Urine pretreatment device for detecting urine iodine as well as preparation method and application of urine pretreatment device

Abstract

The invention provides a urine pretreatment device for detecting urine iodine. The urine pretreatment device comprises a solid-phase extraction small column, a booster and a cleaning solution. The solid-phase extraction small column adopts a urine treatment matrix as a filler, the specific surface area of the matrix is large, and the column efficiency is high; sodium indophenol compounds are bonded on the surface of the activated carbon to serve as a treatment reagent, reduction type substances such as vitamin C can be accurately adsorbed or grabbed, and treatment is cleaner; the extraction column filling technology is mature, and batch production can be realized; the urine pretreatment device is provided with a cleaning solution, the solid-phase extraction small column is washed by the cleaning solution before use, and interference substances in urine can be efficiently adsorbed after a matrix is activated; the selected booster is tightly combined with the column cylinder of the small solid-phase extraction column, so that the pressure tolerance is high, the pushing is smooth, and the urine pretreatment effect is ensured; the method is especially suitable for dry chemical detection and kit detection of iodide ions in urine. The invention has the advantages of short urine treatment time, strong interference removal capability, high efficiency, stable performance, simple preparation process and convenience in use, and is suitable for wide-range application.

Description

Technical Field

[0001] The present invention relates to the technical field of in vitro diagnostic reagents, and particularly to a urine pretreatment device for detecting urinary iodine, a preparation method thereof, and an application thereof. Background Art

[0002] Iodine deficiency disorder is widely distributed worldwide and is a micro-nutrient deficiency disorder that poses the greatest harm to human intellectual health. Clinically, the iodine nutritional status of the human body is usually evaluated by detecting urinary iodine. The classic method for determining urinary iodine is the Sandell-Kolthoff reaction, which is the method recommended by the WTO and the IDDICC for urine iodine determination. The detection principle is that iodide ions catalyze the redox reaction to produce a color change. However, some interfering substances in urine will act as oxidants or reductants to affect the redox reaction. Therefore, in order to remove these interfering substances during urine detection, urine needs to be pretreated.

[0003] In previous urine pretreatment methods, urine had to be digested with perchloric acid at 110°C, or the specimen was ashed in a high-temperature furnace before determination. However, perchloric acid is very unstable. After method improvement, ammonium sulfate was used to digest urine to increase the stability of the reagent. However, this method requires special digestion instruments, has a long pretreatment time, is not convenient to operate, and is not convenient to popularize and apply in grass-roots areas with severe iodine deficiency disorders. In recent years, there have been reports on the application of reagent kits in urine iodine detection. Among them, urine pretreatment is to treat urine through a titration container, and specific adsorption of interfering substances in urine is achieved through an adsorbent. This urine treatment method is time-consuming, inconvenient to use, and not conducive to large-scale use.

[0004] Therefore, it is very necessary to overcome the deficiencies of the previous urine pretreatment methods in urine iodine detection, and to provide a urine pretreatment device for detecting urinary iodine, a preparation method thereof, and an application thereof, which is simple to operate, convenient to use, has a good interference removal effect, and is beneficial to detection. Summary of the Invention

[0005] The object of the present invention is to provide a urine pretreatment device for detecting urinary iodine, a preparation method thereof, and an application thereof. The present invention can simply and quickly remove interfering substances in urine, and has no influence on iodide ions in urine. This method has a short treatment time, a good interference removal effect, stable performance, is convenient to use, is beneficial to detection, and is suitable for large-scale application.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] One aspect of the present invention is to provide a urine pretreatment device for detecting urinary iodine, including a solid-phase extraction cartridge, a booster, and a cleaning solution; wherein,

[0008] The solid-phase extraction cartridge is used to hold the urine treatment matrix and filter urine, and includes an empty cylinder. The top of the empty cylinder is provided with an opening, the bottom is provided with small holes, and the inside is sealed with a filler through a front sieve plate and a rear sieve plate;

[0009] The booster is used to apply pressure to the column after filling with liquid, and is tightly combined with the empty cylinder;

[0010] The cleaning solution is used to clean the solid-phase extraction column and is prepared from a buffer solution.

[0011] Another aspect of the present invention is to provide a preparation method of a urine pretreatment device for detecting urinary iodine, including the following steps:

[0012] 1) Prepare the solid-phase extraction cartridge: Mix activated carbon and a treatment reagent in proportion, add a certain proportion of pure water, control the temperature and stir to react, filter by suction, filter out the filtrate, collect the filter cake, bake the filter cake until dry, collect it, and obtain the urine treatment matrix for standby; Take an empty cylinder, add a front sieve plate at the front end, add the urine treatment matrix, and seal the rear end with a rear sieve plate to form a filler, compact and seal it to obtain the solid-phase extraction cartridge;

[0013] 2) Prepare the booster: The booster can be tightly combined with the empty cylinder and push smoothly;

[0014] 3) Prepare the cleaning solution: Prepare a buffer solution according to a certain molar concentration and pH, filter and collect the filtrate to obtain the cleaning solution.

[0015] Preferably, in step 1), the activated carbon is one of coal-based activated carbon, synthetic material activated carbon, and wood-based activated carbon. The activated carbon has a mesh number of 150-400 and a porosity of 50-80%;

[0016] Preferably, the treatment reagent in the present invention is one of potassium permanganate and sodium nitrite; Research results show that when the treatment reagent is potassium permanganate or sodium nitrite, the substances generated after reacting with the reducing substances in urine cannot be adsorbed by activated carbon and remain in the urine, affecting the determination results of urinary iodine;

[0017] Preferably, the treatment reagent in the present invention is one or several of indophenol compounds; Research results show that the substances generated after the reaction of indophenol compounds with the reducing substances in urine can be adsorbed by activated carbon, and the influence of the reducing substances in urine can be eliminated. However, indophenol compounds have low water solubility and require solvent solubilization, which affects detection and is not suitable for use;

[0018] Preferably, the present invention selects one or several of indophenol sodium salt compounds; Research results show that indophenol sodium salt compounds have good water solubility, can be completely adsorbed by activated carbon, and can be completely bonded with activated carbon at high temperature to form a filler for use in a solid-phase extraction cartridge, and the application effect is good;

[0019] More preferably, compared with sodium chlorophenol indophenol, sodium bromophenol indophenol has stronger adsorption ability and bonding ability with activated carbon, and better application effect;

[0020] The treatment reagent is one or more of sodium indophenol compounds. Further, the treatment reagent in the present invention is one or more of sodium bromophenol indophenol compounds;

[0021] Indophenol is o-chlorophenol indophenol, 3'-chloroindophenol, 5-isopropyl-2-methylindophenol, 2,6-dichloroindophenol, 2,6-dichloroindophenol acetate, 2,3',6-trichloroindophenol, 2,2',6-trichloro-indophenol, 2,6-dichloro-3'-methylindophenol, 2,6-dibromoindophenol, 2,6-dibromoindophenol acetate, 2,3',6-tribromoindophenol, 2,2',6-tribromo-indophenol, 2,6-dibromo-3'-methylindophenol; Further, the bromoindophenol is 2,6-dibromoindophenol, 2,6-dibromoindophenol acetate, 2,3',6-tribromoindophenol, 2,2',6-tribromo-indophenol, 2,6-dibromo-3'-methylindophenol;

[0022] The mass ratio of the activated carbon to the treatment reagent is 100:1 - 15; the volume ratio of the activated carbon to the pure water is 1:1 - 10; the reaction temperature of the temperature-controlled stirring is 20 - 100 °C, and the reaction time of the temperature-controlled stirring is 1 - 12 h;

[0023] Preferably, in step 1), the baking temperature is 90 - 150 °C; the baking time is 1 - 12 h.

[0024] Preferably, in step 1), the mass of the urine treatment matrix in the solid phase extraction column is 0.1 - 2.0 g;

[0025] Preferably, in step 2), the cleaning solution is one of potassium dihydrogen phosphate-disodium hydrogen phosphate buffer solution, potassium citrate-citric acid buffer solution, acetic acid-sodium acetate buffer solution;

[0026] The concentration of the buffer solution is 5 - 200 mM, the pH value of the buffer solution is 5 - 7, and the volume of the buffer solution is 5 - 20 ml;

[0027] Preferably, in step 3), the tolerance pressure of the booster combined with the empty column cylinder is 0.1 - 1.0 MPa;

[0028] One aspect of the present invention is to provide an application of a urine pretreatment device for detecting urinary iodine, including the following steps:

[0029] 1) Solid-phase extraction cartridge cleaning: At room temperature, add the cleaning solution into the cartridge cylinder, push the booster into the cartridge cylinder, push the booster to clean the urine treatment matrix, and push out the excess cleaning solution;

[0030] 2) Urine pretreatment: At room temperature, add the natural urine to be tested into the solid-phase extraction cartridge, push the booster slowly to filter, collect the filtrate to obtain the natural urine sample to be tested;

[0031] 3) Urinary iodine detection:

[0032] When applied to dry chemical detection of urinary iodine, at room temperature, immerse the urinary iodine test strip into the natural urine sample to be tested at room temperature respectively, take it out, react for a certain time, compare the color of the reaction result of the urinary iodine test strip with the standard colorimetric card, and read the value;

[0033] When applied to the detection of urinary iodine kit, at room temperature, add the second reaction solution of the urinary iodine kit into the natural urine sample to be tested, then add the first reaction solution of the urinary iodine kit, shake well, react for a certain time, compare the color of the reaction solution color development result with the standard colorimetric card, and read the value.

[0034] Preferably, in step 1), the room temperature is 20 - 25 °C; the cleaning time of the cleaning solution is 1 - 10 s.

[0035] Preferably, in step 2), the natural urine to be tested is 24-hour urine or mid-stream morning urine, and the urine sample is stored at 4 - 8 °C for later use; the volume of the natural urine to be tested is 1 - 5 ml; the filtration speed of the natural urine to be tested is 1 - 10 s.

[0036] Preferably, in step 3), when applied to dry chemical detection of urinary iodine, the mass-volume ratio of the urine treatment matrix to the natural urine sample to be tested is 0.5 g:1 - 5 ml; the immersion time of the urinary iodine test strip into the urine is 1 - 5 s; the reaction time is 0.5 - 1 min;

[0037] When applied to the detection of urinary iodine kit, the treatment time of the urine treatment matrix is 1 - 10 s; the volume ratio of the natural urine sample to be tested, the second reaction solution and the first reaction solution in the urinary iodine kit is 10:3 - 6:1 - 2; the reaction time is 0.5 - 1 minute.

[0038] The principle adopted by a urine pretreatment device for detecting urinary iodine provided by the present invention, its preparation method and application is as follows: Powdered activated carbon has a microcrystalline structure with irregular arrangement. There are micropores in the crystal, which endows it with a large specific surface area and strong adsorption capacity. Organic substances with relatively large molecular weights such as pigments, proteins and bilirubin in urine are easily adsorbed by activated carbon, thereby excluding the interference of pigments and macromolecular organic substances in urine. Reducing substances such as vitamin C in urine have a certain reducibility, which will cause certain interference to the redox reaction catalyzed by iodide ions. By adding oxidative treatment reagents such as indophenol sodium compounds, which are adsorbed on the surface of activated carbon and further bonded by high-temperature baking, a urine treatment matrix with a large specific surface area is formed. Once it comes into contact with reducing substances such as vitamin C in an aqueous solution, a reaction occurs rapidly, thereby removing the interference of reducing substances such as vitamin C in urine. Iodide ions in urine exist in the form of inorganic salts, are not adsorbed and reacted by the urine treatment matrix, and are not affected. They can be easily filtered out in the urine pretreatment device to obtain a colorless and pure urine sample to be detected that can be accurately measured.

[0039] The present invention has achieved the following beneficial technical effects compared with the prior art:

[0040] A urine pretreatment device for detecting urinary iodine provided by the present invention, firstly, the filler used in the solid-phase extraction cartridge is a urine treatment matrix, which is formed by the uniform dispersion adsorption of activated carbon and treatment reagents in an aqueous phase under controlled temperature and then bonded by high-temperature baking. The matrix has a large specific surface area and high column efficiency; secondly, indophenol sodium compounds are bonded to the surface of activated carbon as treatment reagents, which can accurately adsorb or capture reducing substances such as vitamin C and the treatment is cleaner; thirdly, the filling technology of the extraction cartridge is mature and can be mass-produced; in addition, the urine pretreatment device of the present invention is equipped with a cleaning solution. Before use, the solid-phase extraction cartridge is rinsed with the cleaning solution. After activating the matrix, it can efficiently adsorb the interfering substances in urine; finally, the selected booster is tightly combined with the cylinder of the solid-phase extraction cartridge, has high pressure tolerance and smooth pushing, ensuring the urine pretreatment effect; The present invention provides an application of a urine pretreatment device for detecting urine, which is particularly suitable for the dry chemical detection and kit detection of iodide ions in urine. Therefore, the urine pretreatment device for detecting urinary iodine provided by the present invention has a short urine treatment time, strong interference removal ability, high efficiency, stable performance, and a simple preparation process, is easy to use, and is suitable for wide application. Description of the Drawings

[0041] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0042] Figure 1 Schematic structural diagram of a urine pretreatment device for detecting urinary iodine in the present invention. Specific embodiments

[0043] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0044] The purpose of the present invention is to provide a urine pretreatment device for detecting urinary iodine, its preparation method and application to solve the problems existing in the prior art.

[0045] A urine pretreatment device for detecting urinary iodine, as Figure 1 shown, includes a solid-phase extraction cartridge 1, a booster 2 and a cleaning solution; wherein,

[0046] The solid-phase extraction cartridge 1 is used to hold the urine treatment matrix and filter urine, and includes an empty cylinder 11 with an opening at the top and a small hole at the bottom, and a packing 14 is enclosed inside by a front sieve plate 12 and a rear sieve plate 13;

[0047] The booster 2 is used to apply pressure to the column after loading the liquid, and it cooperates with the empty cylinder 11;

[0048] The cleaning solution is used to clean the solid-phase extraction column and is prepared from a buffer solution.

[0049] A preparation method of a urine pretreatment device for detecting urinary iodine includes the following steps:

[0050] 1) Prepare the solid-phase extraction cartridge: Mix activated carbon and a treatment reagent in proportion, add a certain proportion of pure water, control the temperature, stir, filter, collect the filter cake, dry, and collect to obtain the urine treatment matrix for standby; Take an empty cylinder, add a front sieve plate at the front end, add the urine treatment matrix, and seal the rear end with a rear sieve plate to form a packing, compact and seal to obtain the solid-phase extraction cartridge;

[0051] Among them, the activated carbon is one of coal-based activated carbon, synthetic material activated carbon, and wood-based activated carbon, preferably wood-based activated carbon; The crystal form of the activated carbon is preferably powdery, the mesh number of the activated carbon is 150 - 400 meshes, preferably 250 - 350 meshes, and the porosity is 50 - 80%, preferably 60 - 70%;

[0052] The treatment reagent is one or several of indophenol sodium salt compounds. Further, the treatment reagent in the present invention is one or several of bromoindophenol sodium salt compounds;

[0053] The indophenols mentioned above are o-chlorophenol indophenol, 3'-chloroindophenol, 5-isopropyl-2-methylindophenol, 2,6-dichloroindophenol, 2,6-dichloroindophenol acetate, 2,3',6-trichloroindophenol, 2,2',6-trichloroindophenol, 2,6-dichloro-3'-methylindophenol, 2,6-dibromoindophenol, 2,6-dibromoindophenol acetate, 2,3',6-tribromoindophenol, 2,2',6-tribromoindophenol, 2,6-dibromo-3'-methylindophenol; further, the bromoindophenols are 2,6-dibromoindophenol, 2,6-dibromoindophenol acetate, 2,3',6-tribromoindophenol, 2,2',6-tribromoindophenol, 2,6-dibromo-3'-methylindophenol;

[0054] The mass ratio of activated carbon to treatment reagent is 100:1 - 15, preferably 100:5 - 10; the volume ratio of activated carbon to pure water is 1:1 - 10, preferably 1:2 - 6; the reaction temperature of temperature-controlled stirring is 20 - 100 °C, preferably 50 - 90 °C; the reaction time of temperature-controlled stirring is 1 - 12 h, preferably 2 - 8 h; the baking temperature is 90 - 150 °C, preferably 105 - 130 °C; the drying time is 1 - 12 h, preferably 2 - 8 h;

[0055] The mass of the urine treatment matrix in the solid-phase extraction cartridge is 0.1 - 2.0 g, preferably 0.5 - 1.0 g;

[0056] 2) Prepare the booster: The booster can be tightly combined with the empty column cylinder, and the pressure tolerance of the combination of the booster and the empty column cylinder is 0.1 - 1.0 MPa, preferably 0.2 - 0.7 MPa;

[0057] 3) Prepare the cleaning solution: Prepare a buffer solution according to a certain molar concentration and pH, filter and collect the filtrate to obtain the cleaning solution;

[0058] Among them, the cleaning solution is one of potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution, potassium citrate - citric acid buffer solution, acetic acid - sodium acetate buffer solution, preferably potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution; the concentration of the buffer solution is 5 - 200 mM, preferably 10 - 100 mM, the pH value of the buffer solution is 5 - 7, preferably 5.6 - 6.8, and the volume of the buffer solution is 5 - 20 ml, preferably 5 - 10 ml.

[0059] An application of a urine pretreatment device for detecting urinary iodine includes the following steps:

[0060] 1) Clean the solid-phase extraction cartridge: At room temperature, add the cleaning solution into the column cylinder, push the booster into the column cylinder, push the booster to clean the urine treatment matrix, and push out the excess cleaning solution;

[0061] The cleaning solution is one of potassium dihydrogen phosphate-disodium hydrogen phosphate buffer solution, potassium citrate-citric acid buffer solution, and acetic acid-sodium acetate buffer solution, preferably potassium dihydrogen phosphate-disodium hydrogen phosphate buffer solution; the concentration of the buffer solution is 5-200 mM, preferably 10-100 mM; the pH of the buffer solution is 5-7, preferably 5.6-6.8; the volume of the buffer solution is 5-20 ml, preferably 5-10 ml; the cleaning time of the cleaning solution is 1-10 s, preferably 2-6 s;

[0062] 2) Urine pretreatment: At room temperature, add the natural urine to be tested into the solid-phase extraction column, push the booster to filter slowly, and collect the filtrate to obtain the natural urine to be tested;

[0063] The natural urine is 24-hour urine or mid-stream morning urine, and the urine sample is stored at 4-8 °C for later use; the volume of the pretreated natural urine is 1-5 ml, preferably 1-2.5 ml; the filtration speed of the natural urine is 1-10 s, preferably 2-6 s;

[0064] 3) Urinary iodine detection:

[0065] When applied to dry chemical detection of urinary iodine, at room temperature, immerse the urinary iodine test strip into the natural urine to be tested at room temperature respectively, take it out, react at a fixed time, compare the reaction result of the urinary iodine test strip with the standard colorimetric card for color comparison, and read the value;

[0066] The mass-volume ratio of the urine treatment matrix to the natural urine to be tested is 0.5 g:1-5 ml, preferably 0.5 g:1-2.5 ml; the time for the urinary iodine test strip to be immersed in the urine is 1-5 s, preferably 1-2 s; the reaction time is 0.5-1 min, preferably 1 min;

[0067] The standard colorimetric card has five iodine ion concentrations of 0, 50, 150, 300, and 550 μg / l, corresponding to five color scales respectively; the preparation method of the standard colorimetric card is: take potassium iodide or sodium iodide standard substance, add sodium acetate buffer solution with pH 5.5, and prepare standard solutions with five iodine ion concentrations of 0, 50, 150, 300, and 550 μg / l for standby; clean the solid-phase extraction column with the cleaning solution, add the iodine ion standard solution, push the booster to filter, and collect the filtrate to obtain the iodine ion to be tested; at room temperature, immerse the urinary iodine test strip into the iodine ion to be tested at each concentration respectively, take it out, react at a fixed time for 1 min, read the value, and use the iodine ion concentration to be tested and the corresponding end color of the urinary iodine test strip as color standards to obtain the standard colorimetric card with five color scales correspondingly;

[0068] The preparation method of the urinary iodine test strip is: at room temperature, immerse the filter paper in solution A, dry it, then immerse it in solution B, dry it, cut it into strips to obtain a reagent module; paste one side of the reagent module on a magnetic white sheet, and then cut the magnetic white sheet into strips to obtain the urinary iodine test strip;

[0069] Wherein:

[0070] Solution A contains a buffer, an oxidizing agent, and a stabilizer; the buffer is one of potassium dihydrogen phosphate - disodium hydrogen phosphate buffer or disodium hydrogen phosphate - sodium hydroxide buffer solution, and the concentration of the buffer is 0.1 - 0.5 M; the pH of the buffer is 8.0 - 10.0; the oxidizing agent is one or more of benzenesulfonyl chloramine, benzenesulfonyl bromamine, p - toluenesulfonyl chloramine, p - toluenesulfonyl bromamine, o - toluenesulfonyl chloramine, o - toluenesulfonyl bromamine, N,N - dichlorobenzenesulfonamide, N,N - dibromobenzenesulfonamide, N,N - dichloro - p - toluenesulfonamide, N,N - dibromo - p - toluenesulfonamide, sodium or potassium salts of p - (N,N - dichloroaminosulfonyl)benzoic acid, p - (N,N - dibromoaminosulfonyl)benzoic acid; the stabilizer is one or two of glucose T70 or maltodextrin; the volume - mass ratio of the buffer, the oxidizing agent, and the stabilizer is 1000 mL: 0.5 - 5.0 g: 3.0 - 30.0 g;

[0071] Solution B contains a chromogenic agent and an organic solvent; the chromogenic agent is one of N - (2 - sulfoethyl)-3,3,5,5 - tetramethylbenzidine, N - (3 - sulfopropyl)-3,3,5,5 - tetramethylbenzidine, N - (4 - sulfobutyl)-3,3,5,5 - tetramethylbenzidine; the mass - volume ratio of the chromogenic agent and the organic solvent is 1.0 - 10.0 g: 1000 mL; the organic solvent is one of acetone, dichloromethane, or acetonitrile;

[0072] The filter paper is one of 3MM - type filter paper or 31ET - type filter paper; the room temperature is 20 - 25 °C; the impregnation time of the filter paper in Solution A and Solution B is independently 10 - 30 s; after the filter paper is immersed in Solution A and taken out, it is dried, and the drying temperature is 40 - 70 °C, and the drying speed is 90 - 110 m / h; after the filter paper is immersed in Solution B and taken out, the drying temperature is 30 - 50 °C, and the drying speed is 90 - 110 m / h;

[0073] In the step of cutting the test paper into strips, the width of the strips is 5 - 7 mm. In the step of cutting the porcelain white sheet into strips, the width of the porcelain white sheet is 5 - 7 mm; the length of the porcelain white sheet is 50 - 80 mm; for convenient use, the dried filter paper can be cut to the required size, one side of the filter paper is adhered to the porcelain white sheet through double - sided tape, and then cut to the required size to obtain the urinary iodine test paper;

[0074] When applied to the detection of the urinary iodine test kit, at room temperature, the second reaction solution of the urinary iodine test kit is added to the natural urine sample to be tested, and then the first reaction solution of the urinary iodine test kit is added, shaken well, reacted for a certain time, and the color development result of the reaction solution is compared with the standard colorimetric card for reading;

[0075] The volume ratio of the natural urine sample to be tested, the second reaction solution, and the first reaction solution in the urine iodine test kit is 10:3 - 6:1 - 2; the reaction time is 0.5 - 1 min, preferably 1 min;

[0076] The standard color comparison card has five iodine ion concentrations of 0, 50, 150, 300, and 550 μg / l, corresponding to four color levels respectively; the preparation method of the standard color comparison card is as follows: Take potassium iodide or sodium iodide standard substances, add sodium acetate buffer solution with pH 5.5, and prepare standard solutions with five iodine ion concentrations of 0, 50, 150, 300, and 550 μg / l for standby; Wash the solid-phase extraction column with the cleaning solution, add the iodine ion standard solution, push the booster to filter, collect the filtrate, and separate the iodine ions from the sample to be tested; At room temperature, take 100 μl of the sample to be tested with five iodine ion concentrations respectively, add 40 μl of the second reaction solution, then add 10 μl of the first reaction solution, shake well, and time for 60 s; Repeat each concentration 3 times, and use the color development results of the five concentrations as color standards to obtain the standard color comparison card with four color levels accordingly;

[0077] The preparation method of the urine iodine test kit is as follows: According to a certain mass-volume ratio, add the oxidant and stabilizer to the buffer solution, mix well, filter, and collect the filtrate to prepare the first reaction solution; According to a certain mass-volume ratio, add the color developer and stabilizer to deionized water, mix well, filter, and collect the filtrate to prepare the second reaction solution;

[0078] Among them:

[0079] The oxidant is one or several of sodium or potassium salts of benzenesulfonylchloramide, benzenesulfonylbromamide, p-toluenesulfonylchloramide, p-toluenesulfonylbromamide, o-toluenesulfonylchloramide, o-toluenesulfonylbromamide, N,N-dichlorobenzenesulfonamide, N,N-dibromobenzenesulfonamide, N,N-dichloro-p-toluenesulfonamide, N,N-dibromo-p-toluenesulfonamide, p-(N,N-dichloroaminosulfonyl)benzoic acid, and p-(N,N-dibromoaminosulfonyl)benzoic acid;

[0080] The stabilizer in the first reaction solution is one of dextran T70, sodium alginate, and sodium carboxymethylcellulose; the stabilizer in the second reaction solution is one of polyvinylpyrrolidone and / or sodium carboxymethylcellulose;

[0081] The color developer is one of sodium 3,3',5,5'-tetramethylbenzidine ethanesulfonate, sodium 3,3',5,5'-tetramethylbenzidine propanesulfonate, and sodium 3,3',5,5'-tetramethylbenzidine butanesulfonate;

[0082] The buffer solution in the first reaction solution is potassium dihydrogen phosphate-disodium hydrogen phosphate buffer solution or disodium hydrogen phosphate-sodium hydroxide buffer solution, and the pH of the buffer solution is 9.0 - 9.5;

[0083] The first reaction solution contains 1 - 10 mM of oxidant, 10 - 35 g / L of stabilizer, and 1 L of buffer solution; the second reaction solution contains 1 - 10 g / L of chromogenic agent, 10 - 35 g / L of stabilizer, and 1 L of deionized water;

[0084] The room temperature is 20 - 25 °C.

[0085] A urine pretreatment device for detecting urinary iodine provided by the present invention is applicable to detection reagents with the detection principle of iodide ion catalysis method, especially detection reagents for iodide ion catalytic redox reaction, and further detection reagents using TMB or its derivatives as chromogenic agents.

[0086] The research results show that substances with redox properties in urine are all interfering substances in the determination, such as vitamin C, bilirubin, occult blood (red blood cells), nitrite and other substances. Heme in occult blood (red blood cells) has the function of catalase and interferes with the reaction of substrates with hydrogen peroxide. However, for the urinary iodine detection reagent applicable to the present invention, the substrate is TMB or its derivatives, and the substrate does not have hydrogen peroxide, so there is no interference; nitrite has oxidizing property. The experimental results show that nitrite in urine has no interference with the dry chemical test strip or urine iodine test kit applicable to the present invention. The obvious interfering factors are vitamin C and bilirubin, and the most obvious one is vitamin C.

[0087] Therefore, for a urine pretreatment device for detecting urinary iodine provided by the present invention, the experimental results prove that it can completely eliminate the influence of interfering substances such as vitamin C and bilirubin.

[0088] To make the above objects, features and advantages of the present invention more obvious and understandable, the present invention will be further described in detail below with reference to the drawings and specific embodiments.

[0089] Example 1: Preparation of a urine pretreatment device for detecting urinary iodine:

[0090] 1) Add 50 g of powdered activated carbon (300 mesh, porosity of 65%) to a beaker, add 3 g of 2,6 - dibromoindophenol sodium, add 100 ml of pure water, stir and react at 50 °C for 8 h, perform suction filtration, collect the filter cake, dry it at 105 °C for 8 h, cool to room temperature, and collect to obtain a urine treatment matrix; take multiple empty cylinders, add a sieve plate at the front end of the cylinder, add 0.5 g of the urine treatment matrix to each cylinder, add a sieve plate to seal the rear end of the cylinder, compact it, and seal it to obtain a solid phase extraction small column;

[0091] 2) Prepare a booster that can be tightly combined with the empty cylinder, and the pressure tolerance of the combination of the booster and the empty cylinder is 0.2 MPa;

[0092] 3) Prepare 30 ml of potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution with a concentration of 20 mM and a pH of 6.0, filter it, collect the filtrate, and dispense it into 5 - ml vials to obtain the cleaning solution.

[0093] Example 2:

[0094] Preparation of a urine pretreatment device for detecting urinary iodine:

[0095] 1) Add 50 g of powdered activated carbon (300 mesh, porosity of 65%) to a beaker, add 5 g of sodium 2,6 - dibromoindophenol acetate, add 200 ml of pure water, stir and react at 60 °C for 6 h, perform suction filtration, collect the filter cake, dry it at 110 °C for 6 h, cool it to room temperature, and collect it to obtain the urine treatment matrix; Take multiple empty cylindrical columns, add a sieve plate at the front end of the column, add 1.0 g of the urine treatment matrix to each column, add a sieve plate to seal the rear end of the column, compact it, and seal it to obtain a solid - phase extraction cartridge;

[0096] 2) Prepare a booster that can be tightly combined with the empty cylindrical column, and the tolerance pressure for the combination of the booster and the empty cylindrical column is 0.5 MPa;

[0097] 3) Prepare 60 ml of potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution with a concentration of 50 mM and a pH of 6.8, filter it, collect the filtrate, and dispense it into 10 - ml vials to obtain the cleaning solution.

[0098] Example 3:

[0099] Preparation of a urine pretreatment device for detecting urinary iodine:

[0100] 1) Add 50 g of powdered activated carbon (350 mesh, porosity of 70%) to a beaker, add 5 g of sodium 2,3',6 - tribromoindophenol, add 250 ml of pure water, stir and react at 70 °C for 4 h, perform suction filtration, collect the filter cake, dry it at 120 °C for 5 h, cool it to room temperature, and collect it to obtain the urine treatment matrix; Take multiple empty cylindrical columns, add a sieve plate at the front end of the column, add 0.5 g of the urine treatment matrix to each column, add a sieve plate to seal the rear end of the column, compact it, and seal it to obtain a solid - phase extraction cartridge;

[0101] 2) Prepare a booster that can be tightly combined with the empty cylindrical column, and the tolerance pressure for the combination of the booster and the empty cylindrical column is 0.2 MPa;

[0102] 3) Prepare 30 ml of potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution with a concentration of 10 mM and a pH of 5.6, filter it, collect the filtrate, and dispense it into 5 - ml vials to obtain the cleaning solution.

[0103] Example 4:

[0104] Preparation of a urine pretreatment device for detecting urinary iodine:

[0105] 1) Add 50 g of powdered activated carbon (350 mesh, porosity 70%) to a beaker, add 4 g of sodium 2,2',6-tribromoindophenolate, add 150 ml of pure water, stir and react at 80 °C for 3 h, perform suction filtration, collect the filter cake, dry it at 125 °C for 4 h, cool it to room temperature, and collect to obtain the urine treatment matrix; Take multiple empty cylinders, add a sieve plate at the front end of the cylinder, add 1.0 g of the urine treatment matrix to each cylinder, add a sieve plate to seal the rear end of the cylinder, compact it, and seal it to obtain a solid-phase extraction cartridge;

[0106] 2) Prepare a booster that can be tightly combined with the empty cylinder, and the pressure tolerance for the combination of the booster and the empty cylinder is 0.7 MPa;

[0107] 3) Prepare 50 ml of a potassium dihydrogen phosphate-disodium hydrogen phosphate buffer solution with a concentration of 50 mM and a pH of 6.5, filter it, collect the filtrate, and dispense it into 10 ml / vials to obtain the cleaning solution.

[0108] Example 5:

[0109] Preparation of a urine pretreatment device for detecting urinary iodine:

[0110] 1) Add 50 g of powdered activated carbon (250 mesh, porosity 60%) to a beaker, add 2.5 g of sodium 2,6-dibromo-3'-methylindophenolate, add 100 ml of pure water, stir and react at 90 °C for 2 h, perform suction filtration, collect the filter cake, dry it at 130 °C for 2 h, cool it to room temperature, and collect to obtain the urine treatment matrix; Take multiple empty cylinders, add a sieve plate at the front end of the cylinder, add 0.8 g of the urine treatment matrix to each cylinder, add a sieve plate to seal the rear end of the cylinder, compact it, and seal it to obtain a solid-phase extraction cartridge;

[0111] 2) Prepare a booster that can be tightly combined with the empty cylinder, and the pressure tolerance for the combination of the booster and the empty cylinder is 0.7 MPa;

[0112] 3) Prepare 50 ml of a potassium dihydrogen phosphate-disodium hydrogen phosphate buffer solution with a concentration of 50 mM and a pH of 6.5, filter it, collect the filtrate, and dispense it into 8 ml / vials to obtain the cleaning solution.

[0113] Example 6:

[0114] Preparation of a urine pretreatment device for detecting urinary iodine:

[0115] 1) Add 50 g of powdered activated carbon (250 mesh, porosity 60%) to a beaker, add 5 g of 2,6-dichloroindophenol sodium, add 150 ml of pure water, stir and react at 55 °C for 7 h, perform suction filtration, collect the filter cake, dry it at 110 °C for 7 h, cool it to room temperature, and collect to obtain the urine treatment matrix; Take multiple empty cylinders, add a sieve plate at the front end of the cylinder, add 1 g of the urine treatment matrix to each cylinder, add a sieve plate to seal the rear end of the cylinder, compact it, and seal it to obtain a solid-phase extraction cartridge;

[0116] 2) Prepare a booster that can be tightly combined with the empty cylinder. The pressure tolerance for the combination of the booster and the empty cylinder is 0.6 MPa.

[0117] 3) Prepare 30 ml of potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution with a concentration of 60 mM and a pH of 6.8. Filter it, collect the filtrate, and dispense it into bottles at 5 ml / bottle to obtain the cleaning solution.

[0118] Example 7:

[0119] Preparation of a urine pretreatment device for detecting urinary iodine:

[0120] 1) Add 100 g of powdered activated carbon (350 mesh, porosity of 70%) to a beaker, add 5 g of sodium 2,6 - dichloroindophenol acetate, add 500 ml of pure water, react and stir at 70 °C for 5 h, perform suction filtration, collect the filter cake, dry it at 120 °C for 5 h, cool it to room temperature, and collect it to obtain the urine treatment matrix; Take multiple empty cylinders, add a sieve plate at the front end of the cylinder, add 0.5 g of the urine treatment matrix to each cylinder, add a sieve plate to seal the rear end of the cylinder, compact it, and seal it to obtain a solid - phase extraction small column;

[0121] 2) Prepare a booster that can be tightly combined with the empty cylinder. The pressure tolerance for the combination of the booster and the empty cylinder is 0.3 MPa;

[0122] 3) Prepare 40 ml of potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution with a concentration of 70 mM and a pH of 6.5. Filter it, collect the filtrate, and dispense it into bottles at 2.5 ml / bottle to obtain the cleaning solution.

[0123] Example 8:

[0124] Preparation of a urine pretreatment device for detecting urinary iodine:

[0125] 1) Add 100 g of powdered activated carbon (350 mesh, porosity of 70%) to a beaker, add 10 g of sodium 2,3',6 - trichloroindophenol, add 600 ml of pure water, control the temperature and stir for 8 h, perform suction filtration, collect the filter cake, dry it at 108 °C for 4 h, cool it to room temperature, and collect it to obtain the urine treatment matrix; Take multiple empty cylinders, add a sieve plate at the front end of the cylinder, add 0.5 g of the urine treatment matrix to each cylinder, add a sieve plate to seal the rear end of the cylinder, compact it, and seal it to obtain a solid - phase extraction small column;

[0126] 2) Prepare a booster that can be tightly combined with the empty cylinder. The pressure tolerance for the combination of the booster and the empty cylinder is 0.3 MPa;

[0127] 3) Prepare 40 ml of potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution with a concentration of 50 mM and a pH of 5.6. Filter it, collect the filtrate, and dispense it into bottles at 3.5 ml / bottle to obtain the cleaning solution.

[0128] Example 9:

[0129] Preparation of a urine pretreatment device for detecting urinary iodine:

[0130] 1) Add 100 g of powdered activated carbon (350 mesh, porosity of 70%) to a beaker, add 7.5 g of sodium 2,2',6-trichloroindophenolate, add 400 ml of pure water, stir and react at 85 °C for 3 h, filter by suction, collect the filter cake, dry at 125 °C for 3.5 h, cool to room temperature, collect to obtain the urine treatment matrix; Take multiple empty cylinders, add a sieve plate at the front end of the cylinder, add 0.5 g of the urine treatment matrix to each cylinder, add a sieve plate to seal the rear end of the cylinder, compact and seal to obtain a solid-phase extraction cartridge;

[0131] 2) Prepare a booster that can be tightly combined with the empty cylinder, and the pressure tolerance for the combination of the booster and the empty cylinder is 0.5 MPa;

[0132] 3) Prepare 40 ml of a potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution with a concentration of 50 mM and a pH of 5.6, filter, collect the filtrate, and dispense it into 2.5 ml / vials to obtain the cleaning solution.

[0133] Example 10:

[0134] Preparation of a urine pretreatment device for detecting urinary iodine:

[0135] 1) Add 100 g of powdered activated carbon (350 mesh, porosity of 70%) to a beaker, add 5 g of sodium 2,6-dichloro-3'-methylindophenolate, add 200 ml of pure water, stir and react at 70 °C for 4 h, filter by suction, collect the filter cake, dry at 110 °C for 5 h, cool to room temperature, collect to obtain the urine treatment matrix; Take multiple empty cylinders, add a sieve plate at the front end of the cylinder, add 0.8 g of the urine treatment matrix to each cylinder, add a sieve plate to seal the rear end of the cylinder, compact and seal to obtain a solid-phase extraction cartridge;

[0136] 2) Prepare a booster that can be tightly combined with the empty cylinder, and the pressure tolerance for the combination of the booster and the empty cylinder is 0.2 MPa;

[0137] 3) Prepare 50 ml of a potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution with a concentration of 10 mM and a pH of 6.8, filter, collect the filtrate, and dispense it into 8 ml / vials to obtain the cleaning solution.

[0138] Example 11:

[0139] Preparation of a urine pretreatment device for detecting urinary iodine:

[0140] 1) Add 100 g of powdered activated carbon (300 mesh, porosity of 65%) to a beaker, add 8 g of sodium o-bromophenolindophenol, add 300 ml of pure water, stir and react at 75 °C for 3.5 h, perform suction filtration, collect the filter cake, dry it at 125 °C for 3.5 h, cool to room temperature, and collect to obtain the urine treatment matrix; Take multiple empty cylinders, add a sieve plate at the front end of the cylinder, add 0.75 g of the urine treatment matrix to each cylinder, add a sieve plate to seal the rear end of the cylinder, compact it, and seal it to obtain a solid-phase extraction small column;

[0141] 2) Prepare a booster that can be tightly combined with the empty cylinder, and the tolerance pressure for the combination of the booster and the empty cylinder is 0.3 MPa;

[0142] 3) Prepare 60 ml of potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution with a concentration of 10 mM and a pH of 6.8, filter it, collect the filtrate, and dispense it into bottles at 7.5 ml / bottle to obtain the cleaning solution.

[0143] Example 12:

[0144] Preparation of a urine pretreatment device for detecting urinary iodine:

[0145] 1) Add 100 g of powdered activated carbon (300 mesh, porosity of 65%) to a beaker, add 8 g of sodium 3'-bromophenolindophenol, add 500 ml of pure water, stir and react at 60 °C for 5 h, perform suction filtration, collect the filter cake, dry it at 105 °C for 8 h, cool to room temperature, and collect to obtain the urine treatment matrix; Take multiple empty cylinders, add a sieve plate at the front end of the cylinder, add 1 g of the urine treatment matrix to each cylinder, add a sieve plate to seal the rear end of the cylinder, compact it, and seal it to obtain a solid-phase extraction small column;

[0146] 2) Prepare a booster that can be tightly combined with the empty cylinder, and the tolerance pressure for the combination of the booster and the empty cylinder is 0.6 MPa;

[0147] 3) Prepare 60 ml of potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution with a concentration of 10 mM and a pH of 5.6, filter it, collect the filtrate, and dispense it into bottles at 10 ml / bottle to obtain the cleaning solution.

[0148] Example 13:

[0149] Preparation of a dry chemical test strip for urinary iodine

[0150] 1) Prepare Solution A and Solution B;

[0151] Solution A:

[0152] 1000 mL of 0.1 M potassium dihydrogen phosphate - disodium hydrogen phosphate buffer solution (pH 8.0)

[0153] 25 g of sodium alginate

[0154] 2.5 g of sodium o-toluenesulfonchloramide

[0155] Solution B:

[0156] 5 g of N-(3-sulfopropyl)-3,3,5,5-tetramethylbenzidine

[0157] 1000 mL of acetone

[0158] 2) Preparation of urine iodine test strips: Take 31MM type filter paper, soak it evenly in Solution A for 13 s, dry it in a drying chamber at a temperature of 55 °C with a drying speed of 100 m / h; take the filter paper dried after soaking in Solution A, soak it evenly in Solution B for 25 s, dry it in a drying chamber at a temperature of 35 °C with a drying speed of 100 m / h; cut the dried filter paper into strips with a width of 7 mm, stick one side of the strip on a magnetic white sheet through double-sided tape, cut it into strips with a length of 80 mm and a width of 7 mm, put it in a bucket, seal it and store it in a dry place for later use.

[0159] 3) Preparation of the standard color comparison card package for urine iodine test strips

[0160] Preparation of iodine ion standard solution: Take potassium iodide standard substance, add 0.1 M sodium acetate buffer solution (pH 5.5) to prepare five standard solutions with final iodine ion concentrations of 0 μg / L, 50 μg / L, 150 μg / L, 300 μg / L, and 550 μg / L respectively to obtain iodine ion standard solution;

[0161] Preparation of the standard color comparison card: Wash the solid-phase extraction small column with the cleaning solution, add 1 ml of each of the five iodine ion standard solutions respectively, push the booster to filter for 4 s, collect the filtrate to obtain the iodine ion test solution; at room temperature, immerse the urine iodine test strips into each concentration of iodine ion test solution for 2 s respectively, take them out, react for 1 minute at a fixed time, read the value, use the iodine ion test solution concentration and the corresponding end color of the urine iodine test strip as the color standard to prepare the standard color comparison card, and correspondingly obtain the standard color comparison card with five color scales of 0 μg / L, 50 μg / L, 150 μg / L, 300 μg / L, and 550 μg / L. The color change of the standard color comparison card is yellow-green to blue-green.

[0162] By comparing the color development situation of the natural urine sample to be tested after being treated by the pretreatment device with the urine iodine test strip with the standard color comparison card, the iodine ion concentration in the sample can be semi-quantitatively determined.

[0163] Example 14:

[0164] Preparation of urine iodine test kit

[0165] 1) Solution preparation:

[0166]

[0167]

[0168] 2) Preparation of reaction solution: Filter the above-prepared solution, collect the filtrate, and prepare the first reaction solution and the second reaction solution of the urinary iodine test kit.

[0169] 3) Preparation of the standard colorimetric card package for the urinary iodine test kit

[0170] Preparation of the urinary iodine ion standard solution: Take potassium iodide standard substance, add 0.1M sodium acetate buffer solution (pH 5.5), and prepare five standard solutions with final iodine ion concentrations of 0 μg / L, 50 μg / L, 150 μg / L, 300 μg / L, and 550 μg / L respectively to obtain the iodine ion standard solution;

[0171] Preparation of the standard colorimetric card: Wash the solid-phase extraction small column with the cleaning solution, add 2.5 ml of each of the five iodine ion standard solutions respectively, push the booster to filter for 6 s, collect the filtrate, and obtain the iodine ion test solution; At room temperature, take 100 μl of the artificial urinary iodine test solution at four concentrations respectively, add 40 μl of the second reaction solution, then add 10 μl of the first reaction solution, shake well, time for 1 minute, repeat 3 times for each concentration, and use the color development results of the five concentrations as color standards to prepare the standard colorimetric card, corresponding to obtain the standard colorimetric card with five color levels. The color change of the standard colorimetric card is from yellow-green to blue-green.

[0172] By comparing the color development situation of the natural urine sample to be tested after being processed by the pretreatment device with the urinary iodine test kit with the standard colorimetric card, the iodine ion concentration in the sample can be semi-quantitatively determined.

[0173] Example 15:

[0174] Application of the urine pretreatment device for detecting urinary iodine in dry chemical detection of urinary iodine

[0175] 1) Cleaning of the solid-phase extraction small column: Randomly take the solid-phase extraction small column and the cleaning solution prepared in Examples 1-12 respectively. At room temperature, add the extraction solution into the column tube, push the booster into the column tube, push the booster to clean the urine treatment matrix for 2 s, and push out the excess cleaning solution;

[0176] 2) Urine pretreatment: At room temperature, take 50 portions of the natural urine of the subjects, add 1 ml of the natural urine to be tested into the solid-phase extraction small column respectively, add the booster and filter slowly, collect the filtrate, and obtain the natural urine test solution;

[0177] 3) Application in dry chemical detection of urinary iodine: At room temperature, immerse the urinary iodine test strips into the unpretreated natural urine and the pretreated natural urine test solution of the same person respectively, take them out after timing for 2 s respectively, react for 1 minute at a fixed time, repeat 3 times, compare the reaction results of the urinary iodine test strips with the standard colorimetric card, and read the values.

[0178] The experimental results show that: the color development of the natural urine test strips of untreated subjects is interfered, the color systems of the test strips are inconsistent, and the repeatability is poor; after being treated with the urine pretreatment device for detecting urinary iodine, the color systems of the test strips are consistent, the repeatability is good, the detection results are accurate, and they can fall within the range of 0 μg / L - 550 μg / L, and the implementation effect is good, indicating that the urine pretreatment device is applicable to the detection of urinary iodine in the natural urine of clinical subjects by urinary iodine test strips.

[0179] Example 16:

[0180] The urine pretreatment device for detecting urinary iodine is applied to the detection of urinary iodine kits

[0181] 1) Cleaning of the solid-phase extraction cartridge: Randomly take the solid-phase extraction cartridges and cleaning solutions prepared in Examples 1 - 4 respectively. At room temperature, add the extraction solution into the cartridge, push the booster into the cartridge, push the booster to clean the urine treatment matrix for 6 s, and push out the excess cleaning solution;

[0182] 2) Urine pretreatment: At room temperature, take 2.5 ml of the midstream natural urine of the subject's morning urine, add it to the solid-phase extraction cartridge, and slowly filter it with the booster for 6 s to collect the filtrate to obtain the natural urine test solution of the subject;

[0183] 3) Application to the detection of urinary iodine kits: At room temperature, take 100 μl of the natural urine of the untreated subject and 100 μl of the natural urine test solution of the subject after pretreatment from the same person respectively, add 40 μl of the second reaction solution, then add 10 μl of the first reaction solution, shake well and time for 60 s to observe the end color of the solution. Each sample is repeated 3 times, and compared with the standard colorimetric card to read the value.

[0184] The experimental results show that: for the natural urine of untreated subjects, the color development of the kit is affected, the color systems of the kit are inconsistent, and the repeatability is poor; after being treated with the urine pretreatment device for detecting urinary iodine, the color systems of the kit are consistent, the repeatability is good, the detection results are accurate, and they can fall within the range of 0 μg / L - 550 μg / L, and the implementation effect is good, indicating that the urine pretreatment device is applicable to the detection of urinary iodine in the natural urine of clinical subjects by urinary iodine kits.

[0185] Example 17:

[0186] Performance verification of the urine pretreatment device for detecting urinary iodine applied to urinary iodine test strips

[0187] Four kinds of urine iodine solutions, namely aqueous iodine ion solution, iodine ion standard solution, iodine ion interfering substance standard solution, and natural urine of the subjects, were respectively treated with a urine pretreatment device for detecting urine iodine to obtain four kinds of iodine ion test solutions to be measured after treatment. The four kinds of iodine ion solutions before pretreatment and the four kinds of iodine ion test solutions after pretreatment were simultaneously detected on urine iodine test strips respectively. The following were investigated: ① the adsorption of iodine ions on the urine pretreatment device; ② the treatment ability of the urine pretreatment device for interfering factors such as vitamin C, bilirubin, and pigments; ③ the treatment of real urine by the urine pretreatment device. The specific operation steps are as follows:

[0188] 1) Preparation of aqueous iodine ion solution: A stock solution was prepared with KI and diluted with water to a series of aqueous iodine ion standard solutions with concentrations of 0 μg / L, 50 μg / L, 150 μg / L, 300 μg / L, and 550 μg / L;

[0189] 2) Preparation of iodine ion standard solution: Artificial urine was prepared with urea, creatinine, hippuric acid, disodium hydrogen phosphate, potassium dihydrogen phosphate, dye VII, sodium chloride, potassium chloride, and pure water. An artificial urine stock solution was prepared with KI, and then diluted with artificial urine to a series of artificial urine iodine standard solutions with concentrations of 0 μg / L, 50 μg / L, 150 μg / L, 300 μg / L, and 550 μg / L;

[0190] 3) Iodine ion interfering substance standard solution: A series of iodine ion standard solutions with concentrations of 0 μg / L, 50 μg / L, 150 μg / L, 300 μg / L, and 550 μg / L were prepared with artificial urine, and vitamin C and bilirubin interfering substances were respectively added to prepare iodine ion interfering substance standard solutions containing 20 mM vitamin C and 100 mM bilirubin;

[0191] 4) Preparation of natural urine of the subjects: Urine samples of 50 healthy people collected over 24 hours were stored at 2 - 8 °C.

[0192] A part of each of the above four solutions was taken and treated with a urine pretreatment device for detecting urine iodine. The four kinds of test solutions to be measured after treatment were detected on urine iodine test strips simultaneously, and compared with the above four kinds of iodine ion solutions without pretreatment and the test solutions after pretreatment. The experimental results and conclusions are as follows:

[0193] ① By comparing the detection results of the treated aqueous iodine ion solution and the untreated aqueous iodine ion solution, the colors of the test strips at each gradient were the same, indicating that this urine pretreatment device has no adsorption on iodine ions;

[0194] ② By comparing the detection results of the treated iodine ion standard solution and the untreated iodine ion standard solution, the colors of the test strips at each gradient were the same, indicating that this urine pretreatment device has no adsorption effect on iodine ions. At the same time, the urine pretreatment device can adsorb dye pigments in artificial urine, and the pH of artificial urine has no effect on the detection results;

[0195] ③ After comparing the test results of the processed iodide interference substance solution containing 20 mM vitamin C and 100 mM bilirubin with those of the untreated iodide standard solution, the colors of the test papers at each gradient are the same, indicating that the urine pretreatment device can simultaneously remove the interferences of 20 mM vitamin C and 100 mM bilirubin and has no adsorption on iodide ions;

[0196] ④ After comparing the test results of the processed natural urine of the subjects with those of the processed iodide standard solution, the iodide ion concentration in the urine falls within the range of 0 μg / L - 550 μg / L, and the concentration range of the iodide standard solution is applicable to the detection of the urinary iodine value of the natural urine of clinical subjects.

[0197] Example 18:

[0198] Performance verification of a urine pretreatment device for detecting urinary iodine applied to a urinary iodine test kit

[0199] Four types of urinary iodine solutions, namely an aqueous iodide standard solution, an iodide standard solution, an iodide interference substance standard solution, and the natural urine of the subjects, were respectively processed with the urine pretreatment device for detecting urinary iodine to obtain four processed iodide solution test samples. The four iodide solutions before pretreatment and the four processed iodide test samples were simultaneously detected on the urinary iodine test kit. The following aspects were investigated: ① The adsorption of iodide ions on the urine pretreatment device; ② The processing ability of the urine pretreatment device for interference factors such as vitamin C, bilirubin, and pigments; ③ The processing of real urine by the urine pretreatment device. The specific operation steps are as follows:

[0200] 1) Preparation of the aqueous iodide solution: A stock solution was prepared with KI and diluted with water to a series of aqueous iodide standard solutions with concentrations of 0 μg / L, 50 μg / L, 150 μg / L, 300 μg / L, and 550 μg / L;

[0201] 2) Preparation of the iodide standard solution: Artificial urine was prepared with urea, creatinine, hippuric acid, disodium hydrogen phosphate, potassium dihydrogen phosphate, dye VII, sodium chloride, potassium chloride, and pure water. A stock solution of artificial urine iodide was prepared with KI, and a series of artificial urine iodide standard solutions with concentrations of 0 μg / L, 50 μg / L, 150 μg / L, 300 μg / L, and 550 μg / L were prepared with the artificial urine;

[0202] 3) Iodide interference substance standard solution: A series of iodide standard solutions with concentrations of 0 μg / L, 50 μg / L, 150 μg / L, 300 μg / L, and 550 μg / L were prepared with artificial urine, and vitamin C and bilirubin interference substances were respectively added to prepare iodide interference substance standard solutions containing 20 mM vitamin C and 100 mM bilirubin;

[0203] 4) Preparation of natural urine of subjects: Collect 50 portions of 24-hour urine from healthy individuals and store it at 2-8°C.

[0204] Take a portion of each of the above four solutions and process them with the urine pretreatment device for detecting urinary iodine. The four processed test solutions obtained are compared and detected simultaneously with the above four unpretreated iodide ion solutions and the pretreated test solutions using a urinary iodine kit. The experimental results and conclusions are as follows:

[0205] ① Comparing the test results of the processed aqueous iodide ion solution with the unprocessed iodide ion standard solution, the colors of the kits at each gradient are the same, indicating that this urine pretreatment device has no adsorption on iodide ions;

[0206] ② Comparing the test results of the processed artificial urinary iodide ion standard solution with the unprocessed iodide ion standard solution, the colors of the kits at each gradient are consistent, indicating that this urine pretreatment device has no adsorption effect on iodide ions. At the same time, the urine pretreatment device can adsorb the dye pigments in artificial urine, and the pH of artificial urine has no effect on the test results;

[0207] ③ Comparing the test results of the processed artificial urinary iodide ion interference substance solution containing 20 mM vitamin C and 100 mM bilirubin with the unprocessed iodide ion standard solution, the colors of the kits at each gradient are the same, indicating that the urine pretreatment device can simultaneously remove the interference of 20 mM vitamin C and 100 mM bilirubin and has no adsorption on iodide ions;

[0208] ④ Comparing the test results of the processed natural urine of subjects with the processed iodide ion standard solution, the iodide ion concentration in the urine falls within the range of 0 μg / L - 550 μg / L, and the gradient range of the iodide ion standard solution is applicable to the detection of urinary iodine values in the natural urine of clinical subjects.

[0209] The present invention uses specific examples to elaborate on the principle and implementation mode of the present invention. The description of the above embodiments is only used to help understand the method and its core idea of the present invention; at the same time, for those of ordinary skill in the art, there will be changes in the specific implementation mode and application scope according to the idea of the present invention. In summary, the content of this specification should not be construed as a limitation to the present invention.

Claims

1. A urine pretreatment device for detecting urinary iodine, characterized in that: It includes a solid phase extraction cartridge, a booster and a cleaning solution; wherein, The solid phase extraction column is used to contain urine treatment matrix and filter urine, and comprises an empty column tube, wherein the top of the empty column tube is provided with an opening, the bottom of the empty column tube is provided with a small hole, and the interior of the empty column tube is sealed with a filler through a front sieve plate and a rear sieve plate; The booster is used to provide pressure for passing through the column after the liquid is filled, and is tightly combined with the empty column; The cleaning solution is used for cleaning the solid phase extraction column and is prepared from a buffer solution.

2. A method for preparing a urine pretreatment device for detecting urinary iodine, characterized in that: The following steps are involved: 1) preparing a solid phase extraction column: mixing activated carbon and a treatment reagent in proportion, adding a certain proportion of pure water, stirring under temperature control, filtering, collecting filter cakes, drying, and collecting, to obtain a urine treatment matrix for use; taking an empty column, adding a front sieve plate to the front end, adding a urine treatment matrix, sealing the rear end with a rear sieve plate to form a filler, compacting, and sealing, to obtain a solid phase extraction column; 2) Preparation of booster: The booster can be tightly combined with the empty column, and the pressure resistance of the combination of the booster and the empty column is 0.1-0.5MPa. 3) Prepare a cleaning solution: prepare a buffer solution according to a certain molar concentration and pH, filter and collect the filtrate to obtain a cleaning solution.

3. The method for preparing the urine pretreatment device for detecting urinary iodine according to claim 2, characterized in that: In step 1), the activated carbon is selected from coal-based activated carbon, synthetic activated carbon, and wood-based activated carbon, and the mesh size of the activated carbon is 150-400 meshes, and the porosity is 50-80%; The treatment reagent is one or more sodium salt compounds of indophenol, and further, the treatment reagent is one or more sodium salt compounds of bromoindophenol; The indophenol is o-chlorophenol indophenol, 3'-chloroindophenol, 5-isopropyl-2-methylindophenol, 2,6-dichloroindophenol, 2,6-dichloroindophenol acetate, 2,3',6-trichloroindophenol, 2,2',6-trichloro-indophenol, 2,6-dichloro-3'-methylindophenol, 2,6-dibromoindophenol, 2,6-dibromoindophenol acetate, 2,3',6-tribromoindophenol, 2,2',6-tribromo-indophenol, 2,6-dibromo-3'-methylindophenol; further, the bromoindophenol is 2,6-dibromoindophenol, 2,6-dibromoindophenol acetate, 2,3',6-tribromoindophenol, 2,2',6-tribromo-indophenol, 2,6-dibromo-3'-methylindophenol; The mass ratio of the activated carbon to the treatment reagent is 100:1-15; the volume ratio of the activated carbon to the pure water is 1:1-10; the reaction temperature of the temperature-controlled stirring is 20-100°C, and the reaction time of the temperature-controlled stirring is 1-12h.

4. The method for preparing the urine pretreatment device for detecting urinary iodine according to claim 2, characterized in that: In step 1), the baking temperature is 90-150° C. and the baking time is 1-12 hours.

5. The method for preparing the urine pretreatment device for detecting urinary iodine according to claim 2, characterized in that: In step 1), the mass of the urine treatment matrix in the solid phase extraction column is 0.1-2.0 g.

6. The method for preparing the urine pretreatment device for detecting urinary iodine according to claim 2, characterized in that: In step 2), the cleaning solution is one of potassium dihydrogen phosphate-disodium hydrogen phosphate buffer solution, potassium citrate-citric acid buffer solution, and acetic acid-sodium acetate buffer solution; The concentration of the buffer solution is 5-200 mM, the pH value of the buffer solution is 5-7, and the volume of the buffer solution is 5-20 ml.

7. An application of a urine pretreatment device for detecting urinary iodine, characterized in that: The following steps are involved: 1) Cleaning of solid phase extraction cartridge: at room temperature, add cleaning solution into the cartridge, push the booster into the cartridge, push the booster to clean the urine treatment matrix, and push out excess cleaning solution; 2) Urine pretreatment: Add the natural urine to be tested into the solid phase extraction column at room temperature, push the booster to filter slowly, collect the filtrate, and obtain the natural urine test solution; 3) Urine iodine test: When applied to urine iodine dry chemical detection, the urine iodine test paper is immersed in the natural urine test solution at room temperature, taken out, reacted at a fixed time, and the reaction result of the urine iodine test paper is compared with the color of the standard colorimetric card to read the value; When used for urine iodine test kit detection, at room temperature, add the second reaction solution of the urine iodine test kit into the natural urine test solution, then add the first reaction solution of the urine iodine test kit, shake well, react at a fixed time, compare the color development result of the reaction solution with the color of the standard colorimetric card, and read the value.

8. The use of the urine pretreatment device for detecting urinary iodine according to claim 7, characterized in that: In step 1), the room temperature is 20-25° C.; the cleaning time of the cleaning solution is 1-10 seconds.

9. The use of the urine pretreatment device for detecting urinary iodine according to claim 7, characterized in that: In step 2), the natural urine to be tested is 24-hour urine or mid-morning urine, and the urine sample is stored at 4-8°C for later use; the volume of the natural urine to be tested is 1-5ml; and the filtering speed of the natural urine to be tested is 1-10s.

10. The use of the urine pretreatment device for detecting urinary iodine according to claim 7, characterized in that: In step 3), when applied to urine iodine dry chemical detection, the mass volume ratio of the urine treatment matrix to the natural urine test solution is 0.5g:1-5ml; the time for the urine iodine test paper to be immersed in urine is 1-5s; the reaction time is 0.5-1min; when applied to urine iodine kit detection, the urine treatment matrix treatment time is 1-10s; The volume ratio of the natural urine test solution to the second reaction solution and the first reaction solution in the urine iodine test kit is 10:3-6:1-2; and the reaction time is 0.5-1 minute.

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