CN118130194A - Preparation and detection method of kit for rapidly detecting blood ammonia through headspace fluorescence visualization - Google Patents

Abstract

The invention discloses a preparation and detection method of a kit for rapidly detecting blood ammonia by headspace fluorescence visualization, which comprises the following steps: adding a solid alkaline reactant into the bottom of a test tube, and attaching an inert porous hydrogel sheet to the inner side of the top cover of the test tube; dissolving hydroxytyrosol in secondary distilled water to obtain detection liquid A; dissolving 8-hydroxy julolidine in ethanol with the volume fraction of 75% to obtain detection liquid B; mixing the detection liquid A and the detection liquid B to obtain detection liquid C; and centrifuging the fresh blood sample, and collecting supernatant to obtain a blood ammonia sample to be detected. According to the invention, visual semi-quantitative detection of blood ammonia can be realized by visually identifying and detecting the fluorescence intensity of the base solution. The detection results of fresh whole blood and plasma samples show that the invention has good stability, reproducibility and accuracy; the reaction at room temperature is simple and quick to operate, low in cost and small in interference, and can meet the requirements of clinical blood ammonia and on-site blood ammonia quick detection.

Description

Preparation and detection method of a kit for rapid detection of blood ammonia by headspace fluorescence visualization

Technical Field

The present invention belongs to the field of medical technology, and more specifically, relates to a kit preparation and a detection method for rapid detection of blood ammonia by headspace fluorescence visualization.

Background technique

Blood ammonia (blood ammonia; plasma ammonia; serum ammonia) is ammonia produced by the metabolism of various amino acids in various tissues in the body and ammonia absorbed by the intestines into the blood to form blood ammonia; blood ammonia indicators are generally used to detect liver disease or other acute hepatic encephalopathy; existing clinical blood ammonia meters have large detection errors, many operating reaction steps, and the cost of single sample blood ammonia detection is too high and the detection time is too long. The Chinese invention patent with application number 201910839666.9 discloses a portable visual blood ammonia rapid fluorescence detection device and its preparation and use method. It uses the detection liquid C obtained by mixing dopamine and resorcinol as a detection reagent for detecting blood ammonia. The detection liquid is easy to slip and the alkaline reactant is liquid. The portability, detection speed and visual detection sensitivity of its detection device are still not ideal.

Summary of the invention

An object of the present invention is to solve at least the above-mentioned problems and/or disadvantages and to provide at least the advantages which will be described hereinafter.

In order to achieve these purposes and other advantages according to the present invention, a method for preparing a kit for rapid detection of blood ammonia by headspace fluorescence visualization is provided, comprising:

S1. Add a solid alkaline reactant to the bottom of a test tube and attach an inert porous hydrogel sheet to the inner side of a top cover of the test tube;

S2, dissolving hydroxytyrosol in double distilled water to obtain a detection solution A; dissolving 8-hydroxyjulolidine in 75% by volume ethanol to obtain a detection solution B;

S3, mixing the detection liquid A and the detection liquid B to obtain the detection liquid C;

S4. Centrifuge the fresh blood sample and collect the supernatant to obtain the blood ammonia sample to be tested.

Preferably, the method for preparing the inert porous hydrogel sheet comprises the following steps:

Weigh agarose powder, dissolve it with a mixed solution of high-purity water and anhydrous ethanol, heat in a water bath until the solution is completely transparent, add the agarose solution dropwise to the inside of the top cover of the test tube, close the cover after cooling to room temperature, and put it in a refrigerator for use.

Preferably, the mass volume ratio of the agarose powder to the mixed solution is 0.9-1 g:100-150 mL, the volume ratio of high-purity water to anhydrous ethanol in the mixed solution is 6-8:4-6, and the volume mass ratio of the agarose solution to the solid alkaline reactant is 80-120 μL:0.4-0.6 g.

Preferably, the solid alkaline reactant is NaOH or KOH.

Preferably, the molar concentrations of the detection liquid A and the detection liquid B are both 10.0 mmol/L; the detection liquid A and the detection liquid B are mixed at a molar ratio of 1:4 to 3:2 to obtain the detection liquid C.

Preferably, the detection liquid C is obtained by mixing the detection liquid A and the detection liquid B in a molar ratio of 1:1.

Preferably, in said S4, the fresh blood sample is centrifuged using a fingertip gyro-microcentrifuge, and the supernatant is collected to obtain the blood ammonia sample to be tested.

A detection method of a kit for rapid detection of blood ammonia by headspace fluorescence visualization, comprising the following steps:

Step 1: Add the test solution C dropwise onto the inert porous hydrogel sheet on the inner side of the top cover of the test tube and let it stand at room temperature for 2 to 6 minutes;

Step 2: Place the alkaline reactant at the bottom of the test tube and cover it with the top cover;

Step 3: Inject the ammonium salt standard solution or blood ammonia sample solution into the bottom of the test tube from the middle side of the test tube through a micro-syringe. Mix the ammonium salt standard solution or blood ammonia sample solution with the alkaline reactant. Do not pull out the syringe.

Step 4: Place the bottom of the test tube in hot water and leave it upright for a certain period of time;

Step 5. Use ultraviolet light to vertically irradiate the top cover of the test tube, compare the fluorescence intensity and RGB value of the inert porous hydrogel sheet obtained from the blood ammonia sample test with the fluorescence color card of the ammonium salt standard solution series gel sheet, and realize rapid semi-quantitative detection of blood ammonia by naked eyes based on the fluorescence intensity, and realize rapid quantitative detection of blood ammonia by mobile phone image recognition software Image J based on the RGB value of the inert porous hydrogel sheet.

Preferably, in step 1, the volume ratio of the detection solution C to the agarose solution is 20-40 μL:80-120 μL;

In the step 4, the hot water temperature is 80-90° C., and the vertical static standing time is 1-20 minutes.

Preferably, if the fluorescence brightness of the blood ammonia sample gel sheet exceeds the fluorescence color chart range of the ammonium salt standard solution series gel sheet, the blood ammonia sample is appropriately diluted with double distilled water and then retested.

The present invention includes at least the following beneficial effects: the present invention uses an inert porous hydrogel sheet with an inert ultra-low fluorescence background to fix the detection liquid C, and the gel sheet can also absorb the gas ammonia released by the sample to cause the pH value to increase, triggering the in-situ chemical reaction of hydroxytyrosol and 8-hydroxyljulolidine in the gel sheet, and quickly generating a yellow-green fluorescent product. After the sample reacts for 8 minutes, the fluorescence intensity and RGB value of the gel sheet can be identified by the naked eye and the mobile phone image recognition software Image J, so as to achieve rapid, visual semi-quantitative and quantitative detection of blood ammonia. The present invention uses a fingertip gyro-microcentrifuge to manually or electrically centrifuge blood samples, which is simple and fast to operate. The present invention detects the blood ammonia content in fresh plasma and serum samples, and the comparison with the test results of the commercial blood ammonia detection kit shows that the present invention has good stability, reproducibility and accuracy in detecting blood ammonia; the present invention is convenient to operate for detecting blood ammonia, has low cost and little interference, and can meet the requirements of clinical blood ammonia and on-site rapid detection of blood ammonia.

Other advantages, objectives and features of the present invention will be embodied in part through the following description, and in part will be understood by those skilled in the art through study and practice of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 is a schematic diagram of a specific embodiment of the portable visual blood ammonia rapid fluorescence detection device of the present invention;

Figure 2 is the fluorescence intensity test results of hydroxytyrosol and 8-hydroxyjulolidine in buffer solutions with different pH values, wherein the abscissa is the pH value, the ordinate is the fluorescence intensity, and the unit of the ordinate is the fluorescence intensity unit a.u.;

Figure 3 shows the fluorescence intensity test results of the detection solution C obtained by mixing the detection solution A and the detection solution B according to different volume ratios (molar ratios) and reacting in a pH 9.91BR buffer solution at room temperature for 10 minutes, wherein the abscissa is the volume ratio of the detection solution A to the detection solution B, the ordinate is the fluorescence intensity, and the unit of the ordinate is the fluorescence intensity unit a.u.;

FIG4 is a schematic diagram of a specific embodiment of the ammonium salt standard solution series gel sheet fluorescent color card of the present invention; the numbers in FIG4 represent different concentrations of ammonium salt standard solutions, in micromoles/liter;

FIG5 is a fluorescence photograph of a hydrogel sheet for detecting blood ammonia in a plasma sample in Example 1 and a serum sample in Example 2 of the present invention;

FIG6 is a schematic diagram of a top view of the hydrogel portable kit provided by the present invention;

FIG7 is a schematic diagram of the test tube structure with the top cover closed;

FIG8 is a schematic diagram of the structure of the test tube with the top cover opened;

FIG. 9 is a schematic diagram of the cross-sectional structure of the test tube rack.

Detailed ways

The present invention will be further described in detail below in conjunction with the accompanying drawings so that those skilled in the art can implement the invention with reference to the description.

Example 1

This embodiment provides a method for preparing a kit for rapid detection of blood ammonia by headspace fluorescence visualization, comprising the following steps:

S1. Add 0.5 g of alkaline reactant 3 (analytical grade NaOH) to the bottom of the test tube;

S2. Dissolve hydroxytyrosol in double distilled water and dissolve 8-hydroxyjulolidine in 75% ethanol to obtain detection solution A and detection solution B; the molar concentration of detection solution A and detection solution B are both 10.0 mmol/L;

S3. Mix the detection solution A and the detection solution B at a molar ratio of 1:1 to obtain the detection solution C.

S4. Use a fidget gyro-microcentrifuge to centrifuge the fresh blood sample for 3 minutes, and collect the supernatant to obtain a blood ammonia sample.

As shown in FIG1 , the detection method of the kit for rapid detection of blood ammonia by headspace fluorescence visualization includes:

30.0 μL of detection solution C 4 is added dropwise onto the inert hydrogel 2 on the inner side of the top cover of the test tube 1, and after cooling to room temperature, the top cover 5 is covered;

300.0 μL of blood ammonia sample solution 6 is injected into the solid alkaline reactant 3 at the bottom of the test tube 1 from the side of the middle of the test tube 1 using a micro syringe 7 to form a mixed solution. The bottom of the test tube is placed in 80°C hot water and left to stand vertically for 8 minutes.

The blood ammonia sample reacts with the solid alkaline reactant 3 to release gaseous ammonia 8, which rises and is absorbed by the inert porous hydrogel sheet 2 located on the inner side of the top cover of the test tube 1. The increase in pH value causes hydroxytyrosol and 8-hydroxyjulolidine in the detection solution C4 in the inert porous hydrogel sheet 2 to react rapidly and generate a yellow-green fluorescent product in situ; the blood ammonia sample is a plasma sample.

The specific chemical reaction formula for the generation of fluorescent products by hydroxytyrosol and 8-hydroxyjulolidine is:

The inert porous hydrogel sheet 2 is irradiated vertically by a portable ultraviolet lamp 9 to prepare a series of ammonium salt standard solution gel sheet fluorescence color cards, as shown in FIG4. The plasma or serum sample test gel sheet is compared with the standard fluorescence color card, and the blood ammonia rapid semi-quantitative detection can be achieved by naked eyes according to the fluorescence brightness; the blood ammonia rapid quantitative detection can be achieved by mobile phone image recognition software Image J according to the fluorescence RGB value, as shown in FIG5. The blank sample in FIG5 is 300.0 μL of double distilled water, blood sample 1 is 300.0 μL of plasma sample of healthy volunteers, and blood sample 2 is 300.0 μL of serum sample of healthy volunteers.

As shown in Figure 2, after 50 μL of detection solution C reacted in 1.95 mL of BR buffer solution with different pH values for 8 minutes, the fluorescence intensity of the mixed solution increased with the increase of pH value, and the fluorescence intensity and pH value had a good linear relationship in the range of 7.24 to 10.38. The experimental results show that the higher the pH value, the faster the reaction rate of hydroxytyrosol and 8-hydroxyjulolidine in the detection solution C, and the more yellow-green fluorescent products are generated.

As shown in FIG3 , the effect of different volume ratios of the two solutions of the same molar concentration of the detection solution A and the detection solution B on the fluorescence intensity of the detection solution C under the condition of pH = 9.91 is given. As can be seen from FIG3 , when the molar ratio (volume ratio) of the two reactants of the detection solution A and the detection solution B is 1:1, the fluorescence intensity is the largest. Therefore, the molar ratio of the two reactants of the detection solution A and the detection solution B in the detection solution C of the present invention is preferably 1:1.

Example 2

This embodiment provides a method for preparing a kit for rapid detection of blood ammonia by headspace fluorescence visualization, comprising the following steps:

S1. Add 0.5 g analytical grade NaOH solid 3, an alkaline reactant, directly to the bottom of the test tube;

S2. Dissolve hydroxytyrosol in double distilled water and dissolve 8-hydroxyjulolidine in 75% ethanol to obtain detection solution A and detection solution B; the molar concentration of detection solution A and detection solution B are both 10.0 mmol/L;

S3. Mix the detection solution A and the detection solution B in a volume ratio of 1:1 to obtain the detection solution C.

S4. Use a fidget gyro-microcentrifuge to centrifuge the fresh blood sample without anticoagulant for 3 minutes, and collect the supernatant to obtain the serum sample.

When in use, place 30.0 μL of detection solution C 4 on the inert porous hydrogel sheet 2 on the inner side of the top cover of the test tube 1, and cover it with the top cover 5 after cooling to room temperature;

300.0 μL of serum sample solution 6 is injected into the bottom of the test tube 1 from the outside of the middle of the test tube 1 using a micro syringe 6 to mix with the solid alkaline reactant 3;

The bottom of the test tube was placed in 80-90℃ hot water and left to stand vertically for 8 minutes;

The serum sample reacts with NaOH to release gaseous ammonia 8, which rises and is absorbed by the inert porous hydrogel sheet 2 located on the inner side of the top cover of the test tube 1. The increase in pH value prompts the detection liquid C 4 to react rapidly to generate a yellow-green fluorescent product; the blood ammonia sample is a serum sample.

The present invention uses a transparent plastic tube with a sealable cover and ultra-low fluorescence background as a test tube. The inert porous hydrogel sheet on the inner side of the tube cover is inert, transparent, and non-fluorescent. The bottom of the test tube contains NaOH solid alkaline reactant, which releases gas ammonia 8 after mixing with the sample solution. The gas ammonia 8 rises and is absorbed by the hydrogel sheet on the inner side of the top cover. The increase in pH value triggers the detection liquid C to react rapidly and generate fluorescent products in situ. An ultraviolet lamp 9 is used as an excitation light source to vertically irradiate the gel sheet, and the fluorescence intensity and RGB value of the gel sheet are identified by naked eyes and mobile phone image recognition software Image J.

By comparing with the ammonium salt standard series gel slices, the present invention can achieve rapid, visual semi-quantitative and quantitative detection of blood ammonia. The blood ammonia content in fresh plasma and serum samples was measured three times in parallel, and the test results were compared with the test results of the commercial blood ammonia detection kit, indicating that the blood ammonia content of plasma and serum samples was about 60 μmol/L and 40 μmol/L, respectively. The results show that the detection device and preparation and use method disclosed in the present invention have good reproducibility and accuracy in blood ammonia determination.

The blood ammonia detection device of the present invention integrates two independent sample reaction systems and a blood ammonia detection system into a small test tube by using a plastic tube with a cover; the detection solution C is absorbed and fixed by an inert porous hydrogel sheet before testing; the packaged strong base solid is added to the bottom of the test tube before testing, and the stability and portability are relatively strong; the present invention adopts a fingertip gyro-microcentrifuge to manually or electrically collect plasma or serum samples, and sample collection is not limited by large instruments, testing sites and time; the present invention adopts the naked eye and mobile phone image recognition software Image J to collect gel sheet fluorescence signals, and the practicality is strong; the blood ammonia detection of the present invention is simple and fast to operate, low in cost, and has little interference, and can meet the requirements of clinical blood ammonia and on-site blood ammonia rapid detection.

As shown in FIG. 6 to FIG. 9 , the structure of the kit used for detecting blood ammonia in Example 1 to Example 2 of the present invention includes:

The box body 20 has a detachable test tube rack 21 disposed therein, wherein the test tube rack 21 is provided with a plurality of array-type placement holes 22, in which the test tube 1 is placed, and an inert porous hydrogel sheet 2 is disposed on the inner side of the top cover 5 of the test tube 1;

A syringe placement slot 26, which is disposed in the box body 20, and a micro-syringe is placed in the syringe placement slot 26;

The flashlight placement slot 27 is arranged in the box body 20 , and the syringe placement slot 26 is located between the flashlight placement slot 27 and the test tube rack 21 . An LED ultraviolet flashlight is placed in the flashlight placement slot 27 .

Working principle: A hydrogel portable test kit provided by the present invention realizes the centralized storage of test tubes, reagent bottles, micro-injectors, and LED ultraviolet flashlights, which is convenient for carrying blood ammonia detection. Among them, the test tube is the most important structure of the test kit and the main structure for realizing blood ammonia detection; when performing blood ammonia detection, first add a solid alkaline reactant (analytical pure NaOH or KOH) to the bottom of the test tube 1, mix hydroxytyrosol and 8-hydroxyl-julolidine and then drop them onto the inert porous hydrogel sheet 2 on the inner side of the top cover of the test tube, and then inject the plasma or serum to be tested into the bottom of the test tube through a micro-injector, and the ammonia nitrogen in the blood ammonia or serum is released in the form of gaseous ammonia in a strong alkaline environment, and the pH rises after the inert porous hydrogel sheet 2 absorbs gaseous ammonia in the headspace, which triggers a rapid chemical reaction between hydroxytyrosol and 8-hydroxyl-julolidine in the inert porous hydrogel sheet, and generates a yellow-green fluorescent product in situ; using an LED ultraviolet flashlight as a light source, according to the fluorescence signal of the inert porous hydrogel sheet, blood ammonia is quickly semi-quantitatively and accurately quantitatively detected by naked eyes and smartphone image recognition software, respectively. The higher the blood ammonia content, the more gaseous ammonia is released, the more the pH value of the inert porous hydrogel sheet increases, the stronger the yellow-green fluorescence of the inert porous hydrogel sheet, and the faster the fluorescence signal of the hydrogel sheet responds.

In the above technical solution, an elastic clamp seat 23 is provided in the box body 20, and a reagent bottle 24 is elastically clamped on the elastic clamp seat 23. The reagent bottle 24 is used to contain solid alkaline reactants, such as solid sodium hydroxide and potassium hydroxide.

In the above technical solution, the structure of the test tube 1 includes:

A top cover 5, which is arranged at the upper end of the test tube 1;

A sample reaction pool 101, which is located at the lower end of the test tube 1;

The injection port 102 is arranged on the side wall of the test tube 1, and the injection port 102 is connected with the sample reaction pool 101. The sample reaction pool 101 is used to contain solid alkaline reactants, and is also the place where the blood ammonia to be tested reacts with the solid reactants to release gaseous ammonia. The injection port 102 is used to inject the serum or plasma to be tested through a micro syringe.

In the above technical solution, a mesh convex layer 51 is provided on the inner side of the top cover 5, and the inert porous hydrogel sheet 2 is fixedly attached to the surface of the mesh convex layer 51. The provision of the mesh convex layer 51 increases the adhesion and fixing force to the inert porous hydrogel sheet 2, which can effectively prevent the inert porous hydrogel sheet 2 from falling off.

In the above technical solution, the test tube 1 is sleeved with a rubber ring 30, a fixing tube 31 is arranged on the rubber ring 30, a spring 32 is arranged in the fixing tube 31, a positioning pin 33 is connected to the lower end of the spring 32, and a positioning hole 211 adapted to the positioning pin 33 is opened on the upper surface of the test tube rack 21, the positioning pin 33 is in a stepped shape with a larger upper part and a smaller lower part, and the positioning pin 33 is movably inserted in the positioning hole 211. The cooperation between the positioning pin 33 and the positioning hole 211 realizes the effective position limiting of the test tube 1, and avoids the test tube 1 from rotating and shaking during the carrying process of the reagent kit. At the same time, by arranging the spring 32 in the fixing tube 31 on the positioning pin 33, the positioning pin 33 is given an elastic displacement capacity in the vertical direction, and the hard contact between the test tube 1 and the test tube rack 21 is avoided, and the test tube 1 is provided with position limiting and elastic protection.

In the above technical solution, fan-shaped grooves 28 are respectively arranged on both sides of the syringe placement groove 26 and the flashlight placement groove 27. The fan-shaped grooves 28 are arranged to facilitate the insertion of fingers, so that the micro-syringe and the LED ultraviolet flashlight can be quickly picked up.

In the above technical solution, the box body 20 is also rotatably connected to a cover body 29 , and an elastic clip 291 for clamping and fixing the ammonium salt standard solution series colorimetric card is provided on the inner side of the cover body 29 .

The number of devices and processing scales described here are used to simplify the description of the present invention. Applications, modifications and variations of the present invention will be obvious to those skilled in the art.

Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and the implementation modes, and they can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to the specific details and the illustrations shown and described herein.

Claims (10)

1. The preparation method of the kit for rapidly detecting blood ammonia by headspace fluorescence visualization is characterized by comprising the following steps of:

S1, adding a solid alkaline reactant into the bottom of a test tube, and attaching an inert porous hydrogel sheet to the inner side of a top cover of the test tube;

S2, dissolving hydroxytyrosol in secondary distilled water to obtain detection liquid A; dissolving 8-hydroxy julolidine in ethanol with the volume fraction of 75% to obtain detection liquid B;

S3, mixing the detection liquid A and the detection liquid B to obtain detection liquid C;

s4, centrifuging the fresh blood sample, and collecting supernatant to obtain a blood ammonia sample to be detected.

2. The method for preparing the kit for rapidly detecting ammonia in blood by headspace fluorescence visualization as claimed in claim 1, wherein the method for preparing the inert porous hydrogel sheet comprises the following steps:

Weighing agarose powder, dissolving agarose powder with mixed solution of high-purity water and absolute ethyl alcohol, heating in water bath until the solution is completely transparent, dripping agarose solution into the inner side of a top cover of a test tube, closing the cover after cooling to room temperature, and placing into a refrigerator for standby.

3. The preparation method of the kit for rapidly detecting the blood ammonia by using the headspace fluorescence visualization method according to claim 2, wherein the mass volume ratio of the agarose powder to the mixed solution is 0.9-1 g:100-150 mL, the volume ratio of the high-purity water to the absolute ethyl alcohol in the mixed solution is 6-8:4-6, and the volume mass ratio of the agarose solution to the solid alkaline reactant is 80-120 mu L and 0.4-0.6 g.

4. The method for preparing the kit for rapidly detecting the blood ammonia by using the headspace fluorescence visualization method according to claim 1, wherein the solid alkaline reactant is NaOH or KOH.

5. The method for preparing the kit for rapidly detecting the blood ammonia by using the headspace fluorescence visualization method according to claim 1, wherein the molar concentration of the detection solution A and the detection solution B is 10.0mmol/L; and uniformly mixing the detection liquid A and the detection liquid B according to the molar ratio of 1:4-3:2 to obtain detection liquid C.

6. The method for preparing the kit for rapidly detecting the blood ammonia by using the headspace fluorescence visualization method according to claim 1, wherein the detection solution C is obtained by mixing detection solution A and detection solution B according to a molar ratio of 1:1.

7. The method for preparing the kit for rapidly detecting the blood ammonia by using the headspace fluorescence visualization method according to claim 1, wherein in the step S4, a fingertip gyroscope-micro centrifuge is used for centrifuging a fresh blood sample, and a supernatant is collected to obtain the blood ammonia sample to be detected.

8. A detection method of a kit for rapidly detecting blood ammonia by headspace fluorescence visualization, which is prepared by the preparation method of the kit for rapidly detecting blood ammonia by headspace fluorescence visualization according to any one of claims 1 to 7, and is characterized by comprising the following steps:

Step one, dropwise adding the detection liquid C onto an inert porous hydrogel sheet at the inner side of a top cover of a test tube, and standing at room temperature for 2-6 min;

Step two, placing an alkaline reactant at the bottom of a test tube, and covering a top cover;

step three, injecting an ammonium salt standard solution or an ammonia blood sample solution into the bottom of the test tube from the side surface of the middle part of the test tube through a micro-injector, mixing the ammonium salt standard solution or the ammonia blood sample solution with an alkaline reactant, and not pulling out the injector;

fourthly, placing the bottom of the test tube in hot water, standing vertically and standing for a certain time;

And fifthly, vertically irradiating a top cover of a test tube by using an ultraviolet lamp, comparing the fluorescence intensity and RGB value of the inert porous hydrogel film obtained by detecting the blood ammonia sample with the fluorescence color card of the ammonium salt standard solution series gel film, realizing the rapid semi-quantitative detection of the blood ammonia by naked eyes according to the fluorescence intensity, and realizing the rapid quantitative detection of the blood ammonia by using mobile phone Image recognition software Image J according to the RGB value of the inert porous hydrogel film.

9. The method for detecting the kit for rapidly detecting the blood ammonia by the headspace fluorescence visualization method according to claim 8, wherein in the first step, the volume ratio of the detection solution C to the agarose solution is 20-40. Mu.L and 80-120. Mu.L;

in the fourth step, the temperature of the hot water is 80-90 ℃, and the vertical standing time is 1-20 min.

10. The method for detecting the kit for rapidly detecting the blood ammonia by using the headspace fluorescence visualization according to claim 8, wherein if the fluorescence intensity of the blood ammonia sample gel sheet exceeds the range of the ammonium salt standard solution series gel sheet fluorescence color card, the blood ammonia sample is re-detected after being properly diluted by secondary distilled water.