CN103341360B - Nano material analogue enztme and mercury ion detecting application thereof - Google Patents
Nano material analogue enztme and mercury ion detecting application thereof Download PDFInfo
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- CN103341360B CN103341360B CN201310260670.2A CN201310260670A CN103341360B CN 103341360 B CN103341360 B CN 103341360B CN 201310260670 A CN201310260670 A CN 201310260670A CN 103341360 B CN103341360 B CN 103341360B
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Abstract
The invention provides nano material analogue enztme and the application in mercury ion detecting thereof.After mercury ion mixes with silver nano material there is the various features substrate of lower catalytic oxidation horseradish peroxidase in the aerial oxygen of energy.Thus, can as nano material analogue enztme.The activity of the analogue enztme that mercury ion and silver nano material are formed has direct relation with the concentration of the mercury ion added, and therefore, can measure the concentration of mercury ion according to the spectral signal (absorption spectrum or fluorescence spectrum) of the oxidation product of feature substrate.The present invention provide not only a kind of simulation oxidizing ferment material newly, simultaneously this system can easy, delicately detect mercury ion have selective preferably.The range of linearity that the present invention detects mercury ion is 1.0 × 10
-7mol/L-1.2 × 10
-4mol/L, detects and is limited to 20nM.
Description
Technical field:
The present invention relates to nanosecond science and technology field, particularly relate to novel nano-material analogue enztme and detecting the application in mercury ion.
Background technology:
Enzyme is the basis of all life activity, is the catalyst that in organism, various biochemical reaction carries out, in the physiological activity maintaining organism and metabolic processes, play vital effect.But, the native enzyme purification difficult in organism, expensive, and it is generally to acid, alkali, thermally labile, and easy inactivation sex change, this all limits their extensive use.Therefore, imitating the analogue enztme research of native enzyme to the molecular recognition of substrate and efficient catalytic is a research field highly significant.
[GaoLZ since Yan Xiyun research group Late Cambrian ferroferric oxide nano granules has the catalytic activity of similar peroxidase; ZhuangJ, ZhangJB; ZhangY, GuN, WangTH, FengJ, YangD.L, PerrettS, YanX.NatNanotechnol, 2007,2 (9): 577-583] exploration of nano material mimetic enzyme catalysis activity causes at once and shows great attention to.Afterwards, people found that multiple nano material was as Co successively
3o
4[MuJH, WangY, ZhaoM, ZhangL.Chem.Commun, 2012,48, (2) 540-2542], carbon quantum dot [ShiWB, WangQL, LongYJ, ChengZL, ChenSH, ZhengHZ, HuangYM.Chem.Commun, 2011,47,6695-6697], ceria [JiaoX, SongHJ, ZhaoHH, BaiW, ZhangLC, LvY.Anal.Methods, 2012,4,3261-3267.], FeS [DaiZ, LiuS, BaoJ, Ju, H.Chem.Eur.J, 2009,15,4321-4326.], V
2o
5[Andr é R, Nat á lioF, HumanesM, LeppinJ, HeinzeK, WeverR,
hC, M ü llerWE.G, TremelW.Adv.Funct.Mater.2011,21,501-509.] etc. be also found to have the activity of similar peroxidase, i.e. the feature substrate of catalytic oxidation enzyme in the presence of hydrogen peroxide.Nanoparticle analogue enztme becomes the new study hotspot in one, Analytical Chemistry in Life Science field.The aspects such as the degraded of current bio-sensing, environmental contaminants, electrochemical sensor have potential application widely [JvY, LiB, CaoR.Chem.Commun, 2010,46 (42): 8017-8019; HeW, WuX, LiuJ, etal.ChemMater, 2010,22 (9): 2988-2994; DingN, YanN, RenC, etal.AnalChem, 2010,82 (13): 5897-5899].
Hydrogen peroxide can not be used as oxidant, utilize merely the quasi-enzyme catalytic agent of the dioxygen oxidation feature substrate in air to become simulation oxidizing ferment.Take a hint from the research of fuel cell, it is found that bimetal nano material [LiuJB, HuXN, HouSA, WenT, LiuWQ, ZhuX, WuXC.Chem.Commun, 2011,47, the 10981-10983 based on Pt material; ZhangK, HuXN, LiuJB, YinJJ, HouSA, WenT, HeWW, JiYL, GuoYT, WangQ, WuXC.Langmuir, 2011,27,2796-2803] there is oxidasic characteristic.But this type of catalyst is expensive.And the activity of this type of nano material analogue enztme is lower, many kinds of metal ions can suppress the activity of this type of nano material analogue enztme.The invention provides one more inexpensive and efficient nano material simulation oxidizing ferment.Meanwhile, this nano material analogue enztme can be conveniently used for measuring mercury ion, highly sensitive, selective good.
Summary of the invention:
The object of this invention is to provide a kind of new and effective ground nano material analogue enztme, this system can detect Hg easily simultaneously
2+.
Object of the present invention realizes by following technical measures:
After a, a certain amount of coating material mix with the water-soluble silver salt solution of 20ml0.00lmol/L, add the ultra-pure water of 50ml; Then a certain amount of NaBH is dropwise added
4, magnetic stirring apparatus stirs and spends the night, obtain silver nano material;
B, in 5ml colorimetric cylinder, at a certain temperature, add the silver nano material of certain volume, the mercury ion of variable concentrations, then add the cushioning liquid of the feature substrate of the horseradish peroxidase of certain volume and the 0.2mol/L of certain pH, liquor capacity is settled to 5ml.Silver nano material and mercury ion react the product that formed can the feature substrate of oxygen catalytic oxidation horseradish peroxidase under the existence of oxygen in atmosphere, produces the spectral signal (absorbing or fluorescence) of the oxidation product of feature substrate.Thus, can as nano material analogue enztme.Meanwhile, in this system the active size of analogue enztme and the amount of mercury ion linear.Therefore, can measure mercury ion according to the spectral signal of feature substrate oxidation product, show selective good, highly sensitive advantage.
Object of the present invention also realizes by following technical measures:
The coating material of described silver nano material is PEI, trisodium citrate, sodium polyphosphate, PMMA, and the amount of coating material is 0.5-10 times of the mole of silver; The feature substrate of described horseradish peroxidase has 3,4-dihydroxyphenyl acetic acid, 3,3 ', 5,5 '-tetramethyl benzidine (TMB), o-phenylenediamine (OPD), two (3-ethyl benzo thiazole phenanthroline-6-sulfonic acid) di-ammonium salts (ABTS) of 2,2 '-Lian nitrogen base, feature concentration of substrate is 0.1mM-10mM; The pH of the hac buffer used during catalytic oxidation feature substrate is 2.0-9.0; The temperature used during catalytic oxidation feature substrate is 10-70 DEG C.
accompanying drawing illustrates:
Fig. 1 is mercury ion (a), invents the silver nano material (c) of trisodium citrate modification prepared by the silver nano material (b) of the trisodium citrate modification of preparation and mercury ion and invention as analogue enztme and 1 × 10
-5the situation of MTMB differential responses time.
Fig. 2 be the silver nano material modified of the trisodium citrate of invention preparation and mercury ion reaction before the absorption spectrum of (b) after (a) and reaction.
Fig. 3 is the impact of nitrogen on the analogue enztme performance of system.
Fig. 4 is that system detects the selective of mercury ion.
Embodiment 1:
After the water-soluble silver salt solution mixing of a, 0.5 gram of PMMA and 20ml0.001mol/L, add the ultra-pure water of 50ml; Then a certain amount of NaBH is dropwise added
4, magnetic stirring apparatus stirs and spends the night, obtain silver nano material;
B, in 5mL colorimetric cylinder, at 25 DEG C, add the silver nano material of 1.0mL, add 1.0 × 10
-6the mercury ion of mol/L, then adds 1.0mL1.0 × 10
-3the acetate buffer solution of the 0.2mol/l of feature substrate TMB and 1.0mLpH=6.0 of mol/L, is settled to 5mL by liquor capacity.The absorbance of system is gone out to measure in the characteristic absorption (λ max=650nm) of the oxidation product of TMB.
Embodiment 2:
After a, 0.5 gram of trisodium citrate mix with the water-soluble silver salt solution of 20ml0.001mol/L, add the ultra-pure water of 50ml; Then a certain amount of NaBH is dropwise added
4, magnetic stirring apparatus stirs and spends the night, obtain silver nano material;
B, in 5mL colorimetric cylinder, at 25 DEG C, add the silver nano material of 1.0mL, add 1.0 × 10
-6the mercury ion of mol/L, then adds 1.0mL1.0 × 10
-3the acetate buffer solution of the 0.2mol/l of feature substrate OPD and 1.0mLpH=6.0 of mol/L, is settled to 5mL by liquor capacity.The absorbance of system is gone out to measure in the characteristic absorption (λ max=450nm) of the oxidation product of OPD.
Claims (3)
1., based on a mercury ion detecting method for nano material analogue enztme, it is characterized in that:
The one of the PEI of the mole of a, 0.5-10 silver doubly, trisodium citrate, sodium polyphosphate, PMMA, as coating material, after mixing, adds the ultra-pure water of 50ml with the water-soluble silver salt solution of 20ml0.001mol/L; Then a certain amount of NaBH is dropwise added
4, magnetic stirring apparatus stirs and spends the night, obtain silver nano material;
B, in 5ml colorimetric cylinder, at a certain temperature, add the silver nano material of certain volume, the mercury ion of variable concentrations, what then add 0.1mM-10mM takes from 3,4-dihydroxyphenyl acetic acid, 3,3 ', 5,5 '-tetramethyl benzidine, o-phenylenediamine, 2, a kind of hac buffer as the feature substrate of horseradish peroxidase and the 0.2mol/L of certain pH in two (3-ethyl benzo thiazole phenanthroline-6-sulfonic acid) di-ammonium salts of 2 '-Lian nitrogen base, is settled to 5ml by liquor capacity; Silver nano material and mercury ion react the product that formed can the feature substrate of oxygen catalytic oxidation horseradish peroxidase under the existence of oxygen in atmosphere, produces absorption or the fluorescence spectrum signal of the oxidation product of feature substrate; Thus, can as nano material analogue enztme; Meanwhile, in this system the active size of analogue enztme and the amount of mercury ion linear; Therefore, can measure mercury ion according to the spectral signal of feature substrate oxidation product.
2. a kind of mercury ion detecting method based on nano material analogue enztme according to claim 1, the pH of the hac buffer used when it is characterized in that catalytic oxidation feature substrate is 2.0-9.0.
3. a kind of mercury ion detecting method based on nano material analogue enztme according to claim 1, the temperature used when it is characterized in that catalytic oxidation feature substrate is 10-70 DEG C.
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| CN104308139B (en) * | 2014-09-22 | 2016-03-30 | 福建医科大学 | A kind of platinum nano material with four kinds of analogue enztme activity and preparation method thereof |
| CN104549234B (en) * | 2014-12-11 | 2016-11-02 | 江南大学 | A kind of CDs-Pt nano material with catalase catalysis characteristics |
| CN104897846B (en) * | 2015-06-23 | 2016-08-17 | 江南大学 | A kind of based on the alkaline phosphatase activities detection method being formed in situ non-photoactive nanoparticles material simulation enzyme |
| CN105675519B (en) * | 2016-04-01 | 2018-02-09 | 江南大学 | A kind of mercury ion detecting method |
| CN109482207B (en) * | 2018-11-12 | 2021-08-20 | 国家海洋局第一海洋研究所 | Ag3PO4Mimic enzyme material and application thereof |
| CN109482182A (en) * | 2018-11-12 | 2019-03-19 | 国家海洋局第海洋研究所 | A kind of Ag2WO4Analogue enztme material and its application |
| CN110044860A (en) * | 2019-05-09 | 2019-07-23 | 南宁师范大学 | The method for detecting acetylcholine ester enzyme concentration |
| CN110044859A (en) * | 2019-05-09 | 2019-07-23 | 南宁师范大学 | The method of ratio fluorescent detection acetylcholine |
| CN111337485A (en) * | 2020-03-19 | 2020-06-26 | 江苏科技大学 | Hexavalent chromium colorimetric detection method based on silver nanocluster nanoenzyme |
| CN113029988B (en) * | 2021-04-14 | 2022-09-16 | 贵阳海关综合技术中心(贵州国际旅行卫生保健中心、贵阳海关口岸门诊部) | Based on inhibiting porous Co 3 O 4 Method for detecting multiple heavy metals with nano-disc catalytic activity |
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