CN115161019A - Preparation method of nitrogen-doped luminescent carbon quantum dot and application of nitrogen-doped luminescent carbon quantum dot in rapid detection of lysine content in pig serum - Google Patents
Preparation method of nitrogen-doped luminescent carbon quantum dot and application of nitrogen-doped luminescent carbon quantum dot in rapid detection of lysine content in pig serum Download PDFInfo
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Abstract
The invention discloses a preparation method of nitrogen-doped luminescent carbon quantum dots and application thereof in rapidly detecting lysine content in pig serum, wherein 2, 3-diaminopyridine, acrylic acid and zinc chloride are used as raw materials, and the carbon quantum dots are prepared by an ionothermal method and used as a fluorescent probe for detecting lysine; manufacturing a micro-fluidic chip with two parallel channels; adding a polyethylene glycol diacrylate solution, 2-hydroxy-2-methyl-phenyl acetone and a carbon dot solution into a channel, and forming a hydrogel microstructure embedded with carbon quantum dots in the channel by using a light patterning method as a detection element; adding the serum to be detected into a detection channel of the chip, putting the chip into an imaging device, and taking a picture by using a mobile phone; the photograph was processed, and the relative fluorescence intensity was calculated to determine the lysine content in the serum. The invention has the advantages of high sensitivity, high specificity, simplicity, rapidness, low sample requirement, low cost and the like, and is beneficial to applying an ideal amino acid mode to implement accurate feeding.
Description
Technical Field
The invention relates to the crossing field of carbon nano materials, fluorescence detection and microfluidic technologies, in particular to a preparation method of nitrogen-doped luminescent carbon quantum dots and application of the nitrogen-doped luminescent carbon quantum dots in rapid detection of lysine content in pig serum.
Background
Lysine is one of the essential amino acids, the first limiting amino acid for pigs fed on a corn-soybean meal type ration. If the lysine cannot meet the nutritional requirement, the growth of the pigs is hindered; if the amount of lysine added exceeds the nutritional requirement, it is wasted and the corresponding antagonistic amino acid functions. Therefore, the method can accurately measure the content of lysine in the pig serum and is beneficial to realizing the requirement of accurate nutritional formula by applying an ideal amino acid mode.
Traditional methods for detecting lysine include gas chromatography-mass spectrometry (GC-MS), high Performance Liquid Chromatography (HPLC), liquid chromatography-tandem mass spectrometry (LC-MS/MS), nuclear Magnetic Resonance (NMR) and the like. Although the methods can accurately determine the content of lysine, the methods have the defects of expensive large-scale instruments, complex detection process, long time consumption, requirement of professional operation and the like, and are not suitable for popularization and application in pig breeding sites. Therefore, it is required to develop a portable device and method for detecting the content of lysine in pig serum, which is simple and rapid to operate.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a preparation method of a nitrogen-doped luminescent carbon quantum dot and application thereof in rapidly detecting the lysine content in pig serum. According to the invention, nitrogen-doped luminescent carbon quantum dots are applied to a microfluidic chip, and a portable device for quantitative fluorescence detection of lysine content in pig serum based on a smart phone is designed, so that a detection result can be obtained quickly and accurately.
In order to achieve the purpose, the invention designs a preparation method of a nitrogen-doped luminescent carbon quantum dot, which comprises the following steps:
1) Weighing 2, 3-diaminopyridine, acrylic acid and zinc chloride according to a molar ratio of 1;
2) Adding 2, 3-diaminopyridine, acrylic acid and zinc chloride into tetraethylene glycol, and stirring to mix uniformly; reacting for 50-60 min at the temperature of 90-110 ℃ to fully dissolve reactants; heating to 200-220 ℃ for reaction for 220-260 min, and cooling to obtain a crude product solution;
3) Dissolving the crude product solution in methanol, carrying out centrifugal filtration to obtain a filtered product, and removing the solvent from the product in a rotary evaporator to obtain a crude product;
4) Adding diethyl ether into the crude product, shaking to separate out a black solid, collecting the precipitate, adding methanol again to dissolve, adding diethyl ether, shaking to separate out a solid, repeating for several times, and removing unreacted zinc chloride; and drying in vacuum to obtain solid, namely the nitrogen-doped luminescent carbon quantum dots.
Further, in the step 1), the molar ratio of the 2, 3-diaminopyridine to the acrylic acid to the zinc chloride is 1.
Further, in the step 2), the dissolving temperature is 100 ℃, and the reaction is carried out for 1h; then the temperature is raised to 210 ℃ and the reaction is carried out for 4h.
The invention also provides application of the nitrogen-doped luminescent carbon quantum dot prepared by the method as a fluorescent probe in lysine detection.
As a preferred scheme, the specific method of the application is as follows:
1) Dissolving carbon quantum dots to prepare a carbon dot solution;
2) Respectively taking lysine solutions with different concentrations, correspondingly adding the lysine solutions with different concentrations into respective carbon dot solutions, uniformly mixing to obtain different mixed solutions, detecting the different mixed solutions under an excitation wavelength (380 nm), recording fluorescence spectra, and establishing a linear relation between the lysine concentration and the intensity of a fluorescence peak (436 nm);
3) Adding a sample to be detected into the carbon dot solution, uniformly mixing, recording the intensity of a fluorescence peak (436 nm) at an excitation wavelength (380 nm), and calculating according to a linear relation to obtain the lysine concentration of the sample to be detected.
Preferably, in the step 1), the concentration of the carbon dot solution is 0.01 to 0.05mg/mL.
The invention also provides application of the nitrogen-doped luminescent carbon quantum dot prepared by the method in rapid and portable detection of lysine.
The invention also provides a manufacturing method of the microfluidic chip for detecting the relative fluorescence intensity, which comprises the following steps:
1) Drawing a template drawing of the microfluidic chip, wherein each chip is provided with two identical parallel straight channels (a detection channel and a reference channel), and each channel is provided with an inlet and an outlet; manufacturing a micro-fluidic chip template by a conventional method;
2) Pouring Polydimethylsiloxane (PDMS) prepolymer into a template, and stripping the cured PDMS prepolymer from the template; punching at the inlet and outlet of the channel; plasma bonding a Polydimethylsiloxane (PDMS) component with a glass sheet to form a complete chip;
3) Dissolving the nitrogen-doped luminescent carbon quantum dots prepared by the method of claim 1 to obtain a carbon dot solution; then uniformly mixing a polyethylene glycol diacrylate solution, 2-hydroxy-2-methyl-phenyl acetone and a carbon dot solution to be used as a hydrogel precursor solution, injecting the hydrogel precursor solution into a chip channel, covering a photomask on the chip, and carrying out ultraviolet exposure curing;
4) And washing the chip, removing the non-polymerized and solidified hydrogel precursor solution, and leaving the hydrogel microstructure embedded with the nitrogen-doped luminescent carbon quantum dots in the channel as a detection element to obtain the micro-fluidic chip containing the detection element.
Preferably, in the step 3), the concentration of the carbon dot solution is 0.05-0.2 mg/mL;
the volume ratio of the polyethylene glycol diacrylate solution to the 2-hydroxy-2-methyl-phenyl acetone to the carbon dot solution is (100).
The polyethylene glycol diacrylate solution is formed by dissolving polyethylene glycol diacrylate in phosphate buffer salt solution to form 50% (w/v) solution.
The invention also provides a portable device for quantitatively detecting the content of lysine in pig serum based on the quantitative fluorescence of the smart phone, which comprises an imaging device shell, wherein the top surface of the imaging device shell is provided with a camera shooting port for shooting by the smart phone, the side wall of the imaging device shell is provided with an insertion hole of a chip support, the middle of the chip support is of a hollow structure, the chip support is horizontally inserted into the imaging device shell from the insertion hole, the microfluidic chip prepared by the method of the claim is placed on the chip support, and an imaging lens and a transmitting optical filter are arranged between the camera shooting port and the microfluidic chip from top to bottom; and an exciting light filter, a condensing lens and an ultraviolet LED light source are arranged below the chip support.
The method for detecting the lysine content in the pig serum by using the portable device comprises the following steps:
1) Respectively injecting lysine solutions with different concentrations into a detection channel of the microfluidic chip, and taking a picture by a mobile phone; respectively calculating the relative fluorescence intensity I/I of lysine solutions with different concentrations according to the formula (1) 0 (ii) a Obtaining a linear relation formula of different lysine concentrations and relative fluorescence intensity; wherein the content of the first and second substances,
in the formula, I is the average gray value of the detection elements in the corrected detection channel;
I 0 the corrected average gray value of the detection elements in the reference channel is obtained;
I meas is the average gray value of the detecting elements in the detecting channel;
I ref is the average gray value of the detection elements in the reference channel;
I bg is a backAverage gray value of the scene area;
2) During detection, a sample to be detected is injected into a detection channel of the microfluidic chip, and a picture is taken by a mobile phone; calculating to obtain the relative fluorescence intensity I/I of the sample to be detected 0 (ii) a And calculating the lysine concentration in the sample to be tested according to the determined linear relation formula.
The invention has the beneficial effects that:
1. the carbon dot for detecting lysine is simple to prepare, high in fluorescence quantum yield, wide in linear range (5-300 mu mol/L), high in sensitivity (LOD =1.7 mu mol/L), high in specificity, good in water solubility and light stability, and capable of being well applied to detection of the content of lysine in pig serum, and the standard adding recovery rate is (94.6% -105.2%);
2. the carbon dots are fixed on the microfluidic chip, so that the method has the advantages of miniaturization, less consumption of samples and reagents, simplicity in operation and the like;
3. the portable quantitative fluorescence detection device based on the smart phone avoids the use of instruments such as a fluorescence microscope and the like, is more suitable for field detection, has a good linear relation in the concentration range of 25-300 mu mol/L for the detection of lysine, has the detection limit of 15.8 mu mol/L and requires 3 minutes of reaction time.
Drawings
FIG. 1 is a schematic diagram of a method for preparing a nitrogen-doped luminescent carbon quantum dot in example 1 of the present invention;
FIG. 2 is a graph (A) of fluorescence spectra of nitrogen-doped luminescent carbon quantum dots prepared in example 1 in different concentrations of lysine solution and a linear relationship graph (B) of lysine solubility and fluorescence intensity;
FIG. 3 is a fluorescence selectivity spectrum of nitrogen-doped luminescent carbon quantum dots prepared in example 1;
FIG. 4 is a schematic diagram of the structure of the microfluidic chip (A) and a schematic diagram of the preparation of the detection element in the chip (B) in example 3;
FIG. 5 is a schematic view of the structure of a fluorescence detecting apparatus in example 4;
in the figure, an imaging device shell 1, a camera port 1.1, an insertion port 1.2, a chip support 2, a microfluidic chip 3, an imaging lens 4, an emission light filter 5, an excitation light filter 6, a condensing lens 7, an ultraviolet LED light source 8 and a smart phone 9;
FIG. 6 is a flowchart of the method for detecting the lysine content in pig serum by the portable device in example 5.
Detailed Description
The present invention is described in further detail below with reference to specific examples so that those skilled in the art can understand the invention.
Example 1
The preparation method of the nitrogen-doped luminescent carbon quantum dot shown in fig. 1 comprises the following steps:
1) Weighing 0.5mmol of 2, 3-diaminopyridine, 3mmol of acrylic acid and 2.5 mmol of zinc chloride;
2) Adding 2, 3-diaminopyridine, acrylic acid and zinc chloride into 5mL of tetraethylene glycol, and stirring to mix uniformly; placing in a sand bath, and reacting at 100 deg.C for 60min to fully dissolve the reactant; heating to 210 ℃ for reaction for 240min, and cooling to obtain a crude product solution;
3) Dissolving the crude product solution in 50mL of methanol to obtain a yellow-brown liquid, centrifuging for 20min at the rotation speed of 8000r/min, filtering to remove large-particle residues, and removing the solvent from the filtered product in a rotary evaporator to obtain a crude product;
4) Adding diethyl ether into the crude product, shaking to separate out a black solid, collecting the precipitate, adding 5mL of methanol again to dissolve the precipitate, adding diethyl ether, shaking to separate out the solid, repeating the steps for a plurality of times, removing unreacted zinc chloride, and collecting the black solid; and finally, drying the black solid at 80 ℃ for 6h in vacuum to obtain the solid, namely the nitrogen-doped luminescent carbon quantum dot (carbon dot for short).
Example 2
1. The carbon dots are used as fluorescent probes for detecting the sensitivity of the lysine, and the method comprises the following specific steps:
1) The carbon dots prepared in example 1 were dissolved in ultrapure water to prepare a carbon dot solution having a concentration of 0.025mg/mL
2) Preparing lysine solutions with concentrations of 0, 1, 5, 10, 20, 40, 60, 80, 100, 120, 140, 200, 300 μmol/L with phosphate buffered saline (PBS, 1mmol/L, pH = 6.0), respectively;
3) Respectively taking 100 mu L of lysine solution with different concentrations, respectively adding the lysine solution into the carbon dot solution (0.025 mg/mL) of 2mL respectively, and uniformly mixing; different mixtures were obtained, detected at an excitation wavelength of 380nm and fluorescence spectra were recorded (fig. 2A), as shown in fig. 2 (a): the fluorescence intensity of the carbon dots is enhanced along with the increase of the lysine concentration; then establishing a linear relation of the lysine concentration and the fluorescence peak (436 nm) intensity (figure 2B); as shown in fig. 2B: has linearity in the concentration range of 5-300 mu mol/L, the linear equation is y =0.00458x +1.01319 2 =0.99693, detection limit is 1.7. Mu. Mol/L.
As can be seen from FIG. 2, the carbon dots have high sensitivity when used as fluorescent probes for detecting lysine.
2. The carbon dots are used as fluorescent probes for detecting the specificity of the lysine, and the specific steps are as follows:
1) Solutions of potential interferents were prepared separately in PBS, including alanine (Ala, 500. Mu. Mol/L), histidine (His, 500. Mu. Mol/L), threonine (Thr, 500. Mu. Mol/L), leucine (Leu, 500. Mu. Mol/L), valine (Val, 500. Mu. Mol/L), glycine (Gly, 2.0 mmol/L), methionine (Met, 500. Mu. Mol/L), glutathione (GSH, 2.0 mmol/L), glucose (Glucose, 10.0 mmol/L), K + (0.1mol/L)、 Na + (0.1mol/L)、Fe 2+ (10μmol/L)、Fe 3+ (10μmol/L)、Mg 2+ (10 μmol/L)、Cu 2+ (10. Mu. Mol/L) and Mn 2+ (10μmol/L)；
2) Adding 100 μ L of the above interferent solution into 2mL of the above carbon dot solution (0.025 mg/mL), mixing, detecting and recording fluorescence spectrum under excitation wavelength of 380nm, and calculating fluorescence intensity variation I/I 0 I and I 0 The fluorescence intensity at 436nm for the carbon spot with and without the addition of interferents, respectively.
As shown in FIG. 3, the fluorescence intensity of the carbon spot is not substantially affected by other interferents, and has a specific response to lysine.
3. Carbon dot as fluorescent probe for detecting lysine in pig serum
Porcine serum samples provided by the animal science and technology institute of Huazhong university of agriculture are taken to verify the feasibility of the carbon dot for detecting the actual biological samples. The standard method is adopted to measure lysine in the pig serum sample by a standard method and a standard recovery experiment, and the measurement result is shown in table 1.
TABLE 1 carbon point for lysine in pig serum samples
As can be seen from Table 1, the spiking recovery rate of lysine in the actual serum sample is 94.6-105.2%, and RSD is less than or equal to 4.7%, which indicates that the carbon points can realize the detection of lysine in the actual serum sample.
Example 3
The manufacturing method of the microfluidic chip for detecting relative fluorescence intensity as shown in fig. 4 comprises the following steps:
1) Drawing a template diagram of the microfluidic chip (as shown in fig. 4A), wherein each chip is provided with two identical parallel straight channels (a detection channel and a reference channel), and each channel is provided with an inlet and an outlet; manufacturing a micro-fluidic chip template by adopting a soft lithography method; wherein, the size of each chip is about 12 multiplied by 10mm, the width of the channel is 0.8mm, the height is 0.3mm, and the length of the main channel is about 6.5 mm;
2) Pouring Polydimethylsiloxane (PDMS) prepolymer into a template, heating for 3h at the temperature of 65 ℃, and stripping from the template after curing; dicing into individual chip components; punching at the inlet and outlet of the channel; bonding the PDMS component and a glass sheet in a plasma manner to form a complete chip, and drying at the temperature of 65 ℃ for 2h;
3) As shown in fig. 4B: dissolving polyethylene glycol diacrylate in PBS to form a 50% (w/v) solution; dissolving the carbon dots prepared in example 1 in ultrapure water to obtain a carbon dot solution with a concentration of 0.1 mg/mL;
4) Then adding 200 mu L of carbon dot solution and 10 mu L of 2-hydroxy-2-methyl-phenyl acetone into 1.0ml of polyethylene glycol diacrylate solution, uniformly mixing to obtain hydrogel precursor solution, injecting the hydrogel precursor solution into a chip channel, covering a photomask on the chip, and exposing the chip under an ultraviolet lamp for 15 seconds to polymerize and solidify the hydrogel;
5) And (3) washing the channel for 3 times by using PBS (phosphate buffer solution), removing the non-polymerized and solidified hydrogel precursor solution, and leaving the hydrogel microstructure embedded with the carbon points in the channel as a detection element to obtain the micro-fluidic chip containing the detection element.
Example 4
The portable device for quantitative fluorescence detection of lysine content in pig serum based on smart phone as shown in fig. 5 is made of black light absorbing material; the micro-fluidic chip comprises an imaging device shell 1, wherein a camera port 1.1 used for shooting of a smart phone is formed in the top surface of the imaging device shell 1, an insertion hole 1.2 of a chip support 2 is formed in the side wall of the imaging device shell 1, the middle of the chip support 2 is of a hollow structure, the chip support 2 is horizontally inserted into the imaging device shell 1 through the insertion hole, a micro-fluidic chip 3 prepared by the method of claim 7 is placed on the chip support 2, and an imaging lens 4 and an emission optical filter 5 are arranged between the camera port 1.1 and the micro-fluidic chip 3 from top to bottom; and an exciting light filter 6, a condensing lens 7 and an ultraviolet LED light source 8 are arranged below the chip support 2.
The device and the chip support in the embodiment are manufactured by 3D printing, when in detection, light emitted by the ultraviolet LED irradiates the microfluidic chip after passing through the condensing lens and the exciting light filter, the detection element in the chip emits fluorescence, and after passing through the emitting light filter, the imaging lens and the camera shooting port, a picture can be shot by a mobile phone.
Example 5
The method for detecting the content of lysine in pig serum by using the portable device shown in fig. 6 comprises the following steps:
1) Respectively injecting lysine solutions with different concentrations into a detection channel of the microfluidic chip, and taking a picture by using a mobile phone; respectively calculating the relative fluorescence intensity I/I of lysine solutions with different concentrations according to the formula (1) 0 (ii) a Obtaining a linear relation formula of different lysine concentrations and relative fluorescence intensity; wherein the content of the first and second substances,
in the formula, I is the average gray value of the detection elements in the corrected detection channel;
I 0 the corrected average gray value of the detection elements in the reference channel is obtained;
I meas is the average gray value of the detecting elements in the detecting channel;
I ref is the average gray value of the detection elements in the reference channel;
I bg the average gray value of the background area;
2) During detection, a sample to be detected is injected into a detection channel of the microfluidic chip, the chip is placed into an imaging device after 3 minutes, an ultraviolet LED light source switch is turned on, and a mobile phone is used for taking a picture; after the picture is grayed, average gray value I of the detection element in the detection channel, the detection element in the reference channel and the background area is respectively read meas 、I ref And I bg Calculating to obtain the relative fluorescence intensity I/I of the sample to be detected 0 (ii) a And calculating the lysine concentration in the sample to be tested according to the determined linear relation formula.
The lysine concentration in the sample to be detected is 47.9 mu mol/L, RSD =2.05%, and the result is matched with the result (46.8 mu mol/L) of an automatic amino acid analyzer, thereby proving the accuracy of the method provided by the invention.
Other parts not described in detail are prior art. Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and all of the embodiments are included in the scope of the present invention.
Claims (10)
1. A preparation method of nitrogen-doped luminescent carbon quantum dots is characterized by comprising the following steps: the method comprises the following steps:
1) Weighing 2, 3-diaminopyridine, acrylic acid and zinc chloride according to a molar ratio of 1;
2) Adding 2, 3-diaminopyridine, acrylic acid and zinc chloride into tetraethylene glycol, and stirring to mix uniformly; reacting for 50-60 min at the temperature of 90-110 ℃ to fully dissolve reactants; heating to 200-220 ℃ for reaction for 220-260 min, and cooling to obtain a crude product solution;
3) Dissolving the crude product solution in methanol, carrying out centrifugal filtration to obtain a filtered product, and removing the solvent from the product in a rotary evaporator to obtain a crude product;
4) Adding diethyl ether into the crude product, shaking to separate out a black solid, collecting the precipitate, adding methanol again to dissolve the precipitate, adding diethyl ether, shaking to separate out a solid, repeating the steps for several times, and removing unreacted zinc chloride; and vacuum drying to obtain solid, namely the nitrogen-doped luminescent carbon quantum dots.
2. The method of claim 1, wherein the method comprises: in the step 1), the molar ratio of the 2, 3-diaminopyridine to the acrylic acid to the zinc chloride is 1.
3. The method of claim 1, wherein the method comprises: in the step 2), the dissolving temperature is 100 ℃, and the reaction is carried out for 1h; then the temperature is raised to 210 ℃ and the reaction is carried out for 4h.
4. The application of the nitrogen-doped luminescent carbon quantum dot prepared by the method in claim 1 as a fluorescent probe in lysine detection.
5. The application according to claim 1, characterized in that the specific method of application:
1) Dissolving carbon quantum dots to prepare a carbon dot solution;
2) Respectively taking lysine solutions with different concentrations, correspondingly adding the lysine solutions with different concentrations into respective carbon dot solutions, uniformly mixing to obtain different mixed solutions, detecting the different mixed solutions under exciting light, recording fluorescence spectra, and establishing a linear relation between the lysine concentration and the fluorescence peak intensity;
3) And adding the sample to be detected into the carbon dot solution, uniformly mixing, recording the intensity of a fluorescence peak under exciting light, and calculating according to a linear relation to obtain the lysine concentration of the sample to be detected.
6. The use of claim 5, wherein in step 1), the concentration of the carbon dot solution is 0.01-0.05 mg/mL.
7. Application of the nitrogen-doped luminescent carbon quantum dot prepared by the method in claim 1 in rapid and portable detection of lysine.
8. A manufacturing method of a micro-fluidic chip for detecting relative fluorescence intensity is characterized by comprising the following steps: the method comprises the following steps:
1) Drawing a template diagram of the microfluidic chip, wherein each chip is provided with two identical parallel straight channels, and each channel is provided with an inlet and an outlet; manufacturing a micro-fluidic chip template by a conventional method;
2) Pouring the polydimethylsiloxane prepolymer into a template, and stripping the polydimethylsiloxane prepolymer from the template after curing; punching at the inlet and outlet of the channel; plasma bonding the polydimethylsiloxane component with the glass sheet to form a complete chip;
3) Dissolving the nitrogen-doped luminescent carbon quantum dots prepared by the method of claim 1 to obtain a carbon dot solution; then uniformly mixing a polyethylene glycol diacrylate solution, 2-hydroxy-2-methyl-phenyl acetone and a carbon dot solution to be used as a hydrogel precursor solution, injecting the hydrogel precursor solution into a chip channel, covering a photomask on the chip, and carrying out ultraviolet exposure curing;
4) And washing the chip, removing the hydrogel precursor solution which is not polymerized and solidified, and leaving the hydrogel microstructure embedded with the nitrogen-doped luminescent carbon quantum dots in the channel as a detection element to obtain the micro-fluidic chip containing the detection element.
9. The utility model provides a portable device of quantitative fluorescence detection lysine content in pig serum based on smart mobile phone which characterized in that: the device comprises an imaging device shell (1), wherein a camera port (1.1) for smart phone shooting is formed in the top surface of the imaging device shell (1), an insertion hole (1.2) of a chip support (2) is formed in the side wall of the imaging device shell (1), the middle of the chip support (2) is of a hollow structure, the chip support (2) is horizontally inserted into the imaging device shell (1) through the insertion hole, a microfluidic chip (3) prepared by the method of claim (7) is placed on the chip support (2), and an imaging lens (4) and an emission optical filter (5) are arranged between the camera port (1.1) and the microfluidic chip (3) from top to bottom; an exciting light filter (6), a condensing lens (7) and an ultraviolet LED light source (8) are arranged below the chip support (2).
10. The method for detecting the content of lysine in pig serum by using the portable device of claim 9, wherein the method comprises the following steps: the method comprises the following steps:
1) Respectively injecting lysine solutions with different concentrations into a detection channel of the microfluidic chip, and taking a picture by using a mobile phone; respectively calculating the relative fluorescence intensity I/I of lysine solutions with different concentrations according to the formula (1) 0 (ii) a Obtaining a linear relation formula of different lysine concentrations and relative fluorescence intensity; wherein, the first and the second end of the pipe are connected with each other,
in the formula, I is the average gray value of the detection elements in the corrected detection channel;
I 0 the corrected average gray value of the detection elements in the reference channel is obtained;
I meas is the average gray value of the detecting elements in the detecting channel;
I ref is the average gray value of the detection elements in the reference channel;
I bg the average gray value of the background area is obtained;
2) During detection, a sample to be detected is injected into a detection channel of the microfluidic chip, and a picture is taken by a mobile phone; calculating to obtain the relative fluorescence intensity I/I of the sample to be detected 0 (ii) a Calculating the sample to be measured according to the determined linear relation formulaLysine concentration in (c).
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