CN113340888B - Reagent, kit and detection method for quantitative detection of blood iodine - Google Patents
Reagent, kit and detection method for quantitative detection of blood iodine Download PDFInfo
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- CN113340888B CN113340888B CN202110752886.5A CN202110752886A CN113340888B CN 113340888 B CN113340888 B CN 113340888B CN 202110752886 A CN202110752886 A CN 202110752886A CN 113340888 B CN113340888 B CN 113340888B
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- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 title claims abstract description 50
- 229910052740 iodine Inorganic materials 0.000 title claims abstract description 50
- 239000011630 iodine Substances 0.000 title claims abstract description 50
- 238000001514 detection method Methods 0.000 title claims abstract description 39
- 210000004369 blood Anatomy 0.000 title claims abstract description 15
- 239000008280 blood Substances 0.000 title claims abstract description 15
- 239000003153 chemical reaction reagent Substances 0.000 title abstract description 24
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims abstract description 75
- 210000002966 serum Anatomy 0.000 claims abstract description 56
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 44
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 33
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 27
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 26
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 24
- 239000011780 sodium chloride Substances 0.000 claims abstract description 22
- 230000029087 digestion Effects 0.000 claims abstract description 17
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 claims abstract description 13
- PGJHGXFYDZHMAV-UHFFFAOYSA-K azanium;cerium(3+);disulfate Chemical compound [NH4+].[Ce+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O PGJHGXFYDZHMAV-UHFFFAOYSA-K 0.000 claims abstract description 13
- 239000011790 ferrous sulphate Substances 0.000 claims abstract description 13
- 235000003891 ferrous sulphate Nutrition 0.000 claims abstract description 13
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims abstract description 13
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims abstract description 13
- PTLRDCMBXHILCL-UHFFFAOYSA-M sodium arsenite Chemical compound [Na+].[O-][As]=O PTLRDCMBXHILCL-UHFFFAOYSA-M 0.000 claims abstract description 13
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 12
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 12
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 12
- 239000005708 Sodium hypochlorite Substances 0.000 claims abstract description 10
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims abstract description 10
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000000243 solution Substances 0.000 claims description 135
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 42
- 239000012086 standard solution Substances 0.000 claims description 25
- 230000035484 reaction time Effects 0.000 claims description 22
- OZECDDHOAMNMQI-UHFFFAOYSA-H cerium(3+);trisulfate Chemical compound [Ce+3].[Ce+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O OZECDDHOAMNMQI-UHFFFAOYSA-H 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 10
- 239000000126 substance Substances 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 3
- 238000002405 diagnostic procedure Methods 0.000 claims 1
- 150000002500 ions Chemical class 0.000 claims 1
- 239000000203 mixture Substances 0.000 abstract description 12
- 239000007788 liquid Substances 0.000 abstract description 3
- 230000001360 synchronised effect Effects 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 27
- 238000012360 testing method Methods 0.000 description 21
- 238000004364 calculation method Methods 0.000 description 9
- 238000011084 recovery Methods 0.000 description 7
- 238000001816 cooling Methods 0.000 description 6
- 238000005259 measurement Methods 0.000 description 5
- -1 polyethylene Polymers 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 239000002253 acid Substances 0.000 description 4
- 238000004448 titration Methods 0.000 description 4
- 101001018064 Homo sapiens Lysosomal-trafficking regulator Proteins 0.000 description 3
- 102100033472 Lysosomal-trafficking regulator Human genes 0.000 description 3
- 235000010703 Modiola caroliniana Nutrition 0.000 description 3
- 244000038561 Modiola caroliniana Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- XMBWDFGMSWQBCA-UHFFFAOYSA-M iodide Chemical compound [I-] XMBWDFGMSWQBCA-UHFFFAOYSA-M 0.000 description 3
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 description 3
- 238000010561 standard procedure Methods 0.000 description 3
- 210000002700 urine Anatomy 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 239000003595 mist Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 2
- 239000013582 standard series solution Substances 0.000 description 2
- KZBUYRJDOAKODT-UHFFFAOYSA-N Chlorine Chemical compound ClCl KZBUYRJDOAKODT-UHFFFAOYSA-N 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 208000024799 Thyroid disease Diseases 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- AQLMHYSWFMLWBS-UHFFFAOYSA-N arsenite(1-) Chemical compound O[As](O)[O-] AQLMHYSWFMLWBS-UHFFFAOYSA-N 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 235000021045 dietary change Nutrition 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229940006461 iodide ion Drugs 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- JLKDVMWYMMLWTI-UHFFFAOYSA-M potassium iodate Chemical compound [K+].[O-]I(=O)=O JLKDVMWYMMLWTI-UHFFFAOYSA-M 0.000 description 1
- 239000001230 potassium iodate Substances 0.000 description 1
- 235000006666 potassium iodate Nutrition 0.000 description 1
- 229940093930 potassium iodate Drugs 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000006479 redox reaction Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N2021/775—Indicator and selective membrane
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a reagent, a kit and a detection method for quantitative detection of blood iodine, wherein the reagent comprises the following components: a first composition: acetonitrile and ethyl acetate, or ammonium sulfate and isopropyl alcohol; a second composition: sodium hypochlorite and sodium chloride; third composition: sulfuric acid and hydrochloric acid in combination; fourth composition: sodium chloride and sodium arsenite in combination; a fifth composition: ferrous sulfate, phenanthroline and sulfuric acid in combination; sixth composition: ammonium cerium sulfate, and sulfuric acid. The rapid detection reagent and method for iodine in serum have the advantages of low acidity of digestion liquid, simple and rapid operation, synchronous use in combination with detection equipment, and no influence of manual errors of experimenters, so that the experimental result is accurate and stable.
Description
Technical Field
The invention belongs to the technical field of detection reagents, and particularly relates to a reagent for quantitative detection of blood iodine, a kit and a detection method.
Background
Iodine is one of microelements necessary for maintaining normal thyroid function of human body, and overmuch or overlittle daily intake can cause thyroid diseases, especially is very important for growth and development of fetuses and infants, so that the world health organization and the national children foundation take urine iodine as an important index for evaluating iodine nutrition status, but the urine iodine is greatly influenced by diet, the urine iodine value of an individual is often unstable, serum iodine is relatively stable, and the recent iodine nutrition level of the individual can be more accurately reflected without great change caused by recent diet change.
The existing standard serum iodine test method is WS/T572-2017 determination of iodine in serum-As-Ce catalytic spectrophotometry, which is a thermal digestion method, wherein high-concentration acid is needed to be added into a serum sample as digestion liquid, and digestion is needed to be performed for 12 0min under the condition of accurate control at 130 ℃. During digestion, a large amount of pungent acid mist and chlorine gas can escape, a laboratory is required to be equipped with high-power ventilation equipment, temperature and time are required to be strictly controlled during absorbance measurement, operation is complicated, time required for sample analysis is long, results are easily affected by personal operation methods, accuracy of experimental results cannot be guaranteed, and repeatability is poor. The existing national standard method has the defects of high-temperature heating, long digestion time, and complex operation due to volatilization of a large amount of acid mist in the digestion process, and needs to be further improved.
Disclosure of Invention
Therefore, the invention provides a reagent, a kit and a detection method for quantitative detection of blood iodine, which are used for solving the defects of troublesome detection, high detection cost and low accuracy of the detection method in the prior art.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a reagent for quantitative detection of blood iodine, which comprises the following components:
a first composition: acetonitrile and ethyl acetate, or ammonium sulfate and isopropyl alcohol;
a second composition: sodium hypochlorite and sodium chloride;
third composition: sulfuric acid and hydrochloric acid in combination;
fourth composition: sodium chloride and sodium arsenite in combination;
a fifth composition: ferrous sulfate, phenanthroline and sulfuric acid in combination;
sixth composition: ammonium cerium sulfate, and sulfuric acid.
Preferably, the reagent comprises a solution of which the solvent is water;
the first solution comprises 0.5 to 40 percent of acetonitrile and 0.5 to 40 percent of ethyl acetate by mass percent; or 0.5 to 60 percent of ammonium sulfate and 0.5 to 40 percent of isopropanol by mass percent;
the second solution comprises 0.5-20% of sodium hypochlorite and 0.5-5% of sodium chloride by mass percent;
the third solution comprises 0.5 to 60 percent of sulfuric acid and 0 to 35 percent of hydrochloric acid by mass percent;
the fourth solution comprises 0 to 5 percent of sodium chloride and 0.1 to 10 percent of sodium arsenite by mass percent;
a fifth reagent comprising 0.1 to 16 percent of ferrous sulfate, 0.2 to 19 percent of phenanthroline and 0 to 15 percent of sulfuric acid by mass percent;
and the sixth solution comprises 0-25% of ammonium cerium sulfate, 0-20% of cerium sulfate and 5-30% of sulfuric acid by mass percent.
Preferably, the reagent comprises a solution of which the solvent is water;
a first solution comprising 25% acetonitrile and 10% ethyl acetate by mass; or 20% ammonium sulfate and 8% isopropyl alcohol by mass;
a second solution comprising 10% by mass of sodium hypochlorite and 5% by mass of sodium chloride;
a third solution comprising sulfuric acid with the mass percentage of 15% and hydrochloric acid with the mass percentage of 12%;
a fourth solution comprising 2% of sodium chloride and 1% of sodium arsenite by mass percent;
a fifth reagent comprising 1.2% of ferrous sulfate, 1.2% of phenanthroline and 5% of sulfuric acid by mass percent;
and the sixth solution comprises 0.2% of ammonium cerium sulfate, 0.5% of cerium sulfate and 5% of sulfuric acid by mass percent.
Preferably, the method comprises the steps of,
the first solution comprises 20% of ammonium sulfate and 8% of isopropanol in percentage by mass, and the balance of water;
the second solution comprises 4.0% of sodium hypochlorite and 3.0% of sodium chloride in percentage by mass, and the balance of water;
the third solution comprises, by mass, 20% sulfuric acid and 3.0% hydrochloric acid, with the balance being water;
the fourth solution comprises 2.5% of sodium chloride, 3.0% of sodium arsenite and the balance of water according to mass percent;
the fifth solution comprises 3.0% of ferrous sulfate, 0.2% of phenanthroline and 2.0% of sulfuric acid in percentage by mass, and the balance of water;
the sixth solution comprises, by mass, 0.2% of ammonium cerium sulfate, 1.0% of cerium sulfate, 1.5% of sulfuric acid, and the balance of water.
Preferably, the first solution comprises 40% of acetonitrile and the balance of water in percentage by mass;
the second solution comprises 7.0% of sodium hypochlorite and 4.0% of sodium chloride by mass percent, and the balance of water;
the third solution comprises 18% sulfuric acid and 4.0% hydrochloric acid by mass percent, and the balance is water;
the fourth solution comprises 2.5% of sodium chloride, 5.0% of sodium arsenite and the balance of water according to mass percent;
the fifth solution comprises 2.0% of ferrous sulfate, 0.5% of phenanthroline and 5.0% of sulfuric acid in percentage by mass, and the balance of water;
the sixth solution comprises, by mass, 0.7% of ammonium cerium sulfate, 2.3% of cerium sulfate, 3.0% of sulfuric acid, and the balance of water.
Preferably, the reagent further comprises an iodine standard solution.
The invention also provides a kit for quantitative detection of blood iodine, which comprises the detection reagent.
The invention also provides a method for quantitative detection of blood iodine, the method comprising: detection is performed using the reagents described above.
Preferably, the method further comprises:
respectively adding the first solution into n iodine standard substance solutions with different concentrations and serum to be tested, uniformly mixing, and then digesting for 1-10min to respectively obtain first digestion solutions;
respectively adding a second solution and a third solution into the digested solution, mixing, and then digesting for 1-10min to respectively and correspondingly obtain a second digestion solution;
after the fourth solution, the fifth solution and the sixth solution are respectively added into the second digestion solution, recording the time for changing the blue color of the corresponding second digestion solution into the purple color from the blue color as x seconds;
concentration of the plasma in the serum to be measured y=ax b ，
Wherein,n is the number of iodine standard substance solutions, and n is more than or equal to 6,i and is 1,2,3, … … and n; xi is the reaction time of the ith iodine standard solution, and the unit is s; yi is the concentration of the i-th iodine standard solution, and the unit is mug/L; />For each iodine standard solution, < > for>The average concentration of each iodine standard solution was determined.
Preferably, the volume ratio of the first solution to the serum sample to be tested is 1-8: 1, a step of;
the volume ratio of the second solution to the serum sample to be measured is 1-8: 1, a step of;
the volume ratio of the third solution to the serum sample to be measured is 1-8: 1, a step of;
the volume ratio of the fourth solution to the serum sample to be measured is 1-8: 1, a step of;
the volume ratio of the fifth solution to the serum sample to be measured is 1-8: 1, a step of;
the volume ratio of the sixth solution to the serum sample to be measured is 1-8: 1.
preferably, the volume ratio of the first solution to the serum sample to be tested is 2:1, a step of;
the volume ratio of the second solution to the serum sample to be tested is 2:1, a step of;
the volume ratio of the third solution to the serum sample to be tested is 2:1, a step of;
the volume ratio of the fourth solution to the serum sample to be tested is 7:1, a step of;
the volume ratio of the fifth solution to the serum sample to be tested is 8:1, a step of;
the volume ratio of the sixth solution to the serum sample to be tested is 2:1.
preferably, the serum sample to be tested is obtained by centrifuging 3000r/min of blood to be tested at room temperature for 10min.
The invention has the following advantages:
the rapid detection reagent, the kit and the method for iodine in serum have the advantages that the digestion liquid is low in acidity, the operation is simple and rapid, the rapid detection reagent, the kit and the method can be synchronously used in combination with detection equipment, the rapid detection reagent, the kit and the method are not easily affected by manual errors of experimenters, and the experimental result is accurate and stable.
The test proves that: the invention has the advantages of less sampling amount, less pollution, wide detection range, good repeatability of detection results, high accuracy, no need of complex and high-cost instruments and equipment, simple and quick operation, more than 0.999 of the correlation coefficient r value of the coincidence degree of the reaction power regression curve time and the detection value, real pulling of the reaction time of different concentrations, 0-500 mug/L of the iodine ion detection linear range and 2.1 mug/L of the minimum detection limit.
Detailed Description
Other advantages and advantages of the present invention will become apparent to those skilled in the art from the following detailed description, which, by way of illustration, is to be read in connection with certain specific embodiments, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In the reagent of the invention, deionized pure water has resistivity greater than 18.0M omega.M (according with GB/T6682 primary water).
96% -98% of sodium hydroxide AR (analytical grade), and is measured by a standard titration method; 36% -38% of hydrochloric acid AR (analytical grade), and is measured by a standard titration method; 94% -98% of sulfuric acid AR (analytical grade), and is measured by a standard titration method; acetonitrile AR (analytically pure) 99.9%, isopropanol AR (analytically pure) 99.7%; ethyl acetate AR (analytical grade) 99.5%; ammonium sulfate AR (analytically pure) 99.0%; sodium arsenite AR (analytically pure) 98%; sodium chloride AR (analytically pure) 98%; ferrous sulfate AR (analytical grade) 98%. Determining by standard titration method; phenanthroline (or 1, 10-phenanthroline, C12H8N 2) AR (analytically pure) 98%; ceric ammonium sulfate AR (analytically pure) 98%; cerium sulfate AR (analytically pure) 98%; potassium iodate GR 99.99%; potassium iodide GR 99.99%.
Preparing a solution: 1:1 sulfuric acid solution: adding 50-250ml of pure water into a 500ml volumetric flask, slowly pouring 250ml of concentrated sulfuric acid AR (superior pure), stirring, cooling, diluting to a constant volume of 500ml for later use;
digestion solution: weighing 100ml of acetonitrile, ethyl acetate, isopropanol and the like, and fixing the volume to 500ml by distilled water;
ammonium sulfate solution: taking a 100ml beaker, adding a small amount of pure water, adding ammonium sulfate precisely weighed by using a ten-thousandth balance, stirring and dissolving, then, cooling to room temperature, transferring to a 100ml volumetric flask, and fixing the volume to 100ml by using the pure water;
composite acid solution: taking a 100ml volumetric flask, adding 45ml of pure water, adding perchloric acid AR and a 1:1 sulfuric acid solution, or using sulfuric acid and hydrochloric acid to be independent or mixed, cooling to room temperature, and then using pure water to fix the volume to 100ml;
arsenite solution: taking a 200ml beaker, adding 50ml pure water, adding sodium arsenite AR (high-grade pure) precisely weighed by adopting a ten-thousandth balance, stirring and dissolving, adding sodium chloride, adding a 1:1 sulfuric acid solution, stirring and dissolving and cooling to room temperature, transferring the solution in the beaker into a 100ml volumetric flask, and then fixing the volume to 100ml by using pure water;
ferrous sulfate solution: taking a 100ml volumetric flask, adding 50ml of pure water, adding ferrous sulfate precisely weighed by using a ten-thousandth balance, or adding sulfuric acid solution, stirring for dissolving, adding phenanthroline, cooling to room temperature after dissolving, and fixing the volume to 100ml by using the pure water;
ammonium cerium sulfate solution: taking a 100ml volumetric flask, adding 50ml of pure water, adding ammonium cerium sulfate and cerium sulfate which are precisely weighed by using a ten-thousandth balance, adding sulfuric acid solution, cooling to room temperature after dissolution, and adding pure water to dilute to constant volume to 100ml;
example 1
The present embodiment provides a reagent for quantitative detection of blood iodine, the reagent comprising
The first solution comprises 20% of ammonium sulfate, 8% of isopropanol and the balance of water in percentage by mass;
the second solution comprises 4.0% of sodium hypochlorite, 3.0% of sodium chloride and the balance of water according to mass percent;
the third solution comprises 20% sulfuric acid, 3.0% hydrochloric acid and the balance of water in percentage by mass;
the fourth solution comprises 2.5% of sodium chloride, 3.0% of sodium arsenite and the balance of water according to mass percent;
the fifth solution comprises 3.0% of ferrous sulfate, 0.2% of phenanthroline and 2.0% of sulfuric acid by mass percent, and the balance of water;
the sixth solution comprises, by mass, 0.2% of ammonium cerium sulfate, 1.0% of cerium sulfate, 1.5% of sulfuric acid, and the balance of water.
In this example, iodine content in iodine standard series solution: 5. 20, 50, 100, 200, 300, 400, 500. Mu.g/L.
The method for quantitative detection of blood iodine in the embodiment comprises the following steps:
1. detection of reaction time of iodine standard series solution
Accurately transferring 0.1ml of iodine standard solution into a test tube, adding 0.1-1.0ml of first solution, shaking and mixing the solution in the test tube, digesting for 1-10min at room temperature, adding 0.1-1.0ml of second solution and 0.1-1.0ml of third solution, shaking and mixing the solution in the test tube, and digesting for 1-10min at room temperature. And adding 0.1-1.0ml of fourth solution, 0.1-1.0ml of fifth solution and 0.1-1.0ml of sixth solution into the test tube, shaking and uniformly mixing the solutions in the test tube, timing, standing for observation, stopping timing when the test tube solution changes from blue to mauve, and recording the reaction time x. The specific operation is shown in the following table 1.
TABLE 1
2. Determination of the reaction time in the serum to be tested
Collecting a proper amount of blood by using a disposable vacuum non-anticoagulation blood collection tube, centrifuging for 10min at 3000r/min by using a centrifuge, placing the separated serum into a polyethylene tube with a plug, and preserving at 4 ℃ to obtain a serum sample to be tested;
dividing the serum sample to be tested into two parts, respectively adding 0.1ml of the first solution into a test tube, shaking and mixing the solutions in the test tube, digesting for 1-10min at room temperature, adding 0.2ml of the second solution and 0.1-1.0ml of the third solution, shaking and mixing the solutions in the test tube, and digesting for 1-10min at room temperature. And adding 0.2ml of the fourth solution, 0.2ml of the fifth solution and 0.2ml of the sixth solution, shaking and uniformly mixing the solutions in the test tube, timing, standing for observation, stopping timing when the solution in the test tube is changed from blue to mauve, and recording the color change time from blue to mauve, namely the reaction time x.
3. Calculating the concentration of iodide ions in the serum sample to be tested
Firstly, detecting corresponding reaction time of an iodine standard solution, and obtaining a correlation coefficient according to regression statistics;
calculating a correlation coefficient (r) of the linear regression:
wherein: xi-determining the theoretical concentration of the iodine standard solution; yi-the actual measurement value corresponding to the measured solution concentration; i-1, 2,3, … …, n; the absolute value of the equation correlation coefficient should be more than or equal to 0.999.
Substituting the reaction time of the serum into an iodine element mass calculation formula, and calculating to obtain the iodine ion content in the detection sample. The concentration y of the iodide ions in the serum is calculated according to the reaction time x, and the calculation method of the iodide ion content in the serum is as follows:
calculate the concentration y, y=ax b ；
Average concentration for each iodine standard solution; />Average color change time for iodine standard solution; n is the number of different concentrations of iodine standard solution; xi is the reaction time(s) of the ith iodine standard solution; yi is the concentration of the ith iodine standard solution (μg/L); i is 1,2,3, … …, n.
As shown in table 2, the reaction time of the iodine standard and the calculated a, b values are measured in this example.
TABLE 2
The calculation formula is obtained as y= 6.9472x -1.2453 . According to the calculation formula, the reaction time of the first serum to be measured is 115.431s, and the obtained y values are 34.4. The reaction time of the second serum to be tested was 110.384s and the y values obtained were 36.4, respectively.
Example 2
The difference between this example and example 1 is that the first solution comprises, by mass, 40% acetonitrile, the balance being water; the second solution comprises 7.0% of sodium hypochlorite and 4.0% of sodium chloride by mass percent, and the balance of water; the third solution comprises 18% sulfuric acid and 4.0% hydrochloric acid by mass percent, and the balance is water; the fourth solution comprises 2.5% of sodium chloride, 5.0% of sodium arsenite and the balance of water according to mass percent; the fifth solution comprises 2.0% of ferrous sulfate, 0.5% of phenanthroline and 5.0% of sulfuric acid in percentage by mass, and the balance of water; the sixth solution comprises, by mass, 0.7% of ammonium cerium sulfate, 2.3% of cerium sulfate, 3.0% of sulfuric acid, and the balance of water. The other components are the same as in example 1.
The calculation formula is obtained as y= 7.7929x -1.2548 . According to the calculation formula, the reaction time of the first serum to be detected is 39.323s respectively, and the obtained y values are respectively133.8. The reaction time of the second serum to be tested was 38.654s and the y values obtained were 136.7, respectively.
Example 3
This example differs from example 1 in that the first solution comprises 25% acetonitrile and 10% ethyl acetate by mass; or 20% ammonium sulfate and 8% isopropyl alcohol by mass; a second solution comprising 10% by mass of sodium hypochlorite and 5% by mass of sodium chloride; a third solution comprising sulfuric acid with the mass percentage of 15% and hydrochloric acid with the mass percentage of 12%; a fourth solution comprising 2% of sodium chloride and 1% of sodium arsenite by mass percent; a fifth reagent comprising 1.2% of ferrous sulfate, 1.2% of phenanthroline and 5% of sulfuric acid by mass percent; and the sixth solution comprises 0.2% of ammonium cerium sulfate, 0.5% of cerium sulfate and 5% of sulfuric acid by mass percent. The other components are the same as in example 1.
The calculation formula is obtained as y= 6.4067x -1.2378 . According to the calculation formula, the reaction time of the first serum to be measured is 37.999s, and the obtained y values are 137.3. The reaction time of the second serum to be tested was 38.524s and the y values obtained were 135.0, respectively.
Correlation of test example 1 and Standard Curve
The abscissa of the standard curve of the method is the concentration (mug/L) of the iodine standard solution, the ordinate is the reaction time(s) of the iodine standard solution, the standard curve is continuously and parallelly measured for 6 times under the same temperature and humidity condition, the corresponding correlation coefficient is calculated according to the reaction time measured at each point of each curve, and the variation coefficient of the reaction time of each concentration is calculated.
As shown in Table 3, the detection time of iodine standard products with different concentrations, the corresponding correlation coefficient and the variation coefficient are all above 0.999, and the variation coefficient ranges from 1.82% to 3.79%.
TABLE 3 Table 3
Test example 2, detection limit
The test example was calculated from the standard deviation of 3 times of the blank value according to the detection limit, 100. Mu.L of distilled water was sucked, and the reaction time of the blank tube was measured in parallel for 10 times, with the detection limit being 2.1. Mu.g/L.
Test example 3, repeatability
The test example selects an iodine standard substance solution with the concentration of 125 mug/L, adopts the method to carry out parallel measurement for 10 times, and calculates the average value, standard deviation and variation coefficient. As shown in Table 4, the iodine standard solution having a measured concentration of 125. Mu.g/L had a coefficient of variation of 1.44%.
TABLE 4 Table 4
Test example 4, precision
Precision in batch: serum samples of low, medium and high 3 iodine concentrations were selected and assayed 3 times in parallel using the method of example 1, and the average value and the coefficient of variation were calculated. As shown in Table 5, the average value was found to be 2.41% and the coefficient of variation was found to be 2.18% -2.64%.
TABLE 5
Precision between batches: serum samples with low, medium and high 3 iodine concentrations are selected, each batch is measured in parallel for 3 times by adopting the method of the invention, and the average value and the variation coefficient are calculated. As shown in Table 6, the average value was found to be 2.07% and the coefficient of variation was found to be in the range of 1.12% to 2.94%.
TABLE 6
Test example 5, accuracy
a. As no national standard substance exists in the serum iodine at present, the accuracy of the method for testing the sample labeling recovery rate is verified by adopting the method for testing the sample labeling recovery rate. Serum samples of low, medium and high 3 different concentrations were selected, each sample was assayed 3 times in parallel and the average and recovery were calculated.
According to GBT26124-2011, the recovery rate calculation formula is:
wherein: r-recovery rate; v-adding the volume of the standard solution; v (V) 0 -a volume of a human sample; detecting the concentration of the C-human sample after the standard solution is added; c (C) 0 -a detected concentration of a human sample; c (C) s -concentration of standard solution.
As shown in Table 7, the recovery rate ranges from 96.3% to 106.9%, and the total average recovery rate is 102.4%, which meets the measurement requirements of biological samples.
TABLE 7
b. Alignment with the current standard law (WS/T572-2017)
Serum samples with low, medium and high concentration of 3 iodine are selected, the method is adopted to carry out parallel measurement for 3 times, the current standard method (WS/T572-2017) is adopted to carry out parallel measurement for 3 times, and the current standard method is used as a reference to carry out result comparison and bias analysis on the two measurement methods.
As shown in Table 8, the range of relative bias was measured to be-2.34% -3.20%.
TABLE 8
In the embodiment, a serum sample is digested by adopting chemical reagents such as acetonitrile, ethyl acetate or isopropanol at normal temperature, oxidation-reduction reaction of sodium arsenite and ammonium cerium sulfate is catalyzed by iodine, the reaction temperature is precisely controlled, the color-changing time of the solution is measured, and the content of iodide ions in the serum sample to be measured is calculated by utilizing the linear relationship between the content of the iodide ions and the color-changing time of the solution.
While the invention has been described in detail in the foregoing general description and specific examples, it will be apparent to those skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Claims (3)
1. A non-diagnostic method for quantitative detection of blood iodine, the method comprising the steps of:
a first solution comprising 25% acetonitrile and 10% ethyl acetate by mass; or 20% ammonium sulfate and 8% isopropyl alcohol by mass;
a second solution comprising 10% by mass of sodium hypochlorite and 5% by mass of sodium chloride;
a third solution comprising sulfuric acid with the mass percentage of 15% and hydrochloric acid with the mass percentage of 12%;
a fourth solution comprising 2% of sodium chloride and 1% of sodium arsenite by mass percent;
a fifth solution comprising 1.2% by mass of ferrous sulfate, 1.2% by mass of phenanthroline and 5% by mass of sulfuric acid;
a sixth solution comprising 0.2% by mass of ammonium cerium sulfate, 0.5% by mass of cerium sulfate and 5% by mass of sulfuric acid;
iodine standard solution;
the solvents in the solution are all water;
the method comprises the following steps:
respectively adding the first solution into n iodine standard substance solutions with different concentrations and serum to be tested, uniformly mixing, and then digesting for 1-10min to respectively obtain first digestion solutions;
respectively adding a second solution and a third solution into the digested solution, mixing, and then digesting for 1-10min to respectively and correspondingly obtain a second digestion solution;
after the fourth solution, the fifth solution and the sixth solution are respectively added into the second digestion solution, recording the time for changing the blue color of the corresponding second digestion solution into the purple color from the blue color as x seconds;
concentration of electric ions in serum to be measured
Wherein the method comprises the steps ofN is the number of iodine standard solutions, i is 1,2,3, … …, n; xi is the reaction time of the ith iodine standard solution, and the unit is s; yi is the concentration of the i-th iodine standard solution, and the unit is mug/L; />For each iodine standard solution, < > for>The average concentration of each iodine standard solution;
the volume ratio of the first solution to the serum sample to be tested used in the method is 1-8: 1, a step of;
the volume ratio of the second solution to the serum sample to be measured is 1-8: 1, a step of;
the volume ratio of the third solution to the serum sample to be measured is 1-8: 1, a step of;
the volume ratio of the fourth solution to the serum sample to be measured is 1-8: 1, a step of;
the volume ratio of the fifth solution to the serum sample to be measured is 1-8: 1, a step of;
the volume ratio of the sixth solution to the serum sample to be measured is 1-8: 1.
2. the method of claim 1, wherein,
the serum sample to be detected is obtained by centrifuging 3000r/min of blood to be detected for 10min at room temperature.
3. The method of claim 1, wherein,
the volume ratio of the first solution to the serum sample to be tested used in the method is 2:1, a step of;
the volume ratio of the second solution to the serum sample to be tested is 2:1, a step of;
the volume ratio of the third solution to the serum sample to be tested is 2:1, a step of;
the volume ratio of the fourth solution to the serum sample to be tested is 7:1, a step of;
the volume ratio of the fifth solution to the serum sample to be tested is 8:1, a step of;
the volume ratio of the sixth solution to the serum sample to be tested is 2:1.
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