CN113340888A - 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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- CN113340888A CN113340888A CN202110752886.5A CN202110752886A CN113340888A CN 113340888 A CN113340888 A CN 113340888A CN 202110752886 A CN202110752886 A CN 202110752886A CN 113340888 A CN113340888 A CN 113340888A
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- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 title claims abstract description 57
- 239000011630 iodine Substances 0.000 title claims abstract description 57
- 229910052740 iodine Inorganic materials 0.000 title claims abstract description 57
- 238000001514 detection method Methods 0.000 title claims abstract description 39
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 33
- 210000004369 blood Anatomy 0.000 title claims abstract description 21
- 239000008280 blood Substances 0.000 title claims abstract description 21
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims abstract description 87
- 210000002966 serum Anatomy 0.000 claims abstract description 57
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 52
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 39
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 33
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 28
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims abstract description 26
- 239000011780 sodium chloride Substances 0.000 claims abstract description 26
- 239000000203 mixture Substances 0.000 claims abstract description 19
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 claims abstract description 15
- 230000029087 digestion Effects 0.000 claims abstract description 15
- 239000011790 ferrous sulphate Substances 0.000 claims abstract description 15
- 235000003891 ferrous sulphate Nutrition 0.000 claims abstract description 15
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims abstract description 15
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims abstract description 15
- PTLRDCMBXHILCL-UHFFFAOYSA-M sodium arsenite Chemical compound [Na+].[O-][As]=O PTLRDCMBXHILCL-UHFFFAOYSA-M 0.000 claims abstract description 15
- 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 abstract description 14
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 13
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 13
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 13
- 239000005708 Sodium hypochlorite Substances 0.000 claims abstract description 12
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims abstract description 12
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 claims abstract description 7
- OKJMLYFJRFYBPS-UHFFFAOYSA-J tetraazanium;cerium(4+);tetrasulfate Chemical compound [NH4+].[NH4+].[NH4+].[NH4+].[Ce+4].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O OKJMLYFJRFYBPS-UHFFFAOYSA-J 0.000 claims abstract description 5
- 239000000243 solution Substances 0.000 claims description 146
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 43
- 239000012086 standard solution Substances 0.000 claims description 23
- 230000035484 reaction time Effects 0.000 claims description 22
- 238000002156 mixing Methods 0.000 claims description 10
- 239000000126 substance Substances 0.000 claims description 10
- 238000005259 measurement Methods 0.000 claims description 7
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 claims description 6
- VZDYWEUILIUIDF-UHFFFAOYSA-J cerium(4+);disulfate Chemical compound [Ce+4].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O VZDYWEUILIUIDF-UHFFFAOYSA-J 0.000 claims description 6
- 229910000355 cerium(IV) sulfate Inorganic materials 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 4
- 150000002500 ions Chemical class 0.000 claims description 2
- SWGJCIMEBVHMTA-UHFFFAOYSA-K trisodium;6-oxido-4-sulfo-5-[(4-sulfonatonaphthalen-1-yl)diazenyl]naphthalene-2-sulfonate Chemical compound [Na+].[Na+].[Na+].C1=CC=C2C(N=NC3=C4C(=CC(=CC4=CC=C3O)S([O-])(=O)=O)S([O-])(=O)=O)=CC=C(S([O-])(=O)=O)C2=C1 SWGJCIMEBVHMTA-UHFFFAOYSA-K 0.000 claims description 2
- 239000007788 liquid Substances 0.000 abstract description 2
- 230000001360 synchronised effect Effects 0.000 abstract description 2
- 239000000523 sample Substances 0.000 description 28
- 238000012360 testing method Methods 0.000 description 20
- 238000004364 calculation method Methods 0.000 description 9
- -1 polyethylene Polymers 0.000 description 7
- 238000011084 recovery Methods 0.000 description 6
- 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 description 5
- 230000008859 change Effects 0.000 description 5
- 238000001816 cooling Methods 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 239000002253 acid Substances 0.000 description 4
- 238000010561 standard procedure Methods 0.000 description 4
- 238000004448 titration Methods 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 description 3
- 210000002700 urine Anatomy 0.000 description 3
- 101001018064 Homo sapiens Lysosomal-trafficking regulator Proteins 0.000 description 2
- 102100033472 Lysosomal-trafficking regulator Human genes 0.000 description 2
- 235000010703 Modiola caroliniana Nutrition 0.000 description 2
- 244000038561 Modiola caroliniana Species 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000012153 distilled water Substances 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
- 230000008569 process Effects 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
- 238000002835 absorbance Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- GCPXMJHSNVMWNM-UHFFFAOYSA-N arsenous acid Chemical compound O[As](O)O GCPXMJHSNVMWNM-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
- 230000007547 defect Effects 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
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 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
- XMBWDFGMSWQBCA-UHFFFAOYSA-M iodide Chemical compound [I-] XMBWDFGMSWQBCA-UHFFFAOYSA-M 0.000 description 1
- 229940006461 iodide ion Drugs 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 239000004973 liquid crystal related substance Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 239000011259 mixed solution Substances 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
- 238000006479 redox reaction Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000009423 ventilation 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
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- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention discloses a reagent, a kit and a detection method for quantitative detection of blood iodine, wherein the reagent comprises: a first composition: acetonitrile and ethyl acetate, or ammonium sulfate and isopropanol; a second composition: sodium hypochlorite and sodium chloride; third composition: combining sulfuric acid and hydrochloric acid; fourth composition: sodium chloride and sodium arsenite in combination; a fifth composition: ferrous sulfate, phenanthroline and sulfuric acid; a sixth composition: cerium ammonium sulfate, cerium sulfate, and sulfuric acid. The reagent and the method for rapidly detecting iodine in serum have the advantages of low digestion liquid acidity, simple and rapid operation, synchronous use by combining detection equipment, difficult influence of manual errors of experimenters and accurate and stable experimental results.
Description
Technical Field
The invention belongs to the technical field of detection reagents, and particularly relates to a reagent and a kit for quantitative detection of blood iodine and a detection method.
Background
Iodine is one of trace elements necessary for maintaining normal thyroid gland functions of human bodies, thyroid diseases can be caused by too much or too little daily intake, and is particularly important for growth and development of fetuses and infants, so that the world health organization and the foundation of the United nations for children can use the urine iodine as an important index for evaluating iodine nutrition conditions, but the urine iodine is greatly influenced by diet, the urine iodine value of an individual is often unstable, serum iodine is relatively stable, great change cannot occur due to recent diet change, and the recent iodine nutrition level of the individual can be more accurately reflected.
The existing standard serum iodine detection method is 'determination of iodine in WS/T572-2017 serum-arsenic-cerium catalytic spectrophotometry', the method is a thermal digestion method, high-concentration acid is required to be added into a serum sample to be used as a digestion solution, and digestion is required to be accurately controlled at 130 ℃ for 120 min. A large amount of sharp acid mist and chlorine gas escape in the digestion process, a laboratory is required to be equipped with a high-power ventilation device, the temperature and time must be strictly controlled when the absorbance is measured, the operation is complicated, the time required for analyzing a sample is long, the result is easily influenced by a personal operation method, the accuracy of the experimental result cannot be guaranteed, and the reproducibility is poor. The shortcomings of high-temperature heating, long digestion time, large amount of acid mist volatilization and complex operation in the digestion process of blood samples in the existing national standard method are needed, and further improvement is urgently needed.
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 purpose, the invention provides the following technical scheme:
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 isopropanol;
a second composition: sodium hypochlorite and sodium chloride;
third composition: combining sulfuric acid and hydrochloric acid;
fourth composition: sodium chloride and sodium arsenite in combination;
a fifth composition: ferrous sulfate, phenanthroline and sulfuric acid;
a sixth composition: cerium ammonium sulfate, 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 mass percent of acetonitrile and 0.5 to 40 mass percent of ethyl acetate; 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 to 20 mass percent of sodium hypochlorite and 0.5 to 5 mass percent of sodium chloride;
a third solution, which comprises 0.5 to 60 mass percent of sulfuric acid and 0 to 35 mass percent of hydrochloric acid;
the fourth solution comprises 0 to 5 mass percent of sodium chloride and 0.1 to 10 mass percent of sodium arsenite;
a fifth reagent which comprises 0.1 to 16 mass percent of ferrous sulfate, 0.2 to 19 mass percent of phenanthroline and 0 to 15 mass percent of sulfuric acid;
the sixth solution comprises 0-25% of ammonium ceric sulfate, 0-20% of cerium sulfate and 5-30% of sulfuric acid in percentage by mass.
Preferably, the reagent comprises a solution of which the solvent is water;
a first solution comprising, by mass, 25% acetonitrile and 10% ethyl acetate; or 20 percent of ammonium sulfate and 8 percent of isopropanol by mass percent;
a second solution, which comprises 10% of sodium hypochlorite and 5% of sodium chloride in percentage by mass;
a third solution, which comprises 15 percent of sulfuric acid and 12 percent of hydrochloric acid by mass percent;
a fourth solution, which comprises 2% of sodium chloride and 1% of sodium arsenite in percentage by mass;
a fifth reagent which comprises 1.2 percent of ferrous sulfate, 1.2 percent of phenanthroline and 5 percent of sulfuric acid in percentage by mass;
and the sixth solution comprises 0.2 percent of ammonium ceric sulfate, 0.5 percent of cerium sulfate and 5 percent of sulfuric acid in percentage by mass.
Preferably, the first and second liquid crystal materials are,
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 percent of sodium hypochlorite, 3.0 percent of sodium chloride and the balance of water in percentage by mass;
the third solution comprises, by mass, 20% of sulfuric acid, 3.0% of hydrochloric acid, and the balance of water;
the fourth solution comprises 2.5 percent of sodium chloride, 3.0 percent of sodium arsenite and the balance of water in percentage by mass;
the fifth solution comprises, by mass, 3.0% of ferrous sulfate, 0.2% of phenanthroline, 2.0% of sulfuric acid, and the balance of water;
the sixth solution comprises, in mass percent, 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, by mass, 40% of acetonitrile, and the balance of water;
the second solution comprises 7.0 percent of sodium hypochlorite, 4.0 percent of sodium chloride and the balance of water in percentage by mass;
the third solution comprises 18 percent of sulfuric acid, 4.0 percent of hydrochloric acid and the balance of water in percentage by mass;
the fourth solution comprises 2.5 percent of sodium chloride, 5.0 percent of sodium arsenite and the balance of water in percentage by mass;
the fifth solution comprises, by mass, 2.0% of ferrous sulfate, 0.5% of phenanthroline, 5.0% of sulfuric acid, 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, which comprises the following steps: detection is carried out by using the reagent.
Preferably, the method further comprises:
respectively adding the first solutions into n iodine standard substance solutions with different concentrations and serum to be detected, uniformly mixing, digesting for 1-10min, and respectively and correspondingly obtaining first digestion solutions;
adding a second solution and a third solution into the digested solution respectively, mixing, digesting for 1-10min, and obtaining second digested solutions respectively;
respectively adding a fourth solution, a fifth solution and a sixth solution into the second digestion solution, and recording the time for changing the blue color of the corresponding second digestion solution into the purple red color as x seconds;
the concentration y of the electric ions in the serum to be measured is axb，
Wherein the content of the first and second substances,
n is the number of iodine standard solutions, n is more than or equal to 6, 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 ith iodine standard solution, and the unit is mu g/L;
the solution of each iodine standard substance is used,
is the average concentration of each iodine standard solution.
Preferably, the volume ratio of the first solution to the serum sample to be detected is 1-8: 1;
the volume ratio of the second solution to the serum sample to be detected is 1-8: 1;
the volume ratio of the third solution to the serum sample to be detected is 1-8: 1;
the volume ratio of the fourth solution to the serum sample to be detected is 1-8: 1;
the volume ratio of the fifth solution to the serum sample to be detected is 1-8: 1;
the volume ratio of the sixth solution to the serum sample to be detected is 1-8: 1.
preferably, the volume ratio of the first solution to the serum sample to be tested is 2: 1;
the volume ratio of the second solution to the serum sample to be detected is 2: 1;
the volume ratio of the third solution to the serum sample to be detected is 2: 1;
the volume ratio of the fourth solution to the serum sample to be detected is 7: 1;
the volume ratio of the fifth solution to the serum sample to be detected is 8: 1;
the volume ratio of the sixth solution to the serum sample to be detected is 2: 1.
preferably, the serum sample to be tested is obtained by centrifuging blood to be tested for 10min at room temperature at 3000 r/min.
The invention has the following advantages:
the reagent, the kit and the method for rapidly detecting iodine in serum have the advantages of low digestion liquid acidity, simple and rapid operation, synchronous use by combining detection equipment, difficult influence of manual errors of experimenters and accurate and stable experimental result.
The test proves that: the invention has the advantages of small sample quantity in various reagent detections, less pollution, wide detection range, good repeatability of detection results, high accuracy, no need of complex and expensive instruments and equipment, simple and quick operation, capability of enabling the time of a reaction power regression curve and the related coefficient r value of the detection value goodness of fit to reach over 0.999, truly pulling open the reaction time of different concentrations, capability of enabling the linear range of iodide ion detection to reach 0-500 mug/L and minimum detection limit to be 2.1 mug/L.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. 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.
In the reagent of the invention, deionized pure water has the resistivity larger than 18.0M omega.M (meeting GB/T6682 first-grade water).
96% -98% of sodium hydroxide AR (analytically pure), and measuring by adopting a standard titration method; 36% -38% of hydrochloric acid AR (analytically pure), and measuring by adopting a standard titration method; 94% -98% of sulfuric acid AR (analytically pure), and is measured by a standard titration method; acetonitrile AR (analytical grade) 99.9%, isopropanol AR (analytical grade) 99.7%; ethyl acetate AR (analytical grade) 99.5%; ammonium sulfate AR (analytical grade) 99.0%; 98% of sodium arsenite AR (analytically pure); sodium chloride AR (analytical pure) 98%; ferrous sulfate AR (analytical purity) 98%. Measuring by adopting a standard titration method; 98% of phenanthroline (or called 1, 10-phenanthroline, C12H8N2) AR (analytically pure); cerium ammonium sulfate AR (analytically pure) 98%; cerium sulfate AR (analytical grade) 98%; 99.99 percent of potassium iodate GR; 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 (super pure), stirring simultaneously, cooling, diluting and fixing the volume to 500ml for later use;
digestion solution: measuring 100ml of acetonitrile, ethyl acetate, isopropanol and the like, and fixing the volume to 500ml by using distilled water;
ammonium sulfate solution: adding a small amount of pure water into a 100ml beaker, adding ammonium sulfate precisely weighed by a ten-thousandth balance, stirring and dissolving, then placing to room temperature, transferring to a 100ml volumetric flask, and fixing the volume to 100ml by using the pure water;
and (3) composite acid solution: adding 45ml of pure water into a 100ml volumetric flask, adding perchloric acid AR and a 1:1 sulfuric acid solution, or using a single or mixed solution of sulfuric acid and hydrochloric acid, cooling to room temperature, and then fixing the volume to 100ml by using the pure water;
arsenous acid solution: adding 50ml of pure water into a 200ml beaker, adding sodium arsenite AR (super pure) which is precisely weighed by a ten-thousandth balance, stirring and dissolving, then adding sodium chloride, adding a 1:1 sulfuric acid solution, stirring, dissolving and cooling to room temperature, transferring the solution in the beaker into a 100ml volumetric flask, and then adding pure water to fix the volume to 100 ml;
ferrous sulfate solution: adding 50ml of pure water into a 100ml volumetric flask, adding ferrous sulfate precisely weighed by a one-ten-thousandth balance or adding a sulfuric acid solution, stirring and dissolving, adding phenanthroline, cooling to room temperature after dissolving, and fixing the volume to 100ml by using the pure water;
cerium ammonium sulfate solution: adding 50ml of pure water into a 100ml volumetric flask, adding ammonium cerium sulfate and cerium sulfate which are precisely weighed by a ten-thousandth balance, adding a sulfuric acid solution, cooling to room temperature after dissolution, adding pure water for dilution and fixing the volume to 100 ml;
example 1
This example provides a reagent for quantitative blood iodine detection, which comprises
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 percent of sodium hypochlorite, 3.0 percent of sodium chloride and the balance of water in percentage by mass;
the third solution comprises 20 percent of sulfuric acid, 3.0 percent of hydrochloric acid and the balance of water in percentage by mass;
the fourth solution comprises 2.5 percent of sodium chloride, 3.0 percent of sodium arsenite and the balance of water in percentage by mass;
the fifth solution comprises 3.0 percent of ferrous sulfate, 0.2 percent of phenanthroline, 2.0 percent of sulfuric acid and the balance of water according to mass percentage;
the sixth solution comprises, in mass percent, 0.2% of ammonium cerium sulfate, 1.0% of cerium sulfate, 1.5% of sulfuric acid, and the balance of water.
In this example, the iodine content in the iodine standard line of solutions: 5. 20, 50, 100, 200, 300, 400, 500 mug/L.
The method for quantitatively detecting the blood iodine in the embodiment comprises the following steps of:
1. detection of reaction time of iodine standard line solution
Accurately transferring 0.1ml of iodine standard substance solution into a test tube, adding 0.1-1.0ml of first solution, shaking and uniformly mixing the solution in the test tube, digesting at room temperature for 1-10min, then adding 0.1-1.0ml of second solution and 0.1-1.0ml of third solution, shaking and uniformly mixing the solution in the test tube, and digesting at room temperature for 1-10 min. And then adding 0.1-1.0ml of fourth solution, 0.1-1.0ml of fifth solution and 0.1-1.0ml of sixth solution, shaking and uniformly mixing the solutions in the test tube, timing, standing and observing, stopping timing when the test tube solution is changed from blue to purple, and recording the reaction time x. The specific operation is shown in table 1 below.
TABLE 1
2. Measurement of reaction time in serum to be measured
Collecting appropriate amount of blood with disposable vacuum non-anticoagulant blood collection tube, centrifuging at 3000r/min for 10min, placing the separated serum in polyethylene tube with plug, and storing at 4 deg.C to obtain serum sample to be tested;
dividing the serum sample to be detected into two parts, respectively placing 0.1ml in a test tube, adding 0.2ml of the first solution, shaking and uniformly mixing the solution in the test tube, digesting at room temperature for 1-10min, then adding 0.2ml of the second solution and 0.1-1.0ml of the third solution, shaking and uniformly mixing the solution in the test tube, and digesting at room temperature for 1-10 min. And then adding 0.2ml of fourth solution, 0.2ml of fifth solution and 0.2ml of sixth solution, shaking and uniformly mixing the solutions in the test tube, timing, standing and observing, stopping timing when the solution in the test tube changes 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, after detecting corresponding reaction time of an iodine standard substance solution, calculating according to regression statistics to obtain a correlation coefficient;
calculating the correlation coefficient (r) of the linear regression:
in the formula: xi-measuring the theoretical concentration of the iodine standard solution; yi-the actual measured value corresponding to the concentration of the measured solution; i-1, 2, 3, … …, n; the absolute value of the correlation coefficient of the equation is more than or equal to 0.999.
And substituting the reaction time of the serum into an iodine element mass calculation formula to calculate the content of the iodide ions in the detection sample. And (3) calculating the concentration y of the iodide ions in the serum according to the reaction time x, wherein the calculation method of the content of the iodide ions in the serum comprises the following steps:
calculating the concentration y, y ═ axb；
As shown in Table 2, the reaction time of the iodine standard substance tested in this example and the calculated a, b values are shown.
TABLE 2
The calculation formula is obtained as y-6.9472 x-1.2453. According to the above calculation formula, the reaction time of the first serum to be tested was 115.431s, respectively, and the obtained y values were 34.4, respectively. The reaction time of the second test serum was 110.384s, and the obtained y values were 36.4, respectively.
Example 2
The difference between the embodiment and the embodiment 1 is that the first solution comprises 40% of acetonitrile and the balance of water according to mass percentage; the second solution comprises 7.0 percent of sodium hypochlorite, 4.0 percent of sodium chloride and the balance of water in percentage by mass; the third solution comprises 18 percent of sulfuric acid, 4.0 percent of hydrochloric acid and the balance of water in percentage by mass; the fourth solution comprises 2.5 percent of sodium chloride, 5.0 percent of sodium arsenite and the balance of water in percentage by mass; the fifth solution comprises, by mass, 2.0% of ferrous sulfate, 0.5% of phenanthroline, 5.0% of sulfuric acid, 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 rest of the procedure was the same as in example 1.
The calculation formula is obtained as y-7.7929 x-1.2548. According to the above calculation formula, the reaction time of the first serum to be tested was 39.323s, respectively, and the obtained y values were 133.8, respectively. The reaction time of the second serum to be tested was 38.654s, and the obtained y values were 136.7, respectively.
Example 3
This example differs from example 1 in that the first solution, comprising 25% by mass of acetonitrile and 10% by mass of ethyl acetate; or 20 percent of ammonium sulfate and 8 percent of isopropanol by mass percent; a second solution, which comprises 10% of sodium hypochlorite and 5% of sodium chloride in percentage by mass; a third solution, which comprises 15 percent of sulfuric acid and 12 percent of hydrochloric acid by mass percent; a fourth solution, which comprises 2% of sodium chloride and 1% of sodium arsenite in percentage by mass; a fifth reagent which comprises 1.2 percent of ferrous sulfate, 1.2 percent of phenanthroline and 5 percent of sulfuric acid in percentage by mass; and the sixth solution comprises 0.2 percent of ammonium ceric sulfate, 0.5 percent of cerium sulfate and 5 percent of sulfuric acid in percentage by mass. The rest of the procedure was the same as in example 1.
The calculation formula is obtained as y-6.4067 x-1.2378. According to the above calculation formula, the reaction time of the first serum to be tested was 37.999s, respectively, and the obtained y values were 137.3, respectively. The reaction time of the second serum to be tested was 38.524s, and the obtained y values were 135.0, respectively.
Test example 1 correlation of standard curves
The abscissa of the standard curve of the method is the concentration (mu g/L) of the iodine standard solution, the ordinate is the reaction time(s) of the iodine standard solution, the standard curve is continuously measured in parallel 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 the iodine standards with different concentrations and 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
Test example 2 detection Limit
In this test example, the detection limit was calculated from 3 times the standard deviation of the blank value, and 100. mu.L of distilled water was aspirated, and the reaction time of 10 blank tubes was measured in parallel, and the detection limit was 2.1. mu.g/L.
Test example 3 reproducibility
In the test example, an iodine standard solution with the concentration of 125 mug/L is selected, the method is adopted for parallel measurement for 10 times, and the average value, the standard deviation and the coefficient of variation are calculated. As shown in Table 4, the coefficient of variation of the iodine standard solution having a measured concentration of 125. mu.g/L was 1.44%.
TABLE 4
Test example 4 precision
Internal precision: serum samples with low, medium and high iodine concentrations were selected, and each of the samples was subjected to 3-time parallel measurement using the method of example 1, and the average value and the coefficient of variation were calculated. As shown in Table 5, the average was found to be 2.41%, and the coefficient of variation was found to range from 2.18% to 2.64%.
TABLE 5
Batch precision: serum samples with low, medium and high iodine concentrations are selected, each batch of the serum samples is subjected to parallel determination for 3 times by adopting the method, and the average value and the variation coefficient are calculated. As shown in Table 6, the results showed an average value of 2.07% and a coefficient of variation ranging from 1.12% to 2.94%.
TABLE 6
Test example 5 accuracy
a. Because no national standard substance exists in the serum iodine at present, the method in the embodiment 1 of the invention adopts a method for testing the sample standard adding recovery rate to verify the accuracy. Selecting 3 serum samples with different concentrations, measuring each sample in parallel for 3 times, and calculating the average value and the recovery rate.
According to GBT26124-2011, the recovery rate calculation formula is as follows:
in the formula: r-recovery rate; v-volume of standard solution added; v0-a volume of a human sample; c, detecting the concentration of the human source sample after the human source sample is added into the standard solution; c0-the detected concentration of the human sample; cs-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 the biological samples.
TABLE 7
b. Alignment with Current Standard method (WS/T572-2017)
Selecting serum samples with low, medium and high iodine concentrations, performing parallel determination 3 times by the method, performing parallel determination 3 times by using the current standard method (WS/T572-2017), and performing result comparison and bias analysis on the two measurement methods by using the current standard method as reference.
As shown in Table 8, the relative bias was found to range from-2.34% to-3.20%.
TABLE 8
In the embodiment, a serum sample is digested by chemical reagents such as acetonitrile, ethyl acetate or isopropanol and the like at normal temperature, iodine is used for catalyzing the redox reaction of sodium arsenite and ammonium ceric sulfate, the reaction temperature is precisely controlled, the color change time of the solution is measured, and the content of iodide ions in the serum sample to be detected is calculated by utilizing the linear relation between the content of iodide ions and the color change time of the solution.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. A reagent for quantitative detection of blood iodine, comprising:
a first composition: acetonitrile and ethyl acetate, or ammonium sulfate and isopropanol;
a second composition: sodium hypochlorite and sodium chloride;
third composition: combining sulfuric acid and hydrochloric acid;
fourth composition: sodium chloride and sodium arsenite in combination;
a fifth composition: ferrous sulfate, phenanthroline and sulfuric acid;
a sixth composition: cerium ammonium sulfate, cerium sulfate, and sulfuric acid.
2. The reagent for quantitative blood iodine measurement according to claim 1,
the reagent comprises the following solutions prepared by using the first to sixth compositions, wherein the solvent of the solution is water;
the first solution comprises 0.5 to 40 mass percent of acetonitrile and 0.5 to 40 mass percent of ethyl acetate; 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 to 20 mass percent of sodium hypochlorite and 0.5 to 5 mass percent of sodium chloride;
a third solution, which comprises 0.5 to 60 mass percent of sulfuric acid and 0 to 35 mass percent of hydrochloric acid;
the fourth solution comprises 0 to 5 mass percent of sodium chloride and 0.1 to 10 mass percent of sodium arsenite;
a fifth reagent which comprises 0.1 to 16 mass percent of ferrous sulfate, 0.2 to 19 mass percent of phenanthroline and 0 to 15 mass percent of sulfuric acid;
the sixth solution comprises 0-25% of ammonium ceric sulfate, 0-20% of cerium sulfate and 5-30% of sulfuric acid in percentage by mass.
3. The reagent for quantitative blood iodine measurement according to claim 2,
the reagent comprises the following solution, wherein the solvent of the solution is water;
a first solution comprising, by mass, 25% acetonitrile and 10% ethyl acetate; or 20 percent of ammonium sulfate and 8 percent of isopropanol by mass percent;
a second solution, which comprises 10% of sodium hypochlorite and 5% of sodium chloride in percentage by mass;
a third solution, which comprises 15 percent of sulfuric acid and 12 percent of hydrochloric acid by mass percent;
a fourth solution, which comprises 2% of sodium chloride and 1% of sodium arsenite in percentage by mass;
a fifth reagent which comprises 1.2 percent of ferrous sulfate, 1.2 percent of phenanthroline and 5 percent of sulfuric acid in percentage by mass;
and the sixth solution comprises 0.2 percent of ammonium ceric sulfate, 0.5 percent of cerium sulfate and 5 percent of sulfuric acid in percentage by mass.
4. The reagent for quantitative blood iodine measurement according to claim 3,
the reagent also includes an iodine standard solution.
5. A kit for quantitative detection of blood iodine comprising the reagent of any one of claims 1 to 4.
6. A method for quantitative detection of blood iodine, the method comprising: detection using a reagent according to any one of claims 1 to 4.
7. The method of claim 6,
the method further comprises the following steps:
respectively adding the first solutions into n iodine standard substance solutions with different concentrations and serum to be detected, uniformly mixing, digesting for 1-10min, and respectively and correspondingly obtaining first digestion solutions;
adding a second solution and a third solution into the digested solution respectively, mixing, digesting for 1-10min, and obtaining second digested solutions respectively;
respectively adding a fourth solution, a fifth solution and a sixth solution into the second digestion solution, and recording the time for changing the blue color of the corresponding second digestion solution into the purple red color as x seconds;
the concentration y of the electric ions in the serum to be measured is axb，
Wherein the content of the first and second substances,
n is the amount of the iodine standard solution, 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 ith iodine standard solution, and the unit is mu g/L;
the solution of each iodine standard substance is used,
is the average concentration of each iodine standard solution.
8. The method of claim 7,
the volume ratio of the first solution to the serum sample to be detected is 1-8: 1;
the volume ratio of the second solution to the serum sample to be detected is 1-8: 1;
the volume ratio of the third solution to the serum sample to be detected is 1-8: 1;
the volume ratio of the fourth solution to the serum sample to be detected is 1-8: 1;
the volume ratio of the fifth solution to the serum sample to be detected is 1-8: 1;
the volume ratio of the sixth solution to the serum sample to be detected is 1-8: 1.
9. the method of claim 7,
the serum sample to be tested is obtained by centrifuging blood to be tested for 10min at room temperature at 3000 r/min.
10. The method of claim 7,
the volume ratio of the first solution to the serum sample to be detected is 2: 1;
the volume ratio of the second solution to the serum sample to be detected is 2: 1;
the volume ratio of the third solution to the serum sample to be detected is 2: 1;
the volume ratio of the fourth solution to the serum sample to be detected is 7: 1;
the volume ratio of the fifth solution to the serum sample to be detected is 8: 1;
the volume ratio of the sixth solution to the serum sample to be detected is 2: 1.
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