KR101427988B1 - Smoking means of detecting - Google Patents

Abstract

The present invention relates to smoking detection means for detecting the presence or absence of smoking and detecting the presence of nicotine and its metabolite, cotinine, which remains in the body by urine, A first reagent layer formed on the upper surface of the first reagent layer and equipped with a cyanogen releasing reagent which releases cyanogen by water contained in the urine; a second reagent layer formed on the upper surface of the first reagent layer and reacted with the released cyanogen, A second reagent layer provided with a reagent for forming a cyanogen halide to form a halide; The first reagent layer and the second reagent layer are separated from each other and the pH is maintained before the reaction of the urine, and during the urine reaction, the reagent of the first reagent layer and the reagent of the second reagent layer is mixed well, a buffer solution layer for allowing the pH to be maintained and a second reagent layer or / and a buffer solution layer which reacts quantitatively with the amount of the second reagent layer which reacts with nicotine or cotinine contained in the urine, Change indicator; And a reagent protecting layer formed on an upper surface of the buffer solution layer and allowing the penetration of foreign substances and moisture to the outside of the buffer solution layer. The present invention does not require a separate professional manpower and expensive equipments, so it can be cheaper and simple to grasp the presence or absence of smoking, and can be easily checked at any time, anywhere, so that it is universally usable.

Description

Smoking means of detecting < RTI ID = 0.0 >

The present invention relates to smoking detection means, and more particularly, to smoking detection means for detecting presence or absence of smoking and detecting the presence of nicotine and its metabolite, cotinine, ≪ / RTI >

It is a well-known fact that tobacco (Nicotiann tabacum) is harmful to the human body, and the risk of tobacco smoking is not too much emphasized, and the number of deaths due to smoking, which is a direct cause of the disease, is increasing every year. People who smoke more than 20 cigarettes a day on an adult basis are said to have a six-fold higher risk of developing fever than those who do not. In addition, cigarette smoking in addition to the cardiac muscle to send the blood to the arteries to narrow the heart muscle and oxygen and lack of time and myocardial infarction or angina pectoris is easy to take other risks of smoking and gastric ulcers, duodenal ulcers are more likely to develop, infertility And may be the cause of miscarriage.

Due to the harmful effects of various kinds of tobacco, a variety of national anti - smoking policies and publicity activities are being carried out. Various drugs have been studied to remove harmful components of tobacco. However, the problem is that cigarettes are so addictive that they can not easily break their patience. If smokers attempt to quit smoking, the probability of success is only about 3%, and it is also common for smokers to smoke more if they are exposed to long-term smoking. Therefore, it is best not to smoke, and it is appropriate for public health to induce smoking cessation as early as possible.

In recent years, youth smoking has become a social problem, and various publicity activities have been carried out to prevent such problems. However, there is a problem that it is difficult to actually understand how much young people actually smoke. Therefore, it is necessary to solve the smoking problem of adolescents by accurately grasping the smoking status of adolescents and inducing smoking cessation through continuous care and management of tobacco adolescents.

However, it is not easy to grasp whether or not each individual teen is smoking, and it is even more difficult to enforce it. In addition, the test method for determining whether or not the actual smoking is currently being used is costly, time consuming and manpower consuming, which is not a practical alternative. High performance liquid chromatography (GC) and gas chromatography (GC) methods are used in classical methods to measure the presence or absence of currently used cigarettes. And the measurement time is too long to be utilized properly. Another method is the ELISA (Enzyme Linked Immunoassays) method, which requires expensive specialized equipment and specialists, which is not a general alternative except large hospitals.

Therefore, in order to easily and easily grasp the presence or absence of smoking, it is urgently required to equip the youths with facilities and methods that can simply inspect the facilities at schools, academies, etc., where they live in groups at low cost. However, there is no clear alternative at present.

An object of the present invention to solve such a fundamental problem of the prior art is to provide a means for detecting the presence or absence of smoking by requiring no separate equipment and professional manpower by simply detecting the presence or absence of smoking and the degree of smoking even when the urine is changed to change color.

The objects of the present invention are not limited to the above-mentioned objects, and other objects not mentioned can be clearly understood from the following description.

A first reagent layer formed on one upper surface of the support and having a cyanogen releasing reagent for releasing cyanogen by water contained in the urine; A second reagent layer formed on the upper surface of the reagent layer and having a cyanogen halide forming reagent to react with the released cyanogen to form a cyanogen halide; The first reagent layer and the second reagent layer are separated from each other and the pH is maintained before the reaction of the urine, and during the urine reaction, the reagent of the first reagent layer and the reagent of the second reagent layer is mixed well, a buffer solution layer for allowing the pH to be maintained and a second reagent layer or / and a buffer solution layer which reacts quantitatively with the amount of the second reagent layer which reacts with nicotine or cotinine contained in the urine, Change indicator; And a reagent protecting layer formed on an upper surface of the buffer solution layer to allow foreign substances and water to penetrate therethrough, wherein one side of the support having the first reagent layer and the second reagent layer is contained in the urine to remove nicotine Or cotinine, and detecting the presence or absence of smoking by the concentration of the color changed by the color change indicator.
Preferably, the cyanogen-releasing reagent of the first reagent layer is potassium thiocyanate, the reagent for forming a cyanogen halide of the second reagent layer is chloroamine-T or chloroamine-B, Citrate buffer, the color change indicator is diethylthiobarbituric acid, and the reagent protection layer is a nylon filter or a cellulose filter. On the other hand, the potassium thiocyanate is a solution in which 30 to 70 parts by weight of potassium thiocyanate is mixed with 100 parts by weight of the solvent, and the chloroamine-T or chloroamine- The nylon filter or the cellulose filter has a thickness of 50-150 mu m and a pore size of 20-140 mu m.

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The present invention does not require a separate professional manpower and expensive equipment, so that it is possible to grasp whether smoking is cheap or simple.

In addition, since the present invention does not require a specific facility for inspection, it can be inspected at any time, anywhere, and there are other effects that can be universally used.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a diagram showing the chemical structure of nicotine and cotinine.
FIG. 2 is a schematic diagram showing the formation of glutaconaldehyde by reaction of cyanogen chloride, which is a reagent layer according to the present invention, with nicotine or cotinine. FIG.
Figure 3 shows the reaction of diethylthiobarbituric acid, a color change indicator that reacts with glutaconic aldehyde according to the present invention.
4 is a view showing a state of smoking presence / absence detecting means according to the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention, and the manner of achieving them, will be apparent from and elucidated with reference to the embodiments described hereinafter in conjunction with the accompanying drawings. However, the present invention is not limited to the embodiments described below, but may be embodied in various different forms, and these embodiments are not intended to be exhaustive or to limit the scope of the present invention to the precise form disclosed, The present invention is only defined by the description of the claims. It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In the present specification, the singular form includes plural forms unless otherwise specified in the specification. Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings.

In the case of a smoker, smoking traces remaining in the body, cotinine as a nicotine or nicotine metabolite, is discharged into the urine and quantitatively examined to determine the presence or absence of smoking and the degree of smoking, It is.

First, when smoking is performed, nicotine remains in the blood of the human body, and the chemical structural formula of such nicotine is shown in FIG. These nicotines are converted to various metabolites over time. Normally nicotine has a half-life of about 2 hours in the body, so detection of nicotine in the urine may not be an accurate indicator. However, nicotine is converted into cotinine shown in FIG. 1 by about 70 to 80% through metabolism in the body, and it is more efficient to measure such nicotine since the release half-life of the cotinine is about 17 to 20 hours.

Reaction principle

1. Reactive reagents for reaction with nicotine or cotinine

A reagent capable of quantitatively reacting with nicotine or cotinine by smoking remaining in the urine is needed. To this end, cyanogen halides are used in the present invention. The available cyanogen halides are cyanogen bromide (CNBr), cyanogen chloride (CNCl) and cyanogen iodine (CNI). These cyanogenhalides produce glutaconaldehyde by reaction with nicotine or cotinine. According to the experiments conducted by the present inventors, it was found that the cyanogen bromide among the above-mentioned reactive reagents is not easy to handle relatively, and the cyanogen iodine is the most suitable because the reaction efficiency is not so good, As noted, nicotine or cotinine reacts quantitatively with cyanogen chloride to produce glutaconaldehyde.

2. Detection of clutaconaldehyde

As described above, the amount of nicotine or cotinine in urine is possible by quantitative detection of glutaconic aldehyde. Therefore, in order to visually confirm the glutacondaldehyde, various methods can be used for detecting glutacondialdehyde. However, in order to visually confirm the glutacondaldehyde, the color change caused by the appropriate buffering agent and indicator may be induced and confirmed. As shown in FIG. 3, glutaconic aldehyde can be used as a diethylthiobarbituric acid (diethylthiobarbituric acid) as shown in FIG. 3, since it is preferable to use various indicator agents for color development and most commonly to react with aldehyde groups to induce color expression. It reacts with acid and develops pink color. In addition, the indicators that can be used include diethylthiobarbituric acid, barbituric acid, diethylthiobarbiturate, 5-amino-2-naphthalenesulfonic acid (diethyl thiobarbiturate, 5- amino-2-naphthalenesulfonic acid, 8-amino-2-naphthalenesulfonic acid, 7-amino-1,3-naphthalenesulfonic acid sodium monopotassium salt -amino-1,3-naphthalene sulfonic acid monopotassium salt, 1,4-phenylene diamine dihydrochloride, O-tolidine dihydrochloride, sulfa Sulfanilic acid, sulfanilamide, 4-amino-1-naphthalene sulfonic acid, p-aminobenzoic acid, acid, 4-amino salicylic acid, and the like can be used.

Smoking detection means

In the present invention, the presence or absence of smoking is detected in the form of a test paper by using the reaction principle, and the presence or absence of smoking according to color development can be easily checked by wetting the urine. Hereinafter, Will be described with reference to FIG.

1. Support

The support (1) will suit the size of ordinary litmus test papers, and can be manufactured in various shapes, and various materials such as plastic or paper materials can be used. If you use the rear part by hand, it will suffice.

2. Reagent layer formation

As described above, nicotine or cotinine reacts quantitatively with a cyanogenhalide, preferably cyanogen chloride, and this reactant reacts with the indicator to cause a color change. Therefore, cyanogen chloride and indicator must be contained in the reagent layer, but there are various problems in forming cyanogen chloride itself as a reagent layer. In other words, cyanogen chloride is a highly reactive and unstable compound, which can be easily deteriorated during storage at room temperature, so it is used indirectly. Generally, a cyanogen releasing agent and a cyanogen halide forming agent which are stable at room temperature can be easily reacted in the presence of water to produce cyanogen chlorides.

First, a first reagent layer 2 containing a cyanogen releasing agent is formed on the upper surface of the support. This first reagent layer (2) may suffice to be wetted with a cyanogen releasing agent in the form of a general paper test strip. Therefore, the test paper should be wetted with the cyanogen releasing reagent present in the liquid phase, and then dried.

The cyanogen releasing agent forms a structure containing cyanogen to form a cyanogen halide. Examples of the cyanogen releasing agent include potassium thiocynate, sodium thiocyanate, Potassium cyanide, and sodium cyanide reagents. Potassium thiocynate is suitably used in an amount of 30-70 parts by weight based on 100 parts by weight of the solvent.

Next, a second reagent layer 3 is formed on the upper surface of the first reagent layer 2. The second reagent layer 3 is provided with a cyanogen halide forming agent which reacts with a cyanogen releasing agent contained in the first reagent layer 1. The cyanogen halide forming agent is a reagent containing a halide to form a cyanogen halide. Chloramine-T, chloramine-B, And it is suitably used in an amount of about 40-70 parts by weight based on 100 parts by weight of the solvent. Diethylthiobarbituric acid used as an indicator is made wet on a normal paper test strip.

Next, a buffer solution layer (4) is formed for even mixing of the two reagents. The solution pH of the buffer solution layer is suitably between 4-8.

Examples of the buffer solution include a citrate buffer, an acetate buffer, a citrate-phosphate buffer, a succinate buffer, an aconitate buffer, A carbonate buffer, etc. Any of them can be used. Among them, a citrate buffer which is easy to obtain and easy to handle is most suitable, and it is enough to use an adequate amount of about 2-4 mol . In the buffering reagent layer 4, an indicator may be contained. The indicators may be any of those described above. In this embodiment, diethylthiobarbituric acid is included.

In this embodiment, the indicator is included in the buffer solution layer, but the present invention is not limited thereto. This indicator is effective when the cyanogen releasing agent of the first reagent layer 2 and the cyanogen halide forming agent of the second reagent layer 3 are wetted with urine, Since the reaction occurs in step (4) and cyanogen is formed, it is color-changed. Therefore, it may be included in the second reagent layer, or both the buffer solution and indicator may be mixed together in the second reagent layer. will be. The essential point of the present invention is that when a cyanogen releasing agent and a cyanogen halide forming agent are formed as separate reagent layers and urine is wetted, Since the objective is to release the nogen and cause the color change by the indicator agent, the two reagent layers are separated and the buffer solution and the indicator that maintain the appropriate pH to facilitate the reactivity are mixed together in the second reagent layer It is also acceptable to use it.

Next, the reagent protecting layer 5 is further formed on the upper surface of the first and second reagent layers 2, 3 and the buffer solution layer 4 as described above. The reagent protecting layer 5 is for protecting the reagent layer and is intended to prevent the deterioration of the reagent layer due to external foreign substances or moisture in the air or the like. The reagent protective layer should be transparent for visual observation of color change during the nicotine test and should have a proper thickness and porosity in order to react with the reagent layer due to urine penetration, A reagent protection layer having a pore size of 20-140 mu m may be achieved in various ways. For example, a porous cellulose filter or a nylon filter may be used.

Next, an operation process of the smoking presence detection means constructed as described above will be described. In the case where the reagent is stored at room temperature, the first reagent layer (2) and the second reagent layer (3) are separated from each other by foreign substances or the like due to the reagent protection layer. The urine is passed through the filter of the reagent protection layer 5 to wet the first reagent layer 2, the second reagent layer 3 and the buffer solution layer 4. At this time, the cyanogen halide forming agent reacting with the cyanogen releasing agent reacts with the water contained in the urine to generate cyanogen chloride, and the produced cyanogen chloride is converted into urine Is reacted quantitatively with nicotine or cotinine contained in the genus to convert it into a glucatonaldehyde, which reacts with diethylthiobarbituric acid used as an indicator to develop a pink color. The presence or absence of smoking and the degree of smoking can be visually observed according to the concentration of pink, and the degree of smoking can be grasped by quantitatively calculating the degree of color change using the light absorbance meter.

Example

10 smokers and 10 nonsmokers were tested as a population and tested in order to verify the effectiveness of the smoking detection means of the present invention in comparison with the known ELISA method. For accurate comparison, in the present invention, the quantitative concentration of the developed color density was determined by using a light absorber.

10 non-smokers

Nonsmoker DFI ([mu] g / ml) ELISA ([mu] g / ml) Remarks One - 0.00 2 - 0.00 3 - 0.02 4 - 0.08 5 - 0.06 6 - 0.04 7 - 0.06 8 - 0.00 9 - 0.04 10 - 0.04

As shown in Table 1, in the present invention, there was no reaction when measured using a light absorber, and it was found that both were effective since the ELISA method also showed an error within the error range.

For 10 smokers

smoker DFI ([mu] g / ml) ELISA ([mu] g / ml) Remarks One 2 1.5 2 5 5.4 3 2 2.2 4 2 1.9 5 5 6.0 6 12 11 7 2 1.9 8 2 2.5 9 5 5.8 10 5 4.5

The results are almost the same as those obtained by the conventional ELISA method and the method of the present invention, and this shows that the present invention is effective.

Claims (13)

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A first reagent layer formed on one upper surface of the support and having a cyanogen releasing reagent for releasing cyanogen by water contained in the urine;
A second reagent layer formed on an upper surface of the first reagent layer and having a cyanogen halide forming reagent to react with the released cyanogen to form a cyanogen halide;
The first reagent layer and the second reagent layer are separated from each other and the pH is maintained before the reaction of the urine, and during the urine reaction, the reagent of the first reagent layer and the reagent of the second reagent layer is mixed well, a buffer solution layer for allowing the pH to be maintained:
A color change indicator which is contained in the second reagent layer and / or the buffer solution layer and quantitatively reacts with the amount of the second reagent layer which reacts with nicotine or cotinine in the urine to cause color change; And
And a reagent protecting layer formed on an upper surface of the buffer solution layer and allowing the penetration of foreign matter and moisture to the outside,
One side of the support provided with the first reagent layer and the second reagent layer is contained in the urine and quantitatively reacted with nicotine or cotinine contained in the urine to detect the presence or absence of smoking by the color concentration changed by the color change indicator Wherein the detection means detects the presence or absence of smoking. 8. The method of claim 7,
Wherein the cyanogen releasing reagent of the first reagent layer is potasium thiocyanate and the cyano group halide forming reagent of the second reagent layer is chloroamine-T or chloroamine-B, the buffer solution is a citrate buffer, Wherein the color change indicating agent is diethylthiobarbituric acid, and the reagent protecting layer is a nylon filter or a cellulose filter. 9. The method of claim 8,
The above-mentioned potassium thiocyanate is prepared by mixing 30 to 70 parts by weight of potasium thiocyanate with 100 parts by weight of the solvent,
Wherein the chloramine-T or the chloroamine-B is a mixed solution of 40 to 70 parts by weight based on 100 parts by weight of the solvent. 10. The method of claim 9,
Wherein the nylon filter or the cellulose filter has a thickness of 50 to 150 占 퐉 and a pore size of 20 to 140 占 퐉.
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