AU2019100367A4 - Synthesis of CoFe Cube Nanozymes and a New Method for Ferrous Ions Detection - Google Patents
Synthesis of CoFe Cube Nanozymes and a New Method for Ferrous Ions Detection Download PDFInfo
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- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 title claims abstract description 37
- 229910003321 CoFe Inorganic materials 0.000 title claims abstract description 27
- 238000001514 detection method Methods 0.000 title claims abstract description 21
- 229910001448 ferrous ion Inorganic materials 0.000 title claims abstract description 15
- 238000000034 method Methods 0.000 title description 11
- 230000015572 biosynthetic process Effects 0.000 title description 3
- 238000003786 synthesis reaction Methods 0.000 title description 3
- 238000004737 colorimetric analysis Methods 0.000 claims abstract description 6
- 238000002835 absorbance Methods 0.000 claims description 19
- 238000006243 chemical reaction Methods 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 239000008367 deionised water Substances 0.000 claims description 7
- 229910021641 deionized water Inorganic materials 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 3
- 239000000758 substrate Substances 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 3
- 238000000975 co-precipitation Methods 0.000 abstract description 2
- 230000001419 dependent effect Effects 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 24
- 230000003197 catalytic effect Effects 0.000 description 18
- 102000004190 Enzymes Human genes 0.000 description 10
- 108090000790 Enzymes Proteins 0.000 description 10
- 239000008363 phosphate buffer Substances 0.000 description 8
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 239000000463 material Substances 0.000 description 5
- NUJOXMJBOLGQSY-UHFFFAOYSA-N manganese dioxide Chemical compound O=[Mn]=O NUJOXMJBOLGQSY-UHFFFAOYSA-N 0.000 description 4
- 238000003917 TEM image Methods 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000002086 nanomaterial Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 102000003992 Peroxidases Human genes 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 description 2
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 239000003574 free electron Substances 0.000 description 1
- 239000003673 groundwater Substances 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000002114 nanocomposite Substances 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 238000006479 redox reaction Methods 0.000 description 1
- 230000027756 respiratory electron transport chain Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J23/00—Catalysts comprising metals or metal oxides or hydroxides, not provided for in group B01J21/00
- B01J23/70—Catalysts comprising metals or metal oxides or hydroxides, not provided for in group B01J21/00 of the iron group metals or copper
- B01J23/74—Iron group metals
- B01J23/75—Cobalt
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
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Abstract
The invention relates to a colorimetric method for the detection of ferrous ion (Fe2) with CoFe cube nanozyme. The CoFe cube nanozyme is obtained through coprecipitation synthesisof CoCl 2 - 6H20,K3[Fe(CN)6] andC6H 5Na 3 O7 - 2H 20solution. The intrinsic oxidase-like activity of CoFe cube nanozyme is demonstrated to be dependent on pH, temperature, and concentration of nanozyme and substrate (TMB). The best detection condition as following: pH is equal to 4.0, the temperature is 20 degree centigrade, the metering of nanozyme is 30 pL and TMB concentration is 0.22 mM. Under the best conditions, the detection range of ferrous ion is from 0.01mM to 0.10 mM. The detection of low concentration of Fe2+ by nanozyme is rapid and accurate.
Description
Synthesis of CoFe Cube Nanozymes and a New Method for Ferrous Ions
Detection
FIELD OF THE INVENTION
The present invention relates to colorimetric detection of ferrous ions by CoFe cube nanozyme, which holds significance in environment, medical, biomedical, and food industrial field.
BACKGROUND OF THE INVENTION
Ferrous ion (Fe2+) is a metal cation. The structure that helps it to combine with free electron makes ferrous ion a strong reductant, and then can react with various oxidizer. Iron is one of the most vital components of the environmental system. It is widely distributed in this world, accounting for 4.75% of the crust content. And in groundwater and mineral, ferrous ion occupies a greater proportion. Ferrous ion also appears in human body as an important component of hemoglobin for oxygen transport. Therefore, the detection method of ferrous ions can bring great economic effect by reducing the cost in analyzing and enhancing the quality of products.
Nanozyme is a new generation of artificial analog enzyme. Like natural
2019100367 05 Apr 2019 enzyme, nanozyme is able to efficiently catalyze the substrate of enzyme under mild conditions, offering the catalytic efficiency and enzymatic reaction kinetics which are similar to natural enzyme. But it is better than natural enzyme stability, even in the strong acid/alkali (pH 2 ~ 10) or large temperature range (4 °C ~ 90 °C), still can maintain 85% catalytic activity. Besides, these advanced materials also have some unique physicochemical properties, which provide prerequisites of designing complex catalytic system. Nanozyme also have low cost of controllable preparation, structure design, adjustable components and catalytic activity. Therefore, the artificial peroxidase mimetics with good catalytic properties will possess a brilliant future in enzyme related applications. In addition to peroxidase mimetics, many kinds of hybrid materials were constructed and proved to be highly efficient catalyst systems in various organic transformation reactions. As a result, great efforts should be made to design and fabricate new nanocomposites with peroxidase-like activity.
A fluorescence detection method using nanoprobe has been developed to detect Fe2+, which is mainly used for the redox reaction between manganese dioxide nanomaterials and ferrous ions to degrade manganese dioxide nanomaterials. And currently, atomic absorbance is also used in the detection requirements for low concentration Fe2+. Compared with other methods, colorimetric method has lower requirement of the
2019100367 05 Apr 2019 equipment and takes less time during the detection. This means colorimetric has a wider use in popular detection because of its low cost and high efficiency. Though test strips used for colorimetric detection is available on the market, they only can be available when the concentration of the sample is greater than 2 mg/L. And when the difference of concentration is large, while the difference of the color scale is small, it can easily lead to misjudgment. Therefore, a method that can fast and accurately detect low concentrations of Fe2+ is of great need.
In this report, we prepared CoFe cube nanozyme by a simple method. CoFe cube nanozyme can efficiently catalyze oxygen to produce free radical or promote electron transfer. Besides, this type of nanozyme has the beneficial of fast, timely, sufficient reaction and high efficiency, which is beneficial to environmental monitoring, food industrial and clinical medicine detection. It plays an important role and provides a new method and material for analysis and detection of Fe2+.
SUMMARY OF THE INVENTION
Nanozyme is a new generation of artificial analog enzyme. Similar to natural enzyme, nanozyme is able to efficiently catalyze the enzyme substrates under mild conditions. Compared to natural enzyme, nanozyme has a better stability, even in strong acid/alkali or large temperature range,
2019100367 05 Apr 2019 the material still can maintain a relatively high catalytic activity. Besides, the unique physicochemical structure of this kind of material also provides prerequisites of designing complex catalytic system. In addition, nanozyme also have low cost of controllable preparation, structure design, adjustable components and catalytic activity, which can provide potential economic benefits. Therefore, the nanozyme should be regarded in a high position.
The object of the invention is to provide a colorimetric method by using nanomaterials to detect Fe2+, which are convenient to synthesis. In order to solve problems mentioned above, this invention provides a new method by using CoFe cube nanozyme to detect Fe2+. In this invention, CoFe cube nanozyme is synthesized by a simple coprecipitation method. It has been observed that variables of catalysis include pH, concentration of substrate, temperature, etc. are approved by various experiments. Finally, the chromogenic reaction of Fe2+ succeeded. And it has been demonstrated that CoFe cube nanozyme has chemical stability and it can conveniently detect Fe2+ with high efficiency.
Preparation of CoFe Cube Nanozymes:
0.5 mM C0CI2 · 6H2O and 1 mM CeHsNasCF · 2H2O were dissolved in mF deionized water, denoted as solution A. 0.25 mM K3[Fe(CN)6]
2019100367 05 Apr 2019 were dissolved in 20 mL deionized water, denoted as solution B. Under rapid stirring, solution B was added slowly into the solution A, continuing to stir for 24 hours. After being centrifuged, washed and dried, purple powder could be obtained, which was recorded as sample CoFePBA-1. Keeping other conditions unchanged, only the amount of K3[Fe(CN)6] substance was changed to ImM, preparing sample, which was recorded as sample CoFePBA-2.
Machines
Transmission electron microscopy (TEM) images of CoFe cube nanozymes were obtained by a transmission electron microscope (FEI Tecnai G2 20 S-TWIN) operating at an accelerating voltage of 200 kV. Five pH Meter, Thermo shaker were also used. Centrifuge used centrifugal force to separate liquid and solid particles or liquid and liquid mixtures. Magnetic stirrer used the principle of homogeneity and reciprocal attraction of the magnetic field, the magnetic field was used to push the magnetic stirrer placed in the container for circumferential operation, thereby it used to stir the CoFe cube nanozyme.
Temperature
0.4 mL of phosphate buffer (pH 4.0) was added into a 1.5 mL centrifuge tube, then that 30 pL sample CoFePBA-2 was added in to the same
2019100367 05 Apr 2019 container. Then 10 pL TMB was added into the tube which was kept under constant temperature (0, 20, 30, 40, 50, 60 and 70 °C) for 8 minutes, the absorbance could be determined at 650 nm. These steps were repeated for 3 times.
The concentration of nanozyme
0.4 mL of phosphate buffer (pH = 4.0) was added to the 1.5 mL centrifuge tube, then sample CoFePBA-2 with different volume 10.0, 20.0, 30.0, 40.0, 50.0 pL and 5.0 pL of TMB were added. The solution was kept reacting at the temperature of 20 °C, then color changes were observed. After 10 minutes, the solution could be detected by identifying light absorbance at 650 nm. These steps needed to be repeated 3 times.
The detection of Fe2+ includes following steps:
The optimum components, pH value and reaction temperature were determined. 0.4 mL of the optimal pH solution was placed in a 1.5 mL centrifuge tube, and the optimal volume of the best nanozyme sample and the optimal volume of TMB were added respectively, setting in the optimal reaction temperature for 10 minutes, and Fe2+ in different concentrations were added. Continue the constant temperature reaction, observe the color change, and measure the absorbance of the solution at 650 nm after 2 minutes. Three samples were repeated for each group. The
2019100367 05 Apr 2019 detection limit, detection range and linear detection range of Fe2+ were determined.
DESCRIPTION OF DRAWINGS
Figure 1 illustrates the TEM image of CoFe Nanozyme.
Figure 2 illustrates the different proportions on the catalytic activity of CoFe Nanozyme.
Figure 3 illustrates the impacts of pH value on the catalytic activity of CoFe Nanozyme.
Figure 4 illustrates the impacts of temperature on the catalytic activity of CoFe Nanozyme.
Figure 5 illustrates the impacts of TMB substrate concentration on the catalytic activity of CoFe Nanozyme.
Figure 6 illustrates the impacts of volume on the catalytic activity of CoFe Nanozyme.
Figure 7 illustrates the the detection of absorbance for different concentration of Fe2+. Δ A= Ao-A, A is the absorbance of Fe2+; Ao is the absorbance of the standard, which without adding any substance.
DESCRIPTION OF PREFERRED EMBODIMENTS
The embodiment of the present invention will be explained in details so that the present invention can be more readily understood.
2019100367 05 Apr 2019
The present invention provides a new method to detect Fe2+, includes following:
The Preparation of CoFe cube nanozyme:
> 0.5 mM C0CI2 ·6Η2Ο and ImM C6H5Na3O7 ·2Η2Ο was added into 20 mL Deionized water, marked as solution A. Repeat the above operation and configure another solution A.
2> 0.25 mM K3[Fe(CN)e] was added into 20 mL deionized water, marked as solution Β. 1 mM K3[Fe(CN)6] was added into 20 mL deionized water, marked as solution C.
Solution A was dissolved by Magnetic mixer, then solution B was added into solution A slowly. This stirring last for 24 hours after the solution B was added. Then recording purple powder as sample CoFePBA-1 after centrifuged, washed and dried; Keeping other conditions unchanged, the solution C was added into solution A, record as a sample CoFePBA-2. TEM image of CoFe Nanozyme is shown in Figure 1.
Comparison of catalytic activity with Different Proportions:
0.4 mF of phosphate buffer (pH 4.0) was added into 1.5 mF centrifuge tube with 30 pF water or CoFe cube nanozyme samples and 10 pF TMB. To find the nanozyme sample with highest catalytic activity, the absorbance could be determined at 650 nm after 10 minutes. Each group repeated for 3 times. As experiment shown, sample CoFePBA-2 has the
2019100367 05 Apr 2019 deepest blue color, which reflects that it may have the highest catalytic activity. As showing the highest peak among all samples, Figure 2 indicated that sample CoFePBA-2 has the strongest absorbance. In order for a more apparent reaction phenomenon, sample CoFePBA-2 will be applied in the following experiments.
Impacts of pH Value:
0.4 mL of phosphate buffer with different pH value (pH 3.0-10.0) was added into a 1.5 mL centrifuge tube, adding 30 pL sample CoFePBA-2 and 10 pL TMB, the absorbance could be determined at 650 nm after 10 minutes, determining the best pH Value buffer. Each group repeated for 3 times. Figure 3 could be deduced that in the buffer solution with pH 4.0, the absorbance achieves the stage, it means that the catalytic activity of nanozyme is the strongest when it is in the environment of pH 4.0.
Impacts of Temperature during the Reaction:
0.4 mL of phosphate buffer (pH 4.0) was added into a 1.5 mL centrifuge tube, then that 30 pL sample CoFePBA-2 was added in to the same container. Then 10 pL TMB was added into the tube which was kept under constant temperature(0, 20, 30, 40, 50, 60 and 70 °C) for 8 minutes, the absorbance could be determined at 650 nm. These steps were repeated for 3 times. As shown in Figure 4, it could be seen that in the temperature
2019100367 05 Apr 2019 of 20 Celsius degrees, the absorbance achieves the stage, which means that the catalytic activity of nanozyme is the strongest when it is in the temperature of 20 Celsius degrees. For convenience , 20 °C will be applied in the following experiments.
Impacts of TMB Concentration:
0.4 mF of phosphate buffer (pH 4.0) was put into a 1.5 mF centrifuge tube. After that, TMB with different volume of 1.0, 2.0, 3.0, 4.0, 5.0, 7.5 pF and 30.0 pF sample CoFePBA-2 were added. The solution was kept reacting at the temperature of 20 °C, then the absorbance could be determined at 650 nm after 10 minutes. These steps were repeated for 3 times. As shown in Figure 5, absorbance was similar when the concentration reached 0.21 to 0.43 mM. So, the concentration of 0.21 mM was chosen for CoFePBA-2 in the following experiments.
Impacts of CoFe Cube Nanozyme:
0.4 mF of phosphate buffer (pH 4.0) was added to the 1.5 mF centrifuge tube, then sample CoFePBA-2 with different volume 10.0, 20.0, 30.0, 40.0, 50.0 pF and 5.0 pF of TMB were added. The solution was kept reacting at the temperature of 20 °C, then color changes were observed. After 10 minutes, the solution could be detected by identifying light absorbance at 650 nm. These steps repeated 3 times. Figure 6 shown that io
2019100367 05 Apr 2019 it achieved stage when the metering of 30 gL of CoFe cube nanozyme. Therefore, metering of 30 gL was chosen for CoFe cube nanozyme in the following experiments.
Determine the range of concentration of Fe—:
0.4 mL of phosphate buffer (pH 4.0) was added to the 1.5 mL centrifuge tube with 30 pL of sample CoLePBA-2 and 5 gL of TMB. After 10 minutes, Le2+ with different volume 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 gL were added. Setting a sample which without Le2+ for comparison; observing color changes after 8 minutes, solution can be detected by identifying absorbance at 650nm. Each sample was repeated 3 times. The detection range of concentration of Ee2+ was the metering of 1 gL or more. When the metering achieved 9 gL, color of solution (blue) faded completely. The solution could be detected by identifying light absorbance at 650 nm. As shown in Ligure 7, the detected range of Le2+is from 0.01 mM to 0.10 mM. This is much more accurate than many methods for detecting Le2+ that are now available.
2019100367 05 Apr 2019
Claims (3)
1. A colorimetric method for detecting ferrous ion (Fe2+), is characterized in including following steps:
preparation of CoFe cube nanozyme:
C0CI2 · 6H2O and CeHsNasOv’SFbO are dissolved in 20 mL deionized water, denoted as solution A. K3[Fe(CN)6] is dissolved in 20 mL deionized water, denoted as solution B;
Under rapid stirring, solution B is added slowly into the solution A, continuing to stir for 24 hours. After being centrifuged, washed and dried, purple powder could be obtained.
2. The colorimetric method for detecting ferrous ion (Fe2+) according to claim 1, wherein during the preparation, the optimized mole ratio of C0CI2 · 6H2O to K3[Fe(CN)6] is 1:2.
3. The colorimetric method for detecting ferrous ion (Fe2+) according to claim 1, wherein the detection of ferrous ion (Fe2+) includes following steps:
the optimum components, pH value and reaction temperature are determined;
0.4 mL of the optimal pH solution is placed in a 1.5 mL centrifuge tube, and the optimal volume of the best nanozyme sample and the optimal volume of TMB are added respectively, setting in the optimal reaction temperature for 10 minutes, and Fe2+with different concentrations are added;
continue the constant temperature reaction, observe the color change, and measure the absorbance of the solution at 650 nm after 8 minutes.
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CN114184789A (en) * | 2021-12-23 | 2022-03-15 | 云南大学 | Prostate specific antigen detection probe and prostate specific antigen detection kit |
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