CN110006881B - Detection kit for iodine content in urine and detection method for iodine content in urine - Google Patents
Detection kit for iodine content in urine and detection method for iodine content in urine Download PDFInfo
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- CN110006881B CN110006881B CN201910260923.3A CN201910260923A CN110006881B CN 110006881 B CN110006881 B CN 110006881B CN 201910260923 A CN201910260923 A CN 201910260923A CN 110006881 B CN110006881 B CN 110006881B
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Abstract
The invention relates to a detection kit for iodine content in urine and a detection method for iodine content in urine. The detection kit for the iodine content in urine comprises a diluent, a color developing agent, an oxidant and a purifying agent, wherein the diluent comprises 3.5-4.5% of sodium chloride by mass concentration, the pH value of the diluent is 4.6, the color developing agent comprises 50mg/L of 3,3',5,5' -tetramethyl benzidine, the oxidant comprises peracetic acid and the diluent, and the mass concentration of the peracetic acid is 0.25%. The detection kit for the content of iodine in urine has good repeatability and less pollution to the environment.
Description
Technical Field
The invention relates to the field of detection, in particular to a detection kit for iodine content in urine and a detection method for iodine content in urine.
Background
Iodine is one of the essential trace elements for human body, is called intelligence element, and is an important raw material for synthesizing thyroid hormone. Once a human body is lack of iodine, a series of morbid states, namely iodine deficiency disease, belong to micronutrient malnutrition, and are one of three major micronutrient malnutrition diseases which are mainly prevented, treated and eliminated in limited time by international organizations such as world health organization, United nations' gene society and the like. The iodine deficiency of women during pregnancy can cause abortion, premature labor, stillbirth and congenital malformation of fetus. Once iodine is deficient in the growth process of children, the intelligence development is seriously affected, and even if iodine is slightly deficient, the normal development of the brain is also affected. Iodine deficiency disease can cause intelligence damage, and meanwhile, the iodine deficiency disease directly causes endemic goiter, cretinism and the like, and seriously harms the physical and mental health of human bodies. On the other hand, excessive iodine also affects the health of the body, mainly manifested as thyroid dysfunction. Generally, about 90% of iodine taken by a human body is discharged from urine, and thus, urine iodine is the best index for evaluating the nutritional status of iodine in a human body. The iodine in urine has the characteristics of low content, complex basic components and large individual difference, so that the method for measuring the urine iodine is required to have high sensitivity, accuracy, anti-interference capability, stability and the like.
Plasma chromatography mass spectrometry (ICP-MS) iodine ions in a urine sample are vaporized at a high temperature of approximately 10000 ℃ using a plasma chromatograph. Detecting the content of iodine by an ion chromatography detector according to different charge-to-mass ratios of ions. The method has high accuracy, but the instrument is expensive and is not suitable for wide application.
The traditional urine iodine determination standard method is an arsenic-cerium catalytic spectrophotometry, the detection limit of the method is 3 mu g/L, and the method has the advantages of high sensitivity, strong anti-interference capability, good repeatability and the like, but the used detection kit contains arsenic trioxide, so that the environment is easily polluted and the human health is harmed, and the traditional urine iodine detection kit without arsenic trioxide has poor repeatability of a color reaction system.
Disclosure of Invention
Therefore, there is a need for a detection kit for detecting iodine content in urine, which has good reproducibility and less environmental pollution.
In addition, a method for detecting the iodine content in urine is also provided.
The utility model provides a detection kit of iodine content in urine, includes diluent, colour development agent, oxidant and purification agent, the diluent includes that mass concentration is 3.5% ~ 4.5% sodium chloride, the pH of diluent is 4.6, the colour development agent includes that the concentration is 50 mg/ L's 3,3',5,5 '-tetramethyl benzidine, the oxidant include peracetic acid and the diluent, peracetic acid's mass concentration is 0.25%.
In one embodiment, the diluent further comprises a pH regulator, and the pH regulator is acetic acid-sodium acetate buffer solution.
In one embodiment, the developer further comprises: citric acid with the concentration of 1.9g/L, sodium ethylene diamine tetracetate with the concentration of 0.4g/L, glycerol with the volume ratio of the color developing agent of 0.1 and dimethyl sulfoxide with the volume ratio of the color developing agent of 0.01.
In one embodiment, the purifying agent is activated carbon.
In one embodiment, the kit further comprises a hydrogel chip, wherein the hydrogel chip is provided with a purification tank and a detection tank, the purification tank is internally provided with the purifying agent, the detection tank is internally provided with a detector capable of detecting iodine content, the purification tank is communicated with the detection tank through a first channel, the hydrogel chip is further provided with a second channel and a third channel, the second channel is communicated with the first channel, the third channel is communicated with the first channel, the second channel is closer to the purification tank than the third channel, the hydrogel chip is further provided with a fourth channel communicated with the purification tank, the diluent and urine to be detected can flow in from the fourth channel, the color developing agent can flow in from the second channel, and the oxidizing agent can flow in from the third channel.
In one embodiment, the hydrogel chip includes a detection plate and a cover plate, the purification cell, the detection cell, the first channel, the second channel, the third channel, and the fourth channel are disposed on the detection plate, and the cover plate covers the detection plate and shields the purification cell, the detection cell, the first channel, the second channel, the third channel, and the fourth channel.
A method for detecting iodine content in urine comprises the following steps:
taking urine to be detected, and mixing the urine with diluent to obtain diluted urine, wherein the diluent comprises 3.5-4.5% of sodium chloride by mass concentration, and the pH of the diluent is 4.6;
purifying the diluted urine by using a purifying agent, and sequentially adding a color-developing agent and an oxidizing agent into the purified diluted urine to obtain a mixed solution, wherein the color-developing agent comprises 3,3',5,5' -tetramethylbenzidine with the concentration of 50mg/L, the oxidizing agent comprises peroxyacetic acid and the diluting solution, and the mass concentration of the peroxyacetic acid is 0.25%; and
and detecting the mixed solution to obtain the content of iodide ions in the urine to be detected.
In one embodiment, in the step of taking the urine to be tested and mixing the urine to be tested with the diluent, the volume ratio of the urine to be tested to the diluent is 1: 5.
In one embodiment, in the step of purifying the diluted urine with the purifying agent, the amount ratio of the purifying agent to the diluted urine is 0.111g/mL to 0.222 g/mL.
In one embodiment, in the step of sequentially adding a color-developing agent and an oxidizing agent to the purified diluted urine, the volume ratio of the diluted urine to the oxidizing agent to the color-developing agent is 3: 1.
In one embodiment, the step of purifying the diluted urine with a purifying agent, and the step of sequentially adding a color-developing agent and an oxidizing agent to the purified diluted urine comprises:
providing a hydrogel chip, wherein the hydrogel chip comprises a purification pool and a detection pool, the purification pool is internally loaded with a purifying agent, the detection pool is internally loaded with a detector capable of detecting iodine content, the purification pool is communicated with the detection pool through a first channel, the hydrogel chip is also provided with a second channel and a third channel, the second channel is communicated with the first channel, the third channel is communicated with the first channel, the second channel is closer to the purification pool than the third channel, the hydrogel chip is also provided with a fourth channel communicated with the purification pool, diluent and urine to be detected can flow in from the fourth channel, a color developing agent can flow in from the second channel, and an oxidizing agent can flow in from the third channel;
and taking the diluted urine, the color developing agent and the oxidizing agent, respectively injecting the diluted urine, the color developing agent and the oxidizing agent into the fourth channel, the second channel and the third channel of the hydrogel chip so as to purify the diluted urine in the purification tank, and then mixing the diluted urine, the color developing agent and the oxidizing agent.
In one embodiment, the step of detecting the mixed solution includes: and starting timing when the mixed liquid completely flows into the detection pool of the hydrogel chip, and detecting the detection pool by using the detector after 300 s.
The detection kit for the content of iodine in urine adopts high- concentration 3,3',5,5' -Tetramethylbenzidine (TMB) and peracetic acid as a color reaction system, the color reaction is relatively violent, the time for achieving color change of the color reaction is short, so that fine color deviation can cause large fluctuation of data results, and the detection kit for the content of iodine in urine adopts low-concentration TMB and low-concentration peracetic acid as color reaction systems, the reaction is relatively mild, the color change fluctuation is small, and a diluent contains sodium chloride with specific content, so that when a purifying agent is used for treating urine, the interference of impurity particles is avoided, and iodine ions are kept to the maximum extent and are not adsorbed, so that the repeatability of a test result of the detection kit is good when the iodine ions are detected. The diluted solution is acidic, and an acidic environment can be provided for the reaction of the color-developing agent and the oxidizing agent. And the detection kit does not adopt an arsenic-cerium color development system which causes environmental pollution, so that the environmental pollution is relatively small. Therefore, the detection kit for the content of iodine in urine has good repeatability and less pollution to the environment.
Drawings
FIG. 1 is a schematic diagram illustrating a structure of a hydrogel chip in a urine detection kit according to an embodiment;
FIG. 2 is a process flow diagram of a method for detecting iodine content in urine according to an embodiment;
fig. 3 is a result curve of 8 times of repeated tests of the detection kit for detecting the iodine content in urine and the Qingdao Sanka kit in example 1 on the iodine content in urine.
Detailed Description
In order that the invention may be more fully understood, reference will now be made to the following description taken in conjunction with the accompanying drawings. The detailed description sets forth the preferred embodiments of the invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
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. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
The kit for detecting the content of iodine in urine of one embodiment comprises a diluent, a color developing agent, an oxidizing agent and a purifying agent. Wherein the diluent comprises 3.5 to 4.5 mass percent of sodium chloride, and the pH value of the diluent is 4.6. The color developer comprises 3,3',5,5' -tetramethyl benzidine (TMB) with the concentration of 50 mg/L. The oxidant comprises peroxyacetic acid and diluent, and the mass concentration of the peroxyacetic acid is 0.25%.
Before use, the diluent, the color-developing agent, the oxidizing agent and the purifying agent are independent from each other.
Specifically, the diluent also comprises a pH regulator, and the pH regulator is acetic acid-sodium acetate buffer solution. The pH regulator is used for regulating the pH value of the diluent. The diluent can be used to dilute the urine to be tested and act as a solvent for the oxidizing agent. The sodium chloride is added into the diluent, so that the urine to be detected can selectively adsorb impurity particles when being treated by a purifying agent subsequently, and iodine ions are retained to the maximum extent. The pH value of the diluent is adjusted to be acidic, so that an acidic environment can be provided for the subsequent color development reaction of the oxidant and the color developing agent.
The developer also comprises citric acid, sodium ethylene diamine tetracetate (EDTA-Na), glycerol, dimethyl sulfoxide (DMSO) and water. Wherein the concentration of the citric acid is 1.9 g/L. The concentration of EDTA-Na was 0.4 g/L. The volume ratio of glycerin to the color developing agent is 0.1. The volume of DMSO and the color developing agent is 0.01.
EDTA-Na is an important chelating agent and can chelate metal ions in the solution and prevent the solution from discoloring, deteriorating and becoming turbid caused by the metal ions. The citric acid has similar action to EDTA-Na except that it provides an acidic environment, i.e. it is used as a complexing agent, a masking agent, and can rapidly complex metal ions.
Because TMB is insoluble in water, it is more soluble in DMSO. Therefore, the addition of DMSO to the developer dissolves TMB. However, since a high concentration of the organic solvent reduces the color developing effect, DMSO is added only to the developer in an amount of 0.01 by volume. Meanwhile, as the melting point of DMSO is 18.5 ℃, the DMSO is solid in the storage process at 4 ℃, and the use is inconvenient, the glycerol with the volume content of 0.1 is added, so that the color developing agent is liquid at 4 ℃ and is convenient to use. The glycerol is added into the color developing agent, so that the spontaneous color development of the TMB can be reduced, and the stability of the TMB is improved.
The peroxyacetic acid is dissolved in the diluent to obtain the oxidant. The peroxyacetic acid in the oxidant can perform oxidation reaction with TMB in the color developing agent under the catalytic action of iodide ions to obtain a blue product. Within a certain concentration range, the higher the iodide ion concentration, the faster the reaction speed, and the more blue products, so that when the color developer, the oxidant, and the urine to be tested are contacted, the darker the color of the obtained solution, the higher the iodide ion concentration. Therefore, the concentration of the iodide ions can be obtained by reflecting the absorbance according to the shade of the solution color. Furthermore, the diluent is used as a solvent of the oxidant, so that the oxidant is acidic, an acidic environment is provided for the reaction of the oxidant and the color developing agent, and the color developing reaction of the oxidant and the color developing agent is facilitated.
The purifying agent can purify urine to be detected. In the present embodiment, the purifying agent is activated carbon. The activated carbon can purify diluted urine to remove interfering substances in the urine, such as bilirubin, vitamin C, sodium chloride and the like, but the activated carbon cannot adsorb iodide ions and does not influence the test result of the iodide ions.
Further, the detection kit for the content of iodine in urine also comprises a hydrogel chip. Referring to fig. 1, the hydrogel chip 200 is provided with a purification cell 210 and a detection cell 220, the purification cell 210 carries a purifying agent, the detection cell 220 carries a detector capable of detecting iodine content, the purification cell 210 is communicated with the detection cell 220 through a first channel 230, the hydrogel chip 200 is further provided with a second channel 240 and a third channel 250, the second channel 240 is communicated with the first channel 230, the third channel 250 is communicated with the first channel 230, the second channel 240 is closer to the purification cell 210 than the third channel 250, the hydrogel chip 200 is further provided with a fourth channel 260 communicated with the purification cell 210, a diluent and urine to be detected can flow in from the fourth channel 260, a color developing agent can flow in from the second channel 240, and an oxidizing agent can flow in from the third channel 250.
Specifically, the hydrogel chip 200 includes a cover plate and a detection plate. The detection plate is provided with a purification pool 210, a detection pool 220, a first channel 230, a second channel 240, a third channel 250 and a fourth channel 260, and the cover plate is covered on the detection plate and shields the purification pool 210, the detection pool 220, the first channel 230, the second channel 240, the third channel 250 and the fourth channel 260. The cover plate can protect the detection plate, so that the detection process is not interfered by external impurity particles.
There is no communication between the second channel 240 and the third channel 250. By arranging the second channel 240 and the third channel 250 to be not communicated, the color developing agent and the oxidizing agent can be mixed with the urine to be detected in sequence, so that the starting point of color developing reaction can be conveniently determined.
When the hydrogel chip is used for detection, firstly, urine to be detected and diluent are mixed to obtain diluted urine, then the diluted urine flows in from the fourth channel, is purified by a purifying agent in a purifying tank, then is mixed with a color developing agent flowing in along the first channel and the second channel and an oxidizing agent flowing in along the third channel to obtain a mixed solution, the mixed solution flows in a detection tank, and a detector is used for detecting the mixed solution to obtain the iodine content in the urine to be detected.
The detector can detect the absorbance of the mixed solution flowing into the detection cell. Specifically, in the present embodiment, the detector is a color recognition sensor. The color recognition sensor can recognize colors and detect absorbance, so that the concentration of iodide ions in the urine to be detected is obtained.
The hydrogel chip can integrate the purification and detection of urine to be detected, has small volume and length of only 5-10 cm, is quick to detect and low in cost, and can be used for quickly detecting the content of iodine in the urine. And the introduction of the hydrogel had no effect on the testing of the iodine content of the urine.
It is understood that in other embodiments, the detection device is not limited to a hydrogel chip. Any device capable of purifying urine to be detected, mixing the purified urine to be detected with the color developing agent and the oxidizing agent to obtain a mixed solution, and detecting the mixed solution can be used as a detection device.
Further, the detection kit also comprises an iodine standard solution. The iodine standard solution is used for drawing a standard curve of the concentration and the absorbance of the iodide ions to obtain a functional relation between the concentration and the absorbance of the iodide ions. Specifically, in the present embodiment, the iodine standard solution includes potassium iodide solutions having iodide ion concentrations of 10. mu.g/L, 60. mu.g/L, 80. mu.g/L, 100. mu.g/L, 300. mu.g/L, 400. mu.g/L, and 500. mu.g/L, respectively. The iodine standard solution with the concentration gradient is selected, so that the correlation coefficient of the relation curve of the concentration of the obtained iodide ions and the absorbance is large, and the test result is more accurate.
The detection kit for the content of iodine in urine has at least the following advantages:
(1) the detection kit can utilize TMB in the color developing agent and peracetic acid in the oxidant to generate a blue product under the catalysis of the iodide ion solution, and the shade of the color of the blue product is related to the iodide ion concentration, so that the iodide ion concentration can be obtained according to the shade of the color. Compared with the high-concentration color developing agent and the high-concentration oxidizing agent, the low-concentration color developing agent and the low-concentration oxidizing agent have the advantage that the repeatability of color developing reaction is good.
(2) The detection kit does not contain arsenide, and has small environmental pollution.
(3) When the detection kit is used for urine detection, a pretreatment process is not required, so that the operation is simplified.
(4) The hydrogel chip in the detection kit can realize integration of urine purification and reaction color development. Four channels and two reaction cells are carved in the hydrogel chip. The first channel is communicated with a purification tank filled with a purifying agent and a detection tank. The urine to be detected flows in from the fourth channel, the purified urine is mixed with the color developing agent and the oxidizing agent flowing in from the second channel and the third channel along the first channel and flows into the detection pool, and the method has the advantages of simplicity and easiness in operation, rapidness, flexibility, high automation degree and the like.
(5) The detection kit can detect the concentration of the iodine within the range of 10-500 mug/L, and the detection range meets the requirement of detecting the content of the iodine in human urine.
Referring to fig. 2, a method for detecting iodine content in urine according to an embodiment includes the following steps:
step S110: taking urine to be detected, and mixing the urine with diluent to obtain diluted urine, wherein the diluent comprises 3.5-4.5% of sodium chloride by mass concentration, and the pH of the diluent is 4.6.
Wherein the volume ratio of the urine to be detected to the diluent is 1: 5. And diluting the urine to be detected by using the diluent so that the concentration of the urine to be detected is in a detection range.
Specifically, the diluent also comprises a pH regulator, and the pH regulator is acetic acid-sodium acetate buffer solution. The sodium chloride is added into the diluent, so that the urine to be detected can selectively adsorb impurity particles when being treated by a purifying agent subsequently, and iodine ions are retained to the maximum extent.
Step S120: purifying the diluted urine by using a purifying agent, and sequentially adding a color developing agent and an oxidizing agent into the purified diluted urine to obtain a mixed solution, wherein the color developing agent comprises 3,3',5,5' -tetramethylbenzidine with the concentration of 50mg/L, the oxidizing agent comprises peracetic acid and a diluent, and the mass concentration of the peracetic acid is 0.25%.
Specifically, the dosage ratio of the purifying agent to the diluted urine is 0.111 g/mL-0.222 g/mL. In the present embodiment, the purifying agent is activated carbon. The activated carbon can purify diluted urine and remove interfering substances in the urine, such as bilirubin, vitamin C, sodium chloride and the like, but the activated carbon does not adsorb iodide ions, so that the urine is purified.
The volume ratio of the diluted urine to the oxidant to the color developing agent is 3: 1. By adding the substances according to the proportion, the diluted urine can be purified, and the color reaction is rapid and within the concentration range.
The reason why the color-developing agent and the oxidizing agent are added in sequence to the diluted urine treated by the purifying agent is that if the color-developing agent and the oxidizing agent are added simultaneously, the color-developing reaction starts immediately, the time for starting the reaction is difficult to accurately judge, and the test result is affected. If the oxidizing agent is added first and then the color-developing agent is added, the oxidizing agent reacts with the diluted urine and also affects the test result. Therefore, the method of mixing the developer and the oxidant is adopted.
Specifically, the color developing agent also comprises citric acid, sodium ethylene diamine tetracetate, glycerol, dimethyl sulfoxide and water. Wherein the concentration of the citric acid is 1.9 g/L. The concentration of EDTA-Na was 0.4 g/L. The volume ratio of glycerin to the color developing agent is 0.1. The volume ratio of the DMSO to the color developing agent is 0.01.
EDTA-Na is an important chelating agent, and can chelate metal ions in the solution and prevent the solution from discoloring, deteriorating and becoming turbid caused by metals. The citric acid has similar action to EDTA-Na except that it provides an acidic environment, i.e. it acts as a complexing agent, a masking agent, and can rapidly precipitate metal ions.
TMB is readily soluble in organic solvents and is more soluble in DMSO. Therefore, DMSO was added to the developer. However, since a high concentration of the organic solvent reduces the color developing effect, DMSO is added only to the developer in an amount of 1% by volume. Meanwhile, the melting point of DMSO is 18.5 ℃, the DMSO is solid in the storage process at 4 ℃, and the DMSO is inconvenient to use, so that glycerol with the volume content of 10% is added, and the color developing agent is liquid at 4 ℃ and is convenient to store.
The peroxyacetic acid in the oxidant can perform oxidation reaction with TMB in the color developing agent under the catalytic action of iodide ions to obtain a blue product. The higher the iodide ion concentration, the faster the reaction speed, and the more blue products, so that when the color developer, the oxidizing agent, and the urine to be measured are contacted, the darker the color of the obtained solution, the higher the iodide ion concentration. Therefore, the concentration of iodide ions can be obtained according to the shade of the color.
Further, step S120 includes:
providing a hydrogel chip, wherein the hydrogel chip comprises a purification tank and a detection tank, a purifying agent is loaded in the purification tank, a detector capable of detecting iodine content is loaded in the detection tank, the purification tank is communicated with the detection tank through a first channel, the hydrogel chip is also provided with a second channel and a third channel, the second channel is communicated with the first channel, the third channel is communicated with the first channel, the second channel is close to the purification tank than the third channel, the hydrogel chip is also provided with a fourth channel communicated with the purification tank, diluent and urine to be detected can flow in from the fourth channel, a color developing agent can flow in from the second channel, and an oxidizing agent can flow in from the third channel;
and taking the diluted urine, the oxidant and the color-developing agent, respectively injecting the diluted urine, the oxidant and the color-developing agent into a fourth channel, a second channel and a third channel of the hydrogel chip so as to purify the diluted urine in the purification tank, and then mixing the diluted urine, the oxidant and the color-developing agent.
The structural schematic diagram of the hydrogel chip is shown in fig. 1. The width of four channels in the hydrogel chip is 3mm, and the length of the hydrogel chip is 5 cm-10 cm. In order to ensure the accuracy of the test, the ratio of the length of the channel through which the diluted urine flows, the length of the channel through which the color developing agent flows and the length of the channel through which the oxidizing agent flows is 3:1, which is the same as the volume ratio of the diluted urine, the oxidizing agent and the color developing agent, before mixing.
The hydrogel chip is used as a detection device to detect the urine to be detected, a channel can be provided for purification of the urine to be detected and color development of a color development system, purification and detection of the urine are integrally completed, and the degree of automation is high. Compared with the traditional rigid material, the hydrogel chip has better stretchability, can make the product design more flexible and convenient, and simultaneously has the advantages of convenience in integration and high-channel analysis.
Step S130: and detecting the mixed solution to obtain the iodine content in the urine to be detected.
Specifically, when the hydrogel chip is used as the detection device in step S120, step S130 includes: and (3) timing when the mixed liquid completely flows into the detection pool of the hydrogel chip, and detecting the detection pool by using a detector after 300 s.
In particular, the detector is a color sensor. Adopt color sensor can detect mixed liquid fast, and arrange the detector in the measuring cell, can realize the purification of urine and the integration that detects, and need not take out mixed liquid and recycle the spectrophotometer and test, simplified the operating procedure, degree of automation is high.
Specifically, the method for detecting the iodine content in urine further comprises the following steps: and detecting the iodide ion standard solutions with a plurality of concentration gradients to obtain a relation curve of the concentration of the iodide ions and the absorbance.
The detection of the iodide ion standard solutions with a plurality of concentration gradients can be carried out by adopting a spectrophotometer method, and can also be carried out by adopting a hydrogel chip and utilizing a detector to detect the iodide ion standard solutions.
In step S130, the content of iodide ions in the urine to be measured is obtained according to the relationship curve between the concentration of iodide ions and the absorbance. In the present embodiment, the concentrations of the iodide ion standard solution include: 10. mu.g/L, 60. mu.g/L, 80. mu.g/L, 100. mu.g/L, 300. mu.g/L, 400. mu.g/L and 500. mu.g/L. The standard solution of iodide ions is an aqueous solution of potassium iodide. And substituting the developed absorbance of the mixed solution into the relation curve to obtain the concentration of the iodide ions in the urine to be detected, and further calculating to obtain the iodide ion content in the urine to be detected.
It should be noted that the method for detecting the mixed solution in step S130 should be the same as the method for detecting the standard solution of iodide ions to obtain the standard curve, so as to ensure the accuracy of the test result.
The method for detecting the iodine content in the urine at least has the following advantages:
(1) according to the method for detecting the content of iodine in urine, the diluted urine is treated by the purifying agent, so that interferents in urine can be removed, and sodium chloride in the diluted urine can reserve iodine ions to the maximum extent, so that the purifying agent has no influence on the content of the iodine ions when treating the diluted urine, and the accuracy and the anti-interference performance of a test result are high.
(2) The detection method for the content of iodine in urine is simple to operate and quick to test.
(3) The method for detecting the content of iodine in urine utilizes a specific oxidant and a specific color developing agent as a color developing system, and the repeatability of a detection result is good.
The hydrogel chip of one embodiment is prepared as follows:
1g of sodium alginate and 8g of acrylamide were dissolved with 36g of pure water to obtain a first solution. Ammonium persulfate (initiator) 0.08g and N, N' -methylenebisacrylamide (crosslinking agent) 0.015g were added to 1mL of water and dissolved to obtain a second solution. And mixing the first solution and the second solution, and vacuumizing for 5min to obtain a third solution. Preparing calcium sulfate dihydrate into a calcium sulfate solution at 40 ℃, then taking 5mL of supernatant, adding 100 mu L of tetramethylethylenediamine catalyst, mixing, and pouring into a third solution to obtain a mixed solution.
Pouring the mixture into a culture dish, shaking (no air bubbles are needed during pouring), placing the plate on a heating plate, and heating. Wherein the heating time of the first semi-cured hydrogel is 15min, and the heating time of the second semi-cured hydrogel is 12 min. The heating temperature was 60 ℃.
According to the structural schematic diagram of the hydrogel chip shown in fig. 1, the first semi-cured hydrogel is cut by using a laser cutting machine to manufacture the hydrogel inner pipeline, so as to obtain a cutting plate. And respectively placing a purifying agent and a detector in a purifying pool and a detecting pool of the cutting plate, then covering the cutting plate with second semi-solidified hydrogel, and heating in a water bath at 50 ℃ for 30 min. And then turning on an ultraviolet lamp, and standing in a moisture preservation box for 1 day to obtain the hydrogel chip.
The following are specific examples:
example 1
The method for detecting the iodine content in the urine in the embodiment is as follows:
(1) 490mL of 0.2mol/L sodium acetate solution is mixed with 510mL of 0.2mol/L acetic acid solution, and sodium chloride is added to obtain acetic acid-sodium acetate diluent with pH value of 4.6, wherein the mass concentration of the sodium chloride is 4%. 25mg of 3,3',5,5' -tetramethylbenzidine was dissolved in 5mL of DMSO, and then 0.95g of citric acid, 0.2g of EDTA-Na, 50mL of glycerin were added, and water was added to 500mL to obtain a color developer. And mixing the peroxyacetic acid with the acetic acid-sodium acetate diluent to obtain the oxidant with the mass concentration of the peroxyacetic acid of 0.25%.
(2) Collecting urine to be tested, taking lmL urine (shaking the urine before sampling), adding 5mL of acetic acid-sodium acetate diluent with pH of 4.6, and mixing uniformly to prepare diluted urine.
(3) Respectively taking 900 mu L of diluted urine, 300 mu L of oxidizing agent and 300 mu L of color developing agent, and sequentially injecting the diluted urine, the color developing agent and the oxidizing agent as shown in the schematic diagram of the hydrogel chip in figure 1, wherein the dosage of the purifying agent in the hydrogel chip is 0.1 g.
(4) And (3) when the urine to be diluted, the color developing agent and the oxidant are all mixed in the detection pool, timing and shaking up, and after 300s, detecting the detection pool by using a color identification sensor, thereby obtaining the content of iodide ions in the urine to be detected.
Example 2
The method for detecting the iodine content in the urine in the embodiment is as follows:
(1) mixing acetic acid, sodium acetate and water, and adding sodium chloride to obtain an acetic acid-sodium acetate diluent with a pH value of 4.6, wherein the mass concentration of the sodium chloride is 3.5%. Dissolving 3,3',5,5' -tetramethyl benzidine in DMSO, and adding citric acid, EDTA-Na, glycerol and water to obtain the color developing agent. The mass concentration of TMB in the color developing agent is 50mg/L, the mass concentration of citric acid is 1.9g/L, the mass concentration of EDTA-Na is 0.4g/L, the addition amount of glycerol is 10% of the volume of the color developing agent, and the addition amount of DMSO is 1% of the volume of the color developing agent. And mixing the peroxyacetic acid with the diluent to obtain the oxidant with the mass concentration of the peroxyacetic acid of 0.25%.
(2) Collecting urine to be detected, taking 1mL of urine to be detected (shaking the urine before sampling), adding 5mL of diluent with pH of 4.6, and fully and uniformly mixing to prepare diluted urine.
(3) Respectively taking 900 mu L of diluted urine, 300 mu L of oxidizing agent and 300 mu L of color developing agent, and sequentially injecting the diluted urine, the color developing agent and the oxidizing agent according to the schematic diagram of the hydrogel chip in figure 1, wherein the dosage of the purifying agent in the hydrogel chip is 0.15 g.
(4) And (3) when the urine to be diluted, the color developing agent and the oxidant are all mixed in the detection pool, timing and shaking up, and after 300s, detecting the detection pool by using a color identification sensor to obtain the content of iodide ions in the urine to be detected.
Example 3
The method for detecting the iodine content in the urine in the embodiment is as follows:
(1) mixing acetic acid, sodium acetate and water, and adding sodium chloride to obtain an acetic acid-sodium acetate diluent with a pH value of 4.6, wherein the mass concentration of the sodium chloride is 4.5%. Dissolving 3,3',5,5' -tetramethyl benzidine in DMSO, adding citric acid, EDTA-Na, glycerol and water to obtain the color developing agent. The mass concentration of TMB in the color developing agent is 50mg/L, the mass concentration of citric acid is 1.9g/L, the mass concentration of EDTA-Na is 0.4g/L, the addition amount of glycerol is 10% of the volume of the color developing agent, and the addition amount of DMSO is 1% of the volume of the color developing agent. And mixing the peroxyacetic acid with the diluent to obtain the oxidant with the mass concentration of the peroxyacetic acid of 0.25%.
(2) Collecting urine to be detected, taking 1mL of urine (shaking the urine before sampling), adding 5mL of diluent with pH of 4.6, and fully and uniformly mixing to prepare diluted urine.
(3) Respectively taking 900 mu L of diluted urine, 300 mu L of oxidizing agent and 300 mu L of color developing agent, and sequentially injecting the diluted urine, the color developing agent and the oxidizing agent as shown in the schematic diagram of the hydrogel chip in figure 1, wherein the dosage of the purifying agent in the hydrogel chip is 0.2 g.
(4) And (3) when the urine to be diluted, the color developing agent and the oxidant are all mixed in the detection pool, timing and shaking up, and after 300s, detecting the detection pool by using a color identification sensor to obtain the content of iodide ions in the urine to be detected.
Comparative example 1
The method for detecting the iodine content in the urine of the comparative example is as follows:
(1) mixing acetic acid, sodium acetate and water, and adding sodium chloride to obtain an acetic acid-sodium acetate diluent with a pH value of 4.6, wherein the mass concentration of the sodium chloride is 2.5%. Dissolving 3,3',5,5' -tetramethyl benzidine in DMSO, adding citric acid, EDTA-Na, glycerol and water to obtain the color developing agent. The mass concentration of TMB in the color developing agent is 50mg/L, the mass concentration of citric acid is 1.9g/L, the mass concentration of EDTA-Na is 0.4g/L, the addition amount of glycerol is 10% of the volume of the color developing agent, and the addition amount of DMSO is 1% of the volume of the color developing agent. And mixing the peroxyacetic acid with the diluent to obtain the oxidant with the mass concentration of the peroxyacetic acid of 0.25%.
(2) Collecting urine to be detected, taking 1mL of urine (shaking the urine before sampling), adding 5mL of diluent with pH of 4.6, and fully and uniformly mixing to prepare diluted urine.
(3) The diluted urine, the color-developing agent and the oxidizing agent were sequentially injected into 900. mu.L of the diluted urine, 300. mu.L of the oxidizing agent and 300. mu.L of the color-developing agent, respectively, as shown in the schematic diagram of the hydrogel chip in FIG. 1.
(4) And (3) when the urine to be diluted, the color developing agent and the oxidant are all mixed in the detection pool, timing and shaking up, and after 300s, detecting the detection pool by using a color identification sensor to obtain the content of iodide ions in the urine to be detected.
The urine to be tested was tested for 8 replicates using the test kits of examples 1-3 and comparative example 1 and the commercially available Qingdao Sankai kit. The test results are shown in table 1 below, and the results of 8 repeated tests on the iodine content in urine by using the detection kit for detecting the iodine content in urine in example 1 and the Qingdao Sanka kit are plotted as a curve, as shown in fig. 3. In fig. 3, a curve marked with a circle is a test result curve of the test kit in example 1, and a curve marked with a diamond is a test result curve of the Qingdao Sanka kit.
TABLE 1 comparison of test results of test kits and commercially available kits of examples and comparative examples
Wherein, the calculation formula of the CV value is as follows: CV value is standard deviation/average value × 100%. From the above experimental results, it can be seen that the CV values of the test results using the test kits of examples 1 to 3 are all around 5%, and the reproducibility is good, whereas the CV value of the test result using the test kit of comparative example 1 is large, because the concentration of sodium chloride in the diluent is low, so that the urine purification effect in the purification step using activated carbon is poor. However, the CV value of the detection kit in the comparative example 1 is still lower than that of the Qingdao Sankai kit, which indicates that the repeatability of the chromogenic reaction system in the comparative example is better than that of the chromogenic reaction system in the Qingdao Sankai kit.
The interference of ascorbic acid (Vc), sodium chloride (NaCl), bilirubin, etc. to the test process was examined according to the composition of urine, and the following test samples were tested in triplicate using the test kit of example 1, respectively, and the test results are shown in table 2 below.
TABLE 2 test results of the test kit of example 1 for different urine
Wherein, the relative error is calculated according to the following formula: relative error (%) — measured average of interference sample-measured average of blank sample × 100%/measured average of blank sample. Blank sample is the urine to be detected with the concentration of the interferent being 0.
The experimental results show that after three types of interference substances are respectively added into the blank sample, compared with a blank control group, the relative errors are less than 15%, so that the interference of ascorbic acid, sodium chloride and bilirubin on the detection of the iodine content in urine can be eliminated after the color reaction system is purified by active carbon.
The above experiment results all show that the detection kit for detecting the iodine content in the urine in the embodiment is adopted to detect the urine, the repeatability of the detection result is good, the influence of interference substances can be eliminated, and the interference resistance is good.
The technical features of the above embodiments can be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the above embodiments are not described, but should be considered as the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above examples only show some embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (11)
1. The utility model provides a detection kit of iodine content in urine, its characterized in that, includes diluent, colour-developing agent, oxidant and purification agent, the diluent comprises the sodium chloride and acetic acid-sodium acetate buffer solution that mass concentration is 3.5% ~ 4.5%, the pH of diluent is 4.6, the colour-developing agent includes that the concentration is 50 mg/L3, 3',5,5' -tetramethyl benzidine, the oxidant includes peracetic acid and the diluent, the mass concentration of peracetic acid is 0.25%, the colour-developing agent still includes citric acid, sodium edetate, glycerine, dimethyl sulfoxide and water, the iodine concentration's that detection kit can detect scope is 10 mug/L ~ 500 mug/L.
2. The kit for detecting the content of iodine in urine according to claim 1, wherein said color-developing agent further comprises: citric acid with the concentration of 1.9g/L, sodium ethylene diamine tetracetate with the concentration of 0.4g/L, glycerol with the volume ratio of the color developing agent of 0.1 and dimethyl sulfoxide with the volume ratio of the color developing agent of 0.01.
3. The kit for detecting the content of iodine in urine according to claim 1, wherein said purifying agent is activated carbon.
4. The kit for detecting the content of iodine in urine according to claim 1, further comprising a hydrogel chip, the hydrogel chip is provided with a purification tank and a detection tank, the purification tank is internally loaded with the purifying agent, a detector capable of detecting the content of iodine is arranged in the detection pool, the purification pool is communicated with the detection pool through a first channel, the hydrogel chip is also provided with a second channel and a third channel, the second channel is communicated with the first channel, the third channel is communicated with the first channel, the second channel is closer to the purification tank than the third channel, the hydrogel chip is also provided with a fourth channel communicated with the purification tank, the diluent and the urine to be detected can flow into the fourth channel, the developer is allowed to flow in from the second channel, and the oxidizer is allowed to flow in from the third channel.
5. The kit for detecting the content of iodine in urine according to claim 4, wherein said hydrogel chip comprises a detection plate and a cover plate, said detection plate is provided with said purification reservoir, said detection reservoir, said first channel, said second channel, said third channel and said fourth channel, said cover plate covers said detection plate and shields said purification reservoir, said detection reservoir, said first channel, said second channel, said third channel and said fourth channel.
6. A method for detecting the content of iodine in urine is characterized by comprising the following steps:
taking urine to be detected, and mixing the urine with a diluent to obtain diluted urine, wherein the diluent consists of sodium chloride with the mass concentration of 3.5-4.5% and acetic acid-sodium acetate buffer solution, and the pH of the diluent is 4.6;
purifying the diluted urine by using a purifying agent, and sequentially adding a color-developing agent and an oxidizing agent into the purified diluted urine to obtain a mixed solution, wherein the color-developing agent comprises 3,3',5,5' -tetramethylbenzidine with the concentration of 50mg/L, the oxidizing agent comprises peroxyacetic acid and the diluent, the mass concentration of the peroxyacetic acid is 0.25%, and the color-developing agent further comprises citric acid, sodium ethylene diamine tetracetate, glycerol, dimethyl sulfoxide and water; and
detecting the mixed solution to obtain the iodine content in the urine to be detected;
wherein the volume ratio of the oxidant to the color developing agent is 1:1, and the concentration range of iodine which can be detected by the method for detecting the content of iodine in urine is 10-500 mug/L.
7. The method for detecting the content of iodine in urine according to claim 6, wherein in the step of taking the urine to be detected and mixing the urine to be detected with the diluent, the volume ratio of the urine to be detected to the diluent is 1: 5.
8. The method for detecting iodine content in urine according to claim 6, wherein in said step of purifying said diluted urine with a purifying agent, the amount ratio of said purifying agent to said diluted urine is 0.111g/mL to 0.222 g/mL.
9. The method for detecting the iodine content in the urine according to claim 6, wherein in the step of sequentially adding a color-developing agent and an oxidizing agent to the purified diluted urine, the volume ratio of the diluted urine to the oxidizing agent to the color-developing agent is 3:1: 1.
10. The method for detecting iodine content in urine according to claim 6, wherein said step of purifying said diluted urine with a purifying agent, and sequentially adding a color-developing agent and an oxidizing agent to said purified diluted urine comprises:
providing a hydrogel chip, wherein the hydrogel chip comprises a purification pool and a detection pool, the purification pool is internally loaded with a purifying agent, the detection pool is internally loaded with a detector capable of detecting iodine content, the purification pool is communicated with the detection pool through a first channel, the hydrogel chip is also provided with a second channel and a third channel, the second channel is communicated with the first channel, the third channel is communicated with the first channel, the second channel is closer to the purification pool than the third channel, the hydrogel chip is also provided with a fourth channel communicated with the purification pool, diluent and urine to be detected can flow in from the fourth channel, a color developing agent can flow in from the second channel, and an oxidizing agent can flow in from the third channel;
and taking the diluted urine, the color developing agent and the oxidizing agent, respectively injecting the diluted urine, the color developing agent and the oxidizing agent into the fourth channel, the second channel and the third channel of the hydrogel chip so as to purify the diluted urine in the purification tank, and then mixing the diluted urine, the color developing agent and the oxidizing agent.
11. The method for detecting iodine content in urine according to claim 10, wherein said step of detecting said mixture comprises: and starting timing when the mixed liquid completely flows into the detection pool of the hydrogel chip, and detecting the mixed liquid in the detection pool by using the detector after 300 s.
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| CN112972128B (en) * | 2021-02-03 | 2022-03-29 | 杭州可靠护理用品股份有限公司 | Baby diaper and urine display line thereof |
| CN113959822B (en) * | 2021-10-29 | 2024-02-20 | 珠海市丽拓生物科技股份有限公司 | Diluent and oxidant for measuring iodine content in urine by peroxyacetic acid oxidation method and application |
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| JP4474010B2 (en) * | 2000-03-15 | 2010-06-02 | アークレイ株式会社 | Specimen with solid component separation ability |
| CN100514043C (en) * | 2006-03-07 | 2009-07-15 | 北京中生金域诊断技术有限公司 | Urine iodine test reagent kit |
| CN103272656B (en) * | 2011-12-20 | 2015-05-27 | 国家纳米科学中心 | Bar code micro fluidic chip and purpose thereof |
| EP3584010A1 (en) * | 2012-07-30 | 2019-12-25 | NMI Naturwissenschaftliches und Medizinisches Institut an der Universität Tübingen | Method of investigation of and/or with a biological sample |
| CN102866249B (en) * | 2012-08-15 | 2015-01-21 | 华北制药集团新药研究开发有限责任公司 | Tetramethylbenzidine (TMB) developing system |
| CN106546586A (en) * | 2016-11-09 | 2017-03-29 | 安徽惠邦生物工程股份有限公司 | The detectable of content of iodine in a kind of vitro detection urine |
| CN108890937B (en) * | 2018-06-29 | 2020-07-07 | 山东省科学院能源研究所 | Preparation method of hydrogel chip with dendritic channel structure |
| CN110006881B (en) * | 2019-04-01 | 2020-09-11 | 深圳大学 | Detection kit for iodine content in urine and detection method for iodine content in urine |
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| US5939328A (en) * | 1995-03-04 | 1999-08-17 | Merck Patent Gesellschaft Mit Beschrankter Haftung | Method for the determination of iodide |
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| CN105424629A (en) * | 2015-12-11 | 2016-03-23 | 苏州汶颢芯片科技有限公司 | Micro-fluidic chip and copper ion detection system |
| CN109470849A (en) * | 2018-10-24 | 2019-03-15 | 四川农业大学 | A kind of haemophilus parasuis antibody assay kit and its application method |
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