CN107064128B - Urine detection reagent and urine detection test paper prepared from same - Google Patents
Urine detection reagent and urine detection test paper prepared from same Download PDFInfo
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- CN107064128B CN107064128B CN201710281189.XA CN201710281189A CN107064128B CN 107064128 B CN107064128 B CN 107064128B CN 201710281189 A CN201710281189 A CN 201710281189A CN 107064128 B CN107064128 B CN 107064128B
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Abstract
The invention provides a urine detection reagent and urine detection test paper, which are used for detecting tyrosine metabolites in urine, wherein the urine detection test paper consists of three detection layers, namely a first layer, a second layer and a third layer from top to bottom; the first layer is a filter layer, the second layer is a detection layer, and the third layer is a drainage liquid absorption layer.
Description
Technical Field
The invention relates to the field of in-vitro diagnostic reagents, in particular to a urine detection reagent, and more particularly relates to a urine detection test paper for detecting tyrosine metabolites in urine.
Background
Tumors are diseases with high morbidity and high mortality seriously threatening human health, and a large amount of research and data prove that early diagnosis and treatment are the most effective methods for preventing and treating the tumors and reducing the mortality. At present, the conventional tumor marker detection method has the defects of low single detection rate, incapability of early detection and the like. The diagnosis and treatment of diseases are often delayed, and a marker and a test method which can be found early, are accurate in detection and are simple and convenient to operate are sought, so that the method is the direction of continuous efforts of medical researchers. The p-hydroxyphenylalanine tyrosine urine test reagent is used for early cancer detection. The main principle is as follows: the abnormally increased tyrosine metabolite in the urine of the cancer patient reacts with the detection reagent in a specific way to generate a precipitate with a characteristic color. The test results were judged visually according to the color of the precipitate.
Tyrosine (Tyrosine) belongs to aromatic amino acid, can be hydroxylated from phenylalanine (Phe), can also be directly taken from food, and is an important semi-essential amino acid for human body. Tyrosine metabolic disorder can cause various amino acid metabolic diseases, such as tyrosinemia, homogentisate urinate, and the like, and has certain relevance with liver and kidney diseases, nervous system degenerative diseases, malignant tumor of qi, and the like. Therefore, the research on tyrosine and its metabolites is not only very important in protein chemistry and evaluation of nutritional status of patients, but also important in early diagnosis, therapeutic effect detection, etiology research and the like of various diseases.
The presence of proteins in biological samples can interfere with the determination of tyrosine and its metabolites, which in turn affects the accuracy of the assay.
The invention provides a urine detection reagent and urine detection test paper, which are used for detecting tyrosine metabolites in urine.
Disclosure of Invention
The invention provides a urine detection reagent for detecting tyrosine metabolites in urine, which is characterized by comprising mercurous nitrate, mercuric sulfate, sodium tungstate and nitric acid.
The invention provides urine test paper, which is used for detecting tyrosine metabolites in urine and consists of three detection layers, namely a first layer, a second layer and a third layer from top to bottom; the first layer is a filtering layer, the second layer is a detection layer, and the third layer is a drainage liquid absorption layer;
wherein the preparation raw material of the first layer comprises filter paper and modified glucose;
the preparation raw materials of the second layer comprise filter paper and a urine detection reagent;
the urine detection reagent comprises mercurous nitrate, mercuric sulfate, sodium tungstate and nitric acid;
the preparation raw material of the third layer comprises filter paper and acrylic acid modified chitosan.
In one embodiment of the invention, the modified glucose is glucan modified by benzimidazole derivative and benzoic acid derivative together.
In one embodiment of the present invention, the benzimidazole derivative is one selected from the group consisting of 3- (1H-benzimidazol-2-yl) -2, 3-dihydroxypropionic acid, [ 3-carboxymethyl-2-oxo-2, 3-dihydro-1H-benzimidazole ] acetic acid, 5, 6-dicarboxybenzimidazole and benzimidazole.
In one embodiment of the present invention, the benzoic acid derivative is phthalic acid.
As an embodiment of the invention, the urine detection reagent further comprises a phosphomolybdotungstic acid-chitosan-nano silica composite.
As an embodiment of the invention, the bottom of the drainage liquid body layer is provided with a hydrophobic layer.
As an embodiment of the present invention, a water storage tank is disposed on the hydrophobic layer.
In one embodiment of the present invention, the filter layer and the detection layer are connected by a peelable adhesive.
A method for detecting tyrosine metabolites in urine at least comprises the following steps:
acquiring human urine to be detected;
dropping the human urine on urine test paper to judge whether color reaction occurs.
In one embodiment of the present invention, the detection method further comprises tearing off the filter layer, and further observing the color reaction.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1: schematic diagram of the urine test paper.
FIG. 2: the urine test paper is a three-dimensional schematic diagram.
Description of the symbols: a filter layer: 1. detection layer: 2. draining the liquid absorption layer: 3. hydrophobic layer: 4. a water storage tank: 4-1.
Detailed Description
The disclosure may be understood more readily by reference to the following detailed description of preferred embodiments of the invention and the examples included therein. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.
When an amount, concentration, or other value or parameter is expressed as a range, preferred range, or as a range of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when a range of "1 to 5" is disclosed, the described range should be interpreted to include the ranges "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a range of values is described herein, unless otherwise stated, the range is intended to include the endpoints thereof and all integers and fractions within the range.
The singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. "optional" or "any" means that the subsequently described event or events may or may not occur, and that the description includes instances where the event occurs and instances where it does not.
Approximating language, as used herein throughout the specification and claims, is intended to modify a quantity, such that the invention is not limited to the specific quantity, but includes portions that are literally received for modification without substantial change in the basic function to which the invention is related. Accordingly, the use of "about" to modify a numerical value means that the invention is not limited to the precise value. In some instances, the approximating language may correspond to the precision of an instrument for measuring the value. In the present description and claims, range limitations may be combined and/or interchanged, including all sub-ranges contained therein if not otherwise stated.
In addition, the indefinite articles "a" and "an" preceding an element or component of the invention are not intended to limit the number requirement (i.e., the number of occurrences) of the element or component. Thus, "a" or "an" should be read to include one or at least one, and the singular form of an element or component also includes the plural unless the stated number clearly indicates that the singular form is intended.
As used herein, each of the following terms has the meaning associated with it in this section. The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. For example, "an element" means one element or more than one element.
As used herein, the term "sensitivity" is defined as a statistical measure of the performance of an assay (e.g., method, test) calculated by dividing the number of true positives by the sum of true positives and false negatives.
As used herein, the term "specificity" is defined as a statistical measure of the performance of an assay (e.g., method, test) calculated by dividing the number of true negatives by the sum of true negatives and false positives.
The invention provides urine test paper, which is used for detecting tyrosine metabolites in urine and consists of three detection layers, namely a first layer, a second layer and a third layer from top to bottom; the first layer is a filtering layer, the second layer is a detection layer, and the third layer is a drainage liquid absorption layer;
wherein the preparation raw material of the first layer comprises filter paper and modified glucose;
the preparation raw materials of the second layer comprise filter paper and a urine detection reagent;
the urine detection reagent comprises mercurous nitrate, mercuric sulfate, sodium tungstate and nitric acid;
the preparation raw material of the third layer comprises filter paper and acrylic acid modified chitosan.
Filter layer
The preparation raw materials of the filter layer comprise filter paper and modified glucose.
Modified glucose
The modified glucose is glucan jointly modified by benzimidazole derivatives and benzoic acid derivatives.
In one embodiment of the present invention, the benzimidazole derivative is one selected from the group consisting of 3- (1H-benzimidazol-2-yl) -2, 3-dihydroxypropionic acid, [ 3-carboxymethyl-2-oxo-2, 3-dihydro-1H-benzimidazole ] acetic acid, 5, 6-dicarboxybenzimidazole and benzimidazole.
In a preferred embodiment of the present invention, the benzimidazole derivative is [ 3-carboxymethyl-2-oxo-2, 3-dihydro-1H-benzimidazole ] acetic acid.
In one embodiment of the present invention, the benzoic acid derivative is phthalic acid.
In a preferred embodiment of the present invention, the benzoic acid derivative is terephthalic acid.
As an embodiment of the invention, the preparation method of the benzimidazole derivative and the benzoic acid derivative jointly modified glucan comprises the following steps:
benzimidazole derivative (0.35g,2.160mmol), phthalic acid derivative (0.166g,1mmol), 3- (4, 5-dimethylthiazole-2) -2, 5-diphenyltetrazolium bromide salt (0.825g,4.304mmol), dextran (0.5g,0.0125mmol) and 4-dimethylaminopyridine (0.056g,0.459mmol) were dissolved in a glass vial containing 40 ml of dimethyl sulfoxide. The reaction was carried out at 30 ℃ for 48 hours. Then placed in a dialysis bag for dialysis and impurities are removed.
As an embodiment of the invention, the dextran has a molecular weight of 4500 daltons to 40000 daltons.
In the invention, the preparation method of the filter layer comprises the following steps: placing filter paper on a filter, wetting by ethanol suction, and drying for 15 minutes by air; and then dissolving the modified glucan prepared in the step (a) in a mixed solution of ethanol, performing suction filtration and deposition on filter paper, and then performing air drying for 15 minutes to prepare a first layer, namely a filter layer.
In the invention, the weight part ratio of the modified glucose to the mercuric sulfate is 1.3: 1.
detection layer
In the invention, the preparation raw materials of the detection layer comprise filter paper and a urine detection reagent.
The urine detection reagent comprises mercurous nitrate, mercuric sulfate, sodium tungstate and nitric acid;
the urine detection reagent also comprises a phosphomolybdotungstic acid-chitosan-nano silicon dioxide compound.
As an embodiment of the invention, the urine detection reagent is prepared from the following raw materials in parts by weight:
in the application, the content of the mercurous nitrate is 15mol/L, which means that the content of the mercurous nitrate in the whole detection reagent is 15mol/L, and the volume calculation reference is based on the volume of the detection reagent. The content of the mercuric sulfate is 1 mol/L; the content of mercury sulfate in the whole detection reagent is 1mol/L, and the volume calculation reference is based on the volume of the detection reagent; the content of sodium tungstate is 2.5 mol/L; the content of sodium tungstate in the whole detection reagent is 2.5mol/L, and the volume calculation reference is based on the volume of the detection reagent; the content of the nitric acid is 10 mol/L; the content of nitric acid in the whole detection reagent is 10mol/L, and the volume calculation reference is based on the volume of the detection reagent; the content of the phosphomolybdotungstic acid-chitosan-nano silicon dioxide compound is 0.3g/L, and the volume calculation reference is based on the volume of the detection reagent.
In the application, the term "phosphomolybdotungstic acid-chitosan-nano silica composite" refers to that chitosan and nano silica are grafted to obtain a chitosan-nano silica composite, and then the chitosan-nano silica composite is loaded with phosphomolybdotungstic acid to obtain a phosphomolybdotungstic acid-chitosan-nano silica composite.
In the application, the phosphomolybdotungstic acid, the chitosan and the nano silicon dioxide are all obtained from markets.
The phosphomolybdic tungstic acid is purchased from Xiamen sea Mark science and technology limited;
the chitosan is purchased from the bioscience and technology company Limited of Xianruin;
the nano silicon dioxide is purchased from Yingchuangdegussa;
in a preferred embodiment, the preparation method of the phosphomolybdotungstic acid-chitosan-nano silica composite at least comprises the following steps:
(1) dissolving nano silicon dioxide in an ethanol solution, performing ultrasonic dispersion for 60min at room temperature, adding a silane coupling agent K560, and stirring for 3h at 45 ℃ to obtain a nano silicon dioxide solution; dissolving chitosan in acetic acid solution, and stirring at room temperature for 3h to obtain chitosan solution; adding the chitosan solution into the nano-silica solution, stirring and reacting for 8h, centrifuging and washing for 3 times, washing for 2 times with ethanol, and vacuum drying at 60 ℃ to obtain the chitosan-nano-silica composite.
(2) Dissolving phosphomolybdotungstic acid in deionized water to prepare a phosphomolybdotungstic acid solution with the mass fraction of 10%, adding the chitosan-nano silicon dioxide composite obtained in the step (1), refluxing and stirring at constant temperature for 20 hours, vacuum filtering, and drying to obtain the phosphomolybdotungstic acid-chitosan-nano silicon dioxide composite.
The preparation method of the urine detection reagent comprises the following steps: and uniformly mixing mercurous nitrate, mercuric sulfate, sodium tungstate and nitric acid, and then adding the phosphomolybdotungstic acid-chitosan-nano silicon dioxide compound, and uniformly stirring to obtain the detection reagent for the tyrosine metabolite in the urine.
In the invention, the preparation method of the detection layer comprises the following steps: placing filter paper on a filter, wetting by ethanol suction, and drying for 15 minutes by air; and then, carrying out suction filtration and deposition on the prepared urine detection reagent on filter paper, and then carrying out air drying for 15 minutes to prepare a second layer, namely a detection layer.
Drainage-imbibition layer
The raw materials for preparing the drainage and liquid absorption layer comprise filter paper and acrylic acid modified chitosan.
Acrylic acid modified chitosan
Weighing 1.0g of chitosan, fully dissolving in 6mL of 5 w% formic acid solution, diluting to 50mL with distilled water, transferring to a 150mL three-neck flask after fully swelling, transferring to a conventional oil bath reaction device, continuously stirring, heating to 80 ℃, and keeping constant temperature. Placing a 50mL beaker into an ice water mixture, transferring a certain amount of acrylic acid into the beaker, adjusting the pH value of the acrylic acid to be 6-6.5 by using a 5 w% NaOH solution, slowly adding the acrylic acid into a three-neck flask, reacting for a certain time at a preset temperature to obtain a grafting product solution, adjusting the pH value of the grafting product solution to be alkaline by using the 5 w% NaOH solution, precipitating the product in a flocculent manner, washing the product to be neutral by using a hydrochloric acid solution with the pH value of 4-5, performing suction filtration, drying, crushing, and sieving by using a 100-mesh sieve to obtain the product.
In the invention, the preparation method of the drainage liquid absorbing layer comprises the following steps: placing filter paper on a filter, wetting by ethanol suction, and drying for 15 minutes by air; and then, the prepared acrylic acid modified chitosan is filtered and deposited on filter paper, and is dried for 15 minutes by air, so that a third layer, namely a drainage liquid absorbing layer, can be prepared.
The ratio of the parts by weight of the acrylic acid modified chitosan to the parts by weight of the sodium tungstate is 1: 1.
in the invention, the bottom of the drainage liquid absorption layer is provided with a hydrophobic layer.
Hydrophobic layer
The hydrophobic layer is prepared from polytetrafluoroethylene.
In the invention, a water storage tank is arranged on the hydrophobic layer; when the urine amount is excessive, the urine is stored.
In the invention, the filter layer and the detection layer are connected through a peelable glue, and the peelable glue is a water-based peelable glue which is purchased from a Goods chemical network and has the model of GMY-628.
The second aspect of the present invention provides a method for detecting tyrosine metabolites in urine, comprising at least the following steps:
acquiring human urine to be detected;
dropping the human urine on urine test paper to judge whether color reaction occurs.
In one embodiment of the present invention, the detection method further comprises tearing off the filter layer, and further observing the color reaction.
The mechanism is explained as follows: the urine detection reagent and the urine detection test paper provided by the invention can eliminate the influence of protein in urine, so that the detection result is accurate and the color development effect is obvious; after urine is dripped on the test paper, the filter layer is torn off, so that the positive, weak positive and negative can be accurately judged, and the positive is red; weak positive is pink; the negative color is white or light yellow, so that the technical problem that people misjudge the color difference is solved.
The urine test paper comprises a filter layer 1, a detection layer 2 and a drainage liquid absorption layer 3, wherein a hydrophobic layer 4 is arranged at the bottom of the drainage liquid absorption layer, and a water storage tank 4-1 is arranged on the hydrophobic layer 4.
Embodiment 1: the embodiment provides a urine test paper for detecting tyrosine metabolites in urine, which consists of three detection layers, namely a first layer, a second layer and a third layer from top to bottom; the first layer is a filtering layer, the second layer is a detection layer, and the third layer is a drainage liquid absorption layer;
wherein the preparation raw material of the first layer comprises filter paper and modified glucose;
the preparation raw materials of the second layer comprise filter paper and a urine detection reagent;
the urine detection reagent comprises mercurous nitrate, mercuric sulfate, sodium tungstate and nitric acid;
the preparation raw material of the third layer comprises filter paper and acrylic acid modified chitosan.
Embodiment 2: a urine test strip as in embodiment 1, wherein the modified glucose is a dextran modified by a benzimidazole derivative and a benzoic acid derivative.
Embodiment 3: a urine test strip according to embodiment 2, wherein the benzimidazole derivative is one selected from the group consisting of 3- (1H-benzimidazol-2-yl) -2, 3-dihydroxypropionic acid, [ 3-carboxymethyl-2-oxo-2, 3-dihydro-1H-benzimidazole ] acetic acid, 5, 6-dicarboxybenzimidazole and benzimidazole.
Embodiment 4: a urine test strip according to embodiment 2, wherein said benzoic acid derivative is phthalic acid.
Embodiment 5: a urine test strip according to embodiment 1, wherein the urine test reagent further comprises phosphomolybdotungstic acid-chitosan-nano silica composite.
Embodiment 6: the urine test strip according to embodiment 1, wherein a hydrophobic layer is disposed on a bottom of the drainage-absorbent layer.
Embodiment 7: the urine test strip of embodiment 6, wherein the hydrophobic layer is provided with a water storage tank.
Embodiment 8: the urine test strip of embodiment 1, wherein the filter layer and the detection layer are connected by a peelable adhesive.
Embodiment 9: a method for detecting tyrosine metabolites in urine at least comprises the following steps:
acquiring human urine to be detected;
dropping the human urine on urine test paper to judge whether color reaction occurs.
Embodiment 10: the method for detecting tyrosine metabolites in urine according to embodiment 9, further comprising tearing off the filter layer and observing the color reaction.
The present invention will be specifically described below by way of examples. It should be noted that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention, and that the insubstantial modifications and adaptations of the present invention by those skilled in the art based on the above disclosure are still within the scope of the present invention.
In addition, the starting materials used are commercially available, unless otherwise specified, and the parts used for the following materials are parts by weight.
Example 1: the embodiment provides a urine test paper for detecting tyrosine metabolites in urine, which comprises three detection layers, namely a first layer, a second layer and a third layer from top to bottom; the first layer is a filter layer 1, the second layer is a detection layer 2, the third layer is a drainage liquid absorption layer 3,
wherein the preparation raw material of the first layer comprises filter paper and modified glucose;
the preparation method of the modified glucose comprises the following steps: [ 3-carboxymethyl-2-oxo-2, 3-dihydro-1H-benzimidazole ] acetic acid (0.35g,2.160mmol), terephthalic acid (0.166g,1mmol), 3- (4, 5-dimethylthiazol-2) -2, 5-diphenyltetrazolium bromide salt (0.825g,4.304mmol), dextran (0.5g,0.0125mmol) and 4-dimethylaminopyridine (0.056g,0.459mmol) were dissolved in a glass vial containing 40 ml of dimethyl sulfoxide. The reaction was carried out at 30 ℃ for 48 hours. Then placed in a dialysis bag for dialysis and impurities are removed.
In this example, the dextran has a molecular weight of 10000 daltons.
In the invention, the preparation method of the filter layer comprises the following steps: placing filter paper on a filter, wetting by ethanol suction, and drying for 15 minutes by air; and then dissolving the modified glucan prepared in the step (a) in a mixed solution of ethanol, performing suction filtration and deposition on filter paper, and then performing air drying for 15 minutes to prepare a first layer, namely a filter layer.
In the invention, the weight part ratio of the modified glucose to the mercuric sulfate is 1.3: 1.
in this embodiment, the raw materials for preparing the second layer include filter paper and a urine detection reagent.
The urine detection reagent contains mercurous nitrate, mercuric sulfate, sodium tungstate, nitric acid, phosphomolybdotungstic acid-chitosan-nano silicon dioxide compound.
The urine detection reagent is prepared from the following raw materials in parts by weight:
in this example, the phosphomolybdotungstic acid, chitosan, and nano-silica were all commercially available.
The phosphomolybdic tungstic acid is purchased from Xiamen sea Mark science and technology limited;
the chitosan is purchased from the bioscience and technology company Limited of Xianruin;
the nano silicon dioxide is purchased from winning Chuangdegusai.
The preparation method of the phosphomolybdotungstic acid-chitosan-nano silicon dioxide compound at least comprises the following steps:
(1) dissolving nano silicon dioxide in an ethanol solution, performing ultrasonic dispersion for 60min at room temperature, adding a silane coupling agent K560, and stirring for 3h at 45 ℃ to obtain a nano silicon dioxide solution; dissolving chitosan in acetic acid solution, and stirring at room temperature for 3h to obtain chitosan solution; adding the chitosan solution into the nano-silica solution, stirring and reacting for 8h, centrifuging and washing for 3 times, washing for 2 times with ethanol, and vacuum drying at 60 ℃ to obtain the chitosan-nano-silica composite.
(2) Dissolving phosphomolybdotungstic acid in deionized water to prepare a phosphomolybdotungstic acid solution with the mass fraction of 10%, adding the chitosan-nano silicon dioxide composite obtained in the step (1), refluxing and stirring at constant temperature for 20 hours, vacuum filtering, and drying to obtain the phosphomolybdotungstic acid-chitosan-nano silicon dioxide composite.
The preparation method of the urine detection reagent comprises the following steps: and uniformly mixing mercurous nitrate, mercuric sulfate, sodium tungstate and nitric acid, and then adding the phosphomolybdotungstic acid-chitosan-nano silicon dioxide compound, and uniformly stirring to obtain the detection reagent for the tyrosine metabolite in the urine.
In the invention, the preparation method of the detection layer comprises the following steps: placing filter paper on a filter, wetting by ethanol suction, and drying for 15 minutes by air; and then, carrying out suction filtration and deposition on the prepared urine detection reagent on filter paper, and then carrying out air drying for 15 minutes to prepare a second layer, namely a detection layer.
The raw materials for preparing the drainage and liquid absorption layer comprise filter paper and acrylic acid modified chitosan.
Acrylic acid modified chitosan
Weighing 1.0g of chitosan, fully dissolving in 6mL of 5 w% formic acid solution, diluting to 50mL with distilled water, transferring to a 150mL three-neck flask after fully swelling, transferring to a conventional oil bath reaction device, continuously stirring, heating to 80 ℃, and keeping constant temperature. Placing a 50mL beaker into an ice water mixture, transferring a certain amount of acrylic acid into the beaker, adjusting the pH value of the acrylic acid to be 6-6.5 by using a 5 w% NaOH solution, slowly adding the acrylic acid into a three-neck flask, reacting for a certain time at a preset temperature to obtain a grafting product solution, adjusting the pH value of the grafting product solution to be alkaline by using the 5 w% NaOH solution, precipitating the product in a flocculent manner, washing the product to be neutral by using a hydrochloric acid solution with the pH value of 4-5, performing suction filtration, drying, crushing, and sieving by using a 100-mesh sieve to obtain the product.
In the invention, the preparation method of the drainage liquid absorbing layer comprises the following steps: placing filter paper on a filter, wetting by ethanol suction, and drying for 15 minutes by air; and then, the prepared acrylic acid modified chitosan is filtered and deposited on filter paper, and is dried for 15 minutes by air, so that a third layer, namely a drainage liquid absorbing layer, can be prepared.
The ratio of the parts by weight of the acrylic acid modified chitosan to the parts by weight of the sodium tungstate is 1: 1.
in the invention, the bottom of the drainage liquid absorption layer is provided with a water drainage layer 4.
The hydrophobic layer is prepared from polytetrafluoroethylene.
In the invention, a water storage tank 4-1 is arranged on the hydrophobic layer 4; when the urine amount is excessive, the urine is stored.
In the invention, the filter layer and the detection layer are connected through a peelable glue, and the peelable glue is a water-based peelable glue which is purchased from a Goods chemical network and has the model of GMY-628.
In a second aspect of the present invention, there is provided a method for detecting tyrosine metabolites in urine, comprising the steps of:
acquiring human urine to be detected;
dropping the human urine on urine test paper to judge whether color reaction occurs.
The detection method also comprises the steps of tearing off the filter layer and further observing the color reaction.
Wherein, the volume ratio of the urine to the detection reagent is 1: 3.
example 2: this example differs from example 1 in that the urine detection reagent does not comprise a phosphomolybdotungstic acid-chitosan-nanosilica complex.
Example 3: this example differs from example 1 in that the starting material for the first layer is free of modified glucose.
Example 4: this example is different from example 1 in that the preparation raw material of the first layer modified glucose was changed to unmodified glucose, which was purchased from national drug group chemical agents limited.
Example 5: this example differs from example 1 in that the modified glucose of the first layer is added to the urine test reagent of the second layer, i.e., the raw materials for preparing the urine test reagent of the second layer comprise: modified glucose, mercurous nitrate, mercuric sulfate, sodium tungstate and nitric acid.
Example 6: this example differs from example 1 in that the color reaction was directly observed without tearing off the filter layer during the detection of tyrosine metabolites in urine.
Example 7: the difference between this embodiment and embodiment 1 is that the positions of the first filter layer and the second detection layer are replaced, and the detection test paper sequentially comprises a detection layer, a filter layer, and a drainage liquid absorption layer from top to bottom.
Example 8: the difference between the embodiment and the embodiment 1 is that the modified glucose is glucose modified by benzimidazole derivative, and the preparation method of the modified glucose is as follows: [ 3-carboxymethyl-2-oxo-2, 3-dihydro-1H-benzimidazole ] acetic acid (0.35g,2.160mmol), 3- (4, 5-dimethylthiazol-2) -2, 5-diphenyltetrazolium bromide salt (0.825g,4.304mmol), dextran (0.5g,0.0125mmol) and 4-dimethylaminopyridine (0.056g,0.459mmol) were dissolved in a glass vial containing 40 ml of dimethyl sulfoxide. The reaction was carried out at 30 ℃ for 48 hours. Then placed in a dialysis bag for dialysis and impurities are removed.
Example 9: the difference between this example and example 1 is that the modified glucose is benzoic acid derivative modified glucose, and the preparation method of the modified glucose is as follows: terephthalic acid (0.166g,1mmol), 3- (4, 5-dimethylthiazole-2) -2, 5-diphenyltetrazolium bromide (0.825g,4.304mmol), dextran (0.5g,0.0125mmol) and 4-dimethylaminopyridine (0.056g,0.459mmol) were dissolved in a glass vial containing 40 ml of dimethyl sulfoxide. The reaction was carried out at 30 ℃ for 48 hours. Then placed in a dialysis bag for dialysis and impurities are removed.
Example 10: this example differs from example 1 in that the starting material for the third layer was made without acrylic modified chitosan.
Example 11: this example differs from example 1 in that the starting materials for the preparation of the third layer comprise filter paper and chitosan.
And (3) testing:
1000 cancer patients with an empty stomach in the morning are selected, and 10mL of urine is respectively taken as human urine to be detected.
The detection method comprises the following steps:
1. the human urine to be detected is respectively contacted with the urine test paper in the embodiments 1 to 11, and whether the color reaction occurs is judged.
Wherein a positive is presented in red; weak positive is pink; negatives appeared white or light yellow.
Table 1 characterization test
The detection rates of 100 cervical cancer patients, 100 gastric cancer patients, 100 breast cancer patients, 100 liver cancer patients and 100 lung cancer patients were tested according to the selection of example 1, example 2, example 3, example 5, example 6 and example 7, and the results are shown in table 2.
TABLE 2 cancer detection Rate
The foregoing examples are illustrative only, and serve to explain some of the features of the present disclosure. The appended claims are intended to claim as broad a scope as is contemplated, and the examples presented herein are merely illustrative of selected implementations in accordance with all possible combinations of examples. Accordingly, it is applicants' intention that the appended claims are not to be limited by the choice of examples illustrating features of the invention. And that advances in science and technology will result in possible equivalents or sub-substitutes not currently contemplated for reasons of inaccuracy in language representation, and such changes should also be construed where possible to be covered by the appended claims.
Claims (6)
1. A urine test paper is used for detecting tyrosine metabolites in urine and is characterized by comprising three detection layers, namely a first layer, a second layer and a third layer from top to bottom; the first layer is a filtering layer, the second layer is a detection layer, and the third layer is a drainage liquid absorption layer;
wherein the preparation raw material of the first layer comprises filter paper and modified glucose; the modified glucose is glucan jointly modified by benzimidazole derivatives and benzoic acid derivatives; the benzimidazole derivative is [ 3-carboxymethyl-2-oxo-2, 3-dihydro-1H-benzimidazole ] acetic acid; the benzoic acid derivative is terephthalic acid;
the preparation raw materials of the second layer comprise filter paper and a urine detection reagent;
the urine detection reagent comprises mercurous nitrate, mercuric sulfate, sodium tungstate and nitric acid;
the preparation raw material of the third layer comprises filter paper and acrylic acid modified chitosan.
2. The urine test strip of claim 1, wherein said urine test reagent further comprises phosphomolybdotungstic acid-chitosan-nanosilica complex.
3. The urine test strip of claim 1 wherein the bottom of said drainage fluid layer is provided with a hydrophobic layer.
4. The urine test strip of claim 3 wherein said hydrophobic layer is provided with a reservoir.
5. The urine test strip of claim 1 wherein said filter layer and said detection layer are joined by a peelable adhesive.
6. A method for detecting tyrosine metabolites in urine is characterized by at least comprising the following steps:
acquiring human urine to be detected;
dropping the human urine on urine test paper to judge whether a color reaction occurs;
the detection method also comprises the steps of tearing off the filter layer within 1-3 seconds after the urine is dripped on the urine detection test paper, and further observing the color reaction;
the urine test strip is the urine test strip according to any one of claims 1 to 5.
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