CN106546586A - The detectable of content of iodine in a kind of vitro detection urine - Google Patents
The detectable of content of iodine in a kind of vitro detection urine Download PDFInfo
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Abstract
The invention discloses in a kind of vitro detection urine content of iodine detectable, including oxidant and developer；The oxidant, by mass percentage concentration, consists of the following composition：Glacial acetic acid 10%～30%, hydrogen peroxide 1%～20%, hydrochloric acid 0.1%～10%, sulphuric acid 0.1%～10%, quinolinic acid 0.1%～5%, purification water surplus；The developer is TMB alcoholic solutions that mass percentage concentration is 0.025%～0.2%.It is higher that repeatable and accuracy is surveyed in health check-up of the present invention, and develop the color gradient and color stability it is also very good, using simple to operate, reaction detection is time-consuming few, without the need for doing multiprocessing to urine sample, simple and fast, without heavy metal compound and aldehyde condensate, is difficult to environment, safety and environmental protection, by the detection by quantitative to content of iodine in urine, testing result is used for the Iodine nutrition situation for evaluating detected crowd to the present invention, it is adaptable in the serious basic medical unit popularization and application of iodine deficiency.
Description
Technical field
The present invention relates to detection field, relates in particular to a kind of detectable of content of iodine in vitro detection urine.
Background technology
Iodine is the indispensable important element of synthetic thyroid hormone, and the development to brain plays an important role.If anemia of pregnant woman
In-vivo iodine deficiency can then cause the brain injury of fetus and infant, cause irreversible mental retardation and Psychomotor ability barrier
Hinder, show as staying, it is stupid, deaf and dumb, paralysed, and abstract thinking ability difference defect.
The detection method of existing urine iodine is urine iodine arsenic cerium Spect Rophotometric Catalytic Method For Determination method, and its principle is by reaction
The Ce of yellow4+It is reduced into colourless Ce3+, content of iodine is higher, and response speed is faster, remaining Ce4+Then fewer, control is anti-
Temperature and time is answered, the residue Ce in measure system under 420nm wavelength4+Absorbance.
But, when using the method, urine specimen process needs to add 100 DEG C of digestion urines of high temperature after Ammonium persulfate.
It is measured after 60min again, with certain dangerous and nocuity, process time is oversize, is not easy to operation, time cycle
It is long, and the reagent of the method such as arsenic trioxide etc. is containing heavy metal compound poisonous and harmful, easily to environment, and
Need it is special clear up, semi-automatic biochemical analyzer, inconvenience is in the serious basic unit's popularization and application of iodine deficiency.
The content of the invention
The technical problem to be solved is to provide a kind of safety and environmental protection, and repeatable and accuracy is higher, and
Using simple to operate, reaction detection takes the detectable of content of iodine in few vitro detection urine.
In order to solve above-mentioned technical problem, the present invention is adopted the following technical scheme that：Content of iodine in a kind of vitro detection urine
Detectable, including oxidant and developer；
The oxidant, by mass percentage concentration, consists of the following composition：
The developer is TMB alcoholic solutions that mass percentage concentration is 0.025%～0.2%.
In acid condition, glacial acetic acid generates peracetic acid under the oxidation of strong acid, aoxidize 33 ', 5 ' 5 ' under catalysis of iodine-
Tetramethyl benzidine, in ethanol medium, shows blueness, and in certain scope, the concentration of iodine is directly proportional to colored intensity；Quinoline
Acid contributes to oxidation.
Further, the solvent of the TMB alcoholic solutions is ethanol.In implementing the present invention, it may, inventor has found to use
Solvent of the ethanol as TMB, is conducive to transparency and the definition for developing the color, and lifts color speed effect, while can promote to increase
The color developing effect of strong chromogenic reaction.
Further, also including diluent, the diluent is the citric acid-buffered sodium citrate of pH=3.0～5.0
Liquid or acetate buffer or Tris buffer.
Further, also including urine pretreating agent, the urine pretreating agent is activated carbon.There is activated carbon absorption to make
With, affect during urine can be removed detect impurity.
Further, the activated carbon first passes through following methods in advance and processed：Dehydrated alcohol is added in activated carbon, is stirred
Stood after uniform, activated carbon is fully settled, then abandon supernatant and float, add purified water, stir laggard
Row stands, and activated carbon is fully settled, then supernatant discarded, carries out sucking filtration process afterwards, finally dries the activated carbon after sucking filtration
It is dry.Activated carbon can reach the purpose of the organic compound in absorption urine, obtain pure colourless through above pretreatment
Urine specimen.
Further, also including iodine titer.Iodine titer is used as calculating standard and draws standard curve.
Further, the iodine titer includes the liquor kalii iodide of 50 μ g/L, the liquor kalii iodide of 100 μ g/L, 250 μ
The liquor kalii iodide of g/L, the liquor kalii iodide of 400 μ g/L, the liquor kalii iodide of 500 μ g/L, the liquor kalii iodide of 1000 μ g/L.
Beneficial effects of the present invention are embodied in：
1. in vitro detection urine of the present invention, the detection of the detectable of content of iodine is repeatable and accuracy is higher, and
Colour developing gradient and color stability are also very good.
2. in vitro detection urine of the present invention the detectable of content of iodine using simple to operate, reaction detection is time-consuming few, nothing
Multiprocessing, simple and fast need to be done to urine sample.
3. the present invention does not contain heavy metal compound and aldehyde condensate, is difficult to environment, safety and environmental protection, the present invention
By the detection by quantitative to content of iodine in urine, testing result is used for the Iodine nutrition situation for evaluating detected crowd, it is adaptable to
The serious basic medical unit popularization and application of iodine deficiency.
Specific embodiment
With reference to embodiment, the invention will be further described：
Embodiment 1
The detectable of content of iodine in a kind of vitro detection urine, including oxidant, developer, diluent and urine locate in advance
Reason agent；
The oxidant, by mass percentage concentration, consists of the following composition：Glacial acetic acid 20%, hydrogen peroxide 15%, hydrochloric acid
5%th, sulphuric acid 5%, quinolinic acid 2.5%, purification water surplus, preparation method is：Each raw material is taken, is fully mixed, you can standby；
The developer is the TMB alcoholic solutions that mass percentage concentration is 0.1%, and preparation method is：TMB is taken, is added anhydrous
Ethanol, is sufficiently stirred for dissolving, then seals, lucifuge, away from oxidant, 2～8 DEG C of preservations；
The diluent is the citric acid-sodium citrate buffer solution of pH=4.0, and preparation method is：0.01mol/ is prepared first
The sodium citrate solution of the citric acid solution and 0.01mol/L of L, then by the citric acid solution for preparing and sodium citrate solution
Mix, pH value is surveyed to 4.0,2～8 DEG C of sealing preserves；
The urine pretreating agent is activated carbon, and the activated carbon first passes through following methods in advance and processed：Take activated carbon 50g
Be placed in beaker, be poured slowly into dehydrated alcohol 100ml, then 10min is stirred on magnetic stirring apparatuss, stand, fill activated carbon
Divide sedimentation, abandon supernatant and float, add purified water and be sufficiently stirred for 5min, stand, activated carbon is fully settled, supernatant discarded
After carry out sucking filtration, the activated carbon after sucking filtration is placed in be put into 50 DEG C dry 5 hours, take out sealing preserve.
Embodiment 2
The detectable of content of iodine in a kind of vitro detection urine, including oxidant, developer, diluent and urine locate in advance
Reason agent；
The oxidant, by mass percentage concentration, consists of the following composition：Glacial acetic acid 10%, hydrogen peroxide 20%, hydrochloric acid
0.1%th, sulphuric acid 10%, quinolinic acid 0.1%, purification water surplus, preparation method is：Each raw material is taken, is fully mixed, you can standby；
The developer is the TMB alcoholic solutions that mass percentage concentration is 0.025%, and preparation method is：TMB is taken, nothing is added
Water-ethanol, is sufficiently stirred for dissolving, then seals, lucifuge, away from oxidant, 2～8 DEG C of preservations；
The diluent is the Tris buffer of pH=3.0, and preparation method is：The trihydroxy methyl of 0.01mol/L is prepared first
The hydrochloric acid solution of aminomethane solution and 0.01mol/L, then will be the tris solution for preparing and hydrochloric acid molten
Liquid is mixed, and surveys pH value to 3.0,2～8 DEG C of sealing preserves；
The urine pretreating agent is activated carbon, and the activated carbon first passes through following methods in advance and processed：Take activated carbon 50g
Be placed in beaker, be poured slowly into dehydrated alcohol 100ml, then 10min is stirred on magnetic stirring apparatuss, stand, fill activated carbon
Divide sedimentation, abandon supernatant and float, add purified water and be sufficiently stirred for 5min, stand, activated carbon is fully settled, supernatant discarded
After carry out sucking filtration, the activated carbon after sucking filtration is placed in be put into 50 DEG C dry 5 hours, take out sealing preserve.
Embodiment 3
The detectable of content of iodine in a kind of vitro detection urine, including oxidant, developer, diluent and urine locate in advance
Reason agent；
The oxidant, by mass percentage concentration, consists of the following composition：Glacial acetic acid 30%, hydrogen peroxide 1%, hydrochloric acid
10%th, sulphuric acid 0.1%, quinolinic acid 5%, purification water surplus；Preparation method is：Each raw material is taken, is fully mixed, you can standby；
The developer is the TMB alcoholic solutions that mass percentage concentration is 0.2%, and preparation method is：TMB is taken, is added anhydrous
Ethanol, is sufficiently stirred for dissolving, then seals, lucifuge, away from oxidant, 2～8 DEG C of preservations；
The diluent is the acetate buffer of pH=5.0, and preparation method is：The acetic acid for first preparing 0.01mol/L is molten
Then the acetic acid solution for preparing and sodium acetate solution are mixed by the sodium acetate solution of liquid and 0.01mol/L, survey pH value to 5.0,
2～8 DEG C of sealing preserves；
The urine pretreating agent is activated carbon, and the activated carbon first passes through following methods in advance and processed：Take activated carbon 50g
Be placed in beaker, be poured slowly into dehydrated alcohol 100ml, then 10min is stirred on magnetic stirring apparatuss, stand, fill activated carbon
Divide sedimentation, abandon supernatant and float, add purified water and be sufficiently stirred for 5min, stand, activated carbon is fully settled, supernatant discarded
After carry out sucking filtration, the activated carbon after sucking filtration is placed in be put into 50 DEG C dry 5 hours, take out sealing preserve.
Embodiment 4
The urine iodine detecting step of the detectable of content of iodine in vitro detection urine of the present invention
The preparation of 4.1 iodine titers
A, 1000 μ g/L titers are prepared：0.0131g potassium iodide is weighed, and with 100ml being settled to after purification water dissolution, then is taken
10ml adds purified water to be dissolved to 1L, and 2-8 DEG C keeps in dark place.
B, 50 μ g/L titers are prepared：Taking a liquid 5ml adds purified water to be settled to 100ml, i.e. concentration for 150 μ g/L iodine standards
Solution.
C, 100 μ g/L titers are prepared：Taking a liquid 10ml adds purified water to be settled to 100ml, i.e. concentration for 150 μ g/L iodine marks
Quasi- solution.
D, 250 μ g/L titers are prepared：Taking a liquid 25ml adds purified water to be settled to 100ml, i.e. concentration for 150 μ g/L iodine marks
Quasi- solution.
E, 400 μ g/L titers are prepared：Taking a liquid 40ml adds purified water to be settled to 100ml, i.e. concentration for 150 μ g/L iodine marks
Quasi- solution.
F, 500 μ g/L titers are prepared：Taking a liquid 50ml adds purified water to be settled to 100ml, i.e. concentration for 150 μ g/L iodine marks
Quasi- solution.
4.2 detecting step
1) sample treatment：Take 1ml diluents to be added in 1ml urines (urina sanguinis middle segment urine sample is excellent), add 1.8g urine
Liquid pretreating agent, shakes up, filtration treatment, obtains purification urine, standby；
2) sample measurement：To in purification urine, 150ul developers are added immediately, is mixed, is added 30ul oxidants, mixed
After even, it is immediately placed in analyser detector (such as spectrophotometer), during accurate response 60 seconds, colorimetric determination reads sample
The content of iodine in concentration value, as urine.
Note：If urine iodine concentration is more than 500 μ g/L, then diluted at double, detected that income value should be multiplied by by the above process
Corresponding extension rate.
Embodiment 5
The analysis of the accuracy of the detectable of content of iodine in vitro detection urine of the present invention
Accuracy is calculated as follows：
In formula：M is measurement result average；T is control liquid target
Value；B is relative deviation.
In formula：CV is the coefficient of variation；For the meansigma methodss of measured value；S
For standard deviation.
Using the detectable of content of iodine in the vitro detection urine of the embodiment of the present invention 1 to iodine composition in Lyophilized Human Urine point
Analysis standard substance (GBW09108-GBW09110) (142 μ g/L) carries out replication 10 times, the results are shown in Table shown in 1：
Table 1
| Urine iodine reference material | Survey 1 | Survey 2 | Survey 3 | Survey 4 | Survey 5 | Survey 6 | Survey 7 | Survey 8 | Survey 9 | Survey 10 | Average |
| 142(μg/L) | 142 | 145 | 144 | 138 | 146 | 144 | 152 | 139 | 136 | 141 | 142 |
Using the detectable of content of iodine in the vitro detection urine of the embodiment of the present invention 1 to Lyophilized Human Urine iodine component analyses
National standard material (142 μ g/L) carries out replication 20 times, as a result shown in table 2：
Table 2
Through calculating：The detectable of content of iodine in vitro detection urine of the present invention
As can be seen here, urine iodine can be effectively improved using the detectable of content of iodine in vitro detection urine of the present invention
Detection accuracy, and it is simple to operate, and reaction detection is time-consuming few.
Operated by automatization's detection by quantitative using the present invention, detection is repeatable and accuracy is higher, the present invention：
Minimum detectability：20μg/L.
The range of linearity：50～500 μ g/L, linearly dependent coefficient：r≥0.990.
Withinrun precision：Coefficient of variation CV≤10%.
Betweenrun precision：Coefficient of variation CV≤15%.
Accuracy：Relative deviation B%≤15%.
Specificity：The protein of 25mg/dl, the glucose of 50mg/dl, the bilirubin of 0.5mg/dl, the acetyl of 5mg/dl
The nitrite of acetic acid, 0.03mg/dl, it is noiseless to detecting.
Embodiment 6
Pretreating effect checking test of the urine pretreating agent to urine in the present invention
Test point 3 groups of controls, first group be add the protein of 25mg, the glucose of 50mg, the bilirubin of 0.5mg,
The urine ketone bodies of 5mg, the 50ug/L of the nitrite of 0.03mg, the urine mark of the 158ug/L content of iodine of vitamin C 0.6mmol/L
This；Second group of urine specimen for 158ug/L content of iodine；The 3rd group of aqueous solution specimen for 158ug/L content of iodine.
The detectable of content of iodine in vitro detection urine of the present invention is adopted to upper according to the operational approach that embodiment 4 is provided
Stating three groups of samples carries out the measure of content of iodine respectively, and as a result the result for three groups of detections is identical, determines and show that content of iodine is
158ug/L, in showing the present invention, urine pretreating agent can effectively remove protein, glucose, bilirubin, urine ketone in urine
Body, nitrite, vitamin C etc. affect the composition of detection.
Embodiment 7
The colour developing gradient checking of the detectable of content of iodine in vitro detection urine of the present invention
Using content of iodine in the vitro detection urine of the embodiment of the present invention 1 detectable to 70ug/L, 200ug/L,
The urine specimen of 400ug/L content of iodine carries out multigroup Iodine assay respectively, as a result see the table below 3
Table 3
From the results shown in Table 3, in vitro detection urine of the present invention, the colour developing gradient of the detectable of content of iodine is non-
Chang Hao, and the color stability of this reagent is also very good, and the multiple result of replication is all.
It should be understood that example as herein described and embodiment are not limited to the present invention, this area only for explanation
Technical staff can make various modifications or change according to it, all any modifications within the spirit and principles in the present invention, made,
Equivalent, improvement etc., should be included within the scope of the present invention.
Claims (7)
1. in a kind of vitro detection urine content of iodine detectable, it is characterised in that including oxidant and developer；
The oxidant, by mass percentage concentration, consists of the following composition：
The developer is TMB alcoholic solutions that mass percentage concentration is 0.025%～0.2%.
2. in vitro detection urine as claimed in claim 1 content of iodine detectable, it is characterised in that the TMB alcohol is molten
The solvent of liquid is ethanol.
3. in vitro detection urine as claimed in claim 1 or 2 content of iodine detectable, it is characterised in that also including dilute
Liquid is released, the diluent is the citric acid-sodium citrate buffer solution or acetate buffer of pH=3.0～5.0 or Tris bufferings
Liquid.
4. in vitro detection urine as claimed in claim 1 or 2 content of iodine detectable, it is characterised in that also including urine
Liquid pretreating agent, the urine pretreating agent are activated carbon.
5. in vitro detection urine as claimed in claim 4 content of iodine detectable, it is characterised in that the activated carbon is pre-
First pass through following methods to process：Dehydrated alcohol is added in activated carbon, is stood after stirring, make activated carbon fully sink
Drop, then abandons supernatant and float, adds purified water, stood after stirring, activated carbon is fully settled, then abandon
Supernatant is removed, sucking filtration process is carried out afterwards, finally the activated carbon after sucking filtration is dried.
6. in vitro detection urine as claimed in claim 1 or 2 content of iodine detectable, it is characterised in that also including iodine
Titer.
7. the detectable of content of iodine in the vitro detection urine that such as claim 6 is stated, it is characterised in that the iodine titer bag
Include the liquor kalii iodide of 50 μ g/L, the liquor kalii iodide of 100 μ g/L, the liquor kalii iodide of 250 μ g/L, the potassium iodide of 400 μ g/L
Solution, the liquor kalii iodide of 500 μ g/L, the liquor kalii iodide of 1000 μ g/L.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107831161A (en) * | 2017-12-06 | 2018-03-23 | 广东轻工职业技术学院 | Small molecule containing acetate is used as the new application of catalyst as class peroxidase |
| WO2020199630A1 (en) * | 2019-04-01 | 2020-10-08 | 深圳大学 | Test kit for iodine content in urine and test method for iodine content in urine |
| CN113959822A (en) * | 2021-10-29 | 2022-01-21 | 珠海市丽拓生物科技股份有限公司 | Diluent and oxidant for determining urine iodine content by peroxyacetic acid oxidation method and application |
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| US20030203495A1 (en) * | 2002-04-25 | 2003-10-30 | Rupp Michael E. | Diagnostic test for elemental imbalances |
| DE19507685B4 (en) * | 1995-03-04 | 2005-05-25 | Merck Patent Gmbh | Method and means for the determination of iodide |
| CN1811394A (en) * | 2006-03-07 | 2006-08-02 | 北京中生金域诊断技术有限公司 | Urine iodine test reagent kit |
| CN105004719A (en) * | 2015-07-21 | 2015-10-28 | 杨先锋 | Urine iodine determination kit and applications thereof |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4220046C1 (en) * | 1992-06-22 | 1993-09-02 | Willmen, Gottfried, 47906 Kempen, De | |
| DE19507685B4 (en) * | 1995-03-04 | 2005-05-25 | Merck Patent Gmbh | Method and means for the determination of iodide |
| US20030203495A1 (en) * | 2002-04-25 | 2003-10-30 | Rupp Michael E. | Diagnostic test for elemental imbalances |
| CN1811394A (en) * | 2006-03-07 | 2006-08-02 | 北京中生金域诊断技术有限公司 | Urine iodine test reagent kit |
| CN105004719A (en) * | 2015-07-21 | 2015-10-28 | 杨先锋 | Urine iodine determination kit and applications thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107831161A (en) * | 2017-12-06 | 2018-03-23 | 广东轻工职业技术学院 | Small molecule containing acetate is used as the new application of catalyst as class peroxidase |
| WO2020199630A1 (en) * | 2019-04-01 | 2020-10-08 | 深圳大学 | Test kit for iodine content in urine and test method for iodine content in urine |
| CN113959822A (en) * | 2021-10-29 | 2022-01-21 | 珠海市丽拓生物科技股份有限公司 | Diluent and oxidant for determining urine iodine content by peroxyacetic acid oxidation method and application |
| CN113959822B (en) * | 2021-10-29 | 2024-02-20 | 珠海市丽拓生物科技股份有限公司 | Diluent and oxidant for measuring iodine content in urine by peroxyacetic acid oxidation method and application |
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Address after: 230088 Hefei province high tech Zone, East Road, No. 18 diving Applicant after: Anhui Huibang Biological Engineering Co., Ltd. Address before: 230088 Hefei province high tech Zone, East Road, No. 18 diving Applicant before: Anhui Hui Bang bioengineering incorporated company |
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Application publication date: 20170329 |