CN113899906A - Wine caramel pigment detection robot and detection method - Google Patents
Wine caramel pigment detection robot and detection method Download PDFInfo
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Abstract
The invention discloses a robot and a method for detecting caramel pigment in wine, wherein the robot comprises a conveying device, a first antibody spraying device, a second antibody spraying device, an elution device and a radioactivity detection device; utilizing specific immunoreaction to specifically combine 4-methylimidazole introduced by caramel pigment in the wine liquid to generate an anti-combination body; radiolabelling an anti-conjugate by specific binding of a second antibody having weak radioactivity to the anti-conjugate to produce a secondary antibody conjugate; finally, the content of the caramel pigment in the wine liquid is obtained by calculating through detecting the radioactivity of the secondary antibody combination; by utilizing the method, the wine caramel pigment detection robot disclosed by the invention can quickly and conveniently carry out quantitative detection on the caramel pigment content in the wine, and has the advantages of high detection sensitivity and wide application scene.
Description
Technical Field
The invention relates to the field of food detection, in particular to a robot and a method for detecting caramel pigment of wine.
Background
The caramel pigment is a complex reddish brown or dark brown mixture generated by utilizing dehydration, decomposition and polymerization of saccharides at high temperature and browning reaction, and is widely applied to the fields of food, medicines, beverages and the like; caramel color can be divided into common caramel, caustic sulfite caramel, ammonia caramel and sulfurous acid ammonia caramel according to the production method, wherein ammonia caramel and sulfurous acid ammonia caramel using ammonia salt as catalyst can generate 4-methylimidazole which is a carcinogenic byproduct in the production process;
acid-resistant caramel pigment used in the wine is ammonia sulfite caramel using ammonium salt as a catalyst, and at present, a part of wine manufacturers deepen the color of the wine body by adding excessive caramel pigment into the wine of low-grade, so that the judgment of consumers on the wine grade is influenced; at present, caramel pigment in food is detected by a chromatograph, and the detection method has the advantages of high detection accuracy and capability of quantitative detection, but has the limitations of complex sample preparation, harsh experimental conditions, various detection devices, low detection efficiency and the like, and cannot be applied to a field real-time detection scene with high requirements on rapid flexibility.
Disclosure of Invention
The invention aims to overcome the defects and provide a wine caramel pigment detection robot and a detection method, which can quickly and flexibly detect caramel pigment in wine.
In order to achieve the purpose, the invention adopts the following specific scheme:
a robot for detecting caramel pigment in wine comprises a conveying device, a first antibody spraying device, a second antibody spraying device, an elution device and a radioactivity detection device;
the conveying device comprises a main bracket, a belt wheel, a motor and a conveying belt; a base film with water absorption is adhered on the conveying belt;
the first antibody spraying device is internally provided with a first antibody liquid containing a first antibody; the first antibody is a substance which can perform specific immunoreaction with 4-methylimidazole contained in caramel pigment; the specific immune reaction can generate a 4-methylimidazole-antigen conjugate, namely an anti-conjugate; the first antibody spraying device is used for spraying the first antibody on the basement membrane;
the second antibody spraying device is filled with a second antibody liquid containing a second antibody; said second antibody specifically binds to said first anti-binding agent to produce a second binding agent; the secondary antibody binding body is a colloidal precipitate; the second antibody is radiolabeled and has weak radioactivity; the second antibody spraying device is used for spraying the second antibody on the basement membrane;
the elution device is filled with eluent; the elution solution may elute the first and second antibodies from the base membrane;
the radioactivity detecting device can detect the radioactivity intensity of the basement membrane.
Furthermore, the front surface of the base membrane is made of a flexible porous material, so that the base membrane has good water absorption.
The wine caramel pigment detection robot further comprises a liquid suction device for dehydrating the eluted basement membrane.
The first antibody spraying device and the second antibody spraying device are identical in structure and respectively comprise a liquid box, a quantitative pump and a spraying head; the front end of the sprinkler head is provided with a plurality of atomization units for atomizing the first antibody liquid and the second antibody liquid.
The invention further provides the radioactivity detection device which further comprises a body, a fluorescent sheet, a convex lens and a photoelectric sensor; the fluorescent sheet is fixedly arranged at the front end of the body; the convex lens is fixedly arranged on the rear side of the fluorescent sheet; the photoelectric sensor is fixedly arranged on the rear side of the convex lens; the fluorescent sheet, the convex lens and the photoelectric sensor are coaxially arranged, and the distance between the photoelectric sensor and the convex lens is smaller than the focal length of the convex lens.
The invention has the beneficial effects that: the method comprises the following steps of taking a basement membrane as a specific immunoreaction carrier, carrying out transport of each detection procedure on the basement membrane by using a conveyor belt, spraying a first antibody and a second antibody on the basement membrane carrying wine liquid by using a first antibody spraying device and a second antibody spraying device to carry out specific immunoreaction so as to carry out radioactive labeling on 4-methylimidazole in the wine liquid, removing unreacted first antibody and unreacted second antibody by using an elution device, carrying out radioactive quantitative detection on the basement membrane, and calculating the mass percentage of caramel pigment in the wine liquid by using radioactivity; the caramel pigment in the wine is quantitatively detected without chromatography; the conveying belt circulation detection mode enables the size of the device to be more compact, and the device can be suitable for field detection use scenes with high requirements on flexibility.
The detection method for the caramel pigment in the wine is obtained by utilizing any scheme of the wine caramel pigment detection robot, and comprises the following steps:
s1, adhering the base film to the outer surface of the conveyor belt;
s2, dispersing and dripping a fixed volume of wine liquid to be detected to the surface of the basement membrane and diffusing the wine liquid in the whole basement membrane;
s3, moving the basal membrane added with the wine liquid to a first antibody spraying position corresponding to the first antibody spraying device through a conveyor belt, and spraying excessive first antibody liquid on the surface of the basal membrane;
s4, standing for a period of time to enable the 4-methylimidazole in the wine liquid and the first antibody in the first antibody liquid to generate specific immunoreaction and generate an anti-binding body;
s5, after the specific immunoreaction is completed, the conveyer belt drives the reacted basement membrane to move to a second antibody spraying position corresponding to the second antibody spraying device, and excessive second antibody liquid is sprayed on the surface of the basement membrane;
s6, standing for a period of time to enable the first anti-binding body in the basement membrane to be specifically bound with the second antibody in the second antibody liquid, and generating a secondary anti-binding body of a colloidal precipitate;
s7, moving the basement membrane carrying the secondary antibody combination into the elution device through the conveyor belt, and eluting the first antibody and the second antibody which are not involved in the reaction and are in a free state in the basement membrane through the elution device to separate the first antibody and the second antibody from the basement membrane;
s8, moving the eluted basement membrane to a radioactivity detection position corresponding to a radioactivity detection device through a conveyor belt, and detecting the radioactivity intensity of the basement membrane through the radioactivity detection device; the amount of the secondary antibody conjugate and the primary antibody conjugate in the basement membrane can be judged according to the radioactive intensity, so that the concentration of 4-methylimidazole in the wine is obtained, and the concentration value of caramel pigment in the wine can be obtained by calculating the concentration of 4-methylimidazole in the caramel pigment.
Compared with the existing method for detecting the caramel pigment in the food, the method has the following beneficial effects:
1. the content of 4-methylimidazole introduced by caramel pigment in the wine is detected by utilizing specific immunoreaction, and then the concentration value of the caramel pigment in the wine is obtained by calculation, so that the complex treatment of a sample and a plurality of detection devices in chromatographic detection are avoided, and the detection is faster and more flexible;
2. the fixed 4-methylimidazole combined with the specific immunoreaction is labeled by an isotope labeling method, and the radioactivity of a reaction combined product is detected, so that the interference of a plurality of impurity pigments with similar properties to the caramel pigment in the wine on a detection result in chromatographic detection is avoided, and the accurate quantitative detection of the content of the caramel pigment in the wine is realized.
Drawings
FIG. 1 is a cut-away perspective view of the present invention;
FIG. 2 is a perspective view of a first antibody spraying device of the present invention in half section;
FIG. 3 is a cut-away perspective view of the delivery device of the present invention;
FIG. 4 is a perspective view of an elution apparatus of the present invention;
FIG. 5 is a perspective view of the radioactivity detecting device of the present invention;
FIG. 6 is a perspective view of the wicking apparatus of the present invention;
FIG. 7 is a schematic view of the detection method of the present invention;
description of reference numerals: 1. a conveying device; 2. a radioactivity detecting device; 3. a second antibody spraying device; 4. a first antibody spraying device; 5. an elution device; 6. a liquid suction device; 7. a liquid box; 8. a constant delivery pump; 9. a first bracket; 10. a sprinkler head; 10a, a dispersion chamber; 10b, micropores; 10c, an atomizing unit; 11. A main support; 12. a pulley; 13. a motor; 14. a base film; 15. a conveyor belt; 16. a piezoelectric ultrasonic vibrating piece; 17. an elution tank; 18. a body; 19. a photosensor; 20. a convex lens; 21. a fluorescent sheet; 22. A second bracket; 23. a liquid suction roller; I. a wine feeding position; II. A first antibody fluid spray location; III, second antibody liquid spraying position; IV, elution position; v, liquid suction position; VI, radioactivity detection position.
Detailed Description
The invention will be described in further detail with reference to the following figures and specific examples, without limiting the scope of the invention.
As shown in fig. 1 to 7, the robot for detecting caramel pigment in wine comprises a conveying device 1, a first antibody spraying device 4, a second antibody spraying device 3, an elution device 5 and a radioactivity detection device 2;
the conveying device 1 comprises a conveying belt 15; a base film 14 having water absorption property is stuck on the conveyor belt 15;
the first antibody spraying device 4 contains a first antibody liquid containing a first antibody; the first antibody is a substance which can perform specific immunoreaction with 4-methylimidazole contained in caramel pigment; the specific immune reaction can generate a 4-methylimidazole-antigen conjugate, namely an anti-conjugate; the first antibody spraying device 4 is used to spray the first antibody on the base film 14;
the second antibody spraying device 3 contains a second antibody liquid containing a second antibody; said second antibody specifically binds to said first anti-binding agent to produce a second binding agent; the secondary antibody binding body is a colloidal precipitate; the second antibody is radiolabeled and has weak radioactivity; the second antibody spraying device 3 for spraying the second antibody on the basement membrane 14;
the elution device 5 is filled with eluent; the eluent can elute the first and second antibodies from the base membrane 14;
the radioactivity detecting device 2 can detect the radioactivity intensity of the basement membrane 14.
Specifically, four belt wheels 12 are distributed on a main bracket 11 of the conveying device 1 in a diamond shape, and a conveying belt 15 is wound on the belt wheels 12 and is in a diamond-shaped conveying state; one of the belt wheels 12 is fixedly connected with a motor 13, and the motor 13 provides power to drive a transmission belt to move through the belt wheel 12 so as to realize the transmission of the base film 14; the surface of the conveyor belt 15 is coated with a hydrophobic coating so as to prevent the elution liquid with certain radioactive rays from adhering to the conveyor belt 15 and influencing the radioactive ray detection result;
specifically, the first antibody can be used for obtaining immune splenocytes generating the 4-methylimidazole antibody through a mouse immune reaction by using a 4-methylimidazole-bovine serum albumin artificial antigen, and then preparing a high-specificity 4-methylimidazole monoclonal antibody mouse anti-lgG through a hybridoma technology, namely the first antibody;
specifically, the second antibody can be used as an antigen through a 4-methylimidazole monoclonal antibody, immune spleen cells capable of generating rabbit anti-mouse lgG can be obtained through rabbit immune reaction, then rabbit anti-mouse lgG capable of being specifically immunized and combined with the 4-methylimidazole monoclonal antibody is prepared through a hybridoma technology, and the obtained rabbit anti-mouse lgG is subjected to radioactive labeling to obtain the second antibody;
specifically, the elution device 5 comprises an elution tank 17 and piezoelectric ultrasonic vibration pieces 16 symmetrically arranged on the elution tank 17; the eluent in the elution device 5 is 0.8% sodium chloride solution which can elute the first antibody liquid and the second antibody liquid in free state which do not participate in the reaction;
the specific working mode of the embodiment is as follows: adhering a base film 14 to the outer side surface of a conveyor belt 15 from a wine feeding position I at the top of a main support 11, sucking wine liquid with a fixed volume through a liquid taking pipe, and dripping the wine liquid on the surface of the base film 14 at multiple points; after the dripping, the wine liquid is uniformly diffused in the base film 14 due to the siphonage effect of the small holes in the base film 14; after the dripping and diffusion of the wine liquid are finished, conveying the base membrane 14 to a first antibody spraying position II through a conveying belt 15, spraying excessive first antibody liquid to the surface of the base membrane 14 through a first antibody spraying device 4, standing for 2-3min, and enabling the first antibody and 4-methylimidazole in the wine liquid to fully generate specific immunoreaction to generate a mouse anti-lgG-4-methylimidazole conjugate; continuing to convey the basement membrane 14 to a second antibody spraying position III through a conveyor belt 15, spraying an excessive amount of second antibody liquid to the surface of the basement membrane 14 through a second antibody spraying device 3, and standing for 3-5min to allow the second antibody to perform specific immunoreaction with the 4-methylimidazole-mouse anti-lgG conjugate generated in the previous specific immunoreaction, so that the radiolabeled second antibody with weak radioactivity is combined with the mouse anti-lgG-4-methylimidazole conjugate to generate a colloidal precipitate-shaped 4-methylimidazole-mouse anti-lgG-rabbit anti-mouse lgG conjugate;
after the reaction is finished, conveying the basement membrane 14 into an elution device 5 through a conveyor belt 15, and eluting and removing the first antibody and the second antibody which are not involved in the specific immune combination reaction and are left on the basement membrane 14 under the condition of ultrasonic stirring, so that only 4-methylimidazole-mouse anti-lgG-rabbit anti-mouse lgG combination in a colloid precipitation shape is remained on the basement membrane 14; after the elution is completed, the conveyor belt 15 conveys the base film 14 to the radioactivity detection position VI, and the radioactivity of the base film 14 is quantitatively detected by the radioactivity detection device 2; defining the measured radioactivity value a, the radioactivity of 1mol of second antibody as b, the molecular proportion coefficient of the combination of the first antibody and the second antibody as c, the molecular proportion coefficient of the combination of the first antibody and 4-methylimidazole as d, and the mass fraction of 4-methylimidazole in the caramel pigment as e; the molar mass of the 4-methylimidazole is f, the adding volume of the wine is v, the density of the wine is g, and the mass percentage of the caramel pigment in the wine is h, so that h is acdf/bevg.
The beneficial effect of this embodiment does: carrying out the transfer of each detection process on a base membrane 14 by using a conveyor belt 15 by using the base membrane 14 as a specific immunoreaction carrier, spraying a first antibody and a second antibody on the base membrane 14 carrying wine by using a first antibody spraying device 4 and a second antibody spraying device 3 to generate specific immunoreaction so as to carry out radioactive labeling on 4-methylimidazole in the wine liquid, removing unreacted first antibody and unreacted second antibody by using an elution device 5, then carrying out radioactive quantitative detection on the base membrane 14, and calculating the mass percentage of caramel pigment in the wine liquid by radioactivity; the caramel pigment in the wine is quantitatively detected without chromatography; the circulating detection mode of the conveyor belt 15 enables the volume of the device to be more compact, and the device can be suitable for field detection use scenes with high requirements on flexibility.
Based on the above embodiment, further, the front surface of the base film 14 is made of a flexible porous material, and has good water absorption.
Specifically, the back of the base film 14 is coated with a removable non-setting adhesive coating to be adhered to the conveyor belt 15; the front surface of the base membrane 14 is a water absorption surface, the material of the water absorption surface is preferably a nitrocellulose membrane or a glass fiber membrane, and the thickness is 0.8-1.6mm, so that the base membrane has good solution absorption and retention capacity, and simultaneously, siphon effect liquid generated by micropores 10b in the base membrane is more favorable for the diffusion of the solution in the membrane layer; meanwhile, the internal porous structure can enable the second antibody and the 4-methylimidazole-mouse anti-lgG conjugate to perform specific immunoreaction to generate a colloid precipitate-shaped 4-methylimidazole-mouse anti-lgG-rabbit anti-mouse lgG conjugate which is adsorbed and aggregated by taking fibers in the basement membrane 14 as a framework, so that the colloid precipitate-shaped 4-methylimidazole-mouse anti-lgG-rabbit anti-mouse lgG conjugate is prevented from falling off from the basement membrane 14 in the elution process, the radioactivity loss is caused, and the radioactivity detection result is low.
In actual use, the base film 14 is aligned with the conveyor belt 15, and the front surface of the base film 14 is pressed by a clean absorbent cotton pad, so that the back surface of the base film is firmly adhered to the conveyor belt 15; after the pasting is finished, the fixed volume of the wine liquid is dripped on the front side of the basement membrane 14, and the grapes are uniformly diffused in the basement membrane 14.
Based on the above embodiment, further, the wine caramel color detection robot further comprises a liquid absorbing device 6 for dehydrating the eluted basement membrane 14.
Specifically, the liquid suction device 6 is installed at the dehydration position of the main bracket 11, and comprises a concave second bracket 22 and a plurality of liquid suction rollers 23 which are sequentially arranged in the grooves of the second bracket 22; the surface of the liquid absorbing roller 23 is made of porous flexible materials, has good rapid water absorption, and is preferably absorbent cotton;
during operation, imbibition device 6 is installed in the conveyer belt 15 outside, and there is a certain distance on imbibition roller 23 surface and the conveyer belt 15 surface at a distance, and this distance is less than base film 14 thickness for base film 14 passes through imbibition device 6 under the drive of conveyer belt 15, and imbibition roller 23 surface and base film 14 front contact adsorb the removal with the adnexed residual eluant in base film 14 surface, prevent that the second antibody that has radioactivity that contains in the residual eluant from causing the interference to the radioactivity testing result.
Based on the above embodiment, further, the first antibody spraying device 4 and the second antibody spraying device 3 have the same structure, and each of them comprises a liquid box 7, a quantitative pump 8 and a spraying head 10; the front end of the sprinkler head 10 is provided with a plurality of atomization units 10c for atomizing the first antibody liquid and the second antibody liquid.
Specifically, a first bracket 9 is fixedly installed at the front end of the liquid box 7, a dosing pump 8 with a liquid inlet communicated with the liquid box 7 is fixedly connected to the rear end of the first bracket 9, and a sprinkler head 10 for atomizing a first antibody liquid is fixedly installed at the front end of the first bracket 9; the liquid inlet end of the sprinkler head 10 is communicated with the liquid outlet at the front end of the constant delivery pump 8; a plurality of atomizing units 10c are uniformly distributed on the front side of the sprinkler head 10, and each atomizing unit 10c is provided with a micropore 10b for spraying a first antibody liquid and a second antibody liquid; the middle part of the sprinkler head 10 is also provided with a dispersion cavity 10a connected with each atomization unit 10 c;
when the quantitative pump 8 works, the fixed volume of the first antibody liquid is pumped out from the liquid box 7 and is pressurized, and the high-pressure first antibody liquid enters the dispersion cavity 10a to be dispersed, and then is sprayed out from the micropores 10b of each atomization unit 10c in the form of small liquid drops to be dispersed on the base membrane 14; by spraying in an atomized manner, the droplets of the first antibody are distributed more evenly over the base membrane 14 and diffuse at a faster rate, reducing the time required for the diffusion process and improving detection efficiency.
In this embodiment, the radioactivity detecting device 2 further includes a main body 18, a fluorescent sheet 21, a convex lens 20, and a photoelectric sensor 19; the fluorescent sheet 21 is fixedly arranged at the front end of the body 18; the convex lens 20 is fixedly arranged at the rear side of the fluorescent sheet 21; the photoelectric sensor 19 is fixedly arranged at the rear side of the convex lens 20; the fluorescent sheet 21, the convex lens 20 and the photoelectric sensor 19 are coaxially arranged, and the distance between the photoelectric sensor 19 and the convex lens 20 is smaller than the focal length of the convex lens 20.
When the fluorescent film works, the base film 14 is positioned below the fluorescent sheet 21, and radioactive rays of a 4-methylimidazole-mouse anti-lgG-rabbit anti-mouse lgG combination in the base film 14 can excite a fluorescent substance on the surface of the fluorescent sheet 21 to emit light and generate fluorescent light spots on the surface of the fluorescent sheet 21; light rays emitted by the surface of the fluorescent plate are focused through the convex lens 20, and because the distance between the photoelectric sensor 19 and the convex lens 20 is smaller than the focal length of the convex lens 20, images formed by light spots of the fluorescent plate are focused on a receiving window of the photoelectric sensor 19, the photoelectric sensor 19 converts optical signals into electric signals, and the radioactivity intensity of the base film 14 can be obtained by monitoring the intensity of the electric signals. The fluorescence sheet 21 is used for exciting luminescence, the fluorescence sheet is focused by the convex lens 20 and imaged, the photoelectric sensor 19 is used for converting the image into an electric signal to detect the radioactive intensity of the base film 14, and the optical signal can be amplified through the convex lens 20, so that the detection sensitivity is improved.
As shown in fig. 1 to 7, the method for detecting caramel pigment in wine by using any one of the wine caramel pigment detection robots comprises the following steps:
step S1: adhering the base film 14 to the outer surface of the conveyor belt 15;
in the step, the base film 14 and the conveyor belt 15 are attached to be leveled and pressed tightly by a cotton absorbent pad after the attachment is finished; the thickness of the base film 14 is adaptively adjusted according to the volume of the dropwise added wine liquid;
step S2: a fixed volume of wine liquid to be tested is dispersedly and dropwise added to the surface of the basement membrane 14 and is diffused in the whole basement membrane 14;
in the step, the wine liquid is dispersedly dripped according to the central part and the four corners to increase the diffusion speed and the diffusion uniformity; after the dripping, the wine needs to be completely permeated into the basement membrane 14, and no floating liquid drops can be on the surface;
step S3: moving the basal membrane 14 to which the wine liquid is dripped to a first antibody spraying position (II) corresponding to the first antibody spraying device 4 through a conveyor belt 15, and spraying excessive first antibody liquid on the surface of the basal membrane 14;
in this step, the spraying pressure of the first antibody is adaptively adjusted according to the area of the basement membrane 14, so that the first antibody solution covers the whole surface of the basement membrane 14; the amount of the first antibody diffused into the basement membrane 14 should be greater than the amount of the first antibody required for the specific immune reaction, so that the 4-methylimidazole contained in the wine solution is completely reacted with the first antibody;
step S4: standing for a period of time to enable the 4-methylimidazole in the wine liquid and the first antibody in the first antibody liquid to generate a specific immunoreaction and generate an anti-conjugate;
in the step, the standing time is adaptively selected according to the area of the base film 14 and the environmental temperature and is not less than 2 min;
step S5: after the specific immunoreaction is completed, the conveyer belt 15 drives the reacted basement membrane 14 to move to a second antibody spraying position III corresponding to the second antibody spraying device 3, and excessive second antibody liquid is sprayed on the surface of the basement membrane 14;
in this step, the amount of the secondary antibody spraying pressure is adaptively adjusted according to the area of the basement membrane 14 so that the secondary antibody liquid covers the entire basement membrane 14 surface; the amount of second antibody that diffuses into basement membrane 14 should be greater than that required for specific immunoreactions to allow complete reaction of the 4-methylimidazole-murine anti-lgG conjugate with the second antibody;
step S6: standing for a period of time to allow specific binding of the primary antibody conjugate in the basement membrane 14 to the secondary antibody in the secondary antibody fluid, thereby generating a colloidal precipitate secondary antibody conjugate;
in the step, the standing time is adaptively selected according to the area of the base film 14 and the environmental temperature and is not less than 3 min;
step S7: moving the basement membrane 14 carrying the secondary antibody conjugate into the elution device 5 by the conveyor belt 15, and eluting the first antibody and the second antibody in a free state which do not participate in the reaction in the basement membrane 14 by the elution device 5 to separate the first antibody and the second antibody from the basement membrane 14;
in the step, the solution in the elution tank 17 is 0.8% sodium chloride solution, the elution time is adaptively adjusted according to the thickness of the base film 14 and the width of the base film 14, and the conveying belt 15 can drive the base film 14 to move in the eluent in the elution process so as to enhance the elution effect;
step S8: moving the eluted base film 14 to a radioactivity detecting position (VI) corresponding to the radioactivity detecting device 2 by the conveyor belt 15, detecting the radioactivity intensity of the base film 14 by the radioactivity detecting device 2; the amount of the secondary antibody conjugate and the primary anti-conjugate in the basement membrane 14 can be judged according to the radioactive intensity, so as to obtain the concentration of 4-methylimidazole in the wine, and the concentration of the caramel pigment in the wine can be calculated to obtain the concentration value of the caramel pigment in the wine;
in the step, the measured radioactivity value a, the radioactivity of 1mol of the second antibody is defined as b, the molecular proportion coefficient of the combination of the first antibody and the second antibody is defined as c, the molecular proportion coefficient of the combination of the first antibody and 4-methylimidazole is defined as d, and the mass fraction of 4-methylimidazole in the caramel pigment is defined as e; the molar mass of the 4-methylimidazole is f, the adding volume of the wine is v, the density of the wine is g, and the mass percentage of the caramel pigment in the wine is h, so that h is acdf/bevg.
The above description is only a preferred embodiment of the present invention, and all equivalent changes or modifications of the structure, characteristics and principles described in the present patent application are included in the protection scope of the present patent application.
Claims (7)
1. The utility model provides a grape wine caramel pigment detection robot which characterized in that: comprises a conveying device (1), a first antibody spraying device (4), a second antibody spraying device (3), an elution device (5) and a radioactivity detection device (2);
the conveying device (1) comprises a main bracket (11), a belt wheel (12), a motor (13) and a conveying belt (15); a base film (14) with water absorption is stuck on the conveyor belt (15);
the first antibody spraying device (4) contains a first antibody liquid containing a first antibody; the first antibody is a substance which can perform specific immunoreaction with 4-methylimidazole contained in caramel pigment; the specific immune reaction can generate a 4-methylimidazole-antigen conjugate, namely an anti-conjugate; the first antibody spraying device (4) is used for spraying the first antibody on the basement membrane (14);
the second antibody spraying device (3) is filled with a second antibody liquid containing a second antibody; said second antibody specifically binds to said first anti-binding agent to produce a second binding agent; the secondary antibody binding body is a colloidal precipitate; the second antibody is radiolabeled and has weak radioactivity; the second antibody spraying device (3) is used for spraying the second antibody on the basement membrane (14);
the elution device (5) is filled with eluent; the eluent can elute the first and second antibodies from the basement membrane (14);
the radioactivity detecting device (2) can detect the radioactivity intensity of the basement membrane (14).
2. The wine caramel color detection robot of claim 1, wherein: the front surface of the base membrane (14) is made of flexible porous materials and has good water absorption.
3. The wine caramel color detection robot of claim 2, wherein: the wine caramel pigment detection robot also comprises a liquid suction device (6) used for dehydrating the eluted basement membrane (14).
4. The wine caramel color detection robot of claim 1, wherein: the first antibody spraying device (4) and the second antibody spraying device (3) have the same structure and respectively comprise a liquid box (7), a quantitative pump (8) and a spraying head (10); the front end of the sprinkler head (10) is provided with a plurality of atomization units (10c) for atomizing the first antibody liquid and the second antibody liquid.
5. The wine caramel color detection robot of claim 5, wherein: the radioactivity detection device (2) further comprises a body (18), a fluorescent sheet (21), a convex lens (20) and a photoelectric sensor (19); the fluorescent sheet (21) is fixedly arranged at the front end of the body (18); the convex lens (20) is fixedly arranged on the rear side of the fluorescent sheet (21); the photoelectric sensor (19) is fixedly arranged on the rear side of the convex lens (20); the fluorescent sheet (21), the convex lens (20) and the photoelectric sensor (19) are coaxially arranged, and the distance between the photoelectric sensor (19) and the convex lens (20) is smaller than the focal length of the convex lens (20).
6. The wine caramel color detection robot of claim 5, wherein: the radioactivity detection device (2) further comprises a body (18), a fluorescent sheet (21), a convex lens (20) and a photoelectric sensor (19); the fluorescent sheet (21) is fixedly arranged at the front end of the body (18); the convex lens (20) is fixedly arranged on the rear side of the fluorescent sheet (21); the photoelectric sensor (19) is fixedly arranged on the rear side of the convex lens (20); the fluorescent sheet (21), the convex lens (20) and the photoelectric sensor (19) are coaxially arranged, and the distance between the photoelectric sensor (19) and the convex lens (20) is smaller than the focal length of the convex lens (20).
7. The detection method of the caramel pigment in the wine by using the wine caramel pigment detection robot as claimed in any one of claims 1 to 5, comprising the following steps:
s1, adhering the base film (14) to the outer surface of the conveyor belt (15);
s2, dropwise adding a fixed volume of wine liquid dispersion to be detected to the surface of the basement membrane (14) and allowing the wine liquid dispersion to diffuse in the whole basement membrane (14);
s3, moving the basal membrane (14) to which the wine liquid is dripped to a first antibody spraying position (II) corresponding to the first antibody spraying device (4) through a conveyor belt (15), and spraying excessive first antibody liquid on the surface of the basal membrane (14);
s4, standing for a period of time to enable the 4-methylimidazole in the wine liquid and the first antibody in the first antibody liquid to generate specific immunoreaction and generate an anti-binding body;
s5, after the specific immunoreaction is completed, the conveyer belt (15) drives the reacted basement membrane (14) to move to a second antibody spraying position (III) corresponding to the second antibody spraying device (3), and excessive second antibody liquid is sprayed on the surface of the basement membrane (14);
s6, standing for a period of time to make the first anti-binding body in the basement membrane (14) and the second antibody in the second antibody liquid generate specific binding to generate a secondary anti-binding body of colloidal sediment;
s7, moving the basement membrane (14) carrying the secondary antibody combination into the elution device (5) through the conveyor belt (15), and eluting the free-state first antibody and the free-state second antibody which do not participate in the reaction in the basement membrane (14) through the elution device (5) so as to separate the basement membrane (14);
s8, moving the eluted basal membrane (14) to a radioactivity detection position (VI) corresponding to the radioactivity detection device (2) through a conveyor belt (15), and detecting the radioactivity intensity of the basal membrane (14) through the radioactivity detection device (2); the amount of the secondary antibody conjugate and the primary antibody conjugate in the basement membrane (14) can be judged according to the radioactive intensity, so that the concentration of 4-methylimidazole in the wine is obtained, and the concentration value of the caramel pigment in the wine can be obtained by calculating the concentration of 4-methylimidazole in the caramel pigment.
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| KR20170116358A (en) * | 2016-04-11 | 2017-10-19 | 한국식품연구원 | Analytical method to detect caramel color in red ginseng products |
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| WO2021007738A1 (en) * | 2019-07-15 | 2021-01-21 | 中国科学院苏州生物医学工程技术研究所 | Functionalized biological multilayer porous membrane immune carrier, preparation method, and application |
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| CN1560629A (en) * | 2004-02-19 | 2005-01-05 | 中国农业大学 | Method for quickly detecting 4-methyl iminazole in food |
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