of each sample (A ₇₃₀ /A ₅₂₅ ) was compared. The contents are shown in FIG. 9A, and the comparison result shows the highest absorbance ratio at 30°C.

As for arsenic ions (As ^{3 +} ), after adjusting the pH of the colorimetric sensor solution obtained in Preparation Example 2 to 4.5, the D-penicillamine concentration was set to 5 mM, and then 8 samples were collected and arsenic ions (As ^{3 + )} ) After making the concentration to 2 μM, the reaction temperature was changed to 10, 20, 30, 40, 50, 60, 70, and 80 ℃ for 30 minutes, and the absorbance ratio of each sample (A ₇₃₀ /A ₅₂₅ ) were compared. The contents are shown in FIG. 9b, and the comparison result shows the highest absorbance ratio at 30°C.

Example 7: Reactivity of gold nanoparticles according to reaction time

After adjusting the pH of the colorimetric sensor solution obtained in Preparation Example 2 to 6, it was made to 5 mM DPL. After taking 5 samples and adding 0, 0.4, 0.8, 1.2 and 1.6 μM of ^{mercury ions (Hg 2 +} ) respectively at room temperature, react for 15 minutes, and measure _{the absorbance ratio over time (A 730} /A _{525 )} Thus, the results are shown in Figure 10a. Referring to FIG. 10A , the absorbance ratio (A ₇₃₀ /A ₅₂₅ ) increases very quickly and constantly up to 3 minutes, and after that, the absorbance ratio (A ₇₃₀ /A ₅₂₅ ) is constant and almost no reaction occurs. Therefore, it is considered that the reaction of the nanoparticle colorimetric sensor of the above conditions is completed within 5 minutes.

As for arsenic ions (As ^{3 +} ), the pH of the colorimetric sensor solution obtained in Preparation Example 2 was adjusted to 4.5, and the D-penicillamine concentration was set to 5 mM. After taking 5 samples and adding 0.1, 0.5, 1, 1.5 and 2 μM of ^{arsenic ions (As 3 +} ) respectively at room temperature, react for 15 minutes, and measure _{the absorbance ratio over time (A 730} /A _{525 )} Thus, the results are shown in Figure 10b. As a result, it is considered that the reaction of the nanoparticle colorimetric sensor under the above conditions is completed within 3 minutes.

Example 8: mercury ion ( Hg ^{2 +} ) Sensitivity of colorimetric sensor solution according to concentration and calibration curve

In order to confirm that the quantitative measurement ^{of mercury ions (Hg 2 +} ) is possible in the colorimetric sensor solution (pH 6) obtained in Preparation Example 2 ^{, mercury ions (Hg 2 +} ) were added to increase the concentration of ^{mercury ions (Hg 2 + ).} 0, 0.2, 0.4, 0.6, 0.8, 1, 1.2 and 1.4 μM, respectively. The reaction was carried out at a reaction temperature of 25° C. for 10 minutes and the color change was observed.

A photograph of each sample is shown in FIG. 11A , FIG. 11B is a UV-vis spectrum, and the absorbance ratio (A ₇₃₀ /A ₅₂₅ ^{) according to the mercury ion (Hg 2+} ) concentration is shown in FIG. 11C .

Referring to FIG. 11A , it can be seen that the blue color gradually increases as the amount of ^{mercury ions (Hg 2 + ) added increases.}

11b, it can be seen that the tendency of the spectral peaks of 525 nm and 730 nm according to the amount of mercury ions added is increased, and as the mercury ions are increased, the 730 nm region increases and the 525 nm region decreases. . Through this, a quantitative curve can be drawn through the absorbance ratio in the 525 and 730 nm regions.

Referring to FIG. 11C , the calibration curve (r ² = 0.9689) according to the absorbance ratio is maintained, and the linear equation shows y = 0.6021x-0.0839. Through this, it can be seen that the colorimetric sensor solution according to the present invention ^{can be quantitatively analyzed for mercury ions (Hg 2 + ).} Table 1 below shows the details of the straight line equation in FIG. 11C.

equation y = a + b*x weight instrumental Residual Sum of Squares (RSS) 7.64926 Pearson's r 0.98662 Adj. R-Square 0.96899 value standard error B intercept 0.0839 0.0369 inclination 0.6021 0.0406

Example 9: Arsenic ion ( As ^{3 +} ) Sensitivity of colorimetric sensor solution according to concentration and calibration curve

In order to confirm whether the quantitative measurement ^{of arsenic ions (As 3 +} ) is possible in the colorimetric sensor solution (pH 4.5) obtained in Preparation Example 2 ^{, arsenic ions (As 3 +} ) were added to increase the concentration of ^{arsenic ions (As 3 + ).} 0, 0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75 and 2 μM, respectively. The reaction was carried out at a reaction temperature of 25° C. for 10 minutes, and color change was observed. A photograph of each sample is shown in FIG. 12A, FIG. 12B is a UV-vis spectrum, and an absorbance ratio (A ₇₃₀ /A ₅₂₅ ^{) according to an arsenic ion (As 3 +} ) concentration is shown in FIG. 12C.

Referring to FIG. 12A , it can be seen that the blue color gradually increases as the amount of arsenic ions (As ^{3 + ) added increases.}

12b, it can be seen that the trend of the spectral peaks at 525 nm and 730 nm according to the amount of arsenic ions added is increased, and as the arsenic ions increase, the 730 nm region increases and the 525 nm region decreases. . Through this, a quantitative curve can be drawn through the absorbance ratio in the 525 and 730 nm regions.

Referring to FIG. 12C , the calibration curve (r ² = 0.9328) according to the absorbance ratio is maintained, and the linear equation shows y = 0.5399x-0.0252. Through this, it can be seen that the colorimetric sensor solution according to the present invention ^{is capable of quantitative analysis of arsenic ions (As 3 + ).} Table 2 below shows the details of the straight line equation in FIG. 12C.

equation y = a + b*x weight instrumental Residual Sum of Squares (RSS) 38.5716 Pearson's r 0.96586 Adj. R-Square 0.93288 value standard error B intercept -0.0225 0.0721 inclination 0.5399 0.0547

Example 10: other ionic compounds Selectivity of colorimetric sensor solution for

The pH of the colorimetric sensor solution obtained in Preparation Example 2 was adjusted to 6, and it was made to 5 mM DPL. In order to confirm the selectivity for various cations and anions, 32 samples were prepared and Li ⁺ (1), Na ⁺ (2), K ⁺ (3), Ag ⁺ (4), Ba ^{2 +} (5), ^{Ca 2 + (6), Cd} 2 + (7), Co 2 + (8), Cu 2+ (9), Hg 2 + (10), Mn 2 + (11), Mg 2 + (12), Ni ^{2 +} (13), Pb ^{2 +} (14), Sn ^{2 +} (15), Zn ^{2 +} (16), Al ³⁺ (17), As ^{3 +} (18), Fe ^{3 +} (19), Ga ^{2 +} (20), Ti ^{3 +} (21), Ge ^{4 +} (22), Cr ^{6 +} (23) and 8 anions NO ₂ ^- (24), NO ₃ ^- (25), PO ₄ ^3- (26) , SO ₄ ^2- (27), F ^- (28), Cl ^- (29), Br ^- (30), I ^- (31).

In FIG. 13a , after adding 10 μM to each gold nanoparticle colorimetric sensor sample, reacted at a reaction temperature of 30° C. for 25 minutes, _{and observed a change in the absorbance ratio (A 730} /A ₅₂₅ ). As a result, mercury ions (Hg ²⁺ ) are different cations. , it can be seen that, unlike anions, the gold nanoparticle colorimetric sensor according to the present invention has very good selectivity.

In FIG. 13b , after adding 10 μM to each gold nanoparticle colorimetric sensor solution (pH 4.5) sample, the reaction was performed at a reaction temperature of 30° C. for 25 minutes, and the _{change in the absorbance ratio (A 730} /A ₅₂₅ ) was observed. As a result, arsenic ions (As ³ It can be seen that the gold nanoparticle colorimetric sensor according to the present invention has very good selectivity, unlike other cations and anions in ^{+ ).}

That is, the detection system of the present invention has higher ^{sensitivity to mercury ions (Hg 2 +} ) and arsenic ions (As ^{3 +} ) than other cations and anions, so that it can be discriminated with the naked eye, and other cations, It can be confirmed that there is a certain selectivity that is distinguished from an anion.

Example 11: Evaluation of the effectiveness of a detection system comprising functionalized gold nanoparticles

For mercury ion (Hg ^{2 +} ) and arsenic ion (As ^{3 +} ) detection experiment in mineral water, tap water and pond water were filtered through a 0.2 μm filter and then mercury ion (Hg ^{2 +} ) and arsenic ion (As ^{3 ) +} ) was checked at pH 6 or pH 4.5, and after confirming that mercury ions (Hg ^{2 +} ) and arsenic ions (As ^{3 +} ) were not present, this was used as a blank solution.

^{A sample was prepared in which mercury ions (Hg 2 +} ) and arsenic ions (As ^{3 +} ) were added to the blank solution so as to be 1, 5 to 20 nM, respectively, and then the absorbance was measured by UV-vis, and in Example 5 The added concentration, the average of the detected amount, the recovery (%), and the coefficient of variation (CV) were measured using the prepared calibration curve, and the results are shown in Tables 3 and 4 below.

Mercury ions (Hg ^{2 +} ) (sensor solution: pH 6.0) Addition concentration (nM) Found mean ±SD ^a (nM) Recovery (%) Coefficient of variation (n=7, %) detection limit
(nM) tap water One 0.94±0.056 94.5 7.1 26.3 10 9.87±0.22 98.7 7.3 20 20.11±0.43 100.5 7.2 Pond water One 0.95±0.055 95.5 7.2 25.0 10 9.75±0.24 97.5 7.4 20 19.48±0.68 97.4 7.3

Arsenic ion (As ^{3 +} ) (sensor solution: pH 4.5) Addition concentration (nM) Found mean ±SD ^a (nM) Recovery (%) Coefficient of variation (n=7, %) detection limit
(nM) tap water One 0.94±0.063 94.6 7.2 21.5 5 4.87±0.45 97.4 7.1 10 10.11±0.23 101.1 7.5 Pond water One 0.95±0.098 95.9 7.3 22.5 5 4.75±0.54 95.1 7.7 10 10.48±0.56 104.8 7.5

^{As shown in Tables 3 and 4 above, the limit of detection (LOD) of mercury ions (Hg 2 +} ) using the gold nanoprobe colorimetric sensor solution surface-modified with D-penicillamine was 26.3 nM in tap water and 25.0 nM in pond water. It was. In the samples to which 1 nM, 5 nM, and 10 nM were added in tap water, the detection amounts were 0.94±0.056, 9.87±0.22, and 20.11±0.43, respectively, very close to the actual addition amount, and the recovery rates were excellent at 94.5, 98.7 and 100.5. , and the coefficients of variation were also excellent at 7.1, 7.3, and 7.2.

The limit of detection (LOD) of arsenic ions (As ^{3 +} ) was 21.5 nM in tap water and 22.5 nM in pond water. In the samples to which 1 nM, 5 nM, and 10 nM were added in tap water, the detection amounts were 0.94±0.063, 4.87±0.45 and 10.11±0.23, respectively, which were very close to the actual addition amount, and the recovery rates were excellent at 94.6, 97.4 and 101.1. The coefficient of variation was also excellent at 7.2, 7.1, and 7.5.

In general, although many obstacles may exist in the detection of ^{mercury ions (Hg 2 +} ) and arsenic ions (As ^{3 +} ) for products composed of various compositions such as drinking water and beverages, D-penicillamine according to the present invention It can be seen that the colorimetric sensor solution including the surface-modified nanoprobe has very good performance and high selectivity.

The embodiments of the present invention described above should not be construed as limiting the technical spirit of the present invention. The protection scope of the present invention is limited only by the matters described in the claims, and those skilled in the art can improve and change the technical idea of the present invention in various forms. Accordingly, such improvements and modifications will fall within the protection scope of the present invention as long as it is apparent to those of ordinary skill in the art.

Claims (15)

A colorimetric sensor for detecting mercury ions and arsenic ions, comprising surface modification-nanoparticles,
The surface modification-nanoparticles are gold nanoparticles; and a modifier represented by the following formula (1);
Wherein the modifier is bound to the surface of the gold nanoparticles, a colorimetric sensor for detecting mercury ions and arsenic ions:
[Formula 1]

According to claim 1,
The size of the gold nanoparticles is 20 nm or less, a colorimetric sensor for detecting mercury ions and arsenic ions. According to claim 1,
A colorimetric sensor for detecting mercury ions and arsenic ions, wherein the concentration of the modifier is 1 to 10 mM based on the total volume of the colorimetric sensor solution. According to claim 1,
The color of the colorimetric sensor is red,
A colorimetric sensor for detecting mercury ions and arsenic ions, which changes color to a dark blue series when mercury ions and arsenic ions are added. According to claim 1,
The colorimetric sensor for detecting mercury ions and arsenic ions has a pH of 3 to 4.5 for detecting mercury ions and arsenic ions of the colorimetric sensor. According to claim 1,
The colorimetric sensor is a colorimetric sensor for detecting mercury ions and arsenic ions to simultaneously detect mercury ions and arsenic ions. According to claim 1,
A colorimetric sensor for detecting mercury ions and arsenic ions, wherein the colorimetric sensor has an absorbance ratio of 0.2 to 1.2. According to claim 1,
The colorimetric sensor is a colorimetric sensor for detecting mercury ions and arsenic ions, characterized in that it detects mercury ions and arsenic ions at 20 to 50 °C. According to claim 1,
The color emission wavelength of the colorimetric sensor is 450 to 600 nm,
A colorimetric sensor for detecting mercury ions and arsenic ions, wherein the color emission wavelength is 520 to 620 nm when the colorimetric sensor detects mercury ions. According to claim 1,
The color emission wavelength of the colorimetric sensor is 450 to 600 nm,
A colorimetric sensor for detecting mercury ions and arsenic ions, wherein the color emission wavelength is 650 to 800 nm when the colorimetric sensor detects arsenic ions. According to claim 1,
A colorimetric sensor for detecting mercury ions and arsenic ions, wherein when the colorimetric sensor reacts with mercury ions and arsenic ions, the reaction time is within 10 minutes. According to claim 1,
A colorimetric sensor for detecting mercury ions and arsenic ions, wherein the colorimetric sensor has a detectable concentration of mercury ions and arsenic ions of 25 nM or more. A method for detecting mercury ions and arsenic ions, comprising:
13. An input step of introducing a sample to be detected into the colorimetric sensor for detecting mercury ions and arsenic ions according to any one of claims 1 to 12;
a reaction and waiting step between the colorimetric sensor and the sample to be detected; and
and a sensing step of detecting mercury ions and arsenic ions in the detection target sample by color change of the colorimetric sensor. 14. The method of claim 13,
The method of detecting mercury ions and arsenic ions further comprising; adjusting at least one of pH and temperature of the colorimetric sensor before the inputting step. As a first solution, preparing a solution in which gold nanoparticles are dispersed;
As a second solution, preparing a solution containing a modifier represented by the following formula (1); and
A method for manufacturing a colorimetric sensor for detecting mercury ions and arsenic ions, comprising: mixing the first solution and the second solution to prepare the surface-modified nanoparticles in which the modifier is bonded to the surface of the gold nanoparticles;
[Formula 1]

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