CN1322947A - Mecury compound testing strip based on enzyme inhibiting reaction - Google Patents
Mecury compound testing strip based on enzyme inhibiting reaction Download PDFInfo
- Publication number
- CN1322947A CN1322947A CN 01118580 CN01118580A CN1322947A CN 1322947 A CN1322947 A CN 1322947A CN 01118580 CN01118580 CN 01118580 CN 01118580 A CN01118580 A CN 01118580A CN 1322947 A CN1322947 A CN 1322947A
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- reaction
- detection
- strip
- mercury
- detection strip
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- 238000006243 chemical reaction Methods 0.000 title claims abstract description 27
- 238000012360 testing method Methods 0.000 title claims abstract description 27
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 8
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 8
- 150000001875 compounds Chemical class 0.000 title description 2
- 230000002401 inhibitory effect Effects 0.000 title 1
- 238000001514 detection method Methods 0.000 claims abstract description 25
- 108010046334 Urease Proteins 0.000 claims abstract description 9
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000004202 carbamide Substances 0.000 claims abstract description 8
- 150000002731 mercury compounds Chemical class 0.000 claims abstract description 5
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims abstract description 4
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims abstract description 4
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims abstract description 4
- 238000011161 development Methods 0.000 claims abstract description 3
- 229920003023 plastic Polymers 0.000 claims description 4
- 229920002307 Dextran Polymers 0.000 claims description 2
- 239000000654 additive Substances 0.000 claims description 2
- 230000000996 additive effect Effects 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims 1
- 238000004451 qualitative analysis Methods 0.000 claims 1
- 238000004445 quantitative analysis Methods 0.000 claims 1
- 229910052753 mercury Inorganic materials 0.000 abstract description 12
- 229920002301 cellulose acetate Polymers 0.000 abstract description 9
- 230000007613 environmental effect Effects 0.000 abstract description 7
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 abstract description 7
- -1 mercury ions Chemical class 0.000 abstract description 6
- 239000012488 sample solution Substances 0.000 abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 5
- 239000012528 membrane Substances 0.000 abstract description 4
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- 229920006267 polyester film Polymers 0.000 abstract description 3
- 230000002596 correlated effect Effects 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 13
- 238000000034 method Methods 0.000 description 8
- 229940088598 enzyme Drugs 0.000 description 5
- 239000000243 solution Substances 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000011159 matrix material Substances 0.000 description 3
- 238000002482 cold vapour atomic absorption spectrometry Methods 0.000 description 2
- 238000002530 cold vapour atomic fluorescence spectroscopy Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 229940100892 mercury compound Drugs 0.000 description 2
- 238000004611 spectroscopical analysis Methods 0.000 description 2
- 108010015776 Glucose oxidase Proteins 0.000 description 1
- 239000004366 Glucose oxidase Substances 0.000 description 1
- JJWSNOOGIUMOEE-UHFFFAOYSA-N Monomethylmercury Chemical compound [Hg]C JJWSNOOGIUMOEE-UHFFFAOYSA-N 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 238000001479 atomic absorption spectroscopy Methods 0.000 description 1
- 238000001636 atomic emission spectroscopy Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- PJDVOLYULHZZAG-UHFFFAOYSA-N ethylmercury Chemical compound CC[Hg] PJDVOLYULHZZAG-UHFFFAOYSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000008239 natural water Substances 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
Images
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- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
本发明属于分析测试领域,涉及一种利用酶抑制显色反应进行定量分析汞化合物的检测条的研制。检测条由聚酯膜载体、反应区两部分组成,反应区包括一片固定有脲酶的醋酸纤维素膜和一片吸附了尿素及海藻糖的精密pH试纸。使用时,直接将检测条插入要检测的样品溶液中,使溶液没过反应区,两分钟后,可以在pH试纸的边缘看到蓝色环出现,随着反应的进行,蓝色环逐渐向中心膨胀,反应区完全变蓝的时间与样品溶液中汞化合物的浓度成正相关。本检测条的特点是灵敏、特异性高、成本低廉和使用方便。本检测条适用于环境水样中无机汞离子和有机汞的痕量检测。
The invention belongs to the field of analysis and testing, and relates to the development of a detection strip for quantitatively analyzing mercury compounds by using enzymes to inhibit color reaction. The detection strip is composed of polyester film carrier and reaction area. The reaction area includes a cellulose acetate membrane immobilized with urease and a precision pH test paper adsorbed with urea and trehalose. When in use, directly insert the test strip into the sample solution to be detected, so that the solution does not pass through the reaction area. After two minutes, a blue ring can be seen on the edge of the pH test paper. As the reaction progresses, the blue ring gradually The center swells, and the time for the reaction zone to completely turn blue is positively correlated with the concentration of mercury compounds in the sample solution. The detection strip is characterized by sensitivity, high specificity, low cost and convenient use. This test strip is suitable for the trace detection of inorganic mercury ions and organic mercury in environmental water samples.
Description
The invention belongs to the technical field of analysis and test, and relates to development of a detection strip for quantitatively analyzing mercury compounds by using an enzyme inhibition color reaction. The test strip consists of two parts, namely a polyester film carrier and a reaction area, wherein the reaction area comprises a cellulose acetate film fixed with urease and a piece of precision pH test paper adsorbing urea and trehalose. When the test strip is used, the test strip is directly inserted into a sample solution to be tested, so that the solution is submerged in a reaction area, a blue ring can be seen at the edge of a pH test paper after two minutes, the blue ring gradually expands towards the center along with the reaction, and the time for the reaction area to completely turn blue is in positive correlation with the concentration of mercury ions in the sample solution. The detection strip has the characteristics of sensitivity, high specificity, low cost and convenient use. The detection strip is suitable for trace detection of mercury ions in an environmental water sample.
The background and current state of the art is generally as follows: at present, the detection of mercury ions in an environmental water sample is mainly realized by methods such as atomic absorption spectroscopy, atomic emission spectroscopy and the like. Although these methods have high sensitivity and accuracy, they require expensive large-scale instruments, complicated sample pretreatment processes, and high requirements on operators, which make them difficult to be used for field detection of environmental samples. To address this problem, simple and sensitive enzyme inhibition assays are used for the detection of mercury. Commonly used enzymes are urease, peroxidase, glucose oxidase, etc., wherein urease is the most studied one, and the measurement of mercury in a sample is mainly realized through the change of color, fluorescence and electrochemical properties caused by enzymatic reaction. Generally speaking, these methods have high sensitivity and selectivity, and are simple to operate. However, most of these methods cannot be used for direct detection of environmental samples due to interference of the matrix in the environmental samples, and still require appropriate pretreatment of the samples. The invention does not need to carry out complex pretreatment on the sample, does not need precise instruments and equipment, and does not need to master special professional skills by a user. References to this can be found in:
(1)W.L.Clevenger,B.W.Smith,and J.D.Winefordner,
Crit.Rev.Anal.Chem.,1997,27(1),1-26
(2)L._rgan and G.Johansson,Anal.Chim.Acta.,1981,
125,45-53
(3)C.Preininger,Mikrochim.Acta,1999,130:209-214
(4)T.N.Shekhovtsova,S.V.Chemetskaya,Anal.Lett.,
1994,27(15)2883-2898
(5)Amine A.,Cremisini C.,Palleschi G.,Mikrochim Acta,
1995,121,183-190
the invention aims to provide a sensitive mercury ion detection strip based on the enzyme inhibition reaction and capillary action principle, which can directly detect a liquid sample, and the stability and the repeatability of the detection strip can meet the requirements of quantitative analysis.
The technical scheme of the invention is realized by the following modes: FIG. 1 is a schematic illustration of such a test strip. In the figure, a is a polyester film carrier, b is a reaction zone, and the reaction zone consists of 1 and 3 films. 1 is cellulose acetate film, and a certain amount of urease is adsorbed and fixed on the surface contacting with 3. 3 is a piece of precision pH test paper, on which a certain amount of urea and additive trehalose or dextran are adsorbed in advance. The reaction area is sealed by viscous transparent plastic 4, and a small hole 2 with the diameter of 1-1.5mm is reserved on the viscous transparent plastic on one side of the cellulose acetate membrane as a sample inlet. When the test strip is used, the test strip is inserted into a liquid sample, the solution is rapidly diffused and soaked in the cellulose acetate film through the sample inlet due to the capillary action of the cellulose acetate film, a large-size matrix in the solution is filtered by the cellulose acetate film and cannot enter a reaction area, and small-molecular inorganic or organic mercury enters the reaction area to be combined with a sulfhydryl group on urease, so that the activity of the enzyme is inhibited. The solution diffused to the edge of the cellulose acetate membrane permeates into the capillary of the pH test paper,then the urea adsorbed on the pH test paper is dissolved out by diffusing the urea from the edge to the center, and the dissolved urea is diffused to a cellulose acetate membrane and is catalyzed and hydrolyzed by urease in the following modes: OH-the yield of (a) is inversely proportional to the degree to which urease is inhibited. Since urea is firstly separated out from the edge of the pH test paper, OH is formed-Firstly, the pH test paper is generated at the edge, so that the periphery of the pH test paper is firstly blue, and the phenomenon of yellow spots of blue ring sleeves is formed. FIG. 2 is photographs taken 11 minutes, 13 minutes, and 14 minutes after the test strip was inserted into the sample solution, respectively, and numbered A, B, C in that order. As can be seen from FIG. 2, the blue ring gradually expanded toward the center and the yellow stain gradually shrunk as the reaction proceeded. The reaction area is completely disappeared to be used as a reaction end point, and the time from the insertion of the test strip into the sample to the reaction end point is in positive correlation with the content of mercury in the sample. According to the color reaction time, the content of the mercury ions in the sample can be obtained from a pre-drawn time-concentration curveAmount of the compound (A).
Table 1 shows the comparison between the present invention and conventional spectroscopic methods (cold vapor atomic absorption spectroscopy, CV-AAS; cold vapor atomic fluorescence spectroscopy, CV-AFS), and it can be seen through the comparison that the present invention is simple in operation, easy to popularize, and can complement the spectroscopic methods to effectively monitor the mercury pollution condition in the environmental water sample on site.
In the process of completing the invention, the main indexes of the test strip are tested repeatedly. The test result shows that the pH range of the test strip is 6.5-7.6, and the temperature range is 25-35 ℃. Under optimized experimental conditions, the detection strip is used for detecting Hg in an aqueous solution2+The detection limit can reach 0.2ng/ml, the detection range is 0.2-200ng/ml, and the RSD is less than or equal to 10 percent. The detection results of methyl mercury and ethyl mercury are similar to those of inorganic mercury, andpb2+、Cu2+、Cd2+、Zn2+、Ni2+and the common heavy metal ions have no obvious interference on detection. The experiment of adding drinking water and natural water shows that the matrix in the water sample has no obvious influence on the measurement of the detection strip.
TABLE 1 comparison of the present invention with conventional spectral analysis methods
The technical skill of the detection limit selective field detection capability requires that the pretreatment requires analysis of the speed Cv-AAs 0.02ng/ml good-difference high-height fast Cv-AFs 0.001ng/ml good-difference high-height fast method 0.2ng/ml better,none or little low and fast
Claims (4)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01118580 CN1210556C (en) | 2001-06-05 | 2001-06-05 | Mecury compound testing strip based on enzyme inhibiting reaction |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01118580 CN1210556C (en) | 2001-06-05 | 2001-06-05 | Mecury compound testing strip based on enzyme inhibiting reaction |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1322947A true CN1322947A (en) | 2001-11-21 |
| CN1210556C CN1210556C (en) | 2005-07-13 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 01118580 Expired - Fee Related CN1210556C (en) | 2001-06-05 | 2001-06-05 | Mecury compound testing strip based on enzyme inhibiting reaction |
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| Country | Link |
|---|---|
| CN (1) | CN1210556C (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100451628C (en) * | 2005-11-17 | 2009-01-14 | 中国科学院兰州化学物理研究所 | A kind of colorimetric detection analysis method of Hg2+ ion |
| CN101545900B (en) * | 2009-05-15 | 2012-09-19 | 北京科技大学 | A biodetector for rapid detection of mercury compounds |
| CN102776266A (en) * | 2012-07-26 | 2012-11-14 | 广州绿洲生化科技股份有限公司 | Rapid testing card for heavy metal mercury and testing method thereof |
-
2001
- 2001-06-05 CN CN 01118580 patent/CN1210556C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100451628C (en) * | 2005-11-17 | 2009-01-14 | 中国科学院兰州化学物理研究所 | A kind of colorimetric detection analysis method of Hg2+ ion |
| CN101545900B (en) * | 2009-05-15 | 2012-09-19 | 北京科技大学 | A biodetector for rapid detection of mercury compounds |
| CN102776266A (en) * | 2012-07-26 | 2012-11-14 | 广州绿洲生化科技股份有限公司 | Rapid testing card for heavy metal mercury and testing method thereof |
| CN102776266B (en) * | 2012-07-26 | 2015-03-18 | 广州绿洲生化科技股份有限公司 | Rapid testing card for heavy metal mercury and testing method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1210556C (en) | 2005-07-13 |
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| Date | Code | Title | Description |
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| C06 | Publication | ||
| PB01 | Publication | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: Beijing Primebiotek Company Limited Assignor: Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences Contract record no.: 2011990000880 Denomination of invention: Mecury compound testing strip based on enzyme inhibiting reaction Granted publication date: 20050713 License type: Exclusive License Open date: 20011121 Record date: 20110908 |
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| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20050713 Termination date: 20130605 |