CN102507542A - Method for quickly and quantitatively measuring urine iodine - Google Patents
Method for quickly and quantitatively measuring urine iodine Download PDFInfo
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- CN102507542A CN102507542A CN2011103156766A CN201110315676A CN102507542A CN 102507542 A CN102507542 A CN 102507542A CN 2011103156766 A CN2011103156766 A CN 2011103156766A CN 201110315676 A CN201110315676 A CN 201110315676A CN 102507542 A CN102507542 A CN 102507542A
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Abstract
The invention relates to a method for quickly and quantitatively measuring urine iodine. The method comprises the steps of: adopting polypropylene microwell plates with good heat resistance and chemical inertness as a digestion container and catalytic reaction container; preventing cross contamination of samples and water vapor leakage from the microwell plate by using a self-made sealing plate; and simultaneously measuring light absorbance of a plurality of samples at the same time by using an ELIASA reader. The method realizes batch measurement of micro samples. According to the method provided by the invention, the usage amount of reagents is greatly reduced, the measuring time is greatly shortened, and the workload in a measuring process is reduced, so that the method is suitable for measuring a large batch of samples. The method of the invention requires only simple instrument, and is low in analysis cost. The analysis efficiency is a little higher than that of an automatic analysis method.
Description
Technical field
The invention belongs to the detectable class, belong to the detection method of measuring content of iodine in the urine particularly.
Background technology
Iodine deficiency and excessive iodine have caused people's common concern as the disease of serious harm human health, thereby it is particularly important accurately to estimate crowd's iodine nutritional status.The part (about 50～75 μ g/Ls) of the iodine that human body is taken in except being absorbed by thyroid gland, all the other all are excreted, and wherein 80%～85% through the urine discharge, generally believes that therefore urine iodide level is the righttest index of evaluation and monitoring nutrition status of iodine.
The health industry standard method of China's urine iodine mensuration is an ammonium persulfate digestion-arsenic cerium catalysis AAS at present; This method is employed in the test tube sample is digested one by one; Reagent dosage is more; Operating process is loaded down with trivial details, and strict for the reaction time, therefore can't realize the fast measuring of sample in batches.
Domestic patent CN 1170147C discloses " urine iodine detection by quantitative kit and urine iodine detecting method ", and iodine standard solution, urine sample add earlier and resolve agent or neutralizing agent after clearing up, and add reductive agent again and reduce.But it is easy inadequately and sensitive.
Standard method is because of its higher sensitivity; In urine iodine is measured, adopted by extensive using, but in the method, the digestion of sample and light-catalyzed reaction process need carry out in test tube one by one; Not only cause working sample in batches; And reagent dosage is big, and particularly big consumption of arsenious acid has caused certain environmental pollution.
Summary of the invention
The objective of the invention is according to above-mentioned present situation condition, aim to provide the method for content of iodine in a kind of quick, accurate detection by quantitative urine.
The method of content of iodine in the detection by quantitative urine of the present invention comprises step:
1, adopting 12 roads to move the liquid volley of rifle fire draws in first row's micropore that 7 parts of iodine standard serial solutions, the 50 μ L that draw variable concentrations simultaneously add to 96 microwell plates; Adopt the volley of rifle fire to draw simultaneously in second row's micropore of 12 parts of urine samples, 50 μ L adding microwell plate then; Same draw four batches of urine samples (12 parts every batch), 50 μ L in addition place respectively microwell plate third and fourth, five, six row's micropores, in the application of sample micropore, add the ammonium persulfate of 100 μ L then with the volley of rifle fire.
2, microwell plate sealing is placed in 100 ℃ the thermostatic drying chamber digests 60min.
3, after the digestion bottom of seal box is cooled to room temperature with tap water, prevents the steam coagulation on microwell plate groove top, stop digestion simultaneously.
4, cooled seal pad is opened, added the arsenic chloride solution mixing of 100 μ L,, add the Cericammoniumsulfate solution of 50 μ L fast then with the every row of volley of rifle fire interval 30s.
5, reaction mixture (27 ℃) reaction at room temperature is after 10 minutes, adopts the every row of ELIASA interval 30s to read in each hole solution at the absorbance numerical value at 420nm place.
Advantage of the present invention and effect:
1, the present invention with 96 microwell plates as digestion vessel and catalytic reaction cell; Utilizing 12 roads to move the liquid volley of rifle fire simultaneously carries out batch moving of reagent and adds; When carrying out a plurality of sample absorbance, ELIASA measures; Reduced the mensuration reagent dosage significantly, shortened minute, simplified running program, to improving efficient, the reduction detection cost that urine iodine detects.
2, the present invention adopts the good polypropylene microwell plate of thermotolerance and chemical inertness as digestion vessel and catalytic reaction cell, utilizes homemade seal pad, prevents that water vapor in the microwell plate from spilling the cross pollution with sample; Adopt ELIASA to measure the absorbance of multiple sample simultaneously simultaneously, realized that the batch of micro-sample is measured.
Embodiment
Below in conjunction with specific embodiment the present invention is described further, but the embodiment that lifts not as to qualification of the present invention.If do not specialize the conventional means that used technological means is well known to those skilled in the art among the embodiment.
Embodiment 1 typical curve and sample test
The method of content of iodine in the detection by quantitative urine of the present invention comprises step:
1, adopting 12 roads to move the liquid volley of rifle fire draws in first row's micropore that 7 parts of iodine standard serial solutions, the 50 μ L that draw variable concentrations simultaneously add to 96 microwell plates; Adopt the volley of rifle fire to draw simultaneously in second row's micropore of 12 parts of urine samples, 50 μ L adding microwell plate then; Same draw four batches of urine samples (12 parts every batch), 50 μ L in addition place respectively microwell plate third and fourth, five, six row's micropores, in the application of sample micropore, add the ammonium persulfate of 100 μ L then with the volley of rifle fire.
2, microwell plate sealing is placed in 100 ℃ the thermostatic drying chamber digests 60min.
3, after the digestion bottom of seal box is cooled to room temperature with tap water, prevents the steam coagulation on microwell plate groove top, stop digestion simultaneously.
4, cooled seal pad is opened, added the arsenic chloride solution mixing of 100 μ L,, add the Cericammoniumsulfate solution of 50 μ L fast then with the every row of volley of rifle fire interval 30s.
5, reaction mixture (27 ℃) reaction at room temperature is after 10 minutes, adopts the every row of ELIASA interval 30s to read in each hole solution at the absorbance numerical value at 420nm place.
Same batch is carried out typical curve drafting and the test of 60 duplicate samples.The result is illustrated in the 0-300 μ g//L iodine concentration scope, and the typical curve equation is: C=17.68-367.4lgA, R=0.9985.
It is 132 μ g/L that 60 example urine content of iodine are measured mean value.
Press standard method, after 60 routine samples are digested one by one, use spectrophotometric determination in test tube, record urine content of iodine mean value 129 μ g/L.The result (Y) that microwell plate is measured in batches and the mensuration result (X) of standard method contrast, regression equation be Y=1.0077X-3.6998, R=0.9902 is through t check, P＞0.05, both there was no significant differences.
Embodiment 2 recovery
With recording the urine iodine concentration is the Potassiumiodate mark solution that the urine specimen of 102 μ g/L adds 50 μ g/L, 100 μ g/L, 150 μ g/L respectively; Reclaim experiment by this experimental technique; Get the recovery and be respectively 93.3%, 105.9% and 96.6%, average recovery rate is 98.6%.
Embodiment 3 precision
Withinrun precision: get the urine specimen of the urine specimen of high iodine concentration (256 μ g/L), medium iodine concentration (152 μ g/L), the urine specimen (49 μ g/L) of low iodine concentration respectively, each urine sample sample is parallel gets 12 parts, in microwell plate, measures for same batch.The average coefficient of variation (CV) that gets high, medium and low sample is respectively 3.35%, 2.32% and 5.61%.Betweenrun precision: get the urine specimen METHOD FOR CONTINUOUS DETERMINATION 3 days of concentration (152 μ g/L), every day 3 times, must criticize an average coefficient of variation is 4.50%.
The experiment of embodiment 4 impermeability
Weigh quality, microwell plate and the sealed silicon glued membrane gross mass of microwell plate respectively; In each well of every microwell plate, add 150 μ L water, the gross mass of the microwell plate quality behind the weighing application of sample, application of sample microwell plate and sealed silicon glued membrane then; Then, place 100 ℃ of permanent drying boxes to heat 60min with the sealing of the microwell plate behind the application of sample; After naturally cooling to room temperature, the gross mass of quality, application of sample microwell plate and the diaphragm seal of difference weighing application of sample microwell plate.Recording experimental data is:
Table 1 microwell plate, microwell plate and pellosil are in the quality (g) in each stage
Calculate heating front and back moisture quality respectively through data in the table,, calculate the moisture entrapment rate according to following formula.
Embodiment 5 cross pollutions experiment
Adopt the microwell plate of two same model, one as control board, and one as brassboard, carries out the cross pollution experiment.Concrete grammar is following: in the control board, series standard liquid is got one at the every interval of 50 μ L join in the micropore; In the brassboard, titer joins in the micropore identical with control board, adds the mixed solution of 50 μ L by standard solution and urine preparation at interval in the micropore.Mixed solution is that iodine standard solution and the 0.90mL urine of 100 μ g/L mixes by 0.10mL concentration.According to embodiment 1 method, the absorbance numerical value in the bioassay standard series micropore is drawn the typical curve of control board and brassboard respectively.The typical curve equation that gets control board is: C=16.42-371.3lgA, the typical curve equation of R=0.9985 brassboard is: C=19.15-363.2lgA, R=0.9985.
The absorbance numerical value of finding standard serial solution in the experiment is between the 0.900-0.170; Suppose shown in absorbance numerical value such as the following table; It is following to adopt the typical curve of control board and brassboard gained to calculate the content of iodine data respectively; The result can know from table, and the result of calculation of two typical curves is coincide, and explains not have cross pollution in the mensuration process.
The urine iodine result of calculation of table 2 control board and the corresponding different absorbances of on-gauge plate typical curve equation
*The unit of result of calculation: μ g/L.
Claims (1)
1. the method for quantitative determination urine iodine is characterized in that concrete steps are following:
(1) adopting 12 roads to move the liquid volley of rifle fire draws in first row's micropore that 7 parts of iodine standard serial solutions, the 50 μ L that draw variable concentrations simultaneously add to 96 microwell plates; Adopt the volley of rifle fire to draw simultaneously in second row's micropore of 12 parts of urine samples, 50 μ L adding microwell plate then; Same draw four batches of urine samples (12 parts every batch), 50 μ L in addition place respectively microwell plate third and fourth, five, six row's micropores, in the application of sample micropore, add the ammonium persulfate of 100 μ L then with the volley of rifle fire.
(2) microwell plate sealing is placed in 100 ℃ the thermostatic drying chamber digests 60min.
(3) after the digestion bottom of seal box is cooled to room temperature with tap water, prevents the steam coagulation on microwell plate groove top, stop digestion simultaneously.
(4) cooled seal pad is opened, added the arsenic chloride solution mixing of 100 μ L,, add the Cericammoniumsulfate solution of 50 μ L fast then with the every row of volley of rifle fire interval 30s.
(5) reaction mixture (27 ℃) reaction at room temperature is after 10 minutes, adopts the every row of ELIASA interval 30s to read in each hole solution at the absorbance numerical value at 420nm place.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103156766A CN102507542A (en) | 2011-10-09 | 2011-10-09 | Method for quickly and quantitatively measuring urine iodine |
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| CN2011103156766A CN102507542A (en) | 2011-10-09 | 2011-10-09 | Method for quickly and quantitatively measuring urine iodine |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| US11313790B2 (en) | 2019-12-16 | 2022-04-26 | Harbin Medical University | Method for detecting iodine concentration in water samples |
-
2011
- 2011-10-09 CN CN2011103156766A patent/CN102507542A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
| US11313790B2 (en) | 2019-12-16 | 2022-04-26 | Harbin Medical University | Method for detecting iodine concentration in water samples |
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Application publication date: 20120620 |