KR102633297B1 - Lactate color sensor, lactate discoloration kit, and sensing method using thereof - Google Patents

Abstract

The present invention relates to a lactate color sensor, a lactate discoloration kit, and a sensing method using the same. More specifically, it is equipped with a small array containing transition metals, solvents, and bacteriophages as main components, and measures the presence or absence of analytes. Accordingly, the present invention relates to a bio-lactate color sensor that changes the color of the small array, a lactate discoloration kit, and a sensing method using the same.

Description

Lactate color sensor, lactate discoloration kit, and sensing method using the same {Lactate color sensor, lactate discoloration kit, and sensing method using the same}

The present invention relates to a lactate color sensor, a lactate discoloration kit, and a sensing method using the same. More specifically, it is equipped with a small array containing transition metals, solvents, and bacteriophages as main components, and measures the presence or absence of analytes. Accordingly, the present invention relates to a bio-lactate color sensor that changes the color of the small array, a lactate discoloration kit, and a sensing method using the same.

It is known that lactate is a fatigue substance produced in the body after exercise. For this reason, lactic acid sensors are widely used in the field and field to determine training effects and training conditions, and are required to be able to detect and measure simply and easily.

However, since the unit price of lactate sidase used in the lactate sensor is higher than that of glucose oxidase used in the lactate sensor, there is a problem that the unit cost of the sensor increases.

For this reason, in order to solve this problem, for example, a lactic acid sensor containing phosphate (Patent No. 3498105) has been proposed to improve the activity of lactic acid oxidase in the reaction layer.

Additionally, as a method for manufacturing biosensors such as lactate sensors, for example, a method includes the step of forming an inorganic gel layer containing at least a mediator, a surfactant, a buffer, and a layered inorganic compound on the top surface of a substrate having electrodes. There is (Patent No. 4088312 Publication).

Therefore, in order to solve the above-mentioned problems, the present inventors recognized that the development of a lactate color sensor was urgent and completed the present invention.

Republic of Korea Patent Publication No. 10-1502277

The object of the present invention is a bio-lactate color sensor, a lactate discoloration kit, and a bio-lactate color sensor equipped with a small array containing transition metals, solvents, and bacteriophages as main components, and changing the color of the small array depending on the presence or absence of analytes. The goal is to provide a sensing method using this.

Another object of the present invention is to provide a lactate color sensor, a lactate discoloration kit, and a sensing method using the same that maximize the color contrast effect by improving the discoloration characteristics of the analyte and increasing the amount of color change.

The technical problems to be achieved by the invention are not limited to the technical problems mentioned above, and other technical problems not mentioned can be clearly understood by those skilled in the art from the description of the present invention.

In order to achieve the above object, the present invention provides a lactate color sensor, a lactate discoloration kit, and a sensing method using the same.

Hereinafter, this specification will be described in more detail.

The present invention provides a lactate color sensor comprising a transition metal, solvent and additives.

In the present invention, the lactate color sensor is characterized in that it additionally includes M13 bacteriophage.

In the present invention, the transition metal is characterized in that it is at least one selected from the group consisting of copper (Cu), iron (Fe), cobalt (Co), and vanadium (V).

In the present invention, the solvent is water, acetone, methanol, ethanol, n-propanol, isopropanol, n-butanol, chloroform, dichloromethane (DCM), toluene, tetrahydrofuran, THF), pyridine, diethyl ether, ethyl acetate, nitromethane, acetonitrile, ammonia, N,N-dimethylformamide (N , N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), formic acid, and propylene carbonate.

In the present invention, the additives include halogen, sodium fluoride (NaF), sodium cyanate (NaNCO), sulfuric acid (H ₂ SO ₄ ), and nitric acid (HNO). ₃ ), oxalic acid, hydrogen peroxide (H ₂ O ₂ ), hydrogen cyanide (HCN), sodium hydroxide (NaOH), hydroxylamine, ethylene amine ), 4-nitrophenol, dithionate (S ₂ O ₆ ^2- ), diborane, sodium borohydride, Iodine (I ₂ ), It is characterized in that it is at least one selected from the group consisting of tris-2 carboxyethyl phosphine hydrochloride (TCEP) and dimethylamine borane (DMAB).

In addition, the present invention provides a lactate discoloration kit comprising a plurality of arrays of lactate color sensors containing transition metals, solvents, and additives.

In the present invention, the lactate discoloration kit is characterized in that the color of the array changes depending on the presence or absence of lactate.

In the present invention, the lactate discoloration kit includes the RGB coordinates of the array composed of each lactate color sensor discolored by adding lactate and the initial RGB coordinates of the array composed of each lactate color sensor to which lactate is not added. It is characterized in that lactate is determined to be present when the root mean square (RMS) for ΔRGB quantified based on coordinates is 20 to 250.

Additionally, the present invention provides a lactate sensing method using a lactate color sensor, comprising the following steps.

(S1) adding lactate, an analyte, to a lactate discoloration kit formed in a plurality of arrays; and

(S2) Detecting a change in color of the plurality of arrays onto which the lactate is added.

All matters mentioned in the lactate color sensor, lactate discoloration kit, and sensing method using the same apply equally unless contradictory.

The lactate color sensor, lactate discoloration kit, and sensing method using the same of the present invention are equipped with a small array containing transition metals, solvents, and bacteriophages as main components, and the color of the small array is changed depending on the presence or absence of analytes. This change can be confirmed with the naked eye.

In addition, the lactate color sensor, lactate discoloration kit, and sensing method using the same of the present invention improve the discoloration characteristics of the analyte and increase the amount of color change, thereby maximizing the color contrast effect and improving selectivity, sensitivity, reproducibility, and reaction speed. It can be significantly improved.

The effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description of the claims.

Figure 1 is an image of sensing lactate using a 3×3 lactate discoloration kit including the lactate color sensor according to the present invention in one array.

The terms used in this specification are general terms that are currently widely used as much as possible while considering the function in the present invention, but this may vary depending on the intention or precedent of a person skilled in the art, the emergence of new technology, etc. In addition, in certain cases, there are terms arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the relevant invention. Therefore, the terms used in the present invention should be defined based on the meaning of the term and the overall content of the present invention, rather than simply the name of the term.

Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the technical field to which the present invention pertains. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and unless clearly defined in the present application, should not be interpreted in an ideal or excessively formal sense. No.

The numerical range includes the values defined in the range above. Any maximum numerical limit given throughout this specification includes all lower numerical limits as if the lower numerical limit were clearly written. Every minimum numerical limit given throughout this specification includes every higher numerical limit as if such higher numerical limit were clearly written. All numerical limits given throughout this specification will include all better numerical ranges within the broader numerical range, as if the narrower numerical limits were clearly written.

Hereinafter, examples of the present invention will be described in detail, but it is obvious that the present invention is not limited to the following examples.

Lactate color sensor

The present invention provides a lactate color sensor containing a transition metal, solvent, and additives that can change color depending on the presence or absence of lactate.

Additionally, the lactate color sensor may additionally include M13 bacteriophage.

The M13 bacteriophage is a particle with a length of 880 nm and a width of 6.6 nm. Unlike nanoparticles made through general organic and inorganic synthesis, it is a protein body expressed through a certain gene, so all particles have a perfectly identical shape. This provides great advantages in the material preparation process. In addition, as a nanoparticle with a high surface to volume ratio, each particle contains about 2,700 pairs of proteins (pVIII protein) on the surface and 4 to 5 pairs of proteins (pIII, pVI, pVII, pIX) at each end. has. In particular, in the case of protein 8 (pVIII), which expresses 2,700 pairs of identical peptides, the paired protein molecules are arranged very densely in a helix with an interval of about 3.3 nm. By appropriately recombining the genes in the bacteriophage, a peptide of the desired type can be expressed on each corresponding surface protein, making it easy to apply to various biosensing fields according to the purpose.

The transition metals include copper (Cu), iron (Fe), cobalt (Co), chromium (Cr), manganese (Mn), nickel (Ni), and vanadium (vanadium). V) may be one or more selected from the group consisting of, and preferably may be one or more selected from the group consisting of copper, iron, cobalt and vanadium.

The transition metal is in periods 4 to 7 of the periodic table; and elements from groups 3 to 12. In the case of elements other than the transition metals, the chemical properties of the main group elements in one cycle change significantly as the number of valence electrons changes, but the transition metals show many similarities not only in a given group but also in the same cycle. In addition, the transition metal has luster, electrical conductivity, and thermal conductivity like general metals, and can especially form ionic compounds with non-metals. When forming an ionic compound with a non-metal, unlike common metals, the transition metal may exist in the form of a complex ion with a certain number of ligands. The complex ion refers to an ion generated through a bond between a transition metal and a ligand.

The solvent is water, acetone, methanol, ethanol, n-propanol, isopropanol, n-butanol, chloroform, dichloromethane (DCM), toluene, tetrahydrofuran (THF), pyridine ( pyridine, diethyl ether, ethyl acetate, nitromethane, acetonitrile, ammonia, N,N-dimethylformamide, It may be one or more selected from the group consisting of DMF), dimethyl sulfoxide (DMSO), formic acid, and propylene carbonate, and is preferably water, acetone, methanol, ethanol, isopropanol, It may be one or more selected from the group consisting of pyridine, acetonitrile, N-dimethylformamide, dimethyl sulfoxide, and propylene carbonate.

The additives include halogen, sodium fluoride (NaF), sodium cyanate (NaNCO), sulfuric acid (H ₂ SO ₄ ), nitric acid (HNO ₃ ), oxalic acid ( oxalic acid, hydrogen peroxide (H ₂ O ₂ ), hydrogen cyanide (HCN), sodium hydroxide (NaOH), hydroxylamine, ethylene amine, 4-night 4-nitrophenol, dithionate (S ₂ O ₆ ^2- ), diborane, sodium borohydride, iodine (I ₂ ), tris (2-chloroethyl) ) It may be one or more selected from the group consisting of phosphate (Tris-2 carboxyethyl phosphine hydrochloride, TCEP) and dimethylamine borane (DMAB), preferably halogen, hydrogen peroxide, hydroxylamine, 4-nitrophenol, It may be one or more selected from the group consisting of iodine and dimethylamine borane.

The lactate color sensor may contain transition metals, solvents, and additives mixed at a concentration ratio of 1:10 to 25:0.5 to 1.2.

lactate discoloration kit

The present invention provides a lactate discoloration kit including a plurality of arrays of lactate color sensors containing transition metals, solvents, and additives.

The lactate color sensor is the same as mentioned above.

The lactate discoloration kit can change the individual colors of the array depending on the presence or absence of lactate. More specifically, the absolute value of ΔR in the RGB coordinates of the array color composed of each lactate color sensor to which the lactate was added and discolored and the initial RGB coordinates of the array composed of each lactate color sensor to which the lactate was not added. If it is 20 or more, it can be quantitatively indicated that lactate is present.

In addition, ΔRGB quantified based on the RGB coordinates of the array color composed of each lactate color sensor discolored by adding the lactate and the initial RGB coordinates of the array composed of each lactate color sensor to which the lactate was not added. The presence of lactate can be quantified when the root mean square (RMS) is 20 to 250, preferably the RMS is 20 to 200, and most preferably the RMS is 20 to 200. It could be 150.

The RMS may be the root mean square of ΔR ² +ΔG ² +ΔB ² based on ΔRGB derived from the RGB coordinates of the array composed of the lactate color sensor.

Lactate sensing method

The present invention provides a lactate sensing method comprising the following steps.

(S1) adding lactate, an analyte, to a lactate discoloration kit formed in a plurality of arrays; and

(S2) Detecting a change in color of the plurality of arrays onto which the lactate is added.

The lactate color sensor and lactate discoloration kit are as mentioned above.

The lactate color sensor, lactate discoloration kit, and sensing method using the same of the present invention improve the discoloration characteristics of the analyte and increase the amount of color change, thereby maximizing the color contrast effect and significantly improving selectivity, sensitivity, reproducibility, and reaction speed. You can do it.

The advantages and features of the present invention and methods for achieving them will become clear with reference to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below and may be implemented in various different forms, and only the embodiments are provided to ensure that the disclosure of the present invention is complete, and are provided by those skilled in the art in the technical field to which the present invention pertains. It is provided to fully inform those who have the scope of the invention, and the present invention is only defined by the scope of the claims.

Example 1. Color change kit including lactate color sensor

A 3×3 lactate discoloration kit containing the lactate color sensor according to the present invention as an array was manufactured, and the lactate color sensor constituting the array contained transition metal-solvent-additive at 1mM:18.2mM: It was prepared by mixing at a concentration ratio of 0.8 mM, and the composition of the transition metal-solvent-additive is shown in [Table 1] below.

[Table 1]

Experimental Example 2. Lactate sensing

2.1. Check for discoloration

In order to confirm the color change sensing of the 3 × 3 lactate color change kit containing the lactate color sensor prepared in Example 1 according to the present invention as one array, the 3 × 3 lactate color change kit (Example 1) was used. 0.1 M of lactate was added dropwise, and this is shown in Figure 1.

Referring to Figure 1, (a) a 3×3 lactate discoloration kit without lactate (Example 1) and (b) a 3×3 lactate discoloration kit with 0.1 M lactate added (Example 1). ) can be seen to have a clear color difference.

2.2. ΔRGB and RMS quantification

In order to quantify the color change sensing of the 3 × 3 lactate color change kit containing the lactate color change sensor prepared in Example 1 according to the present invention as one array, the 3 × 3 lactate color change kit (Example 1) was used. 0.1 M lactate was added dropwise. At this time, the RGB coordinates of the 3×3 lactate discoloration kit including the initial lactate color sensor without the lactate droplets as one array and the 3×3 lactate color sensor including the lactate color sensor with the lactate droplets as one array. 3 ΔRGB, which is the difference between the RGB coordinates of the lactate discoloration kit, was derived, and the different RMS were quantified, and this is shown in [Table 2] below.

[Table 2]

Referring to [Table 2], it can be seen that the absolute value of ΔR for all arrays is 20 or more. In addition, ΔRGB quantified based on the RGB coordinates of the array color composed of each lactate color sensor discolored by adding the lactate and the initial RGB coordinates of each array composed of the lactate color sensor to which the lactate was not added. It can be seen that the root mean square (RMS) for has a minimum of 25.5 (array number 9) and a maximum of 126.6 (array number 1).

From the above results, it was demonstrated that the lactate discoloration kit including the lactate color sensor according to the present invention as one array reacts with high sensitivity to lactate.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in other specific forms without changing its technical idea or essential features. In this regard, the embodiments described above should be understood in all respects as illustrative and not restrictive.

Claims (9)

Contains transition metals, solvents and additives,
The transition metal, solvent and additives are mixed at a concentration ratio of 1:18.2:0.8,
Depending on the presence or absence of lactate, the mixture of transition metals, solvents, and additives is discolored, and ΔRGB quantified based on the RGB coordinates when the mixture is discolored by adding lactate and the initial RGB coordinates without adding lactate. It is characterized in that lactate is determined to be present when the root mean square (RMS) for is 80 to 190,
The transition metals, solvents, and additives are
copper, ethanol and 4-nitrophenol; Copper, isopropanol, and 4-nitrophenol; Copper, methanol and 4-nitrophenol; Copper, water and 4-nitrophenol; Vanadium, acetone and halogen (Cl); and copper, acetonitrile, and DMAB; A lactate color sensor comprising a composition of: delete delete delete delete A lactate discoloration kit comprising a plurality of arrays composed of the lactate color sensors according to claim 1. delete delete (S1) adding lactate, an analyte, to the lactate discoloration kit according to claim 6; and
(S2) detecting a change in color of the plurality of arrays onto which the lactate is dropped. A lactate sensing method using a lactate color sensor, comprising: