CN103616420A - Photoelectrochemical determination method of glucose in serum - Google Patents
Photoelectrochemical determination method of glucose in serum Download PDFInfo
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Abstract
The invention provides a photoelectrochemical determination method of glucose in serum. According to the photoelectrochemical determination method, a nickel oxide nanometer material modified electrode sensitized by a water soluble semiconductor sulfide (with a composition general formula of ES) nanometer material is prepared, wherein the electrode has high photogenerated charge transmission efficiency. Glucose oxidase is fixed on the surface of the nickel oxide electrode sensitized by the semiconductor sulfide by utilizing a layer-by-layer assembling method, and the amount of the glucose oxidase can be accurately controlled through the assembling layer number. The electrode fixed with the glucose oxidase has sensitive photoelectrochemical response to the glucose in a solution. Moreover, the electrode has good selectivity to the determination of the glucose. The photoelectrochemical determination method overcomes the disadvantage of poor selectivity of the existing photoelectrochemical determination of the glucose, and has good effect on the determination of the glucose in the serum.
Description
Technical field:
The present invention relates to analyzing and testing field, the Optical Electro-Chemistry that relates in particular to a kind of glucose in serum is measured new method.
Background technology:
As everyone knows, diabetes are one of diseases of serious harm human health in world wide.Within 2011, there are 300,006,006 million peoples to suffer from diabetes, and, expect the year two thousand thirty diabetic and will reach 500,005,005 million peoples [Whiting D R, Guariguata L, Weil C, et al.Diabetes Res.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 during lower than 3.33～3.89mmol/L, and is called hyperglycaemia [Li X, Zhou Y, Zheng Z, et al.Langmuir, 2009,25 (11): 6580-6586.] while surpassing 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 that it can be the diseases such as diabetes, hypertension and cardio-cerebrovascular etc. provides objective basis [Joseph W.Chem.Rev., 2008,108 (2): 814-825.].Develop easy, glucose assays method is significant fast and accurately.
PhotoelectrochemicalMethod Method is a kind of novel analytical approach [Tokudome H, Yamada Y, Sonezaki S, et al.Appl.Phys.Lett., 2005,87 (21): 213901-213901-3 that just grown up recently; Liu S L, Li C, Cheng J, et al.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, thereby its background signal is lower, can reach the high sensitivity suitable with electrogenerated chemiluminescence.And the instrument of Optical Electro-Chemistry is fairly simple, easily microminiaturized; Owing to adopting Electrochemical Detection, with optical detecting method, to compare, its equipment is more inexpensive.Optical Electro-Chemistry detection method very likely develops into portable, high-throughout timely detection system [Wang P, Ge L, Ge S, et al.Chem.Commun., 2013,49 (32): 3294-3296.].Therefore, Optical Electro-Chemistry is a kind of very promising analytical approach, has been subject to increasing concern [Gill R, Zayats M, Willner I.Angew.Chem., Int.Ed., 2008,47 (40): 7602-7625; Haddour N, Chauvin J, Gondran C, et al.J.Am.Chem.Soc., 2006,128 (30): 9693-9698.Wang G L, Xu JJ, Chen H Y, et al.Biosens.Bioelectron., 2009,25 (4): 791-796; Long Y T, Kong C, Li D W, et al.Small, 2011,7 (12): 1624-1628.].
Semiconductor nano material has special and excellent PhotoelectrochemicalProperties Properties [Zhu W, An Y R, Luo X M, et al.Chem.Commun., 2009,19,2682-2684; An Y R, Tang L L, Jiang X L, et al.Chem.-Eur.J., 2010,16,14439-14446; Kang Q, Yang L X, Chen Y F, et al.Anal.Chem., 2010,82 (23): 9749-9754.], in Optical Electro-Chemistry sensor, there is potential broad prospect of application.In earlier stage, had people by PhotoelectrochemicalMethod Method for the detection of glucose [Tanne J,
d, Khalid W, et al.Anal.Chem., 2011,83 (20): 7778-7785; Zheng M, Cui Y, Li X Y, et al.Electroanal.Chem., 2011,656,167-173; Wang W J, Bao L, Lei J P, et al.Anal.Chim.Acta, 2012,744,33-38.], still, 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 as serum be more complicated, the inside is often contained, compared with reducing substanceses such as the ascorbic acid of multiple types, dopamine, sulfhydryl compounds.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 have good antijamming capability as ascorbic acid, dopamine, sulfhydryl compound (halfcystine, glutathione) etc., overcome the drawback of existing photoelectric method poor selectivity, the mensuration of glucose in serum has been obtained to good effect.
Summary of the invention:
The object of this invention is to provide a kind of technique simple, selectivity is good, highly sensitive, measures the method for the glucose of Optical Electro-Chemistry detection fast.
One of object of the present invention can be achieved by the 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, with the pH of the NaOH solution regulator solution of 1mol/L, be neutral; Then pass into after high pure nitrogen 30min, add the Na of 0.1mol/L
2s aqueous solution, continues logical N
2stir, under uniform temperature, react 30 minutes-1.5 hours, obtain water-soluble sulfide (general formula is expressed as ES) nano material;
B, by being immersed in the mixture solution that contains nickel nitrate and organic amine through pretreated ito glass sheet, in the water-bath of uniform temperature, react after 10 minutes and use deionized water rinsing electrode surface; Then electrode is calcined 30 minutes at a certain temperature, obtained the ITO electrode that nickel oxide is modified;
C, the ITO electrode that resulting nickel oxide is modified are immersed in water-soluble sulfide (ES) the nano material solution of synthesized 30 minutes, wash with water, obtain ITO/NiO/ES modified electrode; ITO/NiO/ES is immersed in the solution of the high molecular polymer that contains 0.5%-2% to 30 minutes, after washing with water, drip 10 microlitre glucose oxidase solution reaction 30 minutes, said process can be repeatedly, more glucose oxidase is fixed to ITO/NiO/ES modified electrode surface;
D, the resulting ITO/NiO/ES modified electrode that is fixed with glucose oxidase is put into certain pH the Tris-HCl buffer solution of 0.1mol/L as working electrode, add the to be determined serum solution that contains glucose, under-0.2V current potential (with respect to saturated Ag/AgCl electrode), on homemade Optical Electro-Chemistry instrument, carry out the mensuration of photocurrent.
One of object of the present invention also can be achieved by the following technical measures:
The soluble-salt of the described metallic ion using while preparing water-soluble sulfide nano material is selected from zinc ion, cadmium ion, the soluble-salt of lead ion; Prepare the organic amine that ITO electrode that nickel oxide modifies is used, be selected from hexamethylenetetramine, octadecylamine, tert-butylamine, dimethylamine, triethylamine, triethanolamine, the temperature of described water-bath is 60-100 ℃, the calcining heat of described nickel oxide electrode is 250-450 ℃; The time high molecular polymer that uses that glucose oxidase is fixed to ITO/NiO/ES modified electrode surface is selected from PAMAM, PDDA, PAH, PEI; The pH of the buffer solution using when described Optical Electro-Chemistry is measured glucose is adjusted to 5.0-9.0.
Accompanying drawing explanation:
Fig. 1 is scanning electron microscope (TEM) figure (A) and X-ray diffraction (XRD) figure (B) that the nickel oxide of invention preparation is modified ITO electrode;
Fig. 2 is that the fixedly number of plies of the glucose oxidase of invention preparation is measured the impact of glucose on Optical Electro-Chemistry;
Fig. 3 is the modified electrode of the invention preparation selectivity to different material;
Embodiment:
Embodiment 1:
After the mercaptoacetic acid of a, 10 milliliters of 0.2mol/L is mixed with the zinc nitrate solution of 50 milliliters of 0.02mol/L, with the pH of the NaOH solution regulator solution of 1mol/L, be neutral; Then pass into after high pure nitrogen 30min, add the Na of 0.1mol/L
2s aqueous solution, continues logical N
2stir, react 1 hour at 50 ℃, obtain water-soluble ZnS nano material;
B, by being immersed in the mixture solution that contains nickel nitrate and hexamethylenetetramine through pretreated ito glass sheet, in the water-bath of 50 ℃, react after 10 minutes and use deionized water rinsing electrode surface; Then electrode is calcined 30 minutes at 300 ℃, obtained the ITO electrode that nickel oxide is modified;
C, the ITO electrode that resulting nickel oxide is modified are immersed in the water-soluble ZnS nano material solution of synthesized 30 minutes, wash with water, obtain ITO/NiO/ZnS modified electrode; ITO/NiO/ZnS is immersed in the solution of the PDDA that contains 1% to 30 minutes, after washing with water, drip 10 microlitre glucose oxidase solution reaction 30 minutes, said process in triplicate, is fixed to ITO/NiO/ZnS modified electrode surface by glucose oxidase;
D, the resulting ITO/NiO/ZnS modified electrode that is fixed with glucose oxidase is put into pH is that the Tris-HCl buffer solution of 7.0 0.1mol/L is as working electrode, add the to be determined serum solution that contains glucose, under-0.2V current potential (with respect to saturated Ag/AgCl electrode), on homemade Optical Electro-Chemistry instrument, carry 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, with the pH of the NaOH solution regulator solution of 1mol/L, be neutral; Then pass into after high pure nitrogen 30min, add the Na of 0.1mol/L
2s aqueous solution, continues logical N
2stir, react 30 minutes at 100 ℃, obtain water soluble CdS nanomaterial;
B, by being immersed in the mixture solution that contains nickel nitrate and triethanolamine through pretreated ito glass sheet, in the water-bath of 70 ℃, react after 10 minutes and use deionized water rinsing electrode surface; Then electrode is calcined 30 minutes at 400 ℃, obtained the ITO electrode that nickel oxide is modified;
C, the ITO electrode that resulting nickel oxide is modified are immersed in the water soluble CdS nanomaterial solution of synthesized 30 minutes, wash with water, obtain ITO/NiO/CdS modified electrode; ITO/NiO/CdS is immersed in the solution of the PEI that contains 0.5% to 30 minutes, after washing with water, drip 10 microlitre glucose oxidase solution reaction 30 minutes, said process repeats four times, more glucose oxidase is fixed to ITO/NiO/CdS modified electrode surface;
D, the resulting ITO/NiO/CdS modified electrode that is fixed with glucose oxidase is put into pH is that the Tris-HCl buffer solution of 5.0 0.1mol/L is as working electrode, add the to be determined serum solution that contains glucose, under-0.2V current potential (with respect to saturated Ag/AgCl electrode), on homemade Optical Electro-Chemistry instrument, carry 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, with the pH of the NaOH solution regulator solution of 1mol/L, be neutral; Then pass into after high pure nitrogen 30min, add the Na of 0.1mol/L
2s aqueous solution, continues logical N
2stir, under room temperature, react 30 minutes, obtain water-soluble PbS nano material;
B, by being immersed in the mixture solution that contains nickel nitrate and diethylamine through pretreated ito glass sheet, in the water-bath of 90 ℃, react after 10 minutes and use deionized water rinsing electrode surface; Then electrode is calcined 30 minutes at 350 ℃, obtained the ITO electrode that nickel oxide is modified;
C, the ITO electrode that resulting nickel oxide is modified are immersed in the water-soluble PbS nano material solution of synthesized 30 minutes, wash with water, obtain ITO/NiO/PbS modified electrode; ITO/NiO/PbS is immersed in the solution of the PAH that contains 1% to 30 minutes, after washing with water, drip 10 microlitre glucose oxidase solution reaction 30 minutes, said process can repeat twice, more glucose oxidase is fixed to ITO/NiO/PbS modified electrode surface;
D, the resulting ITO/NiO/PbS modified electrode that is fixed with glucose oxidase is put into pH is that the Tris-HCl buffer solution of 8.0 0.1mol/L is as working electrode, add the to be determined serum solution that contains glucose, under-0.2V current potential (with respect to saturated Ag/AgCl electrode), on homemade Optical Electro-Chemistry instrument, carry 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 water-soluble metal (representing with the E) ion salt solution of 50 milliliters of 0.02mol/L, with the pH of the NaOH solution regulator solution of 1mol/L, be neutral; Then pass into after high pure nitrogen 30min, add the Na of 0.1mol/L
2s aqueous solution, continues logical N
2stir, under uniform temperature, react 30 minutes-1.5 hours, obtain water-soluble sulfide (general formula is expressed as ES) nano material;
B, by being immersed in the mixture solution that contains nickel nitrate and organic amine through pretreated ito glass sheet, in the water-bath of uniform temperature, react after 10 minutes and use deionized water rinsing electrode surface; Then electrode is calcined 30 minutes at a certain temperature, obtained the ITO electrode that nickel oxide is modified;
C, the ITO electrode that resulting nickel oxide is modified are immersed in water-soluble sulfide (ES) the nano material solution of synthesized 30 minutes, wash with water, obtain ITO/NiO/ES modified electrode; ITO/NiO/ES is immersed in the solution of the high molecular polymer that contains 0.5%-2% to 30 minutes, after washing with water, drip 10 microlitre glucose oxidase solution reaction 30 minutes, said process can be repeatedly, more glucose oxidase is fixed to ITO/NiO/ES modified electrode surface;
D, the resulting ITO/NiO/ES modified electrode that is fixed with glucose oxidase is put into certain pH the Tris-HCl buffer solution of 0.1mol/L as working electrode, add the to be determined serum solution that contains glucose, under-0.2V current potential (with respect to saturated Ag/AgCl electrode), on homemade Optical Electro-Chemistry instrument, carry out the mensuration of photocurrent.
2. water-soluble sulfide nano material claimed in claim 1, the soluble-salt of metallic ion used is selected from zinc ion, cadmium ion, the soluble-salt of lead ion.
3. the preparation of the ITO electrode that nickel oxide according to claim 1 is modified, it is characterized in that described organic amine, be selected from hexamethylenetetramine, octadecylamine, tert-butylamine, dimethylamine, triethylamine, triethanolamine, the temperature of described water-bath is 60-100 ℃, the calcining heat of described nickel oxide electrode is 250-450 ℃.
4. glucose oxidase according to claim 1 is fixed to the method on ITO/NiO/ES modified electrode surface, and the high molecular polymer using 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 is measured glucose is adjusted to 5.0-9.0.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105021672A (en) * | 2015-06-23 | 2015-11-04 | 江南大学 | In-situ oxidation reduction reaction-based dopamine photoelectrochemistry detection method |
| 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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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105021672A (en) * | 2015-06-23 | 2015-11-04 | 江南大学 | In-situ oxidation reduction reaction-based dopamine photoelectrochemistry detection method |
| 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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Effective date of registration: 20200904 Address after: Lizhong Industrial Zone, qianduo Town, Xinghua City, Taizhou City, Jiangsu Province (Gu Zhao) Patentee after: Jiangsu tianxie Medical Equipment Co., Ltd Address before: 214122 Jiangsu Province, Wuxi City Lake Road No. 1800, Jiangnan University Patentee before: Jiangnan University |
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