CN116285976A - Material for detecting glucose and dual-mode signal detection method - Google Patents
Material for detecting glucose and dual-mode signal detection method Download PDFInfo
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 title claims abstract description 120
- 239000008103 glucose Substances 0.000 title claims abstract description 120
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Abstract
The invention provides a material for detecting glucose and a dual-mode signal detection method, which can be used for constructing a novel effective proportion fluorescence colorimetric platform for detecting glucose. The glucose oxidase loaded ruthenium doped carbon quantum dot generates hydrogen peroxide in the presence of glucose, and the hydrogen peroxide is catalyzed by the ruthenium doped carbon quantum dot material with peroxidase activity to generate hydroxyl free radicals, so that the composite material with fluorescence property shows fluorescence enhancement at 630nm, is quenched at 410nm due to an internal light filtering effect, and can be applied to colorimetric detection of glucose at the same time. The biosensor has the advantages of simple equipment, low cost, easy miniaturization, higher sensitivity, wider detection range, lower detection limit and the like, and has important scientific significance and clinical application value for detecting glucose.
Description
Technical Field
The invention belongs to the technical fields of novel nano materials, immunoassay and biosensing, and provides a glucose detection method based on quantum dot nano enzyme, in particular to a biosensor constructed by adopting a nano composite material of GOX@Ru-CDs by taking glucose as a target analyte.
Background
Glucose present in human serum is called blood glucose and is a major substance that provides energy for metabolism in humans. When the blood sugar concentration is abnormal, metabolic disorder diseases can be caused, and the method is one of the main public health problems at present, and the current medical level does not have any medical means capable of radically treating the glucose metabolic disorder diseases, so that the blood sugar level can only be strictly controlled through frequent blood sugar monitoring, and the possibility of complications and death is greatly reduced. Therefore, timely and efficient blood glucose monitoring has important practical application value for preventing, diagnosing and controlling diabetes. Therefore, the development of a rapid, simple and low-cost detection method for early diagnosis and later treatment effects of glucose, survival rate of patients and the like has important significance.
The current methods for diagnosing glucose mainly comprise electrochemical sensors, mass spectra, enzyme-linked immunosorbent assay and the like. Although these methods can be used to detect glucose, they suffer from drawbacks such as high cost, time consuming, and the need for complex large-scale instrumentation by trained personnel, which prevents their widespread use.
But the fluorescence/colorimetry has the advantages of high analysis speed, good sensitivity, high selectivity and the like. Compared to single fluorescence/colorimetric measurements, proportional fluorescence/colorimetric measurements can minimize false signals due to environmental impact and exhibit higher sensitivity and accuracy due to the calibration of the intensity ratios of the two different wavelength determinations.
In the invention, the ruthenium doped carbon quantum dot is used as a new generation nano material, has good adsorptivity, can be dissolved in water, and can be used for identifying biomolecules with identification function such as hybrid enzyme, antibody, aptamer and the like. Meanwhile, the composite material has good fluorescence spectrum signals and ultraviolet absorption spectrum signals, and can generate obvious blue and red color reactions and fluorescence emission changes, so that the visual detection of a target object (hydrogen peroxide and glucose) is realized.
Glucose Oxidase (GOX), also known as glucose oxidase, specifically oxidizes glucose to produce gluconic acid and hydrogen peroxide. Here, we used glucose oxidase to specifically recognize glucose and co-act with ruthenium doped carbon quantum dots with enzyme activity to construct a hydrogen peroxide and glucose immunosensor with the advantages of higher sensitivity, wider detection range, faster detection speed, lower detection limit, convenient operation and the like.
Disclosure of Invention
Aiming at the problems of high cost, low sensitivity, poor stability and the like of the current human glucose detection, the invention provides a material for detecting glucose and a dual-mode signal detection method. Has important scientific significance and clinical application value for detecting glucose.
A material for detecting glucose is ruthenium doped carbon quantum dots Ru-CDs.
The preparation method of the material comprises the following steps: step 1) adding o-phenylenediamine and phosphoric acid into an autoclave with a polytetrafluoroethylene lining, and placing the autoclave into a constant-temperature drying oven for reaction; after the solution is naturally cooled to room temperature, filtering by a filter membrane to obtain blue supernatant; dialyzing the supernatant by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
step 2), weighing the CDs prepared in the step 1), dispersing the CDs in deionized water, and uniformly dispersing the CDs by ultrasonic waves; then, ruCl is added to the mixture 3 And reflux stirring to obtain Ru-CDs solution; the precipitate was collected by centrifugation and the solid was washed with deionized water and ethanol to remove excess ruthenium ions, and lyophilized to give Ru-CDs.
The preparation method of the material provided by the invention specifically comprises the following steps: step 1) adding 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid into a 200mL polytetrafluoroethylene-lined autoclave, and placing the autoclave into a constant-temperature drying oven to react for 24 hours at 200 ℃; after the solution is naturally cooled to room temperature, filtering the solution by a 0.22 mu m filter membrane to obtain blue supernatant; dialyzing the supernatant for 2-3d by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
Step 2) weighing 15mg of CDs prepared in the step 1), dispersing in 10mL of deionized water, and uniformly dispersing by ultrasonic waves; then, 26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain a solution Ru-CDs; the precipitate was collected by centrifugation and the excess ruthenium ions were removed by washing the solid with DW and ethanol, and the Ru-CDs were obtained by three freeze-drying.
A material for detecting glucose is a glucose oxidase-loaded ruthenium-doped carbon quantum dot compound GOX-Ru-CDs.
The invention protects the application of the material GOX-Ru-CDs in the detection of glucose in living cells.
The preparation method of the material comprises the following steps: step 1), in the step 1), o-phenylenediamine and phosphoric acid are added into an autoclave with a polytetrafluoroethylene lining, and the autoclave is placed into a constant-temperature drying oven for reaction; after the solution is naturally cooled to room temperature, filtering by a filter membrane to obtain blue supernatant; dialyzing the supernatant by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
step 2), weighing the CDs prepared in the step 1), dispersing the CDs in deionized water, and uniformly dispersing the CDs by ultrasonic waves; then, ruCl is added to the mixture 3 And reflux stirring to obtain Ru-CDs solution; centrifuging to collect precipitate, washing the solid with deionized water and ethanol to remove excessive ruthenium ions, and freeze-drying to obtain Ru-CDs;
Step 3), weighing Ru-CDs prepared in the step 2), dispersing the Ru-CDs in deionized water, and stirring at room temperature; adding glucose oxidase, stirring at room temperature, centrifuging with deionized water, washing, lyophilizing to obtain GOX-Ru-CDs, dispersing in deionized water, and storing at 4deg.C.
The preparation method of the material provided by the invention specifically comprises the following steps: step 1) adding 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid into a 200mL polytetrafluoroethylene-lined autoclave, and placing the autoclave into a constant-temperature drying oven to react for 24 hours at 200 ℃; after the solution is naturally cooled to room temperature, filtering the solution by a 0.22 mu m filter membrane to obtain blue supernatant; dialyzing the supernatant for 2-3d by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
step 2) weighing 15mg of CDs prepared in the step 1), dispersing in 10mL of deionized water, and uniformly dispersing by ultrasonic waves; then, 26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain a solution Ru-CDs; centrifuging to collect precipitate, washing the solid with DW and ethanol to remove excessive ruthenium ions, and freeze-drying for three times to obtain Ru-CDs;
step 3), weighing 20mg of Ru-CDs prepared in the step 2), dispersing in 50mL of DW, and stirring for 1h at room temperature; adding 0.5mg/mL Glucose Oxidase (GOX), stirring at room temperature for 0.5-3h, centrifuging, washing with DW for three times, lyophilizing to obtain GOX-Ru-CDs, dispersing in 100mL DW, and storing at 4deg.C.
A method for dual mode signal detection of glucose using the above-described materials of the present invention, the method comprising:
the part A comprises the following steps:
step 1), dropwise adding GOX-Ru-CDs solution into a centrifuge tube, and then adding BR buffer solution;
step 2), adding glucose solutions with different concentrations into the solution treated in the step 1) to react in a centrifuge tube;
step 3) the solution processed in the step 2) is rapidly added into a micro four-way cuvette and is excited by lambda in a single way ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured, and a working curve was drawn;
the step of the part B is as follows:
step 1), dropwise adding GOX-Ru-CDs solution into a centrifuge tube, and then adding BR buffer solution;
step 2), adding glucose solutions with different concentrations into the solution treated in the step 1) to react in a centrifuge tube;
and 3) rapidly adding the solution processed in the step 2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
The method for carrying out dual-mode signal detection on glucose by using the material provided by the invention comprises the following steps:
the part A comprises the following steps:
step 1), taking 100 mu L of 0.1-1mg/mL GOX-Ru-CDs solution, dripping the solution into a 2mL centrifuge tube, and then adding 800 mu L of BR buffer solution with pH of 2-8;
Step 2) adding 100 mu L of glucose solution with concentration of 10-100000 mu M into the treated solution in the step 1) into a centrifuge tube, and reacting for 5-30min at 37 ℃;
step 3) adding 100 mu L of the solution processed in the step 2) into a micro four-way cuvette rapidly and exciting lambda in a single mode ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured, and a working curve was drawn;
the step of the part B is as follows:
step 1), taking 100 mu L of 0.1-1mg/mL GOX-Ru-CDs solution, dripping the solution into a 2mL centrifuge tube, and then adding 800 mu L of BR buffer solution with pH of 2-9;
step 2) adding 100 mu L of glucose solution with concentration of 10-100000 mu M into the treated solution in the step 1) into a centrifuge tube, and reacting for 5-30min at 37 ℃;
and 3) rapidly adding the 100 mu L of the solution processed in the step 2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
The preparation method of the BR buffer solution comprises the following steps:
step 1), respectively weighing 0.39g, 0.25g and 0.24g of phosphoric acid, boric acid and acetic acid, dissolving in 100mL of DW to form a 0.04M mixed solution of the three acids, and storing at 4 ℃ for later use;
Step 2) adjusting the pH of the mixed solution of the three acids prepared in the step 1) to 2-8 by using a 0.2M NaOH aqueous solution, and storing at 4 ℃ for standby.
The beneficial results of the invention are:
(1) The invention successfully prepares the GOX@Ru-CDs nanocomposite. Glucose oxidase can be combined on the amino-modified carbon quantum dot material, and can effectively detect glucose and generate hydrogen peroxide. The glucose oxidase has the capability of specifically recognizing glucose by the constructed immunosensor under the screening action of glucose, the fluorescence normal detection range of the prepared sensor is 100-10000 mu M, and the lowest detection lower limit is 2.75 mu M; the linear detection range of the colorimetric method is 10-100 mu M and 100-10000 mu M, and the lowest detection lower limit is 1.35 mu M/15.65 mu M.
(2) The invention uses fluorescence colorimetric/dual signals to enable the output signals to have higher sensitivity.
(3) The invention combines a proportional fluorescence method with a colorimetric method to detect hydrogen peroxide and glucose. The method has the advantages of higher sensitivity, wider detection range, higher detection speed, simple operation and the like.
Drawings
FIG. 1 is a transmission electron microscope characterization of the prepared Carbon Dots (CDs) and ruthenium doped carbon dots (Ru-CDs);
FIG. 2 shows Carbon Dots (CDs), ruthenium-doped carbon dots (Ru-CDs), ruthenium-doped carbon dots and H 2 O 2 Reaction (Ru-CDs-H) 2 O 2 ) The characterization of an X-ray diffractometer for the reaction of ruthenium doped carbon points with glucose (Glu) (GOX@Ru-CDs-Glu) is shown as A; x-ray photoelectron spectroscopy characterization of Ru-CDs is shown as B;
FIG. 3 shows fluorescence and emission and colorimetric spectra (A, C) of GOX@Ru-CDs ratio fluorescent probes in glucose, corresponding to quantitative standard curves (n=3) (B, D);
FIG. 4 is a confocal image of GOX@Ru-CDs incubated with glucose.
Detailed Description
The chemicals and solvents used in the examples were all analytically pure; the raw materials can be purchased from chemical reagent companies or biopharmaceutical companies; the stirring adopts a magnetic stirrer stirring mode.
A material for detecting glucose is ruthenium doped carbon quantum dots Ru-CDs.
The preparation method of the material comprises the following steps: step 1) adding o-phenylenediamine and phosphoric acid into an autoclave with a polytetrafluoroethylene lining, and placing the autoclave into a constant-temperature drying oven for reaction; after the solution is naturally cooled to room temperature, filtering by a filter membrane to obtain blue supernatant; dialyzing the supernatant by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
Step 2), weighing the CDs prepared in the step 1), dispersing the CDs in deionized water, and uniformly dispersing the CDs by ultrasonic waves; then, ruCl is added to the mixture 3 And reflux stirring to obtain Ru-CDs solution; the precipitate was collected by centrifugation and the solid was washed with deionized water and ethanol to remove excess ruthenium ions, and lyophilized to give Ru-CDs.
The preparation method of the material provided by the invention specifically comprises the following steps: step 1) adding 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid into a 200mL polytetrafluoroethylene-lined autoclave, and placing the autoclave into a constant-temperature drying oven to react for 24 hours at 200 ℃; after the solution is naturally cooled to room temperature, filtering the solution by a 0.22 mu m filter membrane to obtain blue supernatant; dialyzing the supernatant for 2-3d by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
step 2) weighing 15mg of CDs prepared in the step 1), dispersing in 10mL of deionized water, and uniformly dispersing by ultrasonic waves; then, 26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain a solution Ru-CDs; the precipitate was collected by centrifugation and the excess ruthenium ions were removed by washing the solid with DW and ethanol, and the Ru-CDs were obtained by three freeze-drying.
A material for detecting glucose is a glucose oxidase-loaded ruthenium-doped carbon quantum dot compound GOX-Ru-CDs.
The invention protects the application of the material GOX-Ru-CDs in the detection of glucose in living cells.
The preparation method of the material comprises the following steps: step 1), in the step 1), o-phenylenediamine and phosphoric acid are added into an autoclave with a polytetrafluoroethylene lining, and the autoclave is placed into a constant-temperature drying oven for reaction; after the solution is naturally cooled to room temperature, filtering by a filter membrane to obtain blue supernatant; dialyzing the supernatant by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
step 2) weighing CDs prepared in the step 1) and dispersing the CDs in deionized waterIn the sub water, the ultrasonic dispersion is uniform; then, ruCl is added to the mixture 3 And reflux stirring to obtain Ru-CDs solution; centrifuging to collect precipitate, washing the solid with deionized water and ethanol to remove excessive ruthenium ions, and freeze-drying to obtain Ru-CDs;
step 3), weighing Ru-CDs prepared in the step 2), dispersing the Ru-CDs in deionized water, and stirring at room temperature; adding glucose oxidase, stirring at room temperature, centrifuging with deionized water, washing, lyophilizing to obtain GOX-Ru-CDs, dispersing in deionized water, and storing at 4deg.C.
The preparation method of the material provided by the invention specifically comprises the following steps: step 1) adding 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid into a 200mL polytetrafluoroethylene-lined autoclave, and placing the autoclave into a constant-temperature drying oven to react for 24 hours at 200 ℃; after the solution is naturally cooled to room temperature, filtering the solution by a 0.22 mu m filter membrane to obtain blue supernatant; dialyzing the supernatant for 2-3d by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
Step 2) weighing 15mg of CDs prepared in the step 1), dispersing in 10mL of deionized water, and uniformly dispersing by ultrasonic waves; then, 26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain a solution Ru-CDs; centrifuging to collect precipitate, washing the solid with DW and ethanol to remove excessive ruthenium ions, and freeze-drying for three times to obtain Ru-CDs;
step 3), weighing 20mg of Ru-CDs prepared in the step 2), dispersing in 50mL of DW, and stirring for 1h at room temperature; adding 0.5mg/mL Glucose Oxidase (GOX), stirring at room temperature for 0.5-3h, centrifuging, washing with DW for three times, lyophilizing to obtain GOX-Ru-CDs, dispersing in 100mL DW, and storing at 4deg.C.
A method for dual mode signal detection of glucose using the above-described materials of the present invention, the method comprising:
the part A comprises the following steps:
step 1), dropwise adding GOX-Ru-CDs solution into a centrifuge tube, and then adding BR buffer solution;
step 2), adding glucose solutions with different concentrations into the solution treated in the step 1) to react in a centrifuge tube;
step 3) the solution processed in the step 2) is rapidly added into a micro four-way cuvette and is excited by lambda in a single way ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured, and a working curve was drawn;
The step of the part B is as follows:
step 1), dropwise adding GOX-Ru-CDs solution into a centrifuge tube, and then adding BR buffer solution;
step 2), adding glucose solutions with different concentrations into the solution treated in the step 1) to react in a centrifuge tube;
and 3) rapidly adding the solution processed in the step 2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
The method for carrying out dual-mode signal detection on glucose by using the material provided by the invention comprises the following steps:
the part A comprises the following steps:
step 1), taking 100 mu L of 0.1-1mg/mL GOX-Ru-CDs solution, dripping the solution into a 2mL centrifuge tube, and then adding 800 mu L of BR buffer solution with pH of 2-8;
step 2) adding 100 mu L of glucose solution with concentration of 10-100000 mu M into the treated solution in the step 1) into a centrifuge tube, and reacting for 5-30min at 37 ℃;
step 3) adding 100 mu L of the solution processed in the step 2) into a micro four-way cuvette rapidly and exciting lambda in a single mode ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured, and a working curve was drawn;
the step of the part B is as follows:
step 1), taking 100 mu L of 0.1-1mg/mL GOX-Ru-CDs solution, dripping the solution into a 2mL centrifuge tube, and then adding 800 mu L of BR buffer solution with pH of 2-9;
Step 2) adding 100 mu L of glucose solution with concentration of 10-100000 mu M into the treated solution in the step 1) into a centrifuge tube, and reacting for 5-30min at 37 ℃;
and 3) rapidly adding the 100 mu L of the solution processed in the step 2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
The preparation method of the BR buffer solution comprises the following steps:
step 1), respectively weighing 0.39g, 0.25g and 0.24g of phosphoric acid, boric acid and acetic acid, dissolving in 100mL of DW to form a 0.04M mixed solution of the three acids, and storing at 4 ℃ for later use;
step 2) adjusting the pH of the mixed solution of the three acids prepared in the step 1) to 2-8 by using a 0.2M NaOH aqueous solution, and storing at 4 ℃ for standby.
The following is only a few examples, in order to better understand the technical solution of the present invention.
Example 1:
a glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of GOX-Ru-CDs solution (0.1 mg/mL) was added dropwise to a 2mL centrifuge tube, and 800. Mu. LpH of BR buffer solution (2) was added.
(2) A series of glucose solutions of different concentrations ranging from 10 to 100000. Mu.M were added to the treated solution of step (1) and reacted at 37℃for 5min.
(3) Rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, and exciting lambda in a single mode ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured and the working curve was plotted.
A glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of GOX-Ru-CDs solution (0.1 mg/mL) was added dropwise to a 2mL centrifuge tube, and 800. Mu. LpH of BR buffer solution (2) was added.
(2) A series of glucose solutions of different concentrations ranging from 10 to 100000. Mu.M were added to the treated solution of step (1) and reacted at 37℃for 5min.
(3) And (3) rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
The preparation method of the glucose oxidase loaded ruthenium doped carbon quantum dot compound (GOX-Ru-CDs) dispersion liquid comprises the following specific steps:
(1) 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid are added into a 200mL polytetrafluoroethylene-lined autoclave, and the mixture is placed in a constant temperature drying oven at 200 ℃ for reaction for 24 hours. After the solution cooled naturally to room temperature, it was filtered through a 0.22 μm filter to give a blue supernatant. And (3) passing the supernatant through a dialysis bag (molecular cutoff is 1000 Da), dialyzing for 2-3d to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use.
(2) 15mg of CDs prepared in the step (1) are weighed and dispersed in 10mL of Deionized Water (DW), and the dispersion is uniform by ultrasonic. Then, 26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain the solution Ru-CDs. The precipitate was collected by centrifugation and the excess ruthenium ions were removed by washing the solid with DW and ethanol, and the Ru-CDs were obtained by three freeze-drying.
(3) 20mg of Ru-CDs prepared in the step (2) are weighed and dispersed in 50mL of DW, and stirred for 1h at room temperature. Adding 0.5mg/mL Glucose Oxidase (GOX), stirring at room temperature for 0.5-3h, centrifuging, washing with DW for three times, lyophilizing to obtain GOX-Ru-CDs, dispersing in 100mL DW, and storing at 4deg.C.
The preparation method of the Britton-Robison (BR) buffer solution comprises the following specific steps:
(1) 0.39g, 0.25g and 0.24g of phosphoric acid, boric acid and acetic acid are weighed respectively, dissolved in 100mL of DW to form a 0.04M mixed solution of the three acids, and placed at 4 ℃ for standby.
(2) And (3) adjusting the pH of the mixed solution of the three acids prepared in the step (2) to 2-8 by using a 0.2M NaOH aqueous solution, and storing at 4 ℃ for standby.
The invention relates to a glucose detection method of a dual-mode signal, which comprises the following specific steps:
(1) The GOX@Ru-CDs nanocomposite is successfully prepared. Glucose oxidase can be combined on the amino-modified carbon quantum dot material, and can effectively detect glucose and generate hydrogen peroxide. The glucose oxidase has the capability of specifically recognizing glucose by the constructed immunosensor under the screening action of glucose, the linear detection range of the prepared sensor is 100-10000 mu M, and the lowest detection lower limit is 2.75 mu M.
(2) The invention uses fluorescence/colorimetric signals to enable the output signals to have higher sensitivity.
(3) The invention combines a proportional fluorescence method with a colorimetric method to detect hydrogen peroxide and glucose. The method has the advantages of higher sensitivity, wider detection range, higher detection speed, simple operation and the like.
Example 2:
a glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of GOX-Ru-CDs solution (0.2 mg/mL) was added dropwise to a 2mL centrifuge tube, and 800. Mu. LpH of BR buffer solution (4) was added.
(2) A series of glucose solutions of different concentrations ranging from 10 to 100000. Mu.M were added to the treated solution of step (1) and reacted at 37℃for 10min.
(3) Rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, and exciting lambda in a single mode ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured and the working curve was plotted.
A glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of GOX-Ru-CDs solution (0.2 mg/mL) was added dropwise to a 2mL centrifuge tube, and 800. Mu. LpH of BR buffer solution (4) was added.
(2) A series of glucose solutions of different concentrations ranging from 10 to 100000. Mu.M were added to the treated solution of step (1) and reacted at 37℃for 10min.
(3) And (3) rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
The preparation method of the glucose oxidase loaded ruthenium doped carbon quantum dot compound (GOX-Ru-CDs) dispersion liquid comprises the following specific steps:
(1) 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid are added into a 200mL polytetrafluoroethylene-lined autoclave, and the mixture is placed in a constant temperature drying oven at 200 ℃ for reaction for 24 hours. After the solution cooled naturally to room temperature, it was filtered through a 0.22 μm filter to give a blue supernatant. And (3) passing the supernatant through a dialysis bag (molecular cutoff is 1000 Da), dialyzing for 2-3d to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use.
(2) 15mg of CDs prepared in the step (1) are weighed and dispersed in 10mL of Deionized Water (DW), and the dispersion is uniform by ultrasonic. Then, 26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain the solution Ru-CDs. The precipitate was collected by centrifugation and the excess ruthenium ions were removed by washing the solid with DW and ethanol, and the Ru-CDs were obtained by three freeze-drying.
(3) 20mg of Ru-CDs prepared in the step (2) are weighed and dispersed in 50mL of DW, and stirred for 1h at room temperature. Adding 0.5mg/mL Glucose Oxidase (GOX), stirring at room temperature for 0.5-3h, centrifuging, washing with DW for three times, lyophilizing to obtain GOX-Ru-CDs, dispersing in 100mL DW, and storing at 4deg.C.
The preparation method of the Britton-Robison (BR) buffer solution comprises the following specific steps:
(1) 0.39g, 0.25g and 0.24g of phosphoric acid, boric acid and acetic acid are weighed respectively, dissolved in 100mL of DW to form a 0.04M mixed solution of the three acids, and placed at 4 ℃ for standby.
(2) And (3) adjusting the pH of the mixed solution of the three acids prepared in the step (2) to 2-8 by using a 0.2M NaOH aqueous solution, and storing at 4 ℃ for standby.
The invention discloses a glucose detection method based on a dual-mode signal, which comprises the following specific steps:
(1) The GOX@Ru-CDs nanocomposite is successfully prepared. Glucose oxidase can be combined on the amino-modified carbon quantum dot material, and can effectively detect glucose and generate hydrogen peroxide. The glucose oxidase has the capability of specifically recognizing glucose by the constructed immunosensor under the screening action of glucose, the linear detection range of the prepared sensor is 100-10000 mu M, and the lowest detection lower limit is 2.75 mu M.
(2) The invention uses fluorescence/colorimetric signals to enable the output signals to have higher sensitivity.
(3) The invention combines a proportional fluorescence method with a colorimetric method to detect hydrogen peroxide and glucose. The method has the advantages of higher sensitivity, wider detection range, higher detection speed, simple operation and the like.
Example 3:
a glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of GOX-Ru-CDs solution (0.4 mg/mL) was added dropwise to a 2mL centrifuge tube, and 800. Mu. LpH of BR buffer solution (6) was added.
(2) A series of glucose solutions of different concentrations ranging from 10 to 100000. Mu.M were added to the treated solution of step (1) and reacted at 37℃for 15min.
(3) Rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, and exciting lambda in a single mode ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured and the working curve was plotted.
A glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of GOX-Ru-CDs solution (0.4 mg/mL) was added dropwise to a 2mL centrifuge tube, and 800. Mu. LpH of BR buffer solution (6) was added.
(2) A series of glucose solutions of different concentrations ranging from 10 to 100000. Mu.M were added to the treated solution of step (1) and reacted at 37℃for 15min.
(3) And (3) rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
The preparation method of the glucose oxidase loaded ruthenium doped carbon quantum dot compound (GOX-Ru-CDs) dispersion liquid comprises the following specific steps:
(1) 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid are added into a 200mL polytetrafluoroethylene-lined autoclave, and the mixture is placed in a constant temperature drying oven at 200 ℃ for reaction for 24 hours. After the solution cooled naturally to room temperature, it was filtered through a 0.22 μm filter to give a blue supernatant. And (3) passing the supernatant through a dialysis bag (molecular cutoff is 1000 Da), dialyzing for 2-3d to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use.
(2) 15mg of CDs prepared in the step (1) are weighed and dispersed in 10mL of Deionized Water (DW), and the dispersion is uniform by ultrasonic. Then, 26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain the solution Ru-CDs. The precipitate was collected by centrifugation and the excess ruthenium ions were removed by washing the solid with DW and ethanol, and the Ru-CDs were obtained by three freeze-drying.
(3) 20mg of Ru-CDs prepared in the step (2) are weighed and dispersed in 50mL of DW, and stirred for 1h at room temperature. Adding 0.5mg/mL Glucose Oxidase (GOX), stirring at room temperature for 0.5-3h, centrifuging, washing with DW for three times, lyophilizing to obtain GOX-Ru-CDs, dispersing in 100mL DW, and storing at 4deg.C.
The preparation method of the Britton-Robison (BR) buffer solution comprises the following specific steps:
(1) 0.39g, 0.25g and 0.24g of phosphoric acid, boric acid and acetic acid are weighed respectively, dissolved in 100mL of DW to form a 0.04M mixed solution of the three acids, and placed at 4 ℃ for standby.
(2) And (3) adjusting the pH of the mixed solution of the three acids prepared in the step (2) to 2-8 by using a 0.2M NaOH aqueous solution, and storing at 4 ℃ for standby.
The invention relates to a glucose detection method of a dual-mode signal, which comprises the following specific steps:
(1) The GOX@Ru-CDs nanocomposite is successfully prepared. Glucose oxidase can be combined on the amino-modified carbon quantum dot material, and can effectively detect glucose and generate hydrogen peroxide. The glucose oxidase has the capability of specifically recognizing glucose by the constructed immunosensor under the screening action of glucose, the linear detection range of the prepared sensor is 100-10000 mu M, and the lowest detection lower limit is 2.75 mu M.
(2) The invention uses fluorescence/colorimetric signals to enable the output signals to have higher sensitivity.
(3) The invention combines a proportional fluorescence method with a colorimetric method to detect hydrogen peroxide and glucose. The method has the advantages of higher sensitivity, wider detection range, higher detection speed, simple operation and the like.
Example 4:
a glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of GOX-Ru-CDs solution (0.6 mg/mL) was added dropwise to a 2mL centrifuge tube, and 800. Mu. LpH of BR buffer solution (7) was added.
(2) A series of glucose solutions with different concentrations of 10-100000 mu M are added into the solution treated in the step (1) and reacted for 20min at 37 ℃.
(3) Rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, and exciting lambda in a single mode ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured and the working curve was plotted.
A glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of GOX-Ru-CDs solution (0.6 mg/mL) was added dropwise to a 2mL centrifuge tube, and 800. Mu. LpH of BR buffer solution (7) was added.
(2) A series of glucose solutions with different concentrations of 10-100000 mu M are added into the solution treated in the step (1) and reacted for 20min at 37 ℃.
(3) And (3) rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
The preparation method of the glucose oxidase loaded ruthenium doped carbon quantum dot compound (GOX-Ru-CDs) dispersion liquid comprises the following specific steps:
(1) 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid are added into a 200mL polytetrafluoroethylene-lined autoclave, and the mixture is placed in a constant temperature drying oven at 200 ℃ for reaction for 24 hours. After the solution cooled naturally to room temperature, it was filtered through a 0.22 μm filter to give a blue supernatant. And (3) passing the supernatant through a dialysis bag (molecular cutoff is 1000 Da), dialyzing for 2-3d to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use.
(2) 15mg of CDs prepared in the step (1) are weighed and dispersed in 10mL of Deionized Water (DW), and the dispersion is uniform by ultrasonic. Then, 26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain the solution Ru-CDs. The precipitate was collected by centrifugation and the excess ruthenium ions were removed by washing the solid with DW and ethanol, and the Ru-CDs were obtained by three freeze-drying.
(3) 20mg of Ru-CDs prepared in the step (2) are weighed and dispersed in 50mL of DW, and stirred for 1h at room temperature. Adding 0.5mg/mL Glucose Oxidase (GOX), stirring at room temperature for 0.5-3h, centrifuging, washing with DW for three times, lyophilizing to obtain GOX-Ru-CDs, dispersing in 100mL DW, and storing at 4deg.C.
The preparation method of the Britton-Robison (BR) buffer solution comprises the following specific steps:
(1) 0.39g, 0.25g and 0.24g of phosphoric acid, boric acid and acetic acid are weighed respectively, dissolved in 100mL of DW to form a 0.04M mixed solution of the three acids, and placed at 4 ℃ for standby.
(2) And (3) adjusting the pH of the mixed solution of the three acids prepared in the step (2) to 2-8 by using a 0.2M NaOH aqueous solution, and storing at 4 ℃ for standby.
The invention discloses a glucose detection method based on a dual-mode signal, which comprises the following specific steps:
(1) The GOX@Ru-CDs nanocomposite is successfully prepared. Glucose oxidase can be combined on the amino-modified carbon quantum dot material, and can effectively detect glucose and generate hydrogen peroxide. The glucose oxidase has the capability of specifically recognizing glucose by the constructed immunosensor under the screening action of glucose, the linear detection range of the prepared sensor is 100-10000 mu M, and the lowest detection lower limit is 2.75 mu M.
(2) The invention uses fluorescence/colorimetric signals to enable the output signals to have higher sensitivity.
(3) The invention combines a proportional fluorescence method with a colorimetric method to detect hydrogen peroxide and glucose. The method has the advantages of higher sensitivity, wider detection range, higher detection speed, simple operation and the like.
Example 5:
a glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of GOX-Ru-CDs solution (0.8 mg/mL) was added dropwise to a 2mL centrifuge tube, and 800. Mu. LpH of BR buffer solution (7.5) was added.
(2) To the treated solution of step (1), 100. Mu.L of a series of glucose solutions of different concentrations ranging from 10 to 100000. Mu.M were added to a centrifuge tube and reacted at 37℃for 25min.
(3) Rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, and exciting lambda in a single mode ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured and the working curve was plotted.
A glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of GOX-Ru-CDs solution (0.8 mg/mL) was added dropwise to a 2mL centrifuge tube, and 800. Mu. LpH of BR buffer solution (7.5) was added.
(2) To the treated solution of step (1), 100. Mu.L of a series of glucose solutions of different concentrations ranging from 10 to 100000. Mu.M were added to a centrifuge tube and reacted at 37℃for 25min.
(3) And (3) rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
The preparation method of the glucose oxidase loaded ruthenium doped carbon quantum dot compound (GOX-Ru-CDs) dispersion liquid comprises the following specific steps:
(1) 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid are added into a 200mL polytetrafluoroethylene-lined autoclave, and the mixture is placed in a constant temperature drying oven at 200 ℃ for reaction for 24 hours. After the solution cooled naturally to room temperature, it was filtered through a 0.22 μm filter to give a blue supernatant. And (3) passing the supernatant through a dialysis bag (molecular cutoff is 1000 Da), dialyzing for 2-3d to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use.
(2) 15mg of CDs prepared in the step (1) are weighed and dispersed in 10mL of Deionized Water (DW), and the dispersion is uniform by ultrasonic. Then mix to26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain the solution Ru-CDs. The precipitate was collected by centrifugation and the excess ruthenium ions were removed by washing the solid with DW and ethanol, and the Ru-CDs were obtained by three freeze-drying.
(3) 20mg of Ru-CDs prepared in the step (2) are weighed and dispersed in 50mL of DW, and stirred for 1h at room temperature. Adding 0.5mg/mL Glucose Oxidase (GOX), stirring at room temperature for 0.5-3h, centrifuging, washing with DW for three times, lyophilizing to obtain GOX-Ru-CDs, dispersing in 100mL DW, and storing at 4deg.C.
The preparation method of the Britton-Robison (BR) buffer solution comprises the following specific steps:
(1) 0.39g, 0.25g and 0.24g of phosphoric acid, boric acid and acetic acid are weighed respectively, dissolved in 100mL of DW to form a 0.04M mixed solution of the three acids, and placed at 4 ℃ for standby.
(2) And (3) adjusting the pH of the mixed solution of the three acids prepared in the step (2) to 2-8 by using a 0.2M NaOH aqueous solution, and storing at 4 ℃ for standby.
The invention discloses a glucose detection method based on a dual-mode signal, which comprises the following specific steps:
(1) The GOX@Ru-CDs nanocomposite is successfully prepared. Glucose oxidase can be combined on the amino-modified carbon quantum dot material, and can effectively detect glucose and generate hydrogen peroxide. The glucose oxidase has the capability of specifically recognizing glucose by the constructed immunosensor under the screening action of glucose, the linear detection range of the prepared sensor is 100-10000 mu M, and the lowest detection lower limit is 2.75 mu M.
(2) The invention uses fluorescence/colorimetric signals to enable the output signals to have higher sensitivity.
(3) The invention combines a proportional fluorescence method with a colorimetric method to detect hydrogen peroxide and glucose. The method has the advantages of higher sensitivity, wider detection range, higher detection speed, simple operation and the like.
Example 6:
a glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of 1mg/mL GOX-Ru-CDs solution was added dropwise to a 2mL centrifuge tube, followed by addition of 800. Mu. LpH of 8 BR buffer.
(2) A series of glucose solutions of different concentrations ranging from 10 to 100000. Mu.M were added to the treated solution of step (1) and reacted at 37℃for 30min.
(3) Rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, and exciting lambda in a single mode ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured and the working curve was plotted.
A glucose detection method based on a dual-mode signal comprises the following specific steps:
(1) mu.L of 1mg/mL GOX-Ru-CDs solution was added dropwise to a 2mL centrifuge tube, followed by addition of 800. Mu. LpH of 8 BR buffer.
(2) A series of glucose solutions of different concentrations ranging from 10 to 100000. Mu.M were added to the treated solution of step (1) and reacted at 37℃for 30min.
(3) And (3) rapidly adding 100 mu L of the solution processed in the step (2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
The preparation method of the glucose oxidase loaded ruthenium doped carbon quantum dot compound (GOX-Ru-CDs) dispersion liquid comprises the following specific steps:
(1) 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid are added into a 200mL polytetrafluoroethylene-lined autoclave, and the mixture is placed in a constant temperature drying oven at 200 ℃ for reaction for 24 hours. After the solution cooled naturally to room temperature, it was filtered through a 0.22 μm filter to give a blue supernatant. And (3) passing the supernatant through a dialysis bag (molecular cutoff is 1000 Da), dialyzing for 2-3d to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use.
(2) 15mg of CDs prepared in the step (1) are weighed and dispersed in 10mL of Deionized Water (DW), and the dispersion is uniform by ultrasonic. Then, 26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain the solution Ru-CDs. The precipitate was collected by centrifugation and excess ruthenium ions were removed by washing the solid with DW and ethanol,and freeze-drying for three times to obtain Ru-CDs.
(3) 20mg of Ru-CDs prepared in the step (2) are weighed and dispersed in 50mL of DW, and stirred for 1h at room temperature. Adding 0.5mg/mL Glucose Oxidase (GOX), stirring at room temperature for 0.5-3h, centrifuging, washing with DW for three times, lyophilizing to obtain GOX-Ru-CDs, dispersing in 100mL DW, and storing at 4deg.C.
The preparation method of the Britton-Robison (BR) buffer solution comprises the following specific steps:
(1) 0.39g, 0.25g and 0.24g of phosphoric acid, boric acid and acetic acid are weighed respectively, dissolved in 100mL of DW to form a 0.04M mixed solution of the three acids, and placed at 4 ℃ for standby.
(2) And (3) adjusting the pH of the mixed solution of the three acids prepared in the step (2) to 2-8 by using a 0.2M NaOH aqueous solution, and storing at 4 ℃ for standby.
The invention relates to a glucose detection method of a dual-mode signal, which comprises the following specific steps:
(1) The GOX@Ru-CDs nanocomposite is successfully prepared. Glucose oxidase can be combined on the amino-modified carbon quantum dot material, and can effectively detect glucose and generate hydrogen peroxide. The glucose oxidase has the capability of specifically recognizing glucose by the constructed immunosensor under the screening action of glucose, the linear detection range of the prepared sensor is 100-10000 mu M, and the lowest detection lower limit is 2.75 mu M.
(2) The invention uses fluorescence/colorimetric signals to enable the output signals to have higher sensitivity.
(3) The invention combines a proportional fluorescence method with a colorimetric method to detect hydrogen peroxide and glucose. The method has the advantages of higher sensitivity, wider detection range, higher detection speed, simple operation and the like.
FIG. 1 is a transmission electron microscope image of CDs (A) and Ru-CDs (B) in step (1) of example 1; as shown in figure a, the synthesized CDs particles have a quasi-spherical morphology, are relatively uniform in nano-size and have a lattice spacing of 0.21nm; as shown in FIG. B, the tragic nature of Ru did not change the nano-size of CDs (0.25 nm) and had well-resolved lattice spacing, corresponding to the graphite-like structure, and these results also demonstrate the successful fabrication of Ru-CDs composites.
FIG. 2 shows Ru-CDs, GOX@Ru-CDs-H 2 O 2 As shown in a graph A, the X-ray diffraction pattern shows that a characteristic diffraction peak of the carbon quantum dot exists at about 23 degrees, the crystal face of the characteristic diffraction peak is shown as (O02), and the structure of the composite material is proved to be consistent with TEM imaging when the tragic impurities of Ru on the carbon quantum dot and the load of glucose oxidase are not changed; as shown in fig. B, X-ray co-electron spectroscopy analysis was performed to evaluate the chemical composition of the prepared Ru-CDs, and X-ray co-electron spectroscopy measurement spectra showed three main peaks corresponding to C1s, N1s, O1s and Ru1s, respectively, demonstrating successful preparation of the composite material.
Fig. 3A, B shows uv absorption spectra and standard curves for different concentrations prepared in step (3) of example 1. The constructed method detects hydrogen peroxide and glucose with different concentrations under the conditions of 480 and 610nm, and the ratio of absorbance values gradually increases with the increase of the concentration of the hydrogen peroxide and the concentration of the glucose (figure 3A). As shown in FIG. 3B, the absorbance has good linear relation with concentration, the linear detection range of the sensor is 10-100 mu M and 100-10000 mu M, the lowest detection lower limit is 1.35 mu M/15.65 mu M, and good linearity and lower LOD value are shown. FIG. 3C, D is a graph of fluorescence spectra of glucose at different concentrations prepared in step (3) of example 1 and a standard curve. The fluorescence excitation wavelength is constructed to be lambda ex The different concentrations of glucose were tested at 370nm, with increasing fluorescence emission at 410nm and 630nm fluorescence ratios (fig. 3C). As shown in FIG. 3D, the fluorescence ratio has good linear relation with the glucose concentration, the linear detection range of the sensor is 10-10000 mu M, the lowest detection lower limit is 2.75 mu M, and good linearity and lower LOD value are shown.
FIG. 4 is a confocal microscopy image (scale 10 μm) of HeLa cells of GOX@Ru-CDs. When glucose was added to the cells, the bright field image of the cells showed normal morphology of HeLa cells (panel a), and bright blue, red fluorescence and complex fluorescence were clearly observed in the fluorescent image (8B-D). These results show that GOX@Ru-CDs show good biocompatibility and have great potential in detecting glucose in living cells.
Table 1 shows that the immunosensor constructed in example 3 measures glucose in an artificial serum sample with recovery rates between 96.4% and 102.6%, indicating that the immunosensor is acceptably accurate and reliable in analysis of the artificial serum sample.
Immunosensor constructed in Table 1 detects glucose in artificial serum
| Sample of | Sample (mu M) | Marked value (mu M) | Measurement value (mu M) | Recovery (%) |
| 1 | 0.00 | (S)±500.00 | 482.22 | 96.4 |
| 2 | 0.00 | (S)±1000.00 | 1026.75 | 102.6 |
| 3 | 0.00 | (S)±5000.00 | 4903.91 | 98.1 |
| 4 | 0.00 | (S)±10000.00 | 9814.99 | 98.3 |
The foregoing description is only a few specific embodiments of the present invention (the embodiments are not intended to be exhaustive, and the scope of the invention includes the scope of the invention and other technical points), and the details or common sense of the present invention are not described in any more detail herein (including but not limited to the shorthand, abbreviations, units commonly used in the art). It should be noted that the above embodiments do not limit the present invention in any way, and it is within the scope of the present invention for those skilled in the art to obtain the technical solution by equivalent substitution or equivalent transformation. The protection scope of the present application shall be subject to the content of the claims, and the description of the specific embodiments and the like in the specification can be used for explaining the content of the claims.
Claims (10)
1. A material for detecting glucose is characterized in that the material is ruthenium doped carbon quantum dots Ru-CDs.
2. A material for detecting glucose is characterized in that the material is glucose oxidase loaded ruthenium doped carbon quantum dot compound GOX-Ru-CDs.
3. Use of the material of claim 2 for the detection of glucose in living cells.
4. A method of preparing a material according to claim 1, comprising the steps of: step 1) adding o-phenylenediamine and phosphoric acid into an autoclave with a polytetrafluoroethylene lining, and placing the autoclave into a constant-temperature drying oven for reaction; after the solution is naturally cooled to room temperature, filtering by a filter membrane to obtain blue supernatant; dialyzing the supernatant by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
step 2), weighing the CDs prepared in the step 1), dispersing the CDs in deionized water, and uniformly dispersing the CDs by ultrasonic waves; then, ruCl is added to the mixture 3 And reflux stirring to obtain Ru-CDs solution; the precipitate was collected by centrifugation and the solid was washed with deionized water and ethanol to remove excess ruthenium ions, and lyophilized to give Ru-CDs.
5. The method for preparing the material according to claim 4, wherein the method comprises the steps of: step 1) adding 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid into a 200mL polytetrafluoroethylene-lined autoclave, and placing the autoclave into a constant-temperature drying oven to react for 24 hours at 200 ℃; after the solution is naturally cooled to room temperature, filtering the solution by a 0.22 mu m filter membrane to obtain blue supernatant; dialyzing the supernatant for 2-3d by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
Step 2) weighing 15mg of CDs prepared in the step 1), dispersing in 10mL of deionized water, and uniformly dispersing by ultrasonic waves; then, 26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain a solution Ru-CDs; the precipitate was collected by centrifugation and the excess ruthenium ions were removed by washing the solid with DW and ethanol, and the Ru-CDs were obtained by three freeze-drying.
6. A method of preparing a material as claimed in claim 2 or 3, wherein the method comprises the steps of: step 1), in the step 1), o-phenylenediamine and phosphoric acid are added into an autoclave with a polytetrafluoroethylene lining, and the autoclave is placed into a constant-temperature drying oven for reaction; after the solution is naturally cooled to room temperature, filtering by a filter membrane to obtain blue supernatant; dialyzing the supernatant by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
step 2), weighing the CDs prepared in the step 1), dispersing the CDs in deionized water, and uniformly dispersing the CDs by ultrasonic waves; then, ruCl is added to the mixture 3 And reflux stirring to obtain Ru-CDs solution; centrifuging to collect precipitate, washing the solid with deionized water and ethanol to remove excessive ruthenium ions, and freeze-drying to obtain Ru-CDs;
step 3), weighing Ru-CDs prepared in the step 2), dispersing the Ru-CDs in deionized water, and stirring at room temperature; adding glucose oxidase, stirring at room temperature, centrifuging with deionized water, washing, lyophilizing to obtain GOX-Ru-CDs, dispersing in deionized water, and storing at 4deg.C.
7. The method for preparing the material according to claim 6, wherein the method comprises the steps of: step 1) adding 4.6mmol of o-phenylenediamine and 15mL of phosphoric acid into a 200mL polytetrafluoroethylene-lined autoclave, and placing the autoclave into a constant-temperature drying oven to react for 24 hours at 200 ℃; after the solution is naturally cooled to room temperature, filtering the solution by a 0.22 mu m filter membrane to obtain blue supernatant; dialyzing the supernatant for 2-3d by a dialysis bag to obtain a quantum dot aqueous solution, and freeze-drying to obtain quantum dot CDs for later use;
step 2) weighing 15mg of CDs prepared in the step 1), dispersing in 10mL of deionized water, and uniformly dispersing by ultrasonic waves; then, 26mg of RuCl was added to the mixture 3 And stirring for 4 hours under reflux to obtain a solution Ru-CDs; centrifuging to collect precipitate, washing the solid with DW and ethanol to remove excessive ruthenium ions, and freeze-drying for three times to obtain Ru-CDs;
step 3), weighing 20mg of Ru-CDs prepared in the step 2), dispersing in 50mL of DW, and stirring for 1h at room temperature; adding 0.5mg/mL Glucose Oxidase (GOX), stirring at room temperature for 0.5-3h, centrifuging, washing with DW for three times, lyophilizing to obtain GOX-Ru-CDs, dispersing in 100mL DW, and storing at 4deg.C.
8. A method for dual mode signal detection of glucose using the material of claim 2, comprising:
The part A comprises the following steps:
step 1), dropwise adding GOX-Ru-CDs solution into a centrifuge tube, and then adding BR buffer solution;
step 2), adding glucose solutions with different concentrations into the solution treated in the step 1) to react in a centrifuge tube;
step 3) the solution processed in the step 2) is rapidly added into a micro four-way cuvette and is excited by lambda in a single way ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured, and a working curve was drawn;
the step of the part B is as follows:
step 1), dropwise adding GOX-Ru-CDs solution into a centrifuge tube, and then adding BR buffer solution;
step 2), adding glucose solutions with different concentrations into the solution treated in the step 1) to react in a centrifuge tube;
and 3) rapidly adding the solution processed in the step 2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
9. A method for dual mode signal detection of glucose using the material of claim 2, comprising:
the part A comprises the following steps:
step 1), taking 100 mu L of 0.1-1mg/mL GOX-Ru-CDs solution, dripping the solution into a 2mL centrifuge tube, and then adding 800 mu L of BR buffer solution with pH of 2-8;
step 2) adding 100 mu L of glucose solution with concentration of 10-100000 mu M into the treated solution in the step 1) into a centrifuge tube, and reacting for 5-30min at 37 ℃;
Step 3) adding 100 mu L of the solution processed in the step 2) into a micro four-way cuvette rapidly and exciting lambda in a single mode ex At=370 nm, the ratio of fluorescence values at 630nm to 410nm was measured, and a working curve was drawn;
the step of the part B is as follows:
step 1), taking 100 mu L of 0.1-1mg/mL GOX-Ru-CDs solution, dripping the solution into a 2mL centrifuge tube, and then adding 800 mu L of BR buffer solution with pH of 2-9;
step 2) adding 100 mu L of glucose solution with concentration of 10-100000 mu M into the treated solution in the step 1) into a centrifuge tube, and reacting for 5-30min at 37 ℃;
and 3) rapidly adding the 100 mu L of the solution processed in the step 2) into a micro four-way cuvette, measuring the ratio of absorbance at 480nm and 610nm, and drawing a working curve.
10. The method for detecting glucose according to claim 9, wherein the preparation method of the BR buffer solution comprises the steps of:
step 1), respectively weighing 0.39g, 0.25g and 0.24g of phosphoric acid, boric acid and acetic acid, dissolving in 100mL of DW to form a 0.04M mixed solution of the three acids, and storing at 4 ℃ for later use;
step 2) adjusting the pH of the mixed solution of the three acids prepared in the step 1) to 2-8 by using a 0.2M NaOH aqueous solution, and storing at 4 ℃ for standby.
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Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104089999A (en) * | 2014-06-25 | 2014-10-08 | 复旦大学 | Carbon quantum dot-nanowire array-based cardiomyocyte signal molecule sensor and preparation method thereof |
| CN105647528A (en) * | 2016-02-02 | 2016-06-08 | 中国石油大学(北京) | Zinc-doped carbon quantum dot, preparation method thereof and application thereof to detection field |
| CN105911030A (en) * | 2016-04-05 | 2016-08-31 | 中国科学院合肥物质科学研究院 | Ratio fluorescence sensor and visualized detection method for glucose |
| CN107478621A (en) * | 2017-06-26 | 2017-12-15 | 南京医科大学 | Metabolizable generation H in serum is detected by ratio fluorescent probe quantitative2O2Biomolecule method |
| CN108169198A (en) * | 2017-12-28 | 2018-06-15 | 大连工业大学 | Utilize the method for fluorescent carbon quantum dot detection Determination of Glucose in Food |
| CN108929684A (en) * | 2018-08-29 | 2018-12-04 | 济南大学 | A kind of coconut palm monofilament is the preparation method of carbon source dysprosium doped carbon quantum dot composite material |
| CN109517595A (en) * | 2018-10-18 | 2019-03-26 | 临沂大学 | Double-bang firecracker based on the building of double check mechanism answers ratio fluorescent probe and its construction method |
| CN110095444A (en) * | 2019-05-09 | 2019-08-06 | 南宁师范大学 | Utilize the method for carbon quantum dot detection glucose in solutions concentration |
| KR102052988B1 (en) * | 2018-08-13 | 2019-12-06 | 경북대학교 산학협력단 | Carbon-Dot-Based Ratiometric Fluorescence Glucose Biosensor |
| CN111504956A (en) * | 2019-01-31 | 2020-08-07 | 华东理工大学 | Preparation of carbon quantum dot fluorescent probe and application of carbon quantum dot fluorescent probe in selective detection of active oxygen |
| WO2020172972A1 (en) * | 2019-02-28 | 2020-09-03 | 中国科学院大连化学物理研究所 | Glucose oxidase-based nanocapsule sensor and preparation and application thereof |
| CN113956875A (en) * | 2021-10-28 | 2022-01-21 | 延安大学 | Preparation method and application of zinc-doped carbon quantum dot fluorescent probe |
| EP3970753A1 (en) * | 2020-09-18 | 2022-03-23 | Universitatea Tehnica "Gheorghe Asachi" Din Iasi | Manganese-dopped nanostructured carbon dots with applications in antitumoral treatments and medical imaging |
-
2023
- 2023-03-28 CN CN202310312123.8A patent/CN116285976B/en active Active
Patent Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104089999A (en) * | 2014-06-25 | 2014-10-08 | 复旦大学 | Carbon quantum dot-nanowire array-based cardiomyocyte signal molecule sensor and preparation method thereof |
| CN105647528A (en) * | 2016-02-02 | 2016-06-08 | 中国石油大学(北京) | Zinc-doped carbon quantum dot, preparation method thereof and application thereof to detection field |
| CN105911030A (en) * | 2016-04-05 | 2016-08-31 | 中国科学院合肥物质科学研究院 | Ratio fluorescence sensor and visualized detection method for glucose |
| CN107478621A (en) * | 2017-06-26 | 2017-12-15 | 南京医科大学 | Metabolizable generation H in serum is detected by ratio fluorescent probe quantitative2O2Biomolecule method |
| CN108169198A (en) * | 2017-12-28 | 2018-06-15 | 大连工业大学 | Utilize the method for fluorescent carbon quantum dot detection Determination of Glucose in Food |
| KR102052988B1 (en) * | 2018-08-13 | 2019-12-06 | 경북대학교 산학협력단 | Carbon-Dot-Based Ratiometric Fluorescence Glucose Biosensor |
| CN108929684A (en) * | 2018-08-29 | 2018-12-04 | 济南大学 | A kind of coconut palm monofilament is the preparation method of carbon source dysprosium doped carbon quantum dot composite material |
| CN109517595A (en) * | 2018-10-18 | 2019-03-26 | 临沂大学 | Double-bang firecracker based on the building of double check mechanism answers ratio fluorescent probe and its construction method |
| CN111504956A (en) * | 2019-01-31 | 2020-08-07 | 华东理工大学 | Preparation of carbon quantum dot fluorescent probe and application of carbon quantum dot fluorescent probe in selective detection of active oxygen |
| WO2020172972A1 (en) * | 2019-02-28 | 2020-09-03 | 中国科学院大连化学物理研究所 | Glucose oxidase-based nanocapsule sensor and preparation and application thereof |
| CN110095444A (en) * | 2019-05-09 | 2019-08-06 | 南宁师范大学 | Utilize the method for carbon quantum dot detection glucose in solutions concentration |
| EP3970753A1 (en) * | 2020-09-18 | 2022-03-23 | Universitatea Tehnica "Gheorghe Asachi" Din Iasi | Manganese-dopped nanostructured carbon dots with applications in antitumoral treatments and medical imaging |
| CN113956875A (en) * | 2021-10-28 | 2022-01-21 | 延安大学 | Preparation method and application of zinc-doped carbon quantum dot fluorescent probe |
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