CN102495203A - Reagent used for detecting urinary iodine by biochemistry analyzer and urinary iodine detecting method - Google Patents
Reagent used for detecting urinary iodine by biochemistry analyzer and urinary iodine detecting method Download PDFInfo
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Abstract
The invention discloses a reagent used for detecting urinary iodine by a biochemistry analyzer and a urinary iodine detecting method. The reagent mainly comprises the following components: digestion solution-ammonium persulfate solution, reagent R1-arsenious acid or arsenic trioxide solution, reagent R2-ceric ammonium sulfate or cerous sulfate solution, and iodine standard solution-potassium iodide (sodium iodide) or potassium iodate (sodium iodate) solution. The detecting method mainly includes the following steps: selecting the detecting reagent, digesting urine sample, setting parameters of the biochemistry analyzer, and determining through the biochemistry analyzer. The detecting method has high automation degree, small human error, little reagent dosage and short detecting time. By adopting the biochemistry analyzer to analyze the urinary iodine, the urinary iodine detecting method expands the detecting items of the instrument, is particularly suitable for the medical organizations equipped with the biochemistry analyzers, and meets the requirements of clinical detection on quick issuing of result and detecting automation.
Description
Technical field
The present invention relates to medical detectable and detection method, particularly a kind of reagent and urine iodine detecting method that is used for Biochemical Analyzer detection by quantitative urine iodine.
Background technology
Urine iodide level is the important indicator that evaluation crowd iodine is taken in situation, is extensively adopted both at home and abroad.Current, iodine deficiency and excessive iodine have become the equal public health problem of paying close attention to of people.Urine iodine detects and has become the important evidence that the crowd science of guidance enriches the iodine.
It is the ultramicro-analysis technology of a complicacy that urine iodine detects.At present, can more accurately measure the domestic and international method that adopts of urine iodine and quilt and have following three kinds:
(1). plasma chromatogram mass spectroscopy (ICP-MS), hereinafter to be referred as method (1).This method is used the plasma chromatograph, under nearly 10000 ℃ high temperature, the iodide ion in the urine sample is gasified.Specific charge according to ion is different, detects the content of iodine through the chromatography of ions detecting device.U.S. disease prevention and control center and Europe part country adopt the method.The method accuracy is high, but instrument is expensive, and about 1,600,000 yuans/platform is not suitable for developing country and extensively adopts.
(2). arsenic cerium catalysis AAS, hereinafter to be referred as method (2).This method uses the content of remaining high valence state cerium in the spectrophotometer dynamic measurement test solution to come indirect determination iodine according to the principle of catalysis of iodine arsenic cerium redox reaction.China reaches most countries employing the method in the world, is China's health industry standard method (WS/T 107-2006).But this method digestion process need use the constant temperature digestion appearance; Course of reaction need be used the thermostatic water-circulator bath case, and measure process need and use spectrophotometer, and consistent for the reaction time that makes each sample during operation; Need accurately to add one time in 30 seconds at interval reagent; Need too during mensuration that accurately with spectrophotometric determination once if mistakes and omissions once, then all that has been achieved is spoiled in 30 seconds at interval.Therefore, complex steps, artificial burden is heavy, and detection time is long, detects 60 parts of urine samples and takes 4-5 hour, and reagent dosage is big, and the consumption of particularly hypertoxic chemicals arsenic is (being 20 times of consumption of arsenic of the present invention) greatly, and people's cognition is damaged.
(3). arsenic cerium catalysis chronometric method, hereinafter to be referred as method (3).This method is the mandate patent of invention (patent No. 01133682.X) of Liu's inventor row an ancient unit of weight.This method is cleared up urine sample through using strong oxidizer at normal temperatures, according to the principle of catalysis of iodine arsenic cerium redox reaction, uses indicator Indicator Reaction terminal point, keeps the reaction time in mind with stopwatch, calculates the urine content of iodine according to the reaction time.This method is carried out urine iodine test item for hospital's (particularly large hospital), and because of operation steps is manual operations entirely, detection time is longer, and need with the naked eye judge reaction end, and the objectivity of experimental result is not strong, so most of hospital also is reluctant to accept.
Summary of the invention
The purpose of this invention is to provide the simple and direct reagent and the urine iodine detecting method that are used for Biochemical Analyzer detection by quantitative urine iodine that quick and precisely detects content of iodine in the urine of a kind of ability.
The object of the invention is accomplished by following technical scheme:
The used detectable splendid attire of the present invention is in kit, and its reagent is become to be grouped into by following main agents:
(1) digestion solution-ammonium persulfate solution, concentration range is: 0.3-2.0mol/L.
(2) reagent R1-arsenious acid or arsenic trioxide solution, wherein the arsenic concentration scope is: 0.01-0.1mol/L.
(3) reagent R
2-Cericammoniumsulfate or cerous sulfate solution, wherein the cerium concentration scope is: 0.001-0.1mol/L.
(4) serial iodine standard solution-potassium iodide (sodium) or Potassiumiodate (sodium) solution, content of iodine is: 0-500 μ g/L.
The preferred value of said main agents composition and content is:
(1). digestion solution-ammonium persulfate solution { c [(NH
4)
2S
2O
8]=1.3mol/L};
(2). reagent R
1-arsenic chloride solution [c (H
3AsO
3)=0.037mol/L];
(3). reagent R
2-Cericammoniumsulfate solution [c (Ce
4+)=0.0076mol/L];
(4). serial iodine standard solution-potassium iodate solution, content of iodine is respectively: 0,50,100,200,300,400 μ g/L.
Use detectable of the present invention to urinate the method that iodine detects, its key step is:
1. select above-mentioned detectable for use;
2. clear up;
Accurately draw serial iodine standard solution, each 50-500 μ l of urine sample in test tube, respectively add 100-1000 μ l digestion solution, place electrically heated drying cabinet 100-110 ℃ to clear up 1 hour; To clear up the sample that finishes and be cooled to room temperature;
3. the Biochemical Analyzer location parameter is set,
3.1 wavelength: single wavelength, selected a kind of wavelength in the 400-600nm scope;
3.2 sample, amount of reagent: the serial iodine standard solution after clearing up, sample 5-200 μ L, reagent R
110-300 μ L, reagent R
210-250 μ L, selected a kind of consumption in amount ranges separately;
3.3 temperature of reaction: selected a kind of temperature in 30-40 ℃ of scope;
3.4 analytical approach: end-point method or rate method;
3.4.1 1 end-point method: add reagent R
1Reaction 0-2min measures adding reagent R
2The OD value of selected time point in the afterreaction 3-20min scope;
3.4.2 two point velocity methods: add reagent R
1Reaction 0-2min, METHOD FOR CONTINUOUS DETERMINATION adds reagent R
2The OD value rate of change of back selected time period in the 3-15min scope;
3.5 calibration steps: calibration curve method, use logarithmic mode (logit-log4p) or splines (spline) match typical curve on the instrument;
3.6 the Direction of Reaction: negative reaction;
4. measure,
Serial iodine standard solution after will clearing up, sample, reagent R
1, reagent R
2Be positioned over respectively in Biochemical Analyzer corresponding standard position, sample position and the reagent position, the Biochemical Analyzer location parameter of " 3 " said setting starts instrument and measures automatically set by step.
The preferred value of the sample in the said detection method, reagent dosage and Biochemical Analyzer location parameter is:
(1) clear up,
Accurately draw serial iodine standard solution, each 300 μ l of urine sample in test tube, respectively add 600 μ l digestion solutions, place electrically heated drying cabinet to clear up 1 hour in 100 ℃;
(2) the Biochemical Analyzer location parameter is set,
(2.1) wavelength: 405nm;
(2.2) sample, amount of reagent: the serial iodine standard solution after clearing up, each 25 μ L of sample, reagent R
1120 μ L, reagent R
236 μ L;
(2.3) temperature of reaction: 37 ℃;
(2.4) analytical approach: two point velocity methods add reagent R
1Reaction 0-2min, METHOD FOR CONTINUOUS DETERMINATION adds reagent R
2The rate of change of the OD value in the back 3min time period;
(2.5) calibration steps: calibration curve method, use logarithmic mode (logit-log4p) the match typical curve on the instrument.
Principle of the present invention:
Use ammonium persulfate to clear up urine sample, decompose the material that disturbs iodine to measure in the urine sample, utilize the principle of catalysis of iodine arsenic cerium reaction, the high more reaction velocity of content of iodine is fast more, and remaining yellow cerium (4+) ion is few more in the test solution.Use Biochemical Analyzer after the reaction time of regulation, to measure the OD value (some end-point methods) of remaining cerium (4+) in the test solution or in the variation (two point velocity methods) of the OD value of official hour section continuous monitoring test solution; Become quantitative relationship according to OD value or OD value rate of change with content of iodine, calculate the urine iodine concentration through calibration curve method.
Beneficial effect of the present invention:
The urine sample of clearing up of the present invention is used the temperature control baking box, again without any need for intermediate steps, directly uses Biochemical Analyzer to measure automatically behind the treatments of the sample, and manually-operated has only application of sample to digest a step, and all the other operation stepss are accomplished (comprise providing and measure the result) by instrument entirely.The present invention compares with the method (2) in the background technology, and the instrument of use is different fully with operation steps, and automaticity of the present invention is high, and personal error is little, and reagent dosage is few, and detection time is short, and 60 duplicate samples only need 1 hour 10 minutes.Its innovative point is the ratio through adjustment key reaction reagent arsenic cerium; The effect of auxiliary reagent in addition; The catalytic reaction speed of iodine is carried out according to the intrinsic response curve equation of Biochemical Analyzer; Create arsenic cerium catalytic reaction and Biochemical Analyzer and measured the good interface of program, realized utilizing the Biochemical Analyzer purpose of fast detecting urine iodine automatically.
The present invention compares with the method (3) in the background technology, and except that the chemical principle that uses was identical, other everyways were inequality or similar.The present invention mainly carries out urine iodine test item to hospital and develops, and reason is that Biochemical Analyzer is the conventional instrument that uses of hospital, is used for the detection of clinical other multiple biochemical indicators, and popularity is high.The present invention uses Biochemical Analyzer to detect urine iodine; Expanded the test item of this instrument; Be particularly suitable for having medical institutions' use of Biochemical Analyzer, satisfied Clinical detection and required to go out fast result and the demand that detects robotization, be with a wide range of applications.
Embodiment
Below in conjunction with embodiment the present invention is further described, but embodiment should not be construed as limitation of the present invention.
1. reagent is formed,
(1) digestion solution: ammonium persulfate solution { c [(NH
4)
2S
2O
8]=1.3mol/L};
(2) reagent R
1: arsenic chloride solution [c (H
3AsO
3)=0.037mol/L];
(3) reagent R
2: Cericammoniumsulfate solution [c (Ce
4+)=0.0076mol/L];
(4) serial iodine standard solution: potassium iodate solution, content of iodine is respectively: 0,50,100,200,300,400 μ g/L.
2. clear up,
Accurately draw serial iodine standard solution, each 300 μ l of urine sample in test tube, respectively add 600 μ l digestion solutions, place 100 ℃ of electrically heated drying cabinets to clear up 1 hour; The ventilating opening of electrothermal drying box top should open wide when clearing up, and does not drive fan blower; To clear up the sample that finishes and be cooled to room temperature;
3. use Hitachi's 7180 type automatic clinical chemistry analyzers, following location parameter be set,
(1) wavelength: 405nm;
(2) sample, amount of reagent: the serial iodine standard solution after clearing up, each 25 μ L of sample, reagent R
1120 μ L, reagent R
236 μ L;
(3) temperature of reaction: 37 ℃;
(4) analytical approach: two point velocity methods add reagent R
1Reaction 2min, METHOD FOR CONTINUOUS DETERMINATION adds reagent R
2The OD value rate of change of (starting point of setting and terminal point are respectively and the 20th point at the 7th) in the back 3min time period;
(5) calibration steps: calibration curve method, use the logarithmic mode on the instrument
(logit-log4p) match typical curve.
4. measure,
Serial iodine standard solution after will clearing up, sample, reagent R
1, reagent R
2Be positioned over respectively in normal bit, sample position and the reagent position of Hitachi's 7180 type automatic clinical chemistry analyzers, start instrument and measure.The sample of measuring is 3 parts of country-level urine iodine standard substances (the freeze-drying urine sample of known content of iodine) and 1 part of interior Quality Control urine sample of using national health industry standard method (background method 2) definite value; Sample is from CDC country IDD(Iodine deficiency disorders) reference laboratories (Beijing); Be respectively that standard substance GBW09108j (is called for short " j "; Content of iodine 68 ± 9 μ g/L), GBW09109f (is called for short " f "; Content of iodine 138 ± 10 μ g/L), GBW091101 (being called for short " 1 ", content of iodine 206 ± 10 μ g/L) and interior Quality Control appearance (being called for short " interior ", content of iodine 170 ± 10 μ g/L).The mensuration result of the inventive method shows; The measured value of sample is all in the range of uncertainty of set-point; And and the absolute value of the relative error between the set-point [(measured value-set-point)/set-point x 100%] is much smaller than 10%; Meet the requirement of analytical approach, explain that this method is a method that detects urine iodine accurately accuracy.See table 1.
The serial iodine standard that table 1 Biochemical Analyzer is measured, the OD value rate of change (△ A/t) and the content of iodine (μ g/L) of sample
*: " 0 " of iodine standard does not promptly have the WS of iodine, is blank.Therefore, no measured value and relative error.
Obviously, those skilled in the art can carry out various changes and modification to the present invention and not break away from the spirit and scope of the present invention.Belong within the scope of claim of the present invention and equivalent technologies thereof if of the present invention these are revised with modification, then the present invention also should comprise these changes and modification interior.
If the content of not making detailed description is arranged, then belong to those skilled in the art's technique known in this instructions, repeat no more here.
Claims (4)
1. one kind is used for the reagent that the Biochemical Analyzer detection by quantitative is urinated iodine, it is characterized in that: the detectable splendid attire is in kit, and its reagent is become to be grouped into by following main agents:
(1) digestion solution-ammonium persulfate solution, concentration range is: 0.3-2.0mol/L;
(2) reagent R
1-arsenious acid or arsenic trioxide solution, wherein the arsenic concentration scope is: 0.01-0.1mol/L;
(3) reagent R
2-Cericammoniumsulfate or cerous sulfate solution, wherein the cerium concentration scope is: 0.001-0.1mol/L;
(4) serial iodine standard solution-potassium iodide (sodium) or Potassiumiodate (sodium) solution, content of iodine is: 0-500 μ g/L.
2. the reagent that is used for Biochemical Analyzer detection by quantitative urine iodine as claimed in claim 1, it is characterized in that: the preferred value of said main agents composition and content is:
(1) digestion solution-ammonium persulfate solution { c [(NH
4)
2S
2O
8]=1.3mol/L};
(2) reagent R
1-arsenic chloride solution [c (H
3AsO
3)=0.037mol/L];
(3) reagent R
2-Cericammoniumsulfate solution [c (Ce
4+)=0.0076mol/L];
(4) serial iodine standard solution-potassium iodate solution, content of iodine is respectively: 0,50,100,200,300,400 μ g/L.
3. one kind is used for the detection method that the Biochemical Analyzer detection by quantitative is urinated iodine, and it is characterized in that: the key step of detection method is:
(1) selects above-mentioned detectable for use;
(2) clear up,
Accurately draw serial iodine standard solution, each 20-500 μ l of urine sample in test tube, respectively add 100-1000 μ l digestion solution, place electrically heated drying cabinet 100-110 ℃ to clear up 1 hour; To clear up the sample that finishes and take out, be cooled to room temperature;
(3) the Biochemical Analyzer location parameter is set,
(3.1) wavelength: single wavelength, selected a kind of wavelength in the 400-600nm scope;
(3.2) sample, amount of reagent: serial iodine standard solution, sample 5-200 μ L after cancellation is separated, reagent R
110-300 μ L, reagent R
210-250 μ L, selected a kind of consumption in amount ranges separately;
(3.3) temperature of reaction: selected a kind of temperature in 30-40 ℃ of scope;
(3.4) analytical approach: end-point method or rate method;
(3.4.1) some end-point methods: add reagent R
1Reaction 0-2min measures adding reagent R
2The OD value of selected time point in the afterreaction 3-20min scope;
(3.4.2) two point velocity methods: add reagent R
1Reaction 0-2min, METHOD FOR CONTINUOUS DETERMINATION adds reagent R
2OD value rate of change in the time period that the back is selected in the 3-15min scope;
(3.5) calibration steps: calibration curve method, use logarithmic mode (logit-log4p) or splines (spline) match typical curve on the instrument;
(3.6) the Direction of Reaction: negative reaction;
(4) measure,
Serial iodine standard solution after will clearing up, sample, reagent R
1, reagent R
2Be positioned over respectively in Biochemical Analyzer corresponding standard position, sample position and the reagent position, the Biochemical Analyzer location parameter of (3) said setting starts instrument and measures set by step.
4. the detection method that is used for Biochemical Analyzer detection by quantitative urine iodine as claimed in claim 3, it is characterized in that: the preferred value of the sample in the said detection method, reagent dosage and Biochemical Analyzer location parameter is:
(1) clear up,
Accurately draw serial iodine standard solution, each 300 μ l of urine sample in test tube, respectively add 600 μ l digestion solutions, place electrically heated drying cabinet to clear up 1 hour in 100 ℃;
(2) the Biochemical Analyzer location parameter is set,
(2.1) wavelength: 405nm;
(2.2) sample, amount of reagent: the serial iodine standard solution after clearing up, each 25 μ L of sample, reagent R
1120 μ L, reagent R
236 μ L;
(2.3) temperature of reaction: 37 ℃;
(2.4) analytical approach: two point velocity methods add reagent R
1Reaction 0-2min, METHOD FOR CONTINUOUS DETERMINATION adds reagent R
2The rate of change of the OD value in the back 3min time period;
(2.5) calibration steps: calibration curve method, use logarithmic mode (logit-log4p) the match typical curve on the instrument.
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Cited By (7)
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| CN107677805A (en) * | 2017-09-30 | 2018-02-09 | 安徽伊普诺康生物技术股份有限公司 | A kind of kit for determining urine iodine and its preparation application method |
| CN108776134A (en) * | 2018-08-14 | 2018-11-09 | 厦门市疾病预防控制中心（厦门市卫生检测检验中心） | One kind being based on the iodide ion detection reagent and method of catalysis of iodine hydrazine-[oxidant-Ferroin reagents] system |
| CN108827951A (en) * | 2018-06-28 | 2018-11-16 | 天津中成佳益生物科技有限公司 | A kind of urine iodine immue quantitative detection reagent box |
| CN109540877A (en) * | 2017-09-22 | 2019-03-29 | 王九宏 | Urinate the reagent and detection method that above microplate reader detects after iodine and saliva iodine room temperature pre-treatment |
| CN110320207A (en) * | 2019-07-23 | 2019-10-11 | 黄艳丽 | A kind of iodine detection reagent and detection method |
| CN111830023A (en) * | 2019-08-19 | 2020-10-27 | 杭州爱光医疗器械有限公司 | Sulfhydryl compound detection reagent, detection test paper, kit, test paper box and preparation method thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017012168A1 (en) * | 2015-07-22 | 2017-01-26 | 江苏英诺华医疗技术有限公司 | Optimized biochemical detection method suitable for medical examination |
| CN109540877A (en) * | 2017-09-22 | 2019-03-29 | 王九宏 | Urinate the reagent and detection method that above microplate reader detects after iodine and saliva iodine room temperature pre-treatment |
| CN107677805A (en) * | 2017-09-30 | 2018-02-09 | 安徽伊普诺康生物技术股份有限公司 | A kind of kit for determining urine iodine and its preparation application method |
| CN108827951A (en) * | 2018-06-28 | 2018-11-16 | 天津中成佳益生物科技有限公司 | A kind of urine iodine immue quantitative detection reagent box |
| CN108827951B (en) * | 2018-06-28 | 2020-08-25 | 天津中成佳益生物科技有限公司 | Quantitative detection kit for urine iodine |
| CN108776134A (en) * | 2018-08-14 | 2018-11-09 | 厦门市疾病预防控制中心（厦门市卫生检测检验中心） | One kind being based on the iodide ion detection reagent and method of catalysis of iodine hydrazine-[oxidant-Ferroin reagents] system |
| CN110320207A (en) * | 2019-07-23 | 2019-10-11 | 黄艳丽 | A kind of iodine detection reagent and detection method |
| CN111830023A (en) * | 2019-08-19 | 2020-10-27 | 杭州爱光医疗器械有限公司 | Sulfhydryl compound detection reagent, detection test paper, kit, test paper box and preparation method thereof |
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