CN103616420B - The Optical Electro-Chemistry assay method of glucose in serum - Google Patents
The Optical Electro-Chemistry assay method of glucose in serum Download PDFInfo
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Abstract
The invention provides the Optical Electro-Chemistry assay method of glucose in serum.The method is by preparing the nickel oxide nano material modified electrode of water soluble semiconductor sulfide (composition general formula is ES) nano material sensitization, and this electrode has good photogenerated charge transfer efficiency.Utilize layer assembly method glucose oxidase to be fixed to the nickel oxide electrode surface of semiconductor sulfide sensitization, the amount of glucose oxidase accurately can be controlled by the assembling number of plies.The glucose secured in the electrode pair solution of glucose oxidase has sensitive Optical Electro-Chemistry response.Further, to the mensuration of glucose, there is good selectivity.Present method solves the drawback that existing Optical Electro-Chemistry measures glucose selective difference, to the mensuration of glucose in serum, there is better effects.
Description
Technical field:
The present invention relates to analysis detection field, the Optical Electro-Chemistry particularly relating to a kind of glucose in serum measures new method.
Background technology:
As everyone knows, diabetes are one of diseases of serious harm human health in world wide.300,006,006 million peoples within 2011, are had to suffer from diabetes, and, expect the year two thousand thirty diabetic and will reach 500,005,005 million people [WhitingDR, GuariguataL, WeilC, etal.DiabetesRes.Cli.Pract., 2011,94 (3): 311-321.].The height of serum glucose level is the important indicator of human health status.Limosis vein blood slurry blood sugar concentration is called hypoglycemia lower than during 3.33 ~ 3.89mmol/L, and more than being called hyperglycaemia [LiX, ZhouY, ZhengZ, etal.Langmuir, 2009,25 (11): 6580-6586.] during 7.80mmol/L.Measuring content of blood sugar is one of most important index in clinical examination, diagnosis, observation of curative effect, course of disease detection, prevention from suffering from the diseases etc. that it can be the diseases such as diabetes, hypertension and cardio-cerebrovascular provide objective basis [JosephW.Chem.Rev., 2008,108 (2): 814-825.].Develop easy, glucose assays method is significant fast and accurately.
PhotoelectrochemicalMethod Method is the novel analytical approach of one [TokudomeH, YamadaY, SonezakiS, etal.Appl.Phys.Lett., 2005,87 (21): 213901-213901-3 that have just grown up recently; LiuSL, LiC, ChengJ, etal.Anal.Chem., 2006,78 (13): 4722-4726.].The testing process of Optical Electro-Chemistry and electrogenerated chemiluminescence are just in time contrary, and it adopts light signal to excite generation electrochemical signals.Owing to adopting multi-form exciting and detection signal, thus its background signal is lower, can reach the high sensitivity suitable with electrogenerated chemiluminescence.Further, the instrument compare of Optical Electro-Chemistry is simple, easily microminiaturized; Owing to adopting Electrochemical Detection, compare with optical detecting method, its equipment is more inexpensive.Optical Electro-Chemistry detection method very likely develops into portable, high-throughout timely detection system [WangP, GeL, GeS, etal.Chem.Commun., 2013,49 (32): 3294-3296.].Therefore, Optical Electro-Chemistry is a kind of very promising analytical approach, receives increasing concern [GillR, ZayatsM, WillnerI.Angew.Chem., Int.Ed., 2008,47 (40): 7602-7625; HaddourN, ChauvinJ, GondranC, etal.J.Am.Chem.Soc., 2006,128 (30): 9693-9698.WangGL, XuJJ, ChenHY, etal.Biosens.Bioelectron., 2009,25 (4): 791-796; LongYT, KongC, LiDW, etal.Small, 2011,7 (12): 1624-1628.].
Semiconductor nano material has special and the PhotoelectrochemicalProperties Properties of excellence [ZhuW, AnYR, LuoXM, etal.Chem.Commun., 2009,19,2682-2684; AnYR, TangLL, JiangXL, etal.Chem.-Eur.J., 2010,16,14439-14446; KangQ, YangLX, ChenYF, etal.Anal.Chem., 2010,82 (23): 9749-9754.], in Optical Electro-Chemistry sensor, there is potential broad prospect of application.In earlier stage, had people PhotoelectrochemicalMethod Method is used for glucose detection [TanneJ,
d, KhalidW, etal.Anal.Chem., 2011,83 (20): 7778-7785; ZhengM, CuiY, LiXY, etal.Electroanal.Chem., 2011,656,167-173; WangWJ, BaoL, LeiJP, etal.Anal.Chim.Acta, 2012,744,33-38.], but existing method is often subject to the interference of the reducing substanceses such as ascorbic acid, dopamine, sulfhydryl compound (halfcystine, glutathione).As everyone knows, living things system such as serum is more complicated, and the inside is often contained, compared with the reducing substances such as ascorbic acid, dopamine, sulfhydryl compound of multiple types.Existing method can not realize the mensuration to actual sample preferably.
The present invention has prepared the semiconductor nano material modified electrode with new structure, some common reducing substanceses of this electrode pair such as ascorbic acid, dopamine, sulfhydryl compound (halfcystine, glutathione) etc. have good antijamming capability, overcome the drawback of existing photoelectric method poor selectivity, good effect is achieved to the mensuration of glucose in serum.
Summary of the invention:
The object of this invention is to provide a kind of technique simple, selectivity is good, highly sensitive, measures Optical Electro-Chemistry fast and detects the method for glucose.
An object of the present invention realizes by following technical measures:
After the coating material of a, 10 milliliters of 0.2mol/L mixes with water-soluble metal (representing with the E) ion salt solution of 50 milliliters of 0.02mol/L, the pH of solution is regulated to be neutral by the NaOH solution of 1mol/L; Then, after passing into high pure nitrogen 30min, the Na of 0.1mol/L is added
2s aqueous solution, continues logical N
2stir, react 30 minutes-1.5 hours under uniform temperature, obtain water-soluble sulfide (general formula is expressed as ES) nano material;
B, by the mixture solution that is immersed in through pretreated ito glass sheet containing nickel nitrate and organic amine, react in the water-bath of uniform temperature after 10 minutes and use deionized water rinsing electrode surface; Then electrode is calcined 30 minutes at a certain temperature, obtain the ITO electrode that nickel oxide is modified;
C, the ITO electrode that obtained nickel oxide is modified to be immersed in synthesized water-soluble sulfide (ES) nanomaterial solution 30 minutes, to wash with water, obtain ITO/NiO/ES modified electrode; ITO/NiO/ES to be immersed in the solution of the high molecular polymer containing 0.5%-2% 30 minutes, after washing with water, drip 10 mul glucose oxidase solution reaction 30 minutes, said process can repeatedly, more glucose oxidase to be fixed to ITO/NiO/ES modified electrode surface;
D, the obtained ITO/NiO/ES modified electrode being fixed with glucose oxidase is put into the Tris-HCl buffer solution of the 0.1mol/L of certain pH as working electrode, add the serum solution containing glucose to be determined, under-0.2V current potential (the Ag/AgCl electrode relative to saturated), homemade Optical Electro-Chemistry instrument carries out the mensuration of photocurrent.
An object of the present invention also realizes by following technical measures:
The soluble-salt of the described metallic ion used when preparing water-soluble sulfide nano material is selected from zinc ion, cadmium ion, the soluble-salt of lead ion; The organic amine that the ITO electrode preparing nickel oxide modification uses, is selected from hexamethylenetetramine, octadecylamine, tert-butylamine, dimethylamine, triethylamine, triethanolamine, the temperature of described water-bath is 60-100 DEG C, and the calcining heat of described nickel oxide electrode is 250-450 DEG C; The time high molecular polymer that uses glucose oxidase being fixed to ITO/NiO/ES modified electrode surface is selected from PAMAM, PDDA, PAH, PEI; The pH of the buffer solution that described Optical Electro-Chemistry uses when measuring glucose is adjusted to 5.0-9.0.
Accompanying drawing illustrates:
Fig. 1 is scanning electron microscope (TEM) figure (A) and X-ray diffraction (XRD) figure (B) of the nickel oxide modification ITO electrode of invention preparation;
Fig. 2 is that the fixing number of plies of the glucose oxidase of invention preparation is on the impact of Optical Electro-Chemistry mensuration glucose;
Fig. 3 is that the modified electrode of invention preparation is to the selectivity of different material;
Embodiment:
Embodiment 1:
After the mercaptoacetic acid of a, 10 milliliters of 0.2mol/L mixes with the zinc nitrate solution of 50 milliliters of 0.02mol/L, the pH of solution is regulated to be neutral by the NaOH solution of 1mol/L; Then, after passing into high pure nitrogen 30min, the Na of 0.1mol/L is added
2s aqueous solution, continues logical N
2stir, react 1 hour at 50 DEG C, obtain water-soluble ZnS nano material;
B, by the mixture solution that is immersed in through pretreated ito glass sheet containing nickel nitrate and hexamethylenetetramine, react in the water-bath of 50 DEG C after 10 minutes and use deionized water rinsing electrode surface; Then electrode is calcined 30 minutes at 300 DEG C, obtain the ITO electrode that nickel oxide is modified;
C, the ITO electrode that obtained nickel oxide is modified to be immersed in synthesized water-soluble ZnS nanomaterial solution 30 minutes, to wash with water, obtain ITO/NiO/ZnS modified electrode; ITO/NiO/ZnS is immersed in containing 1% PDDA solution in 30 minutes, after washing with water, drip 10 mul glucose oxidase solution reaction 30 minutes, glucose oxidase in triplicate, is fixed to ITO/NiO/ZnS modified electrode surface by said process;
D, the obtained ITO/NiO/ZnS modified electrode being fixed with glucose oxidase is put into pH is that the Tris-HCl buffer solution of the 0.1mol/L of 7.0 is as working electrode, add the serum solution containing glucose to be determined, under-0.2V current potential (the Ag/AgCl electrode relative to saturated), homemade Optical Electro-Chemistry instrument carries out the mensuration of photocurrent.
Embodiment 2:
After the dimercaptosuccinic acid of a, 10 milliliters of 0.2mol/L mixes with the cadmium chloride solution of 50 milliliters of 0.02mol/L, the pH of solution is regulated to be neutral by the NaOH solution of 1mol/L; Then, after passing into high pure nitrogen 30min, the Na of 0.1mol/L is added
2s aqueous solution, continues logical N
2stir, react 30 minutes at 100 DEG C, obtain water soluble CdS nanomaterial;
B, by the mixture solution that is immersed in through pretreated ito glass sheet containing nickel nitrate and triethanolamine, react in the water-bath of 70 DEG C after 10 minutes and use deionized water rinsing electrode surface; Then electrode is calcined 30 minutes at 400 DEG C, obtain the ITO electrode that nickel oxide is modified;
C, the ITO electrode that obtained nickel oxide is modified to be immersed in synthesized water soluble CdS nanomaterial solution 30 minutes, to wash with water, obtain ITO/NiO/CdS modified electrode; ITO/NiO/CdS is immersed in containing 0.5% PEI solution in 30 minutes, after washing with water, drip 10 mul glucose oxidase solution reaction 30 minutes, said process repeats four times, more glucose oxidase to be fixed to ITO/NiO/CdS modified electrode surface;
D, the obtained ITO/NiO/CdS modified electrode being fixed with glucose oxidase is put into pH is that the Tris-HCl buffer solution of the 0.1mol/L of 5.0 is as working electrode, add the serum solution containing glucose to be determined, under-0.2V current potential (the Ag/AgCl electrode relative to saturated), homemade Optical Electro-Chemistry instrument carries out the mensuration of photocurrent.
Embodiment 3:
After the halfcystine of a, 10 milliliters of 0.2mol/L mixes with the lead nitrate solution of 50 milliliters of 0.02mol/L, the pH of solution is regulated to be neutral by the NaOH solution of 1mol/L; Then, after passing into high pure nitrogen 30min, the Na of 0.1mol/L is added
2s aqueous solution, continues logical N
2stir, react 30 minutes under room temperature, obtain water-soluble PbS nano material;
B, by the mixture solution that is immersed in through pretreated ito glass sheet containing nickel nitrate and diethylamine, react in the water-bath of 90 DEG C after 10 minutes and use deionized water rinsing electrode surface; Then electrode is calcined 30 minutes at 350 DEG C, obtain the ITO electrode that nickel oxide is modified;
C, the ITO electrode that obtained nickel oxide is modified to be immersed in synthesized water-soluble PbS nanomaterial solution 30 minutes, to wash with water, obtain ITO/NiO/PbS modified electrode; ITO/NiO/PbS is immersed in containing 1% PAH solution in 30 minutes, after washing with water, drip 10 mul glucose oxidase solution reaction 30 minutes, said process can repeat twice, more glucose oxidase to be fixed to ITO/NiO/PbS modified electrode surface;
D, the obtained ITO/NiO/PbS modified electrode being fixed with glucose oxidase is put into pH is that the Tris-HCl buffer solution of the 0.1mol/L of 8.0 is as working electrode, add the serum solution containing glucose to be determined, under-0.2V current potential (the Ag/AgCl electrode relative to saturated), homemade Optical Electro-Chemistry instrument carries out the mensuration of photocurrent.
Claims (5)
1. the Optical Electro-Chemistry assay method of glucose in serum, is characterized in that:
After the coating material of a, 10 milliliters of 0.2mol/L mixes with the water-soluble metal ion salt solusion of 50 milliliters of 0.02mol/L, the pH of solution is regulated to be neutral by the NaOH solution of 1mol/L; Then, after passing into high pure nitrogen 30min, the Na of 0.1mol/L is added
2s aqueous solution, continues logical N
2stir, react 30 minutes-1.5 hours under uniform temperature, obtain water-soluble sulfide nano material;
B, by the mixture solution that is immersed in through pretreated ito glass sheet containing nickel nitrate and organic amine, react in the water-bath of uniform temperature after 10 minutes and use deionized water rinsing electrode surface; Then electrode is calcined 30 minutes at a certain temperature, obtain the ITO electrode that nickel oxide is modified;
C, the ITO electrode that obtained nickel oxide is modified to be immersed in synthesized water-soluble sulfide nanomaterial solution 30 minutes, to wash with water, obtain the ITO electrode that NiO/ sulfide nano-material is modified; The ITO electrode that NiO/ sulfide nano-material is modified to be immersed in the solution of the high molecular polymer containing 0.5%-2% 30 minutes, after washing with water, drip 10 mul glucose oxidase solution reaction 30 minutes, said process can repeatedly, more glucose oxidase to be fixed to the ITO electrode surface that NiO/ sulfide nano-material is modified;
D, the ITO electrode that the obtained NiO/ sulfide nano-material being fixed with glucose oxidase is modified is put into the Tris-HCl buffer solution of the 0.1mol/L of certain pH as working electrode, add the serum solution containing glucose to be determined, under voltage relative to saturated Ag/AgCl electrode-0.2V, homemade Optical Electro-Chemistry instrument carries out the mensuration of photocurrent.
2. the Optical Electro-Chemistry assay method of glucose in serum according to claim 1, the soluble-salt of metallic ion used during synthesizing water-solubility sulfide nano-material is selected from the soluble-salt of zinc ion, cadmium ion, lead ion.
3. the Optical Electro-Chemistry assay method of glucose in serum according to claim 1, the organic amine selected when preparing the ITO electrode of nickel oxide modification, be selected from hexamethylenetetramine, octadecylamine, tert-butylamine, dimethylamine, triethylamine, triethanolamine, the temperature of described water-bath is 60-100 DEG C, and the calcining heat of the ITO electrode that described nickel oxide is modified is 250-450 DEG C.
4. the Optical Electro-Chemistry assay method of glucose in serum according to claim 1, the high molecular polymer used when glucose oxidase being fixed to the ITO electrode surface of NiO/ sulfide nano-material modification is selected from PAMAM, PDDA, PAH, PEI.
5. the Optical Electro-Chemistry assay method of glucose in serum according to claim 1, is characterized in that the pH of buffer solution when Optical Electro-Chemistry measures glucose is adjusted to 5.0-9.0.
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| CN105021672B (en) * | 2015-06-23 | 2017-10-27 | 江南大学 | PhotoelectrochemicalMethod Method detection dopamine based on in-situ oxidation reduction reaction |
| CN105929007A (en) * | 2016-06-15 | 2016-09-07 | 河南大学 | Method of construction for photoelectrochemical glucose oxidase sensor with graphite like g-C3N4-TiO2 nanosheet composite as enzymatic molecule immobilization scaffold |
| CN109239155A (en) * | 2018-09-04 | 2019-01-18 | 湖北工程学院 | The detection method of enzyme-free glucose optical electro-chemistry sensor, enzyme-free glucose concentration |
| CN109856209A (en) * | 2018-12-26 | 2019-06-07 | 济南大学 | ZnxBi2S3+xIt is sensitized NiTiO3The preparation method of self energizing glucose detection optical electro-chemistry sensor |
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| CN102645463A (en) * | 2012-03-31 | 2012-08-22 | 无锡百灵传感技术有限公司 | Preparation method for high-sensitivity amperometric glucose sensor |
| CN103424446A (en) * | 2013-08-13 | 2013-12-04 | 常州大学 | High-sensitivity non-enzymatic glucose electrochemical sensor and production method thereof |
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