CN113959822A - Diluent and oxidant for determining urine iodine content by peroxyacetic acid oxidation method and application - Google Patents
Diluent and oxidant for determining urine iodine content by peroxyacetic acid oxidation method and application Download PDFInfo
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- CN113959822A CN113959822A CN202111275912.6A CN202111275912A CN113959822A CN 113959822 A CN113959822 A CN 113959822A CN 202111275912 A CN202111275912 A CN 202111275912A CN 113959822 A CN113959822 A CN 113959822A
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- peroxyacetic acid
- oxidation method
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- KFSLWBXXFJQRDL-UHFFFAOYSA-N Peracetic acid Chemical compound CC(=O)OO KFSLWBXXFJQRDL-UHFFFAOYSA-N 0.000 title claims abstract description 97
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 title claims abstract description 80
- 239000011630 iodine Substances 0.000 title claims abstract description 80
- 229910052740 iodine Inorganic materials 0.000 title claims abstract description 80
- 210000002700 urine Anatomy 0.000 title claims abstract description 60
- 239000003085 diluting agent Substances 0.000 title claims abstract description 46
- 239000007800 oxidant agent Substances 0.000 title claims abstract description 45
- 238000000034 method Methods 0.000 title claims abstract description 39
- 230000003647 oxidation Effects 0.000 title claims abstract description 30
- 238000007254 oxidation reaction Methods 0.000 title claims abstract description 30
- 230000001590 oxidative effect Effects 0.000 title claims abstract description 29
- 239000000243 solution Substances 0.000 claims abstract description 32
- 239000007853 buffer solution Substances 0.000 claims abstract description 20
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims abstract description 16
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 16
- YASYEJJMZJALEJ-UHFFFAOYSA-N Citric acid monohydrate Chemical compound O.OC(=O)CC(O)(C(O)=O)CC(O)=O YASYEJJMZJALEJ-UHFFFAOYSA-N 0.000 claims abstract description 14
- 229960002303 citric acid monohydrate Drugs 0.000 claims abstract description 14
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims abstract description 13
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000005557 antagonist Substances 0.000 claims abstract description 8
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- 229910000403 monosodium phosphate Inorganic materials 0.000 claims abstract description 8
- 235000019799 monosodium phosphate Nutrition 0.000 claims abstract description 8
- 239000001103 potassium chloride Substances 0.000 claims abstract description 8
- 235000011164 potassium chloride Nutrition 0.000 claims abstract description 8
- 239000011780 sodium chloride Substances 0.000 claims abstract description 8
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims abstract description 8
- 238000001179 sorption measurement Methods 0.000 claims abstract description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 7
- 238000010790 dilution Methods 0.000 claims description 16
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- 239000003755 preservative agent Substances 0.000 claims description 9
- 230000002335 preservative effect Effects 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 5
- 229960002816 potassium chloride Drugs 0.000 claims description 3
- 239000008213 purified water Substances 0.000 claims description 3
- 229960002668 sodium chloride Drugs 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 239000012089 stop solution Substances 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 3
- 238000001514 detection method Methods 0.000 abstract description 48
- 230000035945 sensitivity Effects 0.000 abstract description 7
- 231100000053 low toxicity Toxicity 0.000 abstract description 2
- 238000012360 testing method Methods 0.000 description 19
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 12
- 238000011161 development Methods 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- -1 tetramethylbenzidine peroxyacetate Chemical compound 0.000 description 7
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 6
- 238000002798 spectrophotometry method Methods 0.000 description 6
- 230000031700 light absorption Effects 0.000 description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 229960004106 citric acid Drugs 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 238000012417 linear regression Methods 0.000 description 4
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- 230000000694 effects Effects 0.000 description 3
- 238000006479 redox reaction Methods 0.000 description 3
- 238000010561 standard procedure Methods 0.000 description 3
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 3
- 229940033663 thimerosal Drugs 0.000 description 3
- 206010067997 Iodine deficiency Diseases 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
- TUFLVBXBZSNFCZ-UHFFFAOYSA-N arsenic cerium Chemical compound [As].[Ce] TUFLVBXBZSNFCZ-UHFFFAOYSA-N 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
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- 238000010998 test method Methods 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 1
- ZBQCCTCQUCOXBO-UHFFFAOYSA-N 4-(4-aminophenyl)-2,2,6,6-tetramethylcyclohex-3-en-1-amine Chemical compound CC1(C)C(N)C(C)(C)CC(C=2C=CC(N)=CC=2)=C1 ZBQCCTCQUCOXBO-UHFFFAOYSA-N 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- 241001354491 Lasthenia californica Species 0.000 description 1
- 208000036626 Mental retardation Diseases 0.000 description 1
- PQRTVVJRFXOOGJ-UHFFFAOYSA-N O.O.[Na].[Na].[Na].C(CC(O)(C(=O)O)CC(=O)O)(=O)O Chemical compound O.O.[Na].[Na].[Na].C(CC(O)(C(=O)O)CC(=O)O)(=O)O PQRTVVJRFXOOGJ-UHFFFAOYSA-N 0.000 description 1
- 208000024799 Thyroid disease Diseases 0.000 description 1
- AUYYCJSJGJYCDS-LBPRGKRZSA-N Thyrolar Chemical class IC1=CC(C[C@H](N)C(O)=O)=CC(I)=C1OC1=CC=C(O)C(I)=C1 AUYYCJSJGJYCDS-LBPRGKRZSA-N 0.000 description 1
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- 210000003754 fetus Anatomy 0.000 description 1
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- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
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- 210000000653 nervous system Anatomy 0.000 description 1
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Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
Abstract
The invention provides a diluent, an oxidant and application for determining urine iodine content by a peroxyacetic acid oxidation method, wherein the diluent comprises a buffer solution and an iodine adsorption antagonist, and the buffer solution comprises: the mass concentration of the citric acid monohydrate is 20-30 g/L, the trisodium citrate dihydrate is 10-15 g/L, 4-5% of sodium dihydrogen phosphate and 4-5% of disodium hydrogen phosphate; iodine adsorption antagonists include: the mass concentration of the sodium chloride is 2-3% and the mass concentration of the potassium chloride is 2-3%; the pH value of the diluent is 2.5-2.95. The oxidant comprises 4.5-6% of peroxyacetic acid, 28.2-28.65% of hydrogen peroxide and water. The advantages are that: the diluent has good stability and low toxicity, and has better linear relation and higher accuracy for detecting urine with low-concentration iodine content; the oxidant can obtain a standard curve with good linearity, no intermediate solution is needed, and the sensitivity and the accuracy are good in the detection range of 0-500 mu g/L.
Description
Technical Field
The invention relates to the field of in-vitro diagnosis and detection, in particular to a diluent and an oxidant for determining urine iodine content by a peracetic acid oxidation method and application thereof.
Background
Iodine is an essential trace element for synthesis of various systems of a human body, particularly thyroid hormone and development of a nervous system, and can be taken along with food, drinking water, salt and the like when entering the body, wherein 90 percent of iodine is discharged by the kidney. Urine iodine can roughly reflect the amount of iodine taken by a person and the amount of iodine in the blood. Both iodine deficiency and excess may cause thyroid disorders, constituting a series of damages to the human body, especially iodine deficiency in pregnant women may cause irreversible mental retardation and psychomotor disturbance in the fetus or infant. The iodine level of urine is an important index for measuring the iodine nutrition status of people at present, and is widely applied to the aspects of clinical diagnosis and treatment, medical care, disease prevention and treatment and the like in hospitals.
At present, the methods for detecting the urine iodine mainly comprise an inductively coupled plasma-mass spectrometry method, a neutron digestion method, a spectrophotometry method, a gas chromatography method and the like. The current standard method for detecting the urine iodine in China is an arsenic-cerium catalytic spectrophotometry (national standard method) newly issued by the original Ministry of health in 2016, and has the advantages of good repeatability, strong anti-interference capability, high sensitivity and the like.
Luoxixian et al (No. 41, Vol.41, No. 4, 2020, J.Immunol. Med.) examined the method and technique for measuring iodine level in urine by peroxyacetic acid oxidation method, and confirmed that the method for measuring iodine in urine by peroxyacetic acid oxidation method is simple in operation, good in standard curve and linear relation, high in accuracy, good in specificity and good in repeatability, and is a rapid method for detecting urine iodine which is worth popularizing.
The detection kit for detecting the peroxyacetic acid oxidation method in the market generally comprises an adsorbent, a diluent, a color developing agent, an oxidant, a standard substance and the like, the existing kit has the problems of large deviation of a detection value of a low-concentration iodine solution, low detection value of a high concentration, narrow concentration detection range, insufficient detection sensitivity and the like, and meanwhile, thimerosal or sodium azide is usually added into a diluent of the existing kit to serve as a preservative, but the sodium azide is extremely toxic, and the toxicity of the thimerosal is unknown. The accuracy and sensitivity of the detection kit for measuring the peroxyacetic acid oxidation method in the market can not reach the detection effect of arsenic-cerium catalytic spectrophotometry (national standard method), and the popularization of the peroxyacetic acid oxidation method in the urine iodine detection is influenced.
Disclosure of Invention
The invention provides a diluent and an oxidant for measuring the content of iodine in urine and application thereof, aiming at the problems that the detection value of the existing peracetic acid oxidation method detection reagent is large in deviation on the detection value of low-concentration iodine solution, low in detection value on high concentration, narrow in concentration detection range, insufficient in detection sensitivity and the like.
The technical scheme of the invention is as follows:
a dilution for measuring urine iodine by a peroxyacetic acid oxidation method comprises a buffer solution and an iodine adsorption antagonist, wherein the buffer solution comprises: the mass concentration of the citric acid monohydrate is 20-30 g/L, the trisodium citrate dihydrate is 10-15 g/L, 4-5% of sodium dihydrogen phosphate and 4-5% of disodium hydrogen phosphate; iodine adsorption antagonists include: the mass concentration of the sodium chloride is 2-3% and the mass concentration of the potassium chloride is 2-3%; the pH value of the diluent is 2.5-2.95.
Further, the preservative also comprises a preservative, wherein the preservative is proclin-300 with the mass concentration of 1-2%.
Further, citric acid monohydrate or trisodium citrate dihydrate is also used to adjust the pH of the diluent.
The preparation method of the diluent for measuring the urine iodine content by the peroxyacetic acid oxidation method is used for preparing the diluent and comprises the following steps: weighing citric acid monohydrate, sodium chloride, potassium chloride, trisodium citrate dihydrate, sodium dihydrogen phosphate, disodium hydrogen phosphate and proclin-300 in proportion, dissolving with purified water, fixing the volume to the preparation volume, and stirring uniformly; measuring the pH value of the prepared diluent, and adjusting the pH value to be below 3.0 by using citric acid monohydrate or trisodium citrate dihydrate solution.
The oxidant for measuring the urine iodine content by the peroxyacetic acid oxidation method comprises 4.5-6% of peroxyacetic acid, 28.2-28.65% of hydrogen peroxide and the balance of water.
The application of the dilute solution for measuring the urine iodine content by the peroxyacetic acid oxidation method in measuring the urine iodine content.
Furthermore, the oxidizing agent for measuring the iodine content in urine by the peroxyacetic acid oxidation method is used without using a stop solution.
The invention has the advantages that: the buffer system of two kinds of combinations of citric acid and phosphoric acid is adopted to the diluent, the buffer capacity is stronger, stability is good, the toxicity is low, do not react with the composition of urine sample, can not reduce the content of iodine in the urine, after the diluent dilutes the urine sample, filter through the filter equipment, participate in the oxidation reduction reaction of peracetic acid and 3,3,5, 5-tetramethyl benzidine, provide stable buffering environment for oxidation reduction reaction, guarantee going on smoothly of oxidation reduction reaction, especially the urine detection linear relation to low concentration iodine content is higher, the degree of accuracy is higher. The oxidant can keep the original sensitivity of the oxidant and has a good linear standard curve, and meanwhile, a stop solution is not needed, so that the detection steps of the peroxyacetic acid oxidation method are simplified, and the detection is more accurate and quicker; the detection concentration coverage range is wider, and the sensitivity and the accuracy are good in the detection range of 0-500 mu g/L.
Drawings
FIG. 1 is a curve of the OD value detection of the iodine solution with different concentrations by using the diluent of the present invention;
FIG. 2 is a curve of the OD value detection of the diluent with pH not less than 3.0 to iodine solutions of different concentrations;
FIG. 3 is a standard curve of the detection of the oxidant to iodine solutions of different concentrations;
FIG. 4 is a standard curve of detecting different concentrations of iodine solution with oxidant with peroxyacetic acid content less than 0.3%;
FIG. 5 is a standard curve of the detection of oxidant with peroxyacetic acid content > 8% to iodine solutions of different concentrations.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention relates to a diluent and an oxidant for detecting the content of iodine in urine by a tetramethylbenzidine peroxyacetate oxidation color development method.
The diluent comprises: the mass concentration of the citric acid monohydrate is 20-30 g/L, the citric acid trisodium dihydrate is 10-15 g/L, the sodium dihydrogen phosphate is 4-5%, the disodium hydrogen phosphate is 4-5%, the sodium chloride is 2-3%, the potassium chloride is 2-3%, the proclin-300 is 1-2%, and the pH value is 2.5-2.95.
The oxidizing agent comprises: the mass concentration is 4.5-6% of peroxyacetic acid, 28.2-28.65% of hydrogen peroxide and the balance of water. The oxidant has simple components and is less affected by other chemical components.
The diluent of the present invention comprises a buffer, an iodine adsorption antagonist and a preservative.
The buffer solution comprises a buffer system consisting of 20-30 g/L citric acid monohydrate, 10-15 g/L trisodium citrate dihydrate, 4-5% sodium dihydrogen phosphate and 4-5% disodium hydrogen phosphate, the citric acid monohydrate and the trisodium citrate dihydrate of the citric acid buffer system in the buffer solution are higher in use concentration and have the functions of fresh keeping, oxidation resistance and the like, and the buffer solution adopts a buffer system combining citric acid and phosphoric acid, so that the buffer solution is more in ion type, higher in concentration and higher in buffer capacity.
The iodine adsorption antagonist comprises 2-3% of sodium chloride and 2-3% of potassium chloride in mass concentration, and has certain effects of sterilizing and keeping cell osmotic pressure balance.
proclin-300 as the preservative added into the diluent is an efficient, low-toxicity bacteriostatic agent special for in vitro diagnostic reagents, is used for replacing thimerosal, sodium azide and gentamicin preservatives, can preserve the diluent at room temperature for a long time, and has little harm to human health and environment.
The pH value of the diluent is 2.5-2.95, so that the detection accuracy of the peracetic acid detection method on the urine iodine with low content concentration can be improved.
The preparation of the diluent of the invention comprises the following steps: weighing citric acid monohydrate, sodium chloride, potassium chloride, trisodium citrate dihydrate, sodium dihydrogen phosphate, disodium hydrogen phosphate and proclin-300 according to the proportion, dissolving with purified water, fixing the volume to a preparation volume, and uniformly stirring; and (3) measuring the pH value of the prepared diluent, and adjusting the pH value of the diluent to be below 3.0 by using a small amount of citric acid monohydrate or trisodium citrate dihydrate solution.
The oxidant is a key reagent of a tetramethylbenzidine peroxyacetate oxidation color development method, and the proportion of 4.5-6% of peroxyacetic acid is moderate, so that the color development reaction is not too strong or too weak; the oxidizing agent and 3,3',5,5' -tetramethyl benzidine (TMB) in the color developing solution generate blue color developing substances under the oxidation effect of iodine in urine; the blue color development is in direct proportion to the concentration of iodine in urine. In a certain range, the higher the concentration of iodine, the faster the reaction speed, and the more blue products; when the color developing agent and the oxidizing agent are contacted with the urine to be detected, the deeper the color of the obtained solution is, the higher the concentration of the iodide ions is, and according to the depth of the color of the solution, the absorbance is reflected, so that the concentration of the iodide ions is obtained.
Example 1
The following dilutions were used as an example to perform a comparative validation test of standard iodine solutions:
the prepared buffer solution and iodine solutions with different concentrations (0, 50ug/L, 100ug/L, 150ug/L, 200ug/L, 250ug/L and 300ug/L) participate in oxidation color development reaction of the tetramethylbenzidine peroxyacetate after being filtered by a filter device; the iodine of the iodine solution with each concentration reacts with the oxidant and the developing solution added into the cuvette for developing reaction in a buffer system of the diluent, and finally the light absorption value is measured by ultraviolet spectrophotometry. A test is carried out on a diluent in a commercial peroxyacetic acid kit under the same condition, and a comparison test is carried out by taking the diluent of the kit for measuring the urine iodine by the peroxyacetic acid method of Beijing Zhongsheng gold-field diagnostic technique Limited as an example, wherein the diluent comprises citric acid and magnesium chloride, the pH value is 3-6.5, and the diluent with the pH value being more than or equal to 3.0 is hereinafter referred to as the diluent for short.
The test process is completed by the following steps:
s1, placing the diluent, iodine solution or urine with various concentrations, a cuvette, a urine filter, an oxidant and a color developing solution into corresponding positions in a full-automatic urine iodine detector (LTS-ND180, Zhuhaili Tootho Biotech Co., Ltd.);
s2, starting a full-automatic urine iodine detector (LTS-ND180), entering an instrument homepage → clicking to load diluent according to the operation instruction of the urine iodine detector, fully loading the diluent pipeline → pointing a main page icon → entering a next step homepage → debugging the instrument → a liquid circuit module → a vertical progress machine, pointing a sample injection needle to a vertical position, point cleaning an inner wall, point cleaning an outer wall → resetting → returning, entering a main page → sample testing;
s3, after entering a sample test page, adding a number (the number of the detected samples) and a starting sample disk number of '1' → confirmation application → start of detection by clicking;
s4, after completion of the detection, the concentration of iodine detected can be directly read from the detection result.
TABLE 1 test of dilutions of the invention and dilutions having a pH of 3.0 or more on iodine solution
As shown in Table 1, FIG. 1 and FIG. 2, the light absorption values and corresponding iodine concentrations of the iodine solutions of the above concentrations were obtained by fitting a linear regression curve to obtain a regression coefficient R2And a regression equation; it can be seen that the linear regression coefficient r of the diluent detection standard curve of the present invention2R of 0.9996 vs. dilution pH 3.02Higher than 0.9909, better linearity, and more accurate experimental results on iodine solution.
Example 2
The following test experiments were performed using urine standards as an example of the dilution:
adding the prepared buffer solution and urine standard substances (14.6ug/L, 29.2ug/L, 58.5ug/L, 117ug/L and 234ug/L) with different concentrations into a filter device, and filtering by the filter device to participate in oxidation color development reaction of tetramethylbenzidine peroxyacetate; the iodine of the iodine solution with each concentration reacts with the oxidant and the developing solution added into the cuvette for developing reaction in a buffer system of the diluent, and finally the light absorption value is measured by ultraviolet spectrophotometry. A comparison of a commercial dilution of the peracetic acid kit was made under the same conditions and using the same procedure as in example 1.
And substituting the light absorption value measured by the iodine of the urine clinical sample into the linear regression equation to calculate the concentration of the urine.
TABLE 2 dilution of the invention used in the Peroxoacetic acid urine iodine test
TABLE 3 dilution of pH 3.0 or more for iodine in urine test by Peroxoacetic acid method
Example 3
The following dilution is used as an example to perform repeated tests on low concentration urine clinical samples:
adding the prepared buffer solution and a low-concentration urine clinical sample into a filtering device together, and participating in oxidation color development reaction of the tetramethylbenzidine peroxyacetate after filtering by the filtering device; the iodine of the iodine solution with each concentration reacts with the oxidant and the developing solution added into the cuvette for developing reaction in a buffer system of the diluent, and finally the light absorption value is measured by ultraviolet spectrophotometry. A comparison of a commercial dilution of the peracetic acid kit was made under the same conditions and using the same procedure as in example 1.
TABLE 4 test of clinical specimens with dilutions of the invention and dilutions having a pH of 3.0 or more
As can be seen from the test data of examples 1-3:
the detection value of low-concentration iodine in a standard curve detected by a commercially available diluent with the pH value of more than or equal to 3.0 is lower, the value of a detected clinical sample is higher, and finally the detected low-concentration urine standard substance (<50ug/L) and the low-concentration urine clinical sample are higher than those of an experiment.
Example 4
The following shows the accuracy of the test of the oxidizing agent of the present invention and the commercially available oxidizing agent with different peracetic acid contents on standard iodine solutions under the same test conditions, and the test procedure is the same as that of example 1. The detection results are as follows:
TABLE 5 content of different Peroxyacetic acids Standard Curve measurements of iodine content in urine
| Concentration of iodine | Less than 0.3 percent | 4.5～6％ | More than 8 |
| 0 | 0 | 0 | -0.02 |
| 50 | 0.104 | 0.202 | 0.356 |
| 150 | 0.495 | 0.612 | 1.049 |
| 300 | 0.957 | 1.210 | 1.801 |
| 400 | 1.299 | 1.55 | 1.939 |
| 500 | 1.501 | 1.9 | 2.255 |
The specific detection standard curve is shown in figures 3, 4 and 5, and the linear regression system r of the standard curve of the oxidant and the matched reagent thereof can be seen2The detection concentration range can reach 0-500 ug/L when the concentration is 0.9987; and the peroxyacetic acid content in the oxidant is less than 0.3 percent r2The detection range can reach 0-500 ug/L even if the detection range is 0.9949, but the detection value of 50ug/L iodine solution is low; r of peroxyacetic acid content greater than 8% in oxidant20.9562, the linearity is not good, and the detection range can only reach 0-300 ug/L. Therefore, the detection result of the oxidant is optimal.
Example 5
The following is the test accuracy of the oxidizing agent of the present invention and the commercially available oxidizing agent with different peracetic acid contents on the same test conditions for the urine standard substance, and the test procedure is the same as in example 1. The detection results are as follows:
TABLE 6 detection results of different peroxyacetic acid content oxidants on urine standard substance
It can be seen that the detection results of the oxidant of the present invention on BW09108t (89.8ug/L) and BW09110y (228ug/L) standards are uniform and accurate, while the concentration of the standard with peroxyacetic acid content less than 0.3% in the oxidant is higher, and the detection results with peroxyacetic acid content more than 8% in the oxidant are lower and uniform and inaccurate.
Example 6
The following is the testing accuracy of the oxidizing agent of the present invention and the commercially available oxidizing agent with different peracetic acid contents on clinical urine samples under the same testing conditions, and the testing procedure is the same as that of example 1. The detection results are as follows:
TABLE 7 test results of clinical urine specimen with different peroxyacetic acid content oxidants
It can be seen that the detection results of the oxidizing agent of the present invention for urine with different concentrations are relatively stable, the detection results for urine with low concentration are relatively high when the content of peroxyacetic acid in the oxidizing agent is less than 0.3%, the detection results for urine with low concentration are relatively low when the content of peroxyacetic acid in the oxidizing agent is greater than 8%, and the detection results are unstable and uniform.
Although the present invention has been described with reference to a preferred embodiment, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (7)
1. The diluent for determining the urine iodine content by the peroxyacetic acid oxidation method is characterized by comprising a buffer solution and an iodine adsorption antagonist, wherein the buffer solution comprises: the mass concentration of the citric acid monohydrate is 20-30 g/L, the trisodium citrate dihydrate is 10-15 g/L, 4-5% of sodium dihydrogen phosphate and 4-5% of disodium hydrogen phosphate; iodine adsorption antagonists include: the mass concentration of the sodium chloride is 2-3% and the mass concentration of the potassium chloride is 2-3%; the pH value of the diluent is 2.5-2.95.
2. The diluted solution for measuring the content of iodine in urine by the peroxyacetic acid oxidation method according to claim 1, further comprising a preservative, wherein the preservative is proclin-300 with a mass concentration of 1-2%.
3. The dilution for measuring the iodine content in urine by peroxyacetic acid oxidation method as claimed in claim 1 or 2, wherein citric acid monohydrate or trisodium citrate dihydrate is also used for adjusting the pH value of the dilution.
4. A method for preparing a diluent for measuring the iodine content in urine by a peroxyacetic acid oxidation method, which is used for preparing the diluent of claim 3, comprising the following steps: weighing citric acid monohydrate, sodium chloride, potassium chloride, trisodium citrate dihydrate, sodium dihydrogen phosphate, disodium hydrogen phosphate and proclin-300 in proportion, dissolving with purified water, fixing the volume to the preparation volume, and stirring uniformly; measuring the pH value of the prepared diluent, and adjusting the pH value to be below 3.0 by using citric acid monohydrate or trisodium citrate dihydrate solution.
5. The oxidant for determining the content of the iodine in the urine by a peroxyacetic acid oxidation method is characterized in that: comprises 4.5 to 6 mass percent of peroxyacetic acid, 28.2 to 28.65 mass percent of hydrogen peroxide and the balance of water.
6. Use of the dilution for determining the urine iodine content by the peroxyacetic acid oxidation method as set forth in any one of claims 1 to 3 for determining the urine iodine content.
7. Use according to claim 6, characterized in that: the oxidizing agent for measuring the iodine content in urine by the peroxyacetic acid oxidation method according to claim 5 is used without using a stop solution.
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