CN108822005A - A kind of reversible colorimetric probe and its preparation, application based on malachite green and bisulfite addition product - Google Patents
A kind of reversible colorimetric probe and its preparation, application based on malachite green and bisulfite addition product Download PDFInfo
- Publication number
- CN108822005A CN108822005A CN201810606827.5A CN201810606827A CN108822005A CN 108822005 A CN108822005 A CN 108822005A CN 201810606827 A CN201810606827 A CN 201810606827A CN 108822005 A CN108822005 A CN 108822005A
- Authority
- CN
- China
- Prior art keywords
- malachite green
- glucose
- solution
- colorimetric probe
- probe
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000000523 sample Substances 0.000 title claims abstract description 66
- FDZZZRQASAIRJF-UHFFFAOYSA-M malachite green Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](C)C)C=C1 FDZZZRQASAIRJF-UHFFFAOYSA-M 0.000 title claims abstract description 40
- 229940107698 malachite green Drugs 0.000 title claims abstract description 40
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 title claims abstract description 26
- 230000002441 reversible effect Effects 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 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 claims description 50
- 239000008103 glucose Substances 0.000 claims description 49
- 238000006243 chemical reaction Methods 0.000 claims description 20
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical class OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 241000907663 Siproeta stelenes Species 0.000 claims description 7
- 239000004366 Glucose oxidase Substances 0.000 claims description 6
- 108010015776 Glucose oxidase Proteins 0.000 claims description 5
- 229940116332 glucose oxidase Drugs 0.000 claims description 5
- 235000019420 glucose oxidase Nutrition 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 5
- 239000008363 phosphate buffer Substances 0.000 claims description 4
- 238000012360 testing method Methods 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 3
- AOSFMYBATFLTAQ-UHFFFAOYSA-N 1-amino-3-(benzimidazol-1-yl)propan-2-ol Chemical compound C1=CC=C2N(CC(O)CN)C=NC2=C1 AOSFMYBATFLTAQ-UHFFFAOYSA-N 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- DJEHXEMURTVAOE-UHFFFAOYSA-M potassium bisulfite Chemical compound [K+].OS([O-])=O DJEHXEMURTVAOE-UHFFFAOYSA-M 0.000 claims description 2
- 229940099427 potassium bisulfite Drugs 0.000 claims description 2
- 235000010259 potassium hydrogen sulphite Nutrition 0.000 claims description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims 1
- YZCKVEUIGOORGS-IGMARMGPSA-N Protium Chemical compound [1H] YZCKVEUIGOORGS-IGMARMGPSA-N 0.000 claims 1
- 150000002431 hydrogen Chemical class 0.000 claims 1
- 229910052739 hydrogen Inorganic materials 0.000 claims 1
- 239000001257 hydrogen Substances 0.000 claims 1
- 229910052708 sodium Inorganic materials 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 235000001727 glucose Nutrition 0.000 description 47
- 239000000243 solution Substances 0.000 description 45
- 239000000047 product Substances 0.000 description 21
- 230000008859 change Effects 0.000 description 10
- 210000002966 serum Anatomy 0.000 description 10
- 238000002835 absorbance Methods 0.000 description 8
- 238000004847 absorption spectroscopy Methods 0.000 description 8
- 239000007853 buffer solution Substances 0.000 description 7
- 238000001514 detection method Methods 0.000 description 6
- 239000011259 mixed solution Substances 0.000 description 6
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 5
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 4
- 229930006000 Sucrose Natural products 0.000 description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 4
- 238000007259 addition reaction Methods 0.000 description 4
- 238000001819 mass spectrum Methods 0.000 description 4
- 230000001590 oxidative effect Effects 0.000 description 4
- 229940079827 sodium hydrogen sulfite Drugs 0.000 description 4
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000005720 sucrose Substances 0.000 description 4
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 4
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 3
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 206010012601 diabetes mellitus Diseases 0.000 description 3
- 230000002452 interceptive effect Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 239000008055 phosphate buffer solution Substances 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 235000000346 sugar Nutrition 0.000 description 3
- 239000013589 supplement Substances 0.000 description 3
- 229910021642 ultra pure water Inorganic materials 0.000 description 3
- 239000012498 ultrapure water Substances 0.000 description 3
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- 238000004737 colorimetric analysis Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 229960002163 hydrogen peroxide Drugs 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 206010072289 Eye colour change Diseases 0.000 description 1
- 208000013016 Hypoglycemia Diseases 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 241000219095 Vitis Species 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005562 fading Methods 0.000 description 1
- 150000002304 glucoses Chemical class 0.000 description 1
- 235000021384 green leafy vegetables Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 239000006101 laboratory sample Substances 0.000 description 1
- 125000003071 maltose group Chemical group 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000005935 nucleophilic addition reaction Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C309/00—Sulfonic acids; Halides, esters, or anhydrides thereof
- C07C309/01—Sulfonic acids
- C07C309/02—Sulfonic acids having sulfo groups bound to acyclic carbon atoms
- C07C309/24—Sulfonic acids having sulfo groups bound to acyclic carbon atoms of a carbon skeleton containing six-membered aromatic rings
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
-
- G—PHYSICS
- 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
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1007—Non-condensed systems
Landscapes
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Organic Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to colorimetric probe technical fields, and in particular to a kind of reversible colorimetric probe and its preparation and application based on malachite green and bisulfite addition product.The reversible colorimetric probe structure formula is as follows：.Colorimetric probe high sensitivity of the present invention, selectivity are good, simple and convenient, and may be reused, and are expected to promote the use of in field of biomedicine.
Description
Technical field
The invention belongs to colorimetric probe technical fields, and in particular to one kind is produced based on malachite green and bisulfite addition
The reversible colorimetric probe of object and its preparation and application.
Background technique
Glucose is primary carbon source and the energy source of cell metabolism, is played in the Natural growth process of cell heavy to closing
The effect wanted, the diseases such as glucose level in blood and diabetes or hypoglycemia are closely related.And in the past few decades in,
Diabetes have become one of maximum publilc health threat.Therefore, monitoring blood-sugar content for prevent and treat diabetes or
Other related diseases are of great significance.
Currently, people are it has been reported that many glucose sensing approach, for example, fluorescence sense method, colorimetric sensing method,
Chemiluminescence method for sensing and high performance liquid chromatography etc..In these analysis methods, colorimetric method causes the extensive pass of people
Note, in addition to it is accurate, easy to operate, inexpensive the features such as, colorimetric method can also observe by the naked eye color change to analyze determinand,
Less labour is needed in this way, in this embodiment it is not even necessary to detection device.But current colorimetric probe is mostly expendable consumed product, tool
Have and recycle the glucose reversible colorimetric probe of reusable ability not yet, is unfavorable for sustainable development.
Summary of the invention
In view of this, what the purpose of the present invention is to provide a kind of based on malachite green and bisulfite addition product can
Inverse colorimetric probe and its preparation and application, the reversible colorimetric probe may be implemented to reuse.
To achieve the above object, the technical scheme adopted by the invention is as follows：
A kind of reversible colorimetric probe based on malachite green Yu bisulfite addition product, structural formula are as follows：
。
Preparation method based on malachite green Yu the reversible colorimetric probe of bisulfite addition product, steps are as follows：1）
Take raw material malachite green and bisulfites；
2）Phosphate buffer is added in malachite green and bisulfites（Concentration 10mmol/LPBS）In, malachite after mixing
Green concentration is 20 μM, and the concentration of bisulfites is 25 μM, reacts 3min at room temperature, is obtained based on malachite green and bisulfite
The reversible colorimetric probe solution of root addition product.
The mass ratio of the material of the malachite green and bisulfite is 1：（1-1.25）.
The bisulfites is sodium hydrogensulfite, potassium bisulfite or ammonium bisulfite.
Application of the reversible colorimetric probe in test sample in terms of glucose.
When concrete application, sample to be tested is first mixed into incubation with glucose oxidase（It is incubated at 37 DEG C and carries out 30min）
Obtain reaction solution, then by reaction solution be added reversible colorimetric probe solution in, then respectively the color after test reaction and
Uv-vis spectra variation.
Experiment discovery：As glucose content increases in sample to be tested, it is molten that reversible colorimetric probe is added in reaction solution
Gradually become blue from colourless after in liquid, the variation of uv-vis spectra is：Absorbance at 618 nm is with glucose content
Increase and increase.
The present invention occurs the addition product that 1,4- addition reaction obtains using malachite green and bisulfite and visits as colorimetric
Needle, the study found that glucose can generate oxydol H under the catalytic action of glucose oxidase2O2, and H2O2It can aoxidize
The addition reaction product of malachite green and bisulfite makes the product become blue malachite green again again, if will be to test sample
Product, which mix the reaction solution addition probe solution middle probe solution after being incubated for glucose oxidase, becomes blue, explanation from colourless
Addition reaction product becomes malachite green again, shows in reaction solution at this time containing glucose after glucose oxidase is catalyzed
H2O2, it further illustrates in sample to be tested and contains glucose, it so can be according to H2O2It is caused from it is colourless to blue color
Variation or the variation of corresponding ultraviolet-visible absorption spectroscopy to detect glucose indirectly.Colorimetric probe of the invention was detecting
It after glucose, can also continue to that sufficient bisulfite is added into probe solution, by the product malachite after probe reaction
The green addition product for becoming malachite green and bisulfite again again, to realize repeatedly using for probe.
The invention has the advantages that：1）Raw material malachite green of the present invention, bisulfites are inexpensively easy
, cost is not high；2）Method is easy in the preparation for reversible colorimetric probe of the invention, can easily detect solution and human serum
In glucose, the good, high sensitivity of selectivity, lowest detection are limited to 70 nM；3）Middle probe of the present invention was detecting glucose
Afterwards, can also by continuing to detect glucose again after bisulfite reaction is added, only consume during this it is micro,
Very cheap bisulfites, greatly reduces cost；4）Colorimetric probe high sensitivity of the present invention, selectivity be good, it is simple just
Victory, and may be reused, it is expected to promote the use of in field of biomedicine.
Detailed description of the invention
Fig. 1 is the high resolution mass spectrum figure of malachite green.
Fig. 2 is the high resolution mass spectrum figure after malachite green and bisulfite reaction.
Fig. 3 is the high resolution mass spectrum figure after continuously adding hydroperoxidation after malachite green and bisulfite reaction.
Fig. 4 is that different glucose is added in probe solution（0-1000 µM）With the reaction solution of glucose oxidase
Ultraviolet-visible spectrogram and color change photo afterwards（The concentration containing glucose is 0-1000 in the final mixed solution of 1.5 mL
µM）.
Fig. 5 is the Relative Absorbance at 618 nm（A/A0）With the linear relationship chart of concentration of glucose, wherein A and A0Respectively
It represents there are different glucose and there is no contain extinction of the PBS buffer system of probe at 618 nm when glucose
Degree.
Fig. 6 is the Relative Absorbance for the PBS buffer system for containing probe when potential interference object and glucose coexist in serum,
Illustration from left to right respectively corresponds the system color camera when interfering substance of abscissa from left to right coexists with glucose.
Fig. 7 is the PBS buffer system containing probe solution and its is added other carbohydrate interfering substances（Galactolipin, sucrose, wheat
Bud sugar）With the Relative Absorbance after glucose, illustration from left to right respectively correspond without sugar and containing galactolipin, sucrose, maltose,
System color camera when glucose（Galactolipin in the final mixed solution of 1.5 mL, sucrose, maltose concentration be 1200 μM,
The concentration of glucose is 120 μM）.
Fig. 8 is that the Reversible Cycle ability of probe investigates result figure, and illustration corresponds to the color change of Reversible Cycle laboratory sample
Figure.
Fig. 9 is 1 malachite green of embodiment and bisulfite addition product1H NMR spectra.
Specific embodiment
Illustrate a specific embodiment of the invention below with reference to embodiment, but following embodiment is used only to be described in detail
The present invention does not limit the scope of the invention in any way.
Embodiment 1：
The phosphate buffer solution of 300 μ L50 mM is added in the centrifuge tube of 1.5 mL（PBS, pH 6.8）, continuously add 150 μ
The sodium hydrogensulfite of L200 μM of malachite green and 30 μ L1.25mM form mixed liquor, and being supplemented total volume with ultrapure water is 1.5
ML contains 10 mM phosphate buffer solutions, 20 μM of malachite greens and 25 μM of NaHSO in mixed liquor3, mixed liquor reaction 3 minutes
Can be obtained containing（Call addition product in the following text）Probe solution（Call solution B in the following text）.
HRMS（ESI）：[M+H]+（Malachite green C23H25N2 +）calcd 329.2012, found 329.2018; [M-
H]+（The probe C that malachite green is reacted with bisulfite23H26N2 +O3S）calcd 409.1664, found
409.1595；[M+H]+（The malachite green C that probe is retrieved by hydrogen peroxide oxidation23H25N2 +）calcd 329.2012,
found 329.2017。
1, the reversible reaction Exploration of Mechanism of probe
HSO is added by malachite green3 −And then add H2O2What the high resolution mass spectrum variation occurred afterwards demonstrated probe can
Back reaction mechanism.As shown in Figure 1, the mass spectrometric data for measuring malachite green is 329.2018, the molecular weight of corresponding malachite green
329.2012.HSO is added in malachite green3 −Completely after reaction, occurs anion peak mass spectrometric data 409.1595 on mass spectrogram（Figure
2）, the molecular weight 410.1664 of this addition product formed with expected addition reaction（409.1664）It matches；Addition product adds
Enter H2O2And mass spectrometric data 329.2017 is measured after sufficiently reacting（Fig. 3）, it is consistent with the molecular weight of malachite green, furtherly
Bright addition product is by H2O2It is reoxidized at malachite green.Therefore, it by above-mentioned experimental result and analysis, is inhaled in conjunction with UV, visible light
Spectroscopic data is received, can determine malachite green and HSO3 −And H2O2Reduction-oxidation reversible reaction, i.e. HSO has occurred3 −With malachite
Green C=C double bond occurs nucleophilic addition and generates addition product, and addition product can be by H2O2It is oxidized to original malachite
It is green.These are the result shows that probe, that is, malachite green and HSO3 −Addition product with H2O2After reacting, moreover it is possible to pass through addition
HSO3 −Continuation and H2O2Reaction.
2, the color of probe solution and uv-vis spectra with the concentration of glucose of addition variation
Make 40 μ L PBS buffer solutions first（50 mM, pH 6.8）, 20 μ L glucose oxidizing ferment（10 mg/mL）With 140 μ L
Various concentration（0,1.07,5.36,10.71,53.57,107.14,214.29,428.57,642.86,857.14,1071.43,
1500 μM）Glucose be uniformly mixed, then mixed solution is incubated for 30 minutes in 37 °C to generate the H of various concentration2O2, this
When obtain solution A.
Embodiment 1 is added in the solution A of various concentration to obtain reacting 20 minutes in solution B, then with ultrapure water by total volume
Supplement is 1.5 mL and measures and record the color change figure of its ultraviolet-visible absorption spectroscopy and corresponding solution after mixing.Knot
As illustrated in figures 4-5,1-12 number respectively represents 0,0.1,0.5,1,5,10,20,40,60,80,100,140 μM of concentration to fruit in figure
Glucose, with the increase of concentration of glucose, solution gradually from it is colourless become blue；The variation of uv-vis spectra is：618
Absorbance at nm increases with increasing for glucose content.Linear Quasi is carried out to the concentration of Relative Absorbance and glucose
It closes, when the concentration range for finding the glucose in probe solution is 0.1-140 μM, Relative Absorbance and concentration presentation are good
Linear relationship, detection are limited to 70 nM, have very high sensitivity, far below blood-sugar content on an empty stomach（3.89-6.11 mM）,
It can be applied to the detection of glucose in life sample.
3, influence of the disturbance substance to probe
Make 40 μ L PBS buffer solutions first（50 mM, pH 6.8）, 20 μ L glucose oxidizing ferment（10 mg/mL）With 140 μ L
1.29 mM glucose are uniformly mixed, and are then incubated for 30 minutes mixed solution in 37 °C and are obtained solution C.Then it is made to embodiment 1
Potential various chaff interferents in solution C and serum are added in the solution B obtained（The concentration of potential interference object is 1% human serum in serum
In the concentration containing corresponding interfering substance）Reaction 20 minutes, by total volume supplement be 1.5 mL and after mixing measurement with
Record its ultraviolet-visible absorption spectroscopy and color change.As a result as shown in fig. 6, a-u respectively represents in figure abscissa from a left side in Fig. 6
To right sample, the experimental results showed that other chaff interferents only cause almost negligible influence.Then the present invention has been investigated
The ability of glucose and its analog is distinguished, v-z respectively represents the sample in figure in abscissa from left to right in Fig. 7, and Fig. 7 shows
Other common sugar that content is 10 times of concentration of glucose（Galactolipin, sucrose and maltose）The signal of generation is responded well below Portugal
Grape sugar, in addition it is almost low as background signal.In conclusion the method for the present invention has very high specificity to glucose,
Anti-interference ability is very strong.
4, the Reversible Cycle ability of probe is investigated
The phosphate buffer solution of 300 μ L50 mM is added in the centrifuge tube of 10 mL（PBS, pH 6.8）It is ultrapure with 778 μ L
Water, the sodium hydrogensulfite for continuously adding 1.25 mM of malachite green and 72 μ L of 200 μM of 150 μ L, which reacts to be formed for 3 minutes, to be mixed
Liquid D is closed, and measures its ultraviolet-visible absorption spectroscopy and records its color change figure.Then make 40 μ L PBS buffer solutions（50
MM, pH 6.8）, 20 μ L glucose oxidizing ferment（10 mg/mL）It is uniformly mixed with the glucose of 140 μ L, 2.14 mM, then will
Mixed solution is in 37 °C of incubations, 30 minutes generation H2O2, solution E is obtained at this time.Solution E is added in solution D and is reacted 20 minutes
To 1.5 mL solution F, the color change figure of its ultraviolet-visible absorption spectroscopy and corresponding solution is then measured and recorded.Again to solution
The phosphate buffer solution of 300 μ L, 50 mM is added in F（PBS, pH 6.8）, 72 μ L, 1.25 mM sodium hydrogensulfite and
928 μ L ultrapure waters react 3 minutes formation mixed liquor G, and measure its ultraviolet-visible absorption spectroscopy and record its color change figure.
Then above-mentioned solution E is prepared.Solution E is added in solution G to react 20 minutes and obtains 3 mL Solution Hs, then measures and records it
The color change figure of ultraviolet-visible absorption spectroscopy and corresponding solution.So analogize preparation solution I, J, K, L, M, N.Them are recorded to inhale
Receive absorbance of the peak at 618 nm, color change photo when in addition being recycled every time with cameras record.As a result as shown in figure 8,
Probe is by the H of the glycoxidative generation of grape2O2And HSO3 −After repeated oxidation restores 5 times, ultravioletvisible absorption only has small
Weaken, illustration shows that solution colour also only has faint colour fading, this illustrates that probe has good invertibity, and by multiple
Still retain the H to the glycoxidative generation of grape after redox2O2Reactivity and good spectral property.Therefore, of the invention
In probe can be recycled to detect H2O2And glucose, cost is greatly reduced in this way, is conducive to the method for the present invention
Further genralrlization has important application value in biological and medical field.
5, probe application power of glucose in detection human serum is investigated
Make 40 μ L PBS buffer solutions first（50 mM, pH 6.8）, 20 μ L glucose oxidizing ferment（10 mg/mL）, 15 μ L people
Blood serum sample and 125 μ L glucoses（0,60,120,240 μM）It is uniformly mixed, mixed solution is then incubated for 30 points in 37 °C
Clock obtains solution D.Then solution D is added into solution B made from embodiment 1 to react 20 minutes, with ultrapure water by total volume
Supplement is 1.5 mL and measures and record its ultraviolet-visible absorption spectroscopy and color change after mixing.Recovery testu knot
Fruit is as shown in table 1, and for the rate of recovery of recovery experiment between 92.8% and 108.9%, relative standard deviation is lower than 5.36%, shows
It is the stabilization of this method, accurate and reliable.In addition the extension rate of blood serum sample, 3 reality that colorimetric determination obtains are considered
In blood serum sample the content of glucose be 4.523 mM, 4.998 mM and 4.384 mM, it is basic with blood glucose level reported in the literature
It is consistent, these results sufficiently demonstrate actual application ability of the invention, and the detection for glucose in complex biological sample provides
A kind of new selection.
Table 1 uses the analysis result of glucose in 1% human serum of colorimetric determination
6, Fig. 9 is malachite green and bisulfite addition product1H NMR spectra,1H NMR（400 MHz, DMSO-d 6 ）δ
7.34 (t, J = 4 Hz, 2H), 7.24 (s, 3H), 7.15 (d, J =8.8 Hz, 4H), 6.67 (d, J =
9.2 Hz, 4H), 4.26 (s, 12H)；The structure for demonstrating malachite green and bisulfite addition product is shown knot
Structure formula.
Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, this field is common
Other modifications or equivalent replacement that technical staff makes technical solution of the present invention, without departing from technical solution of the present invention
Spirit and scope, be intended to be within the scope of the claims of the invention.
Claims (7)
1. a kind of reversible colorimetric probe based on malachite green Yu bisulfite addition product, which is characterized in that structural formula is such as
Shown in lower：。
2. the preparation side described in claim 1 based on malachite green Yu the reversible colorimetric probe of bisulfite addition product
Method, which is characterized in that steps are as follows：1）Take raw material malachite green and bisulfites；
2）Malachite green and bisulfites are added in phosphate buffer, react 3min at room temperature, is obtained based on malachite
The green reversible colorimetric probe solution with bisulfite addition product.
3. the preparation method of reversible colorimetric probe as claimed in claim 2, which is characterized in that the malachite green and sulfurous acid
The mass ratio of the material of hydrogen radical is 1：（1-1.25）.
4. the preparation method of reversible colorimetric probe as claimed in claim 2, which is characterized in that the bisulfites is sulfurous
Sour hydrogen sodium, potassium bisulfite or ammonium bisulfite；The concentration of the phosphate buffer is 10mmol/L.
5. application of the reversible colorimetric probe in test sample in terms of glucose described in claim 1.
6. application as claimed in claim 5, which is characterized in that when concrete application, first by sample to be tested and glucose oxidase
Mixing is incubated for and obtains reaction solution, and then reaction solution is added in reversible colorimetric probe solution.
7. application as claimed in claim 6, which is characterized in that described be incubated at 37 DEG C carries out 30min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810606827.5A CN108822005B (en) | 2018-06-13 | 2018-06-13 | Reversible colorimetric probe based on malachite green and bisulfite addition product, and preparation and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810606827.5A CN108822005B (en) | 2018-06-13 | 2018-06-13 | Reversible colorimetric probe based on malachite green and bisulfite addition product, and preparation and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108822005A true CN108822005A (en) | 2018-11-16 |
| CN108822005B CN108822005B (en) | 2021-04-09 |
Family
ID=64145090
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810606827.5A Expired - Fee Related CN108822005B (en) | 2018-06-13 | 2018-06-13 | Reversible colorimetric probe based on malachite green and bisulfite addition product, and preparation and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108822005B (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5631641A (en) * | 1979-08-24 | 1981-03-31 | Yatoron:Kk | New quantitative determination method for hydrogen peroxide |
| DD235115A1 (en) * | 1985-03-04 | 1986-04-23 | Akad Wissenschaften Ddr | METHOD FOR QUANTITATIVE TRACE DETECTION OF HYDROGEN PEROXIDE |
| CN1413235A (en) * | 1999-10-27 | 2003-04-23 | 生命扫描有限公司 | 8-(Anilino)-1-naphthalenesulfonate analogs and their use in analyte detection assays |
| US20080057528A1 (en) * | 2006-08-30 | 2008-03-06 | Kimberly-Clark Worldwide, Inc. | Detection of hydrogen peroxide released by enzyme-catalyzed oxidation of an analyte |
| US20090061528A1 (en) * | 2007-08-06 | 2009-03-05 | Glumetrics Inc. | Hpts-mono and bis cys-ma polymerizable fluorescent dyes for use in analyte sensors |
| CN100577741C (en) * | 2003-12-16 | 2010-01-06 | 帕瑞萨森思公司 | Reagent for detecting analyte |
| CN103424545A (en) * | 2013-08-12 | 2013-12-04 | 深圳市康博霖科技有限公司 | Malachite green oxalate ELISA (enzyme-linked immunosorbent assay) detection kit and method |
-
2018
- 2018-06-13 CN CN201810606827.5A patent/CN108822005B/en not_active Expired - Fee Related
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5631641A (en) * | 1979-08-24 | 1981-03-31 | Yatoron:Kk | New quantitative determination method for hydrogen peroxide |
| DD235115A1 (en) * | 1985-03-04 | 1986-04-23 | Akad Wissenschaften Ddr | METHOD FOR QUANTITATIVE TRACE DETECTION OF HYDROGEN PEROXIDE |
| CN1413235A (en) * | 1999-10-27 | 2003-04-23 | 生命扫描有限公司 | 8-(Anilino)-1-naphthalenesulfonate analogs and their use in analyte detection assays |
| CN100577741C (en) * | 2003-12-16 | 2010-01-06 | 帕瑞萨森思公司 | Reagent for detecting analyte |
| US20080057528A1 (en) * | 2006-08-30 | 2008-03-06 | Kimberly-Clark Worldwide, Inc. | Detection of hydrogen peroxide released by enzyme-catalyzed oxidation of an analyte |
| US20090061528A1 (en) * | 2007-08-06 | 2009-03-05 | Glumetrics Inc. | Hpts-mono and bis cys-ma polymerizable fluorescent dyes for use in analyte sensors |
| CN103424545A (en) * | 2013-08-12 | 2013-12-04 | 深圳市康博霖科技有限公司 | Malachite green oxalate ELISA (enzyme-linked immunosorbent assay) detection kit and method |
Non-Patent Citations (2)
| Title |
|---|
| RICHARDN.MACNAIR等: "A Fatigue Mechanism for Thermochromism in Leucosulfites of Triphenylmethane Compounds", 《THE JOURNAL OF ORGANIC CHEMISTRY》 * |
| YIHUI HU等: "Surface-Enhanced Raman Scattering Active Gold Nanoparticles with Enzyme-Mimicking Activities for Measuring G lucose and Lactate in Living Tissues", 《ACS NANO》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108822005B (en) | 2021-04-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sun et al. | Glucose sensing in supramolecular chemistry | |
| CN110057801A (en) | A kind of ratio fluorescent probe and its hydrogen peroxide and glucose detection application based on aggregation-induced emission property | |
| CN101865853B (en) | Stabilized beta-hydroxybutyric acid detection test paper and preparation method thereof | |
| Wang et al. | A specifically triggered turn-on fluorescent probe platform and its visual imaging of HClO in cells, arthritis and tumors | |
| Fereja et al. | Highly sensitive and selective non-enzymatic glucose detection based on indigo carmine/hemin/H 2 O 2 chemiluminescence | |
| Chen et al. | Signal-on bimodal sensing glucose based on enzyme product-etching MnO2 nanosheets for detachment of MoS2 quantum dots | |
| Liu et al. | A pH-responsive fluorometric and colorimetric system based on silicon quantum dots and 4-nitrophenol for urease activity detection | |
| JP2014528086A (en) | Measurement of lactic acid in biological fluids | |
| Mills | Optical sensors for carbon dioxide and their applications | |
| CN109651249A (en) | A kind of fluorescence probe detecting endocytoplasmic reticulum cysteine and its synthesis and application | |
| CN104330393B (en) | Method for determining glucose by using gold nano-cluster as fluorescence probe | |
| Li et al. | Smartphone-assisted fluorescent microfluidic-chip for sensitive detection of sweat glucose via dual-sensing of O2/H2O2 | |
| CN101620184A (en) | Plastic optical fiber sensing system for simultaneously measuring blood sugar concentration and blood oxygen concentration | |
| Liu et al. | Microplate-integrated biosensors for glucose and lactate | |
| CN108752373A (en) | A kind of fluorescence probe identifying hydrogen peroxide based on benzene boron ester | |
| CN110172070A (en) | A kind of fluorescence probe and its synthetic method and application detecting viscosity and hydrogen peroxide | |
| CN112730367B (en) | Method and device for determining alkaline phosphatase by multi-signal spectrum sensing platform based on portable intelligent terminal | |
| CN1737544A (en) | Method for rapid detecting sodium sulfoxylate formaldehyde in foodstuff | |
| CN110981857B (en) | Ultrasensitive ferrous ion fluorescent probe, preparation method and application | |
| CN110669503B (en) | Preparation and application of carbon monoxide near-infrared fluorescent probe | |
| CN108822005A (en) | A kind of reversible colorimetric probe and its preparation, application based on malachite green and bisulfite addition product | |
| CN103115882B (en) | Detect H2O2Method (2) | |
| CN107561070B (en) | A kind of glucose quick visualization detection method and its preparation and application | |
| Huang et al. | A sensitive and noninvasive cyclic peptide-based electrogenerated chemiluminescence biosensing method for the determination of sweat glucose | |
| Rivera et al. | New ketocyanine-based fluorescent optodes for the development of integrated waveguide sensors |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20210409 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |