CN102495203B - 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 detection reagent and detection method, particularly a kind of reagent and urine iodine detecting method that quantitatively detects urine iodine for Biochemical Analyzer.
Background technology
Urine iodide level is the important indicator of evaluation crowd iodine Intake status, is widely used 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 a complicated ultramicro-analysis technology that urine iodine detects.At present, can urinate iodine and the method that adopted has following three kinds both at home and abroad compared with Accurate Determining:
(1). plasma chromatogram mass spectroscopy (ICP-MS), hereinafter to be referred as method (1).The method is used plasma chromatograph, under the high temperature of nearly 10000 ℃, the iodide ion in urine sample is gasified.Different according to the specific charge of ion, by the content of chromatography of ions detecting device detection iodine.U.S. disease prevention and control center and Some European country adopt the method.The method accuracy is high, but instrument is expensive, approximately 1,600,000 yuans/platform, is not suitable for developing country and extensively adopts.
(2). arsenic cerium Catalytic Spectrophotometry, hereinafter to be referred as method (2).The method, according to the principle of catalysis of iodine arsenic cerium redox reaction, is carried out indirect determination iodine with the content of remaining high valence state cerium in spectrophotometer dynamic measurement test solution.China and in the world most countries adopt the method ，Shi China's health industry standard method (WS/T 107-2006).But the method digestion process need to be used constant temperature digestion instrument, course of reaction need to be used thermostatic water-circulator bath case, measure process need and use spectrophotometer, and when operation is in order to make the reaction time of each sample consistent, need accurate interval within 30 seconds, to add reagent one time, during mensuration, need too accurate interval 30 seconds with spectrophotometric determination once, if mistakes and omissions once, all that has been achieved is spoiled.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 large, and particularly the consumption of severe poisonous chemicals arsenic large (being 20 times of consumption of arsenic of the present invention), damages people's cognition.
(3). arsenic cerium catalysis chronometric method, hereinafter to be referred as method (3).The method is the mandate patent of invention (patent No. 01133682.X) of Liu's inventor row an ancient unit of weight.The method is by using at normal temperatures strong oxidizer to clear up urine sample, according to the principle of catalysis of iodine arsenic cerium redox reaction, uses indicator Indicator Reaction terminal, with stopwatch, keeps the reaction time in mind, calculates urine content of iodine according to the reaction time.The method is carried out urine iodine test item for hospital's (particularly large hospital), and because 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, therefore most of hospital is also reluctant to accept.
Summary of the invention
The object of this invention is to provide a kind of energy simple and direct quick and precisely detect content of iodine in urine for Biochemical Analyzer, quantitatively detect reagent and the urine iodine detecting method of urine iodine.
Object of the present invention is completed by following technical scheme:
The present invention's detection reagent splendid attire used 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 arsenic concentration scope is: 0.01-0.1mol/L.
(3) reagent R
2-Cericammoniumsulfate or cerous sulfate solution, wherein 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 described 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 detection reagent of the present invention to urinate the method that iodine detects, its key step is:
1. select above-mentioned detection reagent;
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, be placed in electrically heated drying cabinet 100-110 ℃ and clear up 1 hour; By clearing up complete sample, be cooled to room temperature;
3. Biochemical Analyzer location parameter is set,
3.1 wavelength: single wavelength, selected a kind of wavelength within the scope of 400-600nm;
3.2 samples, 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 within the scope of 30-40 ℃;
3.4 analytical approachs: end-point method or rate method;
3.4.1 1 end-point method: add reagent R
1reaction 0-2min, measures and adds reagent R
2the optical density value of selected time point within the scope of rear reaction 3-20min;
3.4.2 two point rate assay: add reagent R
1reaction 0-2min, METHOD FOR CONTINUOUS DETERMINATION adds reagent R
2after within the scope of 3-15min the optical density value rate of change of selected time period;
3.5 calibration stepss: calibration curve method, is used logarithmic mode (logit-log4p) or splines (spline) matching typical curve on instrument;
3.6 the Direction of Reaction: negative reaction;
4. measure,
By the serial iodine standard solution after clearing up, sample, reagent R
1, reagent R
2be positioned over respectively in the corresponding normal bit of Biochemical Analyzer, sample position and reagent position, by the Biochemical Analyzer location parameter of the described setting of step " 3 ", start instrument and automatically measure.
The preferred value of the sample in described 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, be placed in electrically heated drying cabinet and clear up 1 hour in 100 ℃;
(2) 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 rate assay, adds reagent R
1reaction 0-2min, METHOD FOR CONTINUOUS DETERMINATION adds reagent R
2the rate of change of the optical density value of rear 3min in the time period;
(2.5) calibration steps: calibration curve method, is used logarithmic mode (logit-log4p) the matching typical curve on instrument.
Principle of the present invention:
Use ammonium persulfate to clear up urine sample, decompose the material that disturbs iodine to measure in urine sample, utilize the principle of catalysis of iodine arsenic cerium reaction, the higher reaction velocity of content of iodine is faster, and in test solution, remaining yellow cerium (4+) ion is fewer.Use Biochemical Analyzer after the reaction time of regulation, measure test solution in remaining cerium (4+) optical density value (some end-point methods) or in the variation (two point rate assay) of the optical density value of official hour section continuous monitoring test solution, according to optical density value or optical density value rate of change, become quantitative relationship with content of iodine, by calibration curve method, calculate urine iodine concentration.
Beneficial effect of the present invention:
The urine sample of clearing up of the present invention is used temperature control baking box, after treatments of the sample again without any need for intermediate steps, directly use Biochemical Analyzer automatically to measure, manual operation only has application of sample to digest a step, and all the other operation stepss complete (comprise and provide measurement result) by instrument entirely.The present invention compares with the method (2) in background technology, and the instrument of use and operation steps are completely different, 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 by adjusting the ratio of key reaction reagent arsenic cerium, in addition the effect of auxiliary reagent, the catalytic of iodine is carried out according to the intrinsic response curve equation of Biochemical Analyzer, created arsenic cerium catalytic reaction and Biochemical Analyzer and measured the good interface of program, realized and utilized the Biochemical Analyzer object of fast detecting urine iodine automatically.
The present invention compares with the method (3) in background technology, and except the chemical principle using is identical, other everyways are not identical or similar.The present invention mainly develops for hospital urine iodine test item, and reason is that Biochemical Analyzer is the conventional instrument using of hospital, and for the detection of clinical other multiple biochemical indicators, popularity is high.The present invention detects urine iodine with Biochemical Analyzer, expanded the test item of this instrument, be particularly suitable for having medical institutions' use of Biochemical Analyzer, met the demand that clinical detection requires to go out fast result and detects robotization, be with a wide range of applications.
Embodiment
Below in conjunction with embodiment, the invention will be further described, but embodiment should not be construed as limitation of the present invention.
1. reagent forms,
(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, be placed in 100 ℃ of electrically heated drying cabinets and clear up 1 hour; While clearing up, the ventilating opening of electrothermal drying box top should open wide, and does not drive fan blower; By clearing up complete sample, 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 rate assay, adds reagent R
1reaction 2min, METHOD FOR CONTINUOUS DETERMINATION adds reagent R
2the optical density value rate of change of (starting point of setting and terminal are respectively and the 20th point at the 7th) in the rear 3min time period;
(5) calibration steps: calibration curve method, is used the logarithmic mode on instrument
(logit-log4p) matching typical curve.
4. measure,
By the serial iodine standard solution after 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), 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 (is called for short " 1 ", content of iodine 206 ± 10 μ g/L) and interior Quality Control sample (be called for short " interior ", content of iodine 170 ± 10 μ g/L).The measurement 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 set-point [(measured value-set-point)/set-point x 100%] is much smaller than 10%, meet the requirement of analytical approach to accuracy, illustrate that this method is a method that detects accurately urine iodine.In Table 1.
The serial iodine standard that table 1 Biochemical Analyzer is measured, optical density value rate of change (△ A/t) and the content of iodine (μ g/L) of sample
*: " 0 " of iodine standard, without the aqueous solution of iodine, is blank.Therefore, without measured value and relative error.
Obviously, those skilled in the art can carry out various changes and modification and not depart from the spirit and scope of the present invention the present invention.If of the present invention these are revised and within modification belongs to the scope of the claims in the present invention and equivalent technologies thereof, the present invention also should comprise these changes and modification interior.
If there is the content not being described in detail in this instructions, belong to the known technology of those skilled in the art, repeat no more herein.
Claims (2)
1. for Biochemical Analyzer, quantitatively detect a detection method for urine iodine, it is characterized in that: the key step of detection method is:
(1) select following detection reagent:
(1.1) digestion solution-ammonium persulfate solution, concentration range is: 0.3-2.0mol/L;
(1.2) reagent R
1-arsenious acid or arsenic trioxide solution, wherein arsenic concentration scope is: 0.01-0.1mol/L;
(1.3) reagent R
2-Cericammoniumsulfate or cerous sulfate solution, wherein cerium concentration scope is: 0.001-0.1mol/L;
(1.4) serial iodine standard solution-potassium iodide or potassium iodate solution, content of iodine is: 0-500 μ g/L;
(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, be placed in electrically heated drying cabinet 100-110 ℃ and clear up 1 hour; By clearing up complete sample, take out, be cooled to room temperature;
(3) Biochemical Analyzer location parameter is set,
(3.1) wavelength: single wavelength, selected a kind of mensuration wavelength within the scope of 400-600nm;
(3.2) sample, amount of reagent: cancel serial iodine standard solution, each 5-200 μ L of sample after solution, 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 within the scope of 30-40 ℃;
(3.4) analytical approach: end-point method or rate method;
(3.4.1) some end-point methods: add reagent R
1reaction 0-2min, measures and adds reagent R
2the optical density value of selected time point within the scope of rear reaction 3-20min;
(3.4.2) two point rate assay: add reagent R
1reaction 0-2min, METHOD FOR CONTINUOUS DETERMINATION adds reagent R
2after within the scope of 3-15min the optical density value rate of change in selected time period;
(3.5) calibration steps: calibration curve method, is used logarithmic mode logit-log4p or splines spline matching typical curve on instrument;
(3.6) the Direction of Reaction: negative reaction;
(4) measure,
By the serial iodine standard solution after clearing up, sample, reagent R
1, reagent R
2be positioned over respectively in the corresponding normal bit of Biochemical Analyzer, sample position and reagent position, by the Biochemical Analyzer location parameter of the described setting of step (3), start instrument and measure.
As claimed in claim 1 for Biochemical Analyzer quantitatively detect urine iodine detection method, it is characterized in that: the preferred value of the sample in described 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, be placed in electrically heated drying cabinet and clear up 1 hour in 100 ℃;
(2) 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 rate assay, adds reagent R
1reaction 0-2min, METHOD FOR CONTINUOUS DETERMINATION adds reagent R
2the rate of change of the optical density value of rear 3min in the time period;
(2.5) calibration steps: calibration curve method, is used the logarithmic mode logit-log4p matching typical curve on instrument.
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| CN105116156B (en) * | 2015-07-22 | 2017-12-12 | 江苏英诺华医疗技术有限公司 | A kind of biochemical detection methods of the suitable medical test of optimization |
| 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 |
| 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 |
| CN108776134B (en) * | 2018-08-14 | 2019-11-12 | 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 reagent] system |
| CN110320207A (en) * | 2019-07-23 | 2019-10-11 | 黄艳丽 | A kind of iodine detection reagent and detection method |
| CN110672593A (en) * | 2019-08-19 | 2020-01-10 | 杭州爱光医疗器械有限公司 | Sulfhydryl compound detection reagent, preparation method and kit thereof |
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