KR20140108002A - Ammonia sensing fluorescent sensor membrane including oxazine, method for preparing the same and method for detecting ammonia concentration in water using the same - Google Patents

Abstract

Provided are an ammonia detecting fluorescent sensor membrane including oxazine, a method for manufacturing the same, and a method for detecting a concentration of ammonia in water using the same. The fluorescent sensor film includes a cellulose based polymeric matrix and an oxizine pigment contained in the polymeric matrix to be fixed. Accordingly, an in-use range of oxazine can be widened, ammonia can be more effectively detected, and contact with ammonia can be increased.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a fluorescent sensor film for detecting ammonia containing an oxazine photo, a method for producing the same, and a method for detecting ammonia in water using the same,

The present invention relates to a sensor, and more particularly, to a fluorescence sensor film for detecting ammonia containing an ox picture and a method of manufacturing the same.

Fluorescence sensors have recently been actively studied because they can more easily detect selective substances by using fluorescence change through selective binding with an inclusion compound by connecting a supramolecular compound to a fluorescent substance.

On the other hand, the concentration of aquatic ammonia is an important parameter which is considered in environmental management, and a technique using a fluorescent dye to detect such ammonia by an optical sensor is being developed. As an example, rhodamine, fluorine-based amine compounds, coumarin, 1-hydroxypyrene-3,6,8-trisulphonate, and oxazine perchlorate have been studied for the detection of ammonia. Among them, oxazine 170 perchlorate has excellent fluorescence properties as fluorescent dyes, but it is not known to those skilled in the art, and thus it is hardly used as fluorescent dyes.

Recently, studies have been made on an ammonia detection sensor using jade photographic perchlorate. A conventional technique related to this technique is disclosed in US Patent Publication No. 2012/0288953.

US Patent Publication No. 2012/0288953 discloses a sensor using the absorption characteristics of the jade photographic perchlorate after coating an optical fiber with an optical photographic perchlorate dye. However, there is a problem in that the efficiency of the manufactured sensor is low because the fluorescence characteristic and the fluorescence emission intensity of the jade photographic perchlorate are not used in the production of the sensor.

In addition, when the jade photographic perchlorate fluorescent dye is used alone, the function of the dye may be deteriorated due to external environment such as alkali, salt, visible light and ultraviolet rays, and it has a disadvantage that it has limited contact with ammonia.

Accordingly, it is an object of the present invention to provide a fluorescent sensor film for detecting ammonia containing jade photographic material having a wider range of use by using jade photographic perchlorate as a fluorescent dye.

The present invention also provides a fluorescent sensor film for detecting ammonia, which prevents deterioration of the function of a dye due to alkali, salt and visible light and ultraviolet rays, which occurs when the jade photographic perchlorate is used singly, and which has high contact with ammonia, .

In order to solve the above problems, one aspect of the present invention provides a fluorescent sensor film for detecting ammonia. The fluorescent sensor film may be a cellulose-based polymer matrix; And an oxazine dye immobilized and contained in the polymer matrix.

The polymer matrix may further include silica formed by a sol-gel reaction of an alkoxysilane compound. As a preferable example, the alkoxysilane compound may include 3-aminopropyltrimethoxysilane (APTMS) and 3-glycidoxypropyl And trimethoxysilane (GPTMS).

The cellulose-based polymer may be selected from the group consisting of methylcellulose, ethylcellulose, carboxymethylcellulose, cellulose acetate phthalide, stearylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose, Lt; RTI ID = 0.0 > ethylcellulose. ≪ / RTI >

The jade photographic dye may comprise jade photographic perchlorate.

In addition, the fluorescence sensor film can emit two different fluorescence wavelengths in contact with ammonia.

According to another aspect of the present invention, there is provided a method of fabricating a fluorescent sensor film for detecting ammonia. The method comprises: preparing a mixed solution by mixing a cellulose-based polymer and an oxazine dye in an organic solvent; And coating the mixed solution on a substrate, followed by drying.

The step of preparing the mixed solution may further include adding an alkoxysilane compound to the mixed solution to prepare the mixed solution as a sol-gel reactionable sol solution.

The cellulose-based polymer may be selected from the group consisting of methylcellulose, ethylcellulose, carboxymethylcellulose, cellulose acetate phthalide, stearylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose, Lt; RTI ID = 0.0 > ethylcellulose. ≪ / RTI >

The alkoxysilane compound may include at least one of 3-aminopropyltrimethoxysilane (APTMS) and 3-glycidoxypropyltrimethoxysilane (GPTMS). As a preferable example, -Aminopropyltrimethoxysilane (APTMS) and the 3-glycidoxypropyltrimethoxysilane (GPTMS) in a volume ratio of 4: 1.

According to another aspect of the present invention, there is provided a method for detecting ammonia in water. Preparing a fluorescent sensor film for detecting ammonia comprising a cellulose-based polymer matrix and an oxazine dye immobilized on the polymer matrix; Measuring two different fluorescence emission wavelengths in a contact condition between ammonia and the fluorescence sensor film; Setting a concentration range of ammonia detectable by the fluorescence sensor film; Calculating a ratio of fluorescence intensity at the two fluorescence wavelengths according to the concentration within the concentration range; And measuring an unknown ammonia concentration contained in the water based on the calculated ratio of the fluorescence intensity.

The ammonia concentration range may include 1 to 60 ppm.

According to the present invention, since the fluorescence sensor film for detecting ammonia containing the jade photograph can broaden the use range of the jade photograph by using the jade photograph as the fluorescent dye, the sensitivity can be higher than that conventionally used only as the absorbance dye The detection of ammonia can be more effectively performed.

Further, by forming the fluorescent sensor film by combining the cellulose-based polymer and the jade photographic perchlorate, the dye can be stabilized from alkali, salt, visible light and ultraviolet rays, and can improve the contact with ammonia.

1 is a graph showing the fluorescence spectrum of jade photographic perchlorate used in the present invention.
FIG. 2 is a photograph showing the color change according to the ammonia concentration change of the fluorescent sensor film prepared in Production Example 1. FIG.
FIG. 3 is a graph showing changes in fluorescence according to the concentration of ammonia in the fluorescence sensor film prepared in Production Example 1. FIG.
4 is a calibration curve (a) showing the fluorescence intensity according to the ammonia concentration of the fluorescent sensor film prepared in Production Example 3 and a calibration curve (b) calculated as a ratio between two emission wavelengths.
5 is a graph showing the reversibility of the fluorescent sensor film according to Production Example 2. Fig.
6A is a graph showing the fluorescence intensity according to the pH change of the fluorescent sensor membrane prepared in Production Example 5. FIG.
FIG. 6B is a graph showing changes in fluorescence intensity according to pH when 10 ppm of ammonia is added to the production example. FIG.
7 is a graph (a) showing fluorescence intensity at emission wavelengths of 630 nm and 565 nm and a graph (b) showing fluorescence intensity after calculating the ratio of emission wavelengths of 630 nm and 565 nm, according to salinity of the fluorescence sensor membrane prepared in Production Example 5. FIG.
8 is a graph showing fluorescence intensity of a fluorescent sensor membrane prepared in Preparation Example 4 according to optical rotation and time.
9 is a graph showing the change in fluorescence intensity of the fluorescence sensor film prepared in Production Example 3 after the initial use and one month's use.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Like reference numerals designate like elements throughout the specification.

According to an embodiment of the present invention, there is provided a fluorescent sensor film for detecting ammonia comprising a cellulose-based polymer matrix and an oxazine dye immobilized in the polymer matrix.

The jade photographic dye may contain jade photographic perchlorate, and the jade photographic perillaite has a broad spectrum and has high fluorescence efficiency, so that it can have excellent properties as a fluorescent dye. Also, the wavelength of fluorescence is longer than the wavelength of the absorbed light, and the intensity of the fluorescence emitted by the fluorescent material is stronger as the concentration of the fluorescent material in the sample is higher. Therefore, it is possible to quantitatively analyze the fluorescent substance in the sample using these characteristics. This method has a very high sensitivity as compared with the detection method using absorbance, and it is possible to selectively detect only a component intended for detection. Furthermore, substances which do not generate fluorescence have the advantage that they can be analyzed after they are converted into a fluorescent substance using a reductive enzyme or a chemical reaction.

The cellulose-based polymer is a matrix-forming substance serving as a support of a fluorescent sensor membrane. Examples of the matrix polymer include methylcellulose, ethylcellulose, carboxymethylcellulose, cellulose acetate phthalide, stearylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, ≪ / RTI > hydroxypropylcellulose, and hydroxypropylcellulose. Among them, ethyl cellulose can be a most suitable candidate material in that an oxazine dye is effectively fixed while an oxazine dye provides a good environment for ammonia contact. The ethylcellulose has durability and is soluble in a wide range of solvents and is resistant to alkaline and saline solutions. Also, it does not change color even when exposed to a wide range of light including visible light and ultraviolet light.

Meanwhile, the polymer matrix may further include silica formed by a sol-gel reaction of an alkoxysilane compound. The sol-gel reaction is a method for producing sol-gel ceramics. The sol is a solid-liquid state in which colloidal or inorganic monomolecular solid molecules are dispersed. As the reaction continues, the dispersed solid molecules are polymerized to form continuous It may become a gel state that has lost its fluidity by forming a solid network structure. The fluorescence sensor membrane using the sol-gel reaction has a merit that it is simple to operate, prevents the penetration of foreign substances, reduces the possibility of contamination, and does not require a separate sample collection step in the sample measurement. Further, in the hydrolysis reaction during the sol-gel reaction, the alkoxysilane compound reacts with water to form a hydroxyl group (-OH), and the hydroxyl group generated by hydrolysis reacts with the hydroxyl group or alkoxy group of another alkoxysilane compound, It is possible to produce metal oxide bonds through the piers. The sol particles formed by these reactions can form a network structure by polycondensation. Such a sol-gel reaction can be used for a long time when immobilized to a fluorescent dye, and it is possible to decrease the activity of the fluorescent dye, thereby increasing the precision of the fluorescent sensor film. The alkoxysilane compound is not particularly limited as long as it is a substance capable of sol-gel reaction, but preferably at least one of 3-aminopropyltrimethoxysilane (APTMS) and 3-glycidoxypropyltrimethoxysilane (GPTMS) . ≪ / RTI > When 3-aminopropyltrimethoxysilane (APTMS) and 3-glycidoxypropyltrimethoxysilane GPTMS are both used, they are mixed at a volume ratio of 4: 1 to form a dense film by blocking phase separation can do.

According to another embodiment of the present invention, there is provided a method of detecting ammonia using the above-described fluorescence sensor film. The ammonia detection method according to this embodiment is as follows. First, the fluorescent sensor membrane is prepared, and then two different fluorescence emission wavelengths emitted from the fluorescent sensor membrane are measured under the condition of contact between ammonia and the fluorescent sensor membrane. Here, two different fluorescence emission wavelengths appear because the spectra of the jade photons changed as the dyes, which were in the anion state, combine with ammonium ions.

After this, the concentration range of ammonia that can be detected is set. At this time, the concentration of free ammonia can be calculated by the following Henderson-Hasselbalch formula.

log [NH ₃ ] = log [NH ₄ ⁺ ] - 2.44

Then, the ratio of the fluorescence intensities at the two fluorescence wavelengths according to the concentration within the set concentration range is calculated. The ratio calculation can be calculated by the ratio of the fluorescence intensities produced by the combination of the ammonia ion and the jade photographic perchlorate at two emission wavelengths.

Finally, the unknown ammonia concentration contained in the water is measured based on the calculated fluorescence intensity ratio.

Hereinafter, preferred examples for the understanding of the present invention will be described. It should be understood, however, that the following examples are intended to aid in the understanding of the present invention and are not intended to limit the scope of the present invention.

≪ Sol-gel solution Manufacturing example >

Aminopropyltrimethoxysilane (APTMS) and 3-glycidoxypropyltrimethoxysilane (GPTMS) were mixed in a ratio of 25: 1 to prepare a sol-gel solution used as a support for a fluorescent sensor film for detecting ammonia. And mixed in a 99% ethanol solution at a volume ratio of 6.25. To this, a certain amount of distilled water and 35% HCl serving as an acid catalyst were added at a volume ratio of 4% to proceed hydrolysis and left at room temperature for at least 2 hours.

<For ammonia detection Fluorescence sensor membrane Manufacturing example >

The sol-gel solution prepared in the preparation example of sol-gel solution and ethyl cellulose, which is a fluorescent dye for detecting ammonia (oxazine 170 perchlorate), and ethyl cellulose were mixed in respective amounts as shown in the following table, and aged at room temperature for 4 hours . The mixture was then coated on a 24-well microplate (NUNC Co., Denmark) and dried at 60 for 12 hours.

Further, in order to know the effect of the sol-gel solution, experiments were also conducted on the production example (Production Example 1) in which only the above-mentioned jade photographic 170 perchlorate and ethyl cellulose were mixed.

Classification Ophthalmic photograph 170 Perchlorate (ug) Sol-gel solution (ul) Ethylcellulose (ug) Production Example 1 20 0 30 Production Example 2 20 100 25 Production Example 3 20 100 20 Production Example 4 10 100 10 Production Example 5 10 50 50 Production Example 6 10 80 20

<For ammonia detection Fluorescence sensor membrane  characteristic Analysis 1 >

In order to monitor the ammonia detecting fluorescent sensor membrane prepared in the above production example, the fluorescent spectrum and the optical characteristics of the ammonia fluorescent sensor membrane were investigated using a multiplex fluorescence analyzer after being coated on a 24-well microplate (Nunc co., Denmark). Six fluorescence sensor films prepared in the above Preparation Example were subjected to 2-D fluorescence spectroscopy using a fluorescence spectrophotometer (F-4500, Hitachi Japan) to determine the optimum detection wavelength.

<Fluorescence sensor membrane characteristics for ammonia detection Analysis 2 >

A multifunctional fluorescent microplate reader (Safire2, Tecan Austria GmbH, Austria) was used to analyze the ammonia detection of the fabricated fluorescence sensor membrane. The concentration range of ammonia was set to 1-60ppm and the intensity of fluorescence was measured according to the concentration of ammonia at 470nm excitation wavelength (λ _ex ) and two emission wavelengths (λ _em = 565, 630).

The concentration range of the ammonia was calculated by the following Henderson-Hasselbalch formula.

log [NH ₃ ] = log [NH ₄ ⁺ ] - 2.44

On the other hand, the reason that the excitation wavelength is different from the emission wavelength is that when the fluorescence is emitted, heat is released from the vibration level and only the remaining energy is released, resulting in a stroke shift.

Fluorescence intensity according to the pH was measured by adding a solution having a pH range of 1-11 to the prepared fluorescence sensor membranes and fluorescence intensity was measured according to salinity by adding 0 mM, 50 mM, 100 mM, 300 mM, and 500 mM sodium chloride solution .

Finally, we experimented with the whitening of ammonia over exposure time using LED with λ = 470nm. Analysis of the data was analyzed by one-way analysis of variance (ANOZVA). Statistical tests were performed using software InStat (version 3.01, GraphPad Software Inc., Sandiego, USA) with a p-value <0.05 between samples.

1 is a graph showing the fluorescence spectrum of jade photographic perchlorate used in the present invention.

Referring to FIG. 1, it can be seen that the jade photographic perchlorate has a wide range of excitation spectrum and high fluorescence efficiency. In particular, it can be seen that it strongly absorbs light having a wavelength larger than 550 nm, and emits fluorescence around 640 nm.

Jade Photographic Perchlorate has the highest fluorescence efficiency among the photographic derivatives of oxazines, and the electron attracting effect of the nitrogen atoms in the molecular structure makes the amino group more acidic, and the jade photographic image is weakly acidic.

Fig. 2 is a photograph showing the reaction between Preparation Example 1 and ammonia before and after the reaction.

Referring to FIG. 2, when ammonia is added to the sensor film containing jade photographic perchlorate, the sensor film, which was blue, turns red. This is because the slightly acidic jade photographic image can give a proton to the weak base, so when adding the ammonium hydroxide solution to the dye of the jade photographic perchlorate, the proton of one of the amino groups is lost and the color changes from blue to red. The order of the reaction of the above-mentioned oxazine dye and ammonia is based on the following formula.

NH ₃ + H ₂ O ↔ NH ₄ ⁺ OH ^- (Equation 1)

NH ₄ ⁺ OH ^- + H ⁺ Dye ^- ↔ NH ₄ ⁺ Dye ^- + H ₂ O (Equation 2)

NH ₄ ⁺ Dye ^- ↔ H ⁺ Dye ^- + NH ₃ (Equation 3)

This reaction results in a significant fluorescence band change as described by a semi-classic oscillator model that predicts two dichroic electronic band systems associated with the jade photographic dye.

When water and ammonia are reacted, ammonium ions and hydroxide ions are produced as shown in the above formula (1). The resulting ammonium ions combine with the dye ions and the sensor membrane turns red (see FIG. 2 a). When the concentration of ammonia is reduced (for example, when the sensor membrane is washed with water), the reaction as shown in Equation 3 occurs and the sensor membrane turns blue again (see FIG. 2 b).

FIG. 3 is a graph showing changes in fluorescence according to the concentration of ammonia according to Production Example 1. FIG.

Referring to FIG. 3, peaks can be found at emission wavelengths of 565 nm and 630 nm when the excitation wavelength is 470 nm. It can be seen that at an ammonia concentration of 1-60 ppm, the jade photographic perchlorate fluorescent dye emits fluorescence at emission wavelengths of 565 nm and 630 nm.

At 1 ppm ammonia concentration, peak 630 nm showed the highest fluorescence intensity, while peak 565 nm showed the lowest fluorescence intensity. However, when the ammonia concentration was increased from 1 ppm to 60 ppm, the fluorescence intensity was exchanged at peaks 630 nm and 565 nm, so that the fluorescence intensity decreased at the peak 630 nm and the fluorescence intensity increased at the peak 565 nm.

On the other hand, the band shift of about 10 nm between the emission wavelength (? _Em = 640 nm) of the jade photographic perchlorate dye in FIG. 1 and the emission wavelength (? _Em = 630 nm) The reason for the peak at 565 nm is due to the change in the spectra of the jade photographic image as the dyes were combined with ammonium ions.

4 is a calibration curve (a) showing the fluorescence intensity according to the ammonia concentration of the fluorescent sensor film prepared in Production Example 3 and a calibration curve (b) calculated as a ratio between two emission wavelengths. Referring to FIG. 4, the fluorescence intensity is higher at an emission wavelength of 630 nm as the concentration of ammonia is lowered, and the fluorescence intensity is higher at an emission wavelength of 565 nm as ammonia concentration is increased.

Referring to FIG. 4 (a), it can be seen that the fluorescence sensor membrane according to the present invention has a high sensitivity in the ammonia concentration range of 1-60 ppm. The regression coefficient (r ² ) = 0.96-0.97 was obtained based on the emission wavelength of 630 nm, and the linear detection range is divided into two ranges. The limit of detection (LOD) value was 0.227ppm in the range of ammonia concentration of 1-20ppm, and the detection limit was 0.179ppm in the range of ammonia concentration of 20-60ppm.

On the other hand, the regression coefficient (r ² ) was 0.92 based on the emission wavelength of 565 nm. The linear detection range was 0.047 ppm in the range of ammonia concentration of 1-10 ppm and detected in the range of ammonia concentration of 10-50 ppm The limit was 0.125 ppm.

Referring to FIG. 4 (b), the calibration curve obtained through the ratio calculation can be confirmed. This ratio calculation can be performed to obtain a high degree of confidence in the ammonia sensor, since the spectra of the jade photons that bind with ammonium ions change in the range of two emission wavelengths (? _Em = 565 nm, 630 nm). The ratio calculation was calculated by the ratio of the fluorescence intensity produced by binding to the photographic perchlorate at the emission wavelengths of 630 nm and 565 nm of the ammonia ion.

(λ_em=630nm(I630)),λ_em=565nm(I565))

(? _em = 630 nm (I630)),? _em = 565 nm (I565)

The advantage of the ratio calculation method is that it can normalize the change in fluorescence intensity not related to the target concentration.

For example, the temporal and spatial distribution of the measured fluorescence intensity can generally be varied due to the uneven distribution of the fluorescent end in the sensor, the dynamics of the fluorescent end in another medium, or the noise of the measurement system, such as a change in illumination intensity . However, according to the fluorescence sensor film of the present invention, not only the ammonia concentration but also factors that may affect the fluorescence intensity can be removed by the ratio calculation measurement, and also the background fluorescence intensity can be deducted.

Thus, fluorescence intensity ratio measurements using two emission wavelengths of 565 nm and 630 nm can provide better calibration curves than when using a single emission wavelength. With this calibration curve, any concentration range can be taken within the range of 1 to 60 ppm (LOD 1-60 ppm = 0.012 ppm) to calculate the linear sensing range with a regression coefficient of 0.95 or greater. That is, when the fluorescent sensor film according to the present invention is used, the concentration of ammonia can be detected with high reliability through the above-described ratio calculation measurement method.

FIG. 5 is a graph showing the reversibility of the fluorescent sensor membrane according to Production Example 2, wherein the change in fluorescence intensity was observed while alternating between 1 ppm ammonia and 10 ppm ammonia. As a result, while 1 ppm of ammonia was added, the fluorescence intensity was kept at a low value, but the fluorescence intensity was increased immediately after addition of 100 ppm of ammonia. This high reversibility was also observed at emission wavelengths of 630 nm and 565 nm, respectively, and at least 95% of the maximum fluorescence intensity was observed for 10 seconds when the initial fluorescence intensity was measured. This means that ethyl cellulose not only provides a good environment for the contact between the oxazine dye and ammonia, but also effectively immobilizes the jade photographic dye despite repeated measurements.

FIG. 6A is a graph showing the fluorescence intensities of the fluorescent sensor films prepared in Production Example 5 according to pH changes, and FIG. 6B is a graph showing fluorescence changes according to pH when 10 ppm of ammonia is added to the production example.

Referring to FIG. 6A, the fluorescence intensity was increased by 1-5% at an emission wavelength of 630 nm in the pH range of 5.5-7.5 without ammonia, but the intensity was not significantly changed when the pH was increased to pH 7.5-11 (p-value = 0.8805). Therefore, it can be seen that the fluorescent sensor film is stable at an emission wavelength of 630 nm when exposed to an alkali solution. In addition, since the emission wavelength of 565 nm is generated by the contact between ammonia and the dye, no peak was observed at the above wavelength in the absence of ammonia.

However, referring to FIG. 6B, when 10 ppm of ammonia was added to the pH solution, a peak appeared at 565 nm, and the fluorescence intensity rapidly increased particularly in the range of pH 5.5-7.5. This corresponds to the dissociation reaction of ammonia in water (Equation 1). If the pH is low, more ammonia molecules can be converted to ammonium ions, so the equilibrium shifts to the right and the amount of ammonium and dye ion pairs increases until the intensity of the jade photographic image in the alkaline environment is reached (Equation 2 ) To increase the fluorescence intensity at 565 nm.

As a result, when the ammonia was not added, the fluorescence intensity change at 630 nm in the pH range of 5.5-7.5 was 1-5%, but when ammonia was added, the emission wavelength was 630 nm and It can be seen that a peak occurs at 565 nm. This means that the fluorescent sensor membrane of the present invention is stable against pH change.

7 is a graph (a) showing fluorescence intensity at emission wavelengths of 630 nm and 565 nm and a graph (b) showing fluorescence intensity after calculating the ratio of emission wavelengths of 630 nm and 565 nm, according to salinity of the fluorescence sensor membrane prepared in Production Example 5. FIG.

When detecting ammonia in water, ammonia exists as ammonia or ammonium ion. In addition, the ratio of ammonia to ammonium ion depends on salinity in addition to pH. The proportion of non-ionized ammonia increases with increasing pH, but decreases with increasing salinity. Referring to FIG. 7, it can be seen that the peak increases within the range of 0-300 mM NaCl solution at an emission wavelength of 565 nm. This means that as the salinity increases, the proportion of ammonia decreases slightly, and therefore NH ₄ ⁺ ions can be used more than NH _{3 at} high salinity. However, as a result of graphing the data values through the above-mentioned ratio calculation, no significant effect was observed on the measurement of ammonia in the fluorescent sensor film even in the presence of salt.

8 is a graph showing fluorescence intensity of a fluorescent sensor membrane prepared in Preparation Example 4 according to optical rotation and time.

Theoretically, the jade photographic perchlorate dyes are light fade resistant in the visible light spectrum region. This property did not change much when the jade photographic perchlorate was immobilized on the support.

8, the fluorescence sensor film according to the present invention was exposed to LED 470 (λ = 470) for one month. As a result, the fluorescence intensity was slightly changed for two days from the exposure start date, There was no. There was a decrease in the fluorescence intensity of the fluorescence sensor membrane (~ 17.5%) after 4 weeks of continuous exposure, but this shows significantly better fade resistance than other fluorescent dyes.

9 is a graph showing changes in fluorescence intensity of the fluorescence sensor film prepared in Production Example 3 after initial use and one month's use. At this time, the fluorescence sensor film was irradiated with LED 470 (? = 470) for several seconds only at 150-200 times of measurement for one month, unlike the experiment shown in Fig.

Referring to FIG. 9, it can be seen that the fluorescence intensity and sensitivity of the fluorescence sensor film are stably maintained even after one month.

In summary, a fluorophore sensor membrane comprising a combination of jade photographic perchlorate and a cellulosic polymer has high sensitivity to ammonia detection, fast response time, high reversibility, resistance to alkalinity, stability to dye, stability to whitening, Lt; / RTI &gt; Furthermore, using the ratio calculation method based on the fluorescence intensity ratio between the two emission wavelengths of the fluorescence sensor film according to the present invention, linear detection characteristics can be realized at any ammonia concentration in the range of 1 to 60 ppm.

Claims (15)

Cellulosic polymer matrix; And
And a fluorescent dye sensor for detecting ammonia containing an oxazine dye immobilized on the polymer matrix. The method according to claim 1,
Wherein the polymer matrix further comprises silica formed by a sol-gel reaction of an alkoxysilane compound. 3. The method of claim 2,
Wherein the alkoxysilane compound comprises at least one of 3-aminopropyltrimethoxysilane (APTMS) and 3-glycidoxypropyltrimethoxysilane (GPTMS). The method according to claim 1,
Wherein the cellulose-based polymer is selected from the group consisting of methyl cellulose, ethyl cellulose, carboxymethyl cellulose, cellulose acetate phthalide, stearyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, and hydroxypropyl cellulose. Fluorescence sensor membrane for. The method according to claim 1,
Wherein the cellulosic polymer comprises ethyl cellulose. The method according to claim 1,
Wherein the photographic dye is a fluorescent sensor film for detecting ammonia containing an oxazine photo-perchlorate. The method according to claim 1,
Wherein the fluorescence sensor film emits two different fluorescence wavelengths under the condition of contact with ammonia. Preparing a mixed solution by mixing a cellulose-based polymer and an oxazine dye in an organic solvent; And
Coating the mixed solution on a substrate, and drying the mixed solution. 9. The method of claim 8,
Wherein the step of preparing the mixed solution further comprises adding an alkoxysilane compound to the mixed solution to prepare the mixed solution as a sol solution capable of sol-gel reaction. 9. The method of claim 8,
Wherein the cellulose-based polymer is selected from the group consisting of methyl cellulose, ethyl cellulose, carboxymethyl cellulose, cellulose acetate phthalide, stearyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, and hydroxypropyl cellulose. For producing a fluorescent sensor film. 9. The method of claim 8,
Wherein the cellulosic polymer comprises ethyl cellulose. 10. The method of claim 9,
Wherein the alkoxysilane compound comprises at least one of 3-aminopropyltrimethoxysilane (APTMS) and 3-glycidoxypropyltrimethoxysilane (GPTMS). 13. The method of claim 12,
Wherein the alkoxysilane compound comprises the 3-aminopropyltrimethoxysilane (APTMS) and the 3-glycidoxypropyltrimethoxysilane (GPTMS) in a volume ratio of 4: 1. In the method for detecting ammonia in water,
Preparing a fluorescent sensor film for detecting ammonia comprising a cellulose-based polymer matrix and an oxazine dye immobilized on the polymer matrix;
Measuring two different fluorescence emission wavelengths in a contact condition between ammonia and the fluorescence sensor film;
Setting a concentration range of ammonia detectable by the fluorescence sensor film;
Calculating a ratio of fluorescence intensity at the two fluorescence wavelengths according to the concentration within the concentration range; And
And measuring an unknown ammonia concentration contained in the water based on the ratio of the calculated fluorescence intensity. 15. The method of claim 14,
Wherein the concentration range of the ammonia is between 1 and 60 ppm.

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