CN104914102A - Micro-fluidic chip and applications thereof - Google Patents
Micro-fluidic chip and applications thereof Download PDFInfo
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- CN104914102A CN104914102A CN201510330599.XA CN201510330599A CN104914102A CN 104914102 A CN104914102 A CN 104914102A CN 201510330599 A CN201510330599 A CN 201510330599A CN 104914102 A CN104914102 A CN 104914102A
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Abstract
The invention provides a micro-fluidic chip and applications thereof. The micro-fluidic chip comprises a chip body made of a transparent material, the chip body comprises a first mixing tank and a second mixing tank which are connected with each other through a micro-channel, and an eighth valve is arranged on the micro-channel; the first mixing tank is connected with a first sample introduction micro-channel, a second sample introduction micro-channel, a third sample introduction micro-channel and a first sample discharge micro-channel, a first valve is arranged on the first sample introduction micro-channel, a second valve is arranged on the second sample introduction micro-channel, a third valve is arranged on the third sample introduction micro-channel, and a fourth valve is arranged on the first sample discharge micro-channel; the second mixing tank is connected with a fourth sample introduction micro-channel, a fifth sample introduction micro-channel and a second sample discharge micro-channel, a fifth valve is arranged on the fourth sample introduction micro-channel, a sixth valve is arranged on the fifth sample introduction micro-channel, and a seventh valve is arranged on the second sample discharge micro-channel. The micro-fluidic chip has good effect when used for detecting lactic acid by adopting a colorimetric method, and thus the prospect of the micro-fluidic chip in application field of the colorimetric method is disclosed.
Description
Technical field
The present invention relates to a kind of micro-fluidic chip, specifically refer to a kind of micro-fluidic chip that may be used for colorimetric determination formulation material, especially may be used for modifying thia Graphene/γ-Fe
2o
3by colorimetric determination lactic acid after nano composite material.
Background technology
In recent years, the microminiaturization of analytical equipment has become an important research direction, and thus the development of microfluidic chip technology also earns widespread respect.Micro-fluidic chip on chip, builds by Micrometer-Nanometer Processing Technology the microflow path system be made up of liquid storage tank, micro-reaction chamber, microchannel etc. micro-function element, after loading biological sample and reactant liquor, microfluidic circuit is formed under forcing pump or electric field action, on chip, carry out a kind of or multiple continuously reaction, reach the object to sample high flux, low consumption, express-analysis.Microfluidic analysis chip is owing to having high integration, the several functions such as sampling, dilution, reagent adding, reaction, separation, detection can be completed on a chip, be otherwise known as micro-full analytical system (Micro total analysis system, μ-TAS).Microfluidic chip technology through the development of nearly ten years, has now become one of sciemtifec and technical sphere of forefront on our times after setting up.At present, micro-fluidic chip has been widely used in the various fields such as biomedicine, high-throughput drug synthesis and screening, environmental monitoring, biological agent detection.
In material with carbon element, graphite is rare is by sp
2the ultra-thin two-dimension web frame that hydbridized carbon atoms is formed, as a kind of material with carbon element of bi-dimensional cellular crystalline network become by monolayer carbon atom dense arrangement, Graphene has advantages such as conducting electricity very well, the large and electron transfer rate of specific surface area is fast.The elements such as N, B be easily doped to graphite rare in, but as compared to B with N, S doping is very special, S atomic radius is far longer than C atom, and the electronegativity between C (2.55) and S (2.58) differs too little, obvious Charger transfer can not be provided in C-S compound, therefore, during chemical doping S is rare to graphite, seem quite difficult.But theory calculate shows, the structure that after doping S atom, meeting distortion graphite is rare, thus promote the performance of Graphene, so it is vital for mixing S atom in graphite is rare.Chinese patent (CN201310080785.3) discloses a kind of method of Graphene being carried out to sulfur doping: provide Graphene, and described Graphene is placed in chemical vapour deposition reaction chamber; Inert gas is adopted to ventilate and pump-down process to described reaction chamber; At 500 ~ 1050 DEG C, pass into sulphur source gas sulfur doping is carried out to described Graphene; In hydrogen and inert gas atmosphere, described reaction chamber is lowered the temperature.The method operation is more loaded down with trivial details, and the chemical vapour deposition technique adopted is high to equipment requirement.Occurring in nature Fe
2o
3polymorphic form mutation there will be a known two kinds, i.e. α-Fe
2o
3with γ-Fe
2o
3.The former stablizes under field conditions (factors), is called haematite; The latter is under field conditions (factors) not as α-Fe
2o
3stable, be in metastable condition, be referred to as maghemite.γ-Fe
2o
3nano particle enjoys favor because of it in the widespread use in the fields such as magnetic, catalysis, air-sensitive, biomedicine, but its instability again limit its application, and therefore preparation can the γ-Fe of long period stable existence
2o
3significant.Prepare magnetic γ-Fe in recent years
2o
3method have a lot, the such as method such as sol-gel process (Chinese patent application CN200410086479.1), sintering process (Chinese patent application CN200710036501.5), these methods are calcined at a certain temperature and obtain γ-Fe
2o
3.Prepare in the magnetic composite of gained except comprising γ-Fe at some
2o
3phase, also comprises the α-Fe of some
2o
3.In addition, these γ-Fe
2o
3poor heat stability, at about 400 DEG C γ-Fe
2o
3α-Fe will be changed into
2o
3.
At present, the method for quantitative measurement content of material is broadly divided into two large classes: bioassay method and physical chemistry determination method.Biologicall test ratio juris realizes the sensitive reaction of material concentration to be measured based on some biological tissue.But the sensitivity of bioassay method and specificity poor.Thus, nearly decades, the development of bioassay method does not have made marked progress.Physical chemistry measures ratio juris to be had a lot, concrete assay method has multiple, as gas chromatography and flame ionic detector coupling method (GC – FID) and Gao Xiao Ye Xiang – electrochemical detector coupling (HLPC – ED) method and Ye Xiang Se Pu – charged spray ionogenic mass spectrometer coupling method (LC/ESI – MS/MS) etc.Traditional chromatography reality with in all have that complicated operation, sample preparation are time-consuming, sensitivity and specificity not, the shortcoming such as apparatus expensive.Therefore, simple, quick, economic, the effective new detecting method of development has become extremely urgent important subject.Colorimetric detection is an importance in analysis science field, and it is crucial is exactly become color to change detecting event transition.Colorimetric detection has very large advantage compared with traditional detection, especially in operability, economy and practicality etc.Colourimetry by the change of naked eyes Direct Recognition reaction solution color, can not need the instrument of costliness or complexity, operates very easy, therefore can be applicable to on-the site analysis and care diagnostic.
Lactic acid is extensively present in biosome as a kind of important organic acid, is the glycometabolic important products of biosome anaerobic.Lactic acid is the carbohydate metabolism material in musculature, and after animal slaughtering, meat generates lactic acid at low temperature ripening period Muscle glycogen through anerobic glycolysis, thus causes the decline of pH.The accumulating amount of lactic acid and the decline degree of pH are proportionate.Lactic acid in a large number generation can cause the decline in various degree of Chilled Meats water-retaining property and tenderness, has substantial connection with the generation of PSE meat, DFD meat.Analyze mensuration lactic acid significant in medical treatment and food industry, and the quantitative of lactic acid and qualitative analysis are very important for the character of overall understanding food and the physiological function of further research lactic acid.At present, the detection method of lactic acid mainly comprises EDTA compleximetry, enzyme process, Enzyme Electrode, liquid phase chromatography etc.The scope of each self adaptation own of these traditional detection methods and precision, also all receive the impact of cost, but these methods with the naked eye can not identify intuitively.Colorimetric detection is an importance in analysis science field, and it is crucial is exactly become color to change detecting event transition.Colorimetric detection has very large advantage compared with traditional detection, especially in operability, economy and practicality etc.Colourimetry by the change of naked eyes Direct Recognition reaction solution color, can not need the instrument of costliness or complexity, operates very easy, therefore can be applicable to on-the site analysis and care diagnostic.
For prior art, the present invention devises a kind of micro-fluidic chip and is applied to the detection field of material, this chip may be used for quick, sensitive, quantitatively detect lactic acid; Further, this micro-fluidic chip, by after increase strong magnets, can conveniently be modified ferromagnetic material, present invention utilizes a kind of stable graphene composite material thia Graphene/γ-Fe
2o
3to be modified on micro-fluidic chip, because γ is-Fe as decorative material
2o
3there is ferromagnetism, therefore facilitate the modification of material; Further experimental studies have found that, modified thia Graphene/γ-Fe
2o
3after, reach the effect better detecting lactic acid.The micro-fluidic chip of the present invention's design and experimental technique, simple, quick, sensitive, effective, and cost is lower, the more important thing is that this chip can reuse; By the use of microflow control technique, in experimentation, reduce the consumption of medicine used and compound substance, save resource.
Summary of the invention
The object of this invention is to provide a kind of micro-fluidic chip that may be used for lactate detection, this chip is at modification thia Graphene/γ-Fe
2o
3can Sensitive Detection lactic acid more after nano composite material.
The present invention is achieved by the following technical solutions:
A kind of micro-fluidic chip, the chip comprising transparent material is overall, and described chip entirety comprises the first mixing pit and the second mixing pit, and the first mixing pit is connected by microchannel with the second mixing pit, and microchannel is provided with the 8th valve;
First mixing pit connects the first sample introduction microchannel, the second sample introduction microchannel, the 3rd sample introduction microchannel and the first stock layout microchannel, first sample introduction microchannel is provided with the first valve, second sample introduction microchannel is provided with the second valve, 3rd sample introduction microchannel is provided with the 3rd valve, and the first stock layout microchannel is provided with the 4th valve;
Second mixing pit connects the 4th sample introduction microchannel, the 5th sample introduction microchannel and the second stock layout microchannel, and the 4th sample introduction microchannel is provided with the 5th valve, and the 5th sample introduction microchannel is provided with the 6th valve, and the second stock layout microchannel is provided with the 7th valve.
In such scheme, the material of described chip entirety is dimethyl silicone polymer, polymethylmethacrylate or ITO material.
In such scheme, the overall lower surface of described chip and be positioned at immediately below the first mixing pit and be provided with dismountable strong magnets.
In such scheme, described first mixing pit and the second mixing pit are equipped with anti-spilled cover plate, and anti-spilled cover plate is dimethyl silicone polymer, polymethylmethacrylate or ITO material.
Described micro-fluidic chip is used for colorimetric determination lactic acid.
Described micro-fluidic chip is for modifying thia Graphene/γ-Fe
2o
3nano composite material is by colorimetric determination lactic acid.
Described micro-fluidic chip is for modifying thia Graphene/γ-Fe
2o
3nano composite material is by the purposes of colorimetric determination lactic acid, and using method comprises the steps:
Step 1, open the first valve, close other valves, by pump by thia Graphene/γ-Fe
2o
3the aqueous dispersions of nano composite material sends into the first mixing pit, closes the first valve, thia Graphene/γ-Fe
2o
3nano composite material is fixed on after bottom the first mixing pit by strong magnets, opens the 4th valve, is discharged by water by pump, closes the 4th valve, dry first mixing pit;
Step 2, open the second valve, after the phosphate buffer solution containing Lactate Oxidase being sent into the first mixing pit by pump, close the second valve; Open the 3rd valve, after the phosphate buffer solution containing lactic acid being sent into the first mixing pit by pump, close the 3rd valve, incubation reaction;
Step 3, open the 8th valve, by pump, the mixed liquor after incubation reaction in step 2 is delivered to the second mixing pit, close the 8th valve;
Step 4, open the 5th valve, by pump, hac buffer is sent into the second mixing pit, close the 5th valve; Open the 6th valve, will containing 3,3 ' by pump, the alcoholic solution of 5,5 ’ – tetramethyl benzidine sends into the second mixing pit, closes the 6th valve, incubation reaction;
Step 5, light irradiate the second mixing pit, scanning absorbance curve, record experimental data;
Step 6, open the 7th valve, by pump, reacted mixed liquor is discharged;
Step 7, cleaning and dry micro-fluidic chip.
The first mixing pit in chip entirety is connected by microchannel with the second mixing pit, and described microchannel is provided with valve; First mixing pit is used for compound substance catalysis Lactate Oxidase decomposing lactic acid and produces H
2o
2, the second mixing pit is used for the H entered from the first mixing pit
2o
2catalysis 3,3 ', 5,5 ’ – tetramethyl benzidine (TMB) and light irradiate with the detection carrying out absorbance; The overall lower surface of chip and be positioned at immediately below the first mixing pit and be provided with dismountable strong magnets, strong magnets is used for magnetic absorption thia Graphene/γ-Fe
2o
3nano composite material, when cleaning the first mixing pit, strong magnets of can dismantling is so that cleaning; First mixing pit connects three sample introduction microchannels and the first stock layout microchannel, wherein the first sample introduction microchannel is used for the sample introduction of compound substance dispersion liquid, second sample introduction microchannel is used for the sample introduction of Lactate Oxidase solution, 3rd sample introduction microchannel is used for the sample introduction of lactic acid solution, and the first stock layout microchannel is for discharging the liquid in compound substance dispersion liquid; Second mixing pit connects two sample introduction microchannels and the second stock layout microchannel, wherein, 4th sample introduction microchannel is used for the sample introduction of hac buffer (ABS) solution, and the 5th sample introduction microchannel is used for the sample introduction of TMB, and the second stock layout microchannel is used for final reactant liquor to discharge.All microchannels are equipped with valve and connect pump, the pump that all sample introduction microchannels connect is equipped with micro syringe.
Micro-fluidic chip is placed on temperature-controlled cabinet, and the pump that sample introduction microchannel is equipped with and micro syringe, for compound substance dispersion liquid, Lactate Oxidase solution, lactic acid solution, TMB solution and ABS sample introduction.Micro-fluidic chip is placed on transmission Bracket for Inspection, and be provided with light launching fiber probe above transmissive support, transmitting probe detects mouth upper surface by collimating mirror and micro-fluidic chip and is affixed.Below support, detection fiber is housed, this detection fiber is affixed by collimating mirror and chip lower surface.In addition, launching fiber connects accurate light source, and detection fiber probe is connected with spectrometer, and the probe of light launching fiber connects with accurate light source.
Beneficial effect of the present invention is:
(1) the micro-fluidic chip line clear designed by, simple and cost is lower, this chip can reuse.
(2) micro-fluidic chip designed by can be advantageously used in colorimetric determination lactic acid, this micro-fluidic chip is by after increase strong magnets, ferromagnetic material can be modified, present invention utilizes a kind of stable graphene composite material thia Graphene/γ-Fe
2o
3to be modified on micro-fluidic chip, because γ is-Fe as decorative material
2o
3there is ferromagnetism, therefore facilitate the modification of material, further experimental studies have found that, modified thia Graphene/γ-Fe
2o
3after, reach the effect better detecting lactic acid.Micro-fluidic chip designed by the present invention has easy and simple to handle, that sample preparation is simple, highly sensitive advantage.
(3) by the use of microflow control technique, in experimentation, reduce the consumption of medicine used and compound substance, save resource.
Accompanying drawing explanation
Fig. 1 is plan structure schematic diagram of the present invention;
Fig. 2 is the abosrption spectrogram of the present invention for colorimetric determination lactic acid.
In figure: 1-first mixing pit; 2-second mixing pit; 3-first sample introduction microchannel; 4-second sample introduction microchannel; 5-the 3rd sample introduction microchannel; 6-first stock layout microchannel; 7-the 4th sample introduction microchannel; 8-the 5th sample introduction microchannel; 9-second stock layout microchannel; 10-microchannel; 11-first valve; 12-second valve; 13-the 3rd valve; 14-the 4th valve; 15-the 5th valve; 16-the 6th valve; 17-the 7th valve; 18-the 8th valve; 19-strong magnets
Embodiment
Below in conjunction with accompanying drawing, the invention will be further described:
As shown in Figure 1, a kind of micro-fluidic chip, the chip comprising dimethyl silicone polymer material, polymethylmethacrylate material or ITO material is overall, described chip entirety comprises the first mixing pit 1 and the second mixing pit 2, first mixing pit 1 is connected by microchannel 10 with the second mixing pit 2, and microchannel 10 is provided with the 8th valve 18; First mixing pit 1 connects the first sample introduction microchannel 4, sample introduction microchannel 3, second, the 3rd sample introduction microchannel 5 and the first stock layout microchannel 6, first sample introduction microchannel 3 is provided with the first valve 11, second sample introduction microchannel 4 is provided with the second valve 12,3rd sample introduction microchannel 4 is provided with the 3rd valve 13, first stock layout microchannel 6 and is provided with the 4th valve 14; Second mixing pit 2 connects the 4th sample introduction microchannel 7, the 5th sample introduction microchannel 8 and the second stock layout microchannel 9,4th sample introduction microchannel 7 is provided with the 5th valve 15,5th sample introduction microchannel 8 is provided with the 6th valve 16, second stock layout microchannel 9 and is provided with the 7th valve 17; The overall lower surface of described chip and be positioned at immediately below the first mixing pit 1 and be provided with strong magnets 19, strong magnets 19 is for magnetic absorption thia Graphene/γ-Fe
2o
3nano composite material; Described first mixing pit 1 and the second mixing pit 2 are equipped with anti-spilled cover plate, and anti-spilled cover plate is dimethyl silicone polymer, polymethylmethacrylate or ITO material.
Here is the specific embodiment of the present invention for colorimetric determination lactic acid:
Embodiment 1
Step 1, open the first valve (11), close other valves, by the thia Graphene/γ-Fe of pump by 200 μ L 2mg/mL
2o
3the aqueous dispersions of nano composite material sends into the first mixing pit (1), closes the first valve (11), thia Graphene/γ-Fe
2o
3after nano composite material is fixed on the first mixing pit (1) bottom by strong magnets (19), open the 4th valve (14), by pump, water is discharged, close the 4th valve (14), dry first mixing pit (1);
Step 2, open the second valve (12), close other valves, after the phosphate buffer solution (10mM, pH=7.4) that 100 μ L contain 0.5 unit Lactate Oxidase being sent into the first mixing pit (1) by pump, close the second valve (12); Open the 3rd valve (13), after the phosphate buffer solution (10mM, pH=7.4) that 100 μ L contain the lactic acid of 10 μm of ol being sent into the first mixing pit (1) by pump, close the 3rd valve (13), incubation reaction;
Step 3, open the 8th valve (18), by pump, the mixed liquor after incubation reaction in step 2 is delivered to the second mixing pit (2), close the 8th valve (18);
Step 4, open the 5th valve (15), by pump, 200 μ L 0.2M hac buffer (pH=4.0) are sent into the second mixing pit (2), close the 5th valve (15); Open the 6th valve (16), by pump, 100 μ L are contained 0.5mg 3,3 ', 5, the alcoholic solution of 5 ’ – tetramethyl benzidine sends into the second mixing pit (2), closes the 6th valve (16), incubation reaction;
Step 5, light irradiate the second mixing pit (2), at λ=652nm place scanning absorbance curve, and record experimental data;
Step 6, open the 7th valve (17), by pump, reacted mixed liquor is discharged;
Step 7, cleaning and dry micro-fluidic chip.
Embodiment 2
Step 1, open the second valve (12), by pump, 100 μ L are contained the phosphate buffer solution (10mM of 0.5 unit Lactate Oxidase, pH=7.4), after sending into the first mixing pit (1), the second valve (12) is closed; Open the 3rd valve (13), after the phosphate buffer solution (10mM, pH=7.4) that 100 μ L contain the lactic acid of 10 μm of ol being sent into the first mixing pit (1) by pump, close the 3rd valve (13), incubation reaction;
Step 2, open the 8th valve (18), by pump, the mixed liquor after incubation reaction in step 1 is delivered to the second mixing pit (2), close the 8th valve (18);
Step 3, open the 5th valve (15), by pump, 200 μ L 0.2M hac buffer (pH=4.0) are sent into the second mixing pit (2), close the 5th valve (15); Open the 6th valve (16), by pump, 100 μ L are contained 0.5mg 3,3 ', 5, the alcoholic solution of 5 ’ – tetramethyl benzidine sends into the second mixing pit (2), closes the 6th valve (16), incubation reaction;
Step 4, light irradiate the second mixing pit (2), at λ=652nm place scanning absorbance curve, and record experimental data;
Step 5, open the 7th valve (17), by pump, reacted mixed liquor is discharged;
Step 6, cleaning and dry micro-fluidic chip.
In Fig. 2, a curve is the scanning result of embodiment 1, and b curve is the scanning result of embodiment 2, can find out, micro-fluidic chip of the present invention can well be used for colorimetric determination lactic acid, and modifies thia Graphene/γ-Fe
2o
3after, absorbance obviously increases, and illustrates that this micro-fluidic chip has good effect at colorimetric determination lactic acid, thus discloses its prospect in colourimetry application.
Foregoing is only the preferred embodiments of the present invention; not for limiting embodiment of the present invention; those of ordinary skill in the art are according to central scope of the present invention and spirit; corresponding flexible or amendment can be carried out very easily; certainly; these amendments made according to the present invention's spirit, all should be included within the present invention's scope required for protection.
Claims (7)
1. a micro-fluidic chip, it is characterized in that, the chip comprising transparent material is overall, described chip entirety comprises the first mixing pit (1) and the second mixing pit (2), first mixing pit (1) is connected by microchannel (10) with the second mixing pit (2), and microchannel (10) are provided with the 8th valve (18);
First mixing pit (1) connects the first sample introduction microchannel (3), the second sample introduction microchannel (4), the 3rd sample introduction microchannel (5) and the first stock layout microchannel (6), first sample introduction microchannel (3) is provided with the first valve (11), second sample introduction microchannel (4) is provided with the second valve (12), 3rd sample introduction microchannel (4) is provided with the 3rd valve (13), and the first stock layout microchannel (6) is provided with the 4th valve (14);
Second mixing pit (2) connects the 4th sample introduction microchannel (7), the 5th sample introduction microchannel (8) and the second stock layout microchannel (9), 4th sample introduction microchannel (7) is provided with the 5th valve (15), 5th sample introduction microchannel (8) is provided with the 6th valve (16), and the second stock layout microchannel (9) is provided with the 7th valve (17).
2. a kind of micro-fluidic chip according to claim 1, is characterized in that, the material of described chip entirety is dimethyl silicone polymer, polymethylmethacrylate or ITO material.
3. a kind of micro-fluidic chip according to claim 1, is characterized in that, the overall lower surface of described chip and be positioned at immediately below the first mixing pit (1) and be provided with dismountable strong magnets (19).
4. a kind of micro-fluidic chip according to claim 1, it is characterized in that, described first mixing pit (1) and the second mixing pit (2) are equipped with anti-spilled cover plate, and anti-spilled cover plate is dimethyl silicone polymer, polymethylmethacrylate or ITO material.
5. the purposes of a kind of micro-fluidic chip according to claim 1,2,3 or 4, is characterized in that, described micro-fluidic chip is used for colorimetric determination lactic acid.
6. the purposes of a kind of micro-fluidic chip according to claim 5, is characterized in that, described micro-fluidic chip is for modifying thia Graphene/γ-Fe
2o
3nano composite material is by colorimetric determination lactic acid.
7. the purposes of a kind of micro-fluidic chip according to claim 6, it is characterized in that, using method comprises the steps:
Step 1, open the first valve (11), close other valves, by pump by thia Graphene/γ-Fe
2o
3the aqueous dispersions of nano composite material sends into the first mixing pit (1), closes the first valve (11), thia Graphene/γ-Fe
2o
3after nano composite material is fixed on the first mixing pit (1) bottom by strong magnets (19), open the 4th valve (14), by pump, water is discharged, close the 4th valve (14), dry first mixing pit (1);
Step 2, open the second valve (12), after the phosphate buffer solution containing Lactate Oxidase being sent into the first mixing pit (1) by pump, close the second valve (12); Open the 3rd valve (13), after the phosphate buffer solution containing lactic acid being sent into the first mixing pit (1) by pump, close the 3rd valve (13), incubation reaction;
Step 3, open the 8th valve (18), by pump, the mixed liquor after incubation reaction in step 2 is delivered to the second mixing pit (2), close the 8th valve (18);
Step 4, open the 5th valve (15), by pump, hac buffer is sent into the second mixing pit (2), close the 5th valve (15); Open the 6th valve (16), will containing 3,3 ' by pump, the alcoholic solution of 5,5 ’ – tetramethyl benzidine sends into the second mixing pit (2), closes the 6th valve (16), incubation reaction;
Step 5, light irradiate the second mixing pit (2), scanning absorbance curve, record experimental data;
Step 6, open the 7th valve (17), by pump, reacted mixed liquor is discharged;
Step 7, cleaning and dry micro-fluidic chip.
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