Analysis method and device for detecting activity of acid phosphatase in soil

Abstract

The invention discloses an analysis method and device for detecting the activity of acid phosphatase in soil, and belongs to the field of acid phosphatase activity analysis. An analysis method for detecting the activity of acid phosphatase in soil mainly comprises the following steps: step 1, preparing a soil raw material, an acetate buffer solution with the pH value of 5, a disodium nitrophenyl phosphate solution and a NaOH solution, then spraying the soil wet, fully stirring the sprayed soil until the humidity of the soil is basically consistent, and taking out the soil; meanwhile, the weight of sprayed water is recorded, the actual specific gravity of the soil raw materials is measured, and the disodium nitrophenyl phosphate solution serves as a substrate; by controlling the humidity of the soil to be consistent, the detection accuracy can be improved, the mixing effect of the acetate buffer solution and the soil is effectively improved through the mixing barrel, and the problem that sampling is difficult in a large-caliber mixing device at present is effectively solved.

Description

Technical Field

[0001] The invention relates to the technical field of acid phosphatase activity analysis, and in particular to an analysis method and device for detecting acid phosphatase activity in soil. Background Art

[0002] Soil phosphorus is an essential nutrient for plant growth, and most of it exists in the form of organic phosphorus. Organic phosphorus usually needs the enzymatic action of soil acid phosphatase to decompose and convert into a form that can be absorbed and utilized by plants. Acid phosphatase is an important hydrolase in soil, which can affect soil fertility and plant metabolic processes, so it is necessary to analyze the activity of acid phosphatase in the soil.

[0003] At present, when testing acid phosphatase in soil, the sampled soil is directly used for testing. Since the humidity of the upper and lower layers of the soil may be different, it may affect the final test results. In addition, during the experimental analysis, the volume of the mixture of soil and acetate buffer is small and it is necessary to keep it in a mixed state at all times. If it is mixed in a large-diameter container, the liquid depth is small, which may affect its sampling. If it is mixed in a small-diameter container, it may affect the mixing of the soil and acetate buffer.

[0004] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention and should not be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art. Summary of the invention

[0005] The technical problem to be solved by the present invention is to overcome the deficiencies of the prior art and provide an analytical method and device for detecting acid phosphatase activity in soil that can overcome the above problems or at least partially solve the above problems.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] An analytical method for detecting acid phosphatase activity in soil mainly comprises the following steps:

[0008] Step 1, prepare a soil raw material, an acetate buffer solution with a pH value of 5, a disodium nitrobenzene phosphate solution and a NaOH solution, then spray the soil with water, and fully stir the sprayed soil to make its humidity basically consistent, and record the weight of the sprayed water to measure the actual specific gravity of the soil raw material, wherein the disodium nitrobenzene phosphate solution is used as a substrate;

[0009] Step 2: Take 10 grams of soil, dry it, and calculate the moisture content of the soil;

[0010] Step 3: Take 30 grams of soil raw material and add it into a mixing device, add 80 ml of acetate buffer to the soil and stir evenly to form a crude enzyme solution, and keep stirring it continuously so that the soil and acetate buffer are always in a stirring state;

[0011] Step 4, take ELISA plate G, set 8 groups of samples and 16 groups of controls in ELISA plate G, use a sampler to take samples, each group of samples uses 100ul crude enzyme solution and 150ul substrate as culture medium; 16 groups of controls include 8 groups of substrate controls and 8 groups of crude enzyme solution controls, the substrate controls use 150ul substrate and 100ul acetate buffer as culture medium, and the crude enzyme solution controls use 100ul crude enzyme solution and 150ul acetate buffer as culture medium;

[0012] Step 5: Place the ELISA plate G filled with the culture solution in step 4 in a 30°C incubator and culture for 60 minutes;

[0013] Step 6: Take out the ELISA plate G after the culture is completed, transfer 100ul of the culture solution in the corresponding well of the ELISA plate H to the corresponding well of the ELISA plate G, and then add 10ul of NaOH solution to each well to terminate the reaction. Finally, place the plate in an ELISA reader for colorimetry at a wavelength of 405nm to obtain the sample Abs, substrate control Abs and crude enzyme solution control Abs;

[0014] Step 7: Calculate the acid phosphatase activity using the p-nitrophenol standard curve.

[0015] An analytical device used in an analytical method for detecting acid phosphatase activity in soil, comprising a mixing device, an ELISA plate, an ELISA instrument, a sampler and an incubator, wherein the mixing device is used for mixing the soil with the acetate buffer in step three, the stirring device comprises a mixing barrel, a fixing ring is fixedly connected in the mixing barrel, a plurality of connecting grooves are circumferentially arranged on the fixing ring, a rotating shaft is rotatably arranged in the mixing barrel, and the fixing ring is sealingly rotatably connected to the lower end of the rotating shaft, a sampling groove is arranged in the rotating shaft, a plurality of groups of through holes are circumferentially arranged on the rotating shaft, a plurality of first fixing grooves are fixedly connected in the mixing barrel in a circumferential manner, and a plurality of first fixing grooves are fixedly connected in the mixing barrel in a circumferential manner. Block, the first fixed block is provided with a first grinding surface and a second grinding surface, and the first fixed block matches the connecting groove, a plurality of second fixed blocks are fixedly connected to the rotating shaft in a circumferential manner, a third grinding surface and a fourth grinding surface are provided on the second fixed block, the third grinding surface matches the first grinding surface, and the fourth grinding surface matches the second grinding surface, a sealing plate matching the first fixed block is fixedly connected to the upper side of the second fixed block, a motor is fixedly connected to the lower side of the mixing barrel, the output end of the motor is fixedly connected to the rotating shaft, and a plurality of buffer components matching the second fixed blocks are provided on the rotating shaft.

[0016] Preferably, the buffer assembly comprises a plurality of limiting grooves circumferentially arranged on the rotating shaft, a slider is slidably arranged in the limiting groove, a spring is symmetrically fixedly connected between the slider and the limiting groove, and a connecting plate is fixedly connected between the slider and the sealing plate.

[0017] Furthermore, a corrugated plate is fixedly connected to a side of the sealing plate away from the second fixed block, and a first pressure-bearing surface, a guide surface, and a second pressure-bearing surface are provided on the corrugated plate, and the guide surface matches the first fixed block.

[0018] In order to provide pressure to the end of the sealing plate, further, the end of the first pressure-bearing surface close to the rotating shaft is inclined toward the end close to the mixing barrel.

[0019] Preferably, the plurality of connecting plates are fixedly connected via connecting rings.

[0020] Preferably, a plurality of fixing rods are fixedly connected to the rotating shaft in a circle, an elastic rod is fixedly connected to the lower end of the fixing rod, and the elastic rod is arranged between the wave plate and the second fixed block. When the elastic rod is squeezed and deformed by the first fixed block and then shakes, the shaking elastic rod hits the wave plate.

[0021] In order to facilitate multi-dimensional stirring of the crude enzyme solution in the mixing barrel, preferably, an elastic sheet is fixedly connected to the end of the elastic rod.

[0022] In order to prevent soil particles from entering the sampling groove, a first gap is further provided between the through hole and the second fixing block.

[0023] In order to increase the liquid level depth, further, an embedded cylinder is sealingly and slidably connected in the sampling groove, a magnetic ring is fixedly connected to the top of the embedded cylinder, and a connecting cavity is arranged in the embedded cylinder.

[0024] Compared with the prior art, the present invention provides an analytical method for detecting acid phosphatase activity in soil, which has the following beneficial effects:

[0025] 1. The analytical method for detecting acid phosphatase activity in soil can improve the detection accuracy by spraying the soil with water and fully stirring the sprayed soil to make the moisture content of the soil basically consistent.

[0026] 2. The analytical method for detecting acid phosphatase activity in soil drives multiple second fixed blocks to rotate forwardly through a motor, and the rotating second fixed blocks mix the soil and acetate buffer in the mixing barrel, and when the fourth grinding surface contacts the second grinding surface, the soil between the fourth grinding surface and the second grinding surface is ground to prevent the soil from sticking together and affecting the mixing effect with the acetate buffer, thereby effectively preventing the acetate buffer from being unevenly mixed with the soil and affecting the accuracy of its analysis.

[0027] 3. The analytical method for detecting acid phosphatase activity in soil has been passed. The motor is controlled to reverse, and the rotating shaft drives the second fixed block to reverse at this time. When the sealing plate rotates to contact and seal with the upper end of the first fixed block, the connecting groove connects the two through holes, and the crude enzyme liquid between the first fixed block and the second fixed block is squeezed into the sampling groove under the action of pressure. When the second fixed block contacts the first fixed block, the connecting groove and the through hole are staggered, and the crude enzyme liquid in the sampling groove can be prevented from flowing back. After multiple squeezing, the crude enzyme liquid level in the sampling groove rises, and each time the crude enzyme liquid enters the sampling groove through the through hole, it will impact the crude enzyme liquid in the sampling groove, so that it is fully mixed, thereby effectively preventing the crude enzyme liquid in the sampling groove from precipitating. When the liquid level in the mixing barrel is lower than the first fixed block, part of the gas is squeezed into the sampling groove, thereby further improving the mixing effect of the crude enzyme liquid in the sampling groove. Finally, the motor is turned off, and the rotating shaft stops when the second fixed block is attached to the first fixed block. At this time, the liquid level in the sampling groove rises, which is convenient for sampling with a sampler, and effectively solves the problem of sampling difficulty in large-caliber mixing devices.

[0028] The parts not involved in the device are the same as the existing technology or can be implemented by the existing technology. The present invention can improve its detection accuracy by controlling the moisture of the soil to achieve consistency, effectively improves the mixing effect of acetate buffer and soil through the mixing barrel, and effectively solves the current problem of difficulty in sampling in large-caliber mixing devices. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is a structural schematic diagram of the present invention;

[0030] Figure 2 It is a cross-sectional schematic diagram of the present invention;

[0031] Figure 3 For the present invention Figure 2 The enlarged schematic diagram of point A in the middle;

[0032] Figure 4 It is a schematic diagram of the structure of the rotating shaft and the elastic rod in the present invention;

[0033] Figure 5 It is a schematic diagram of the unfolded structure of the embedded barrel and the rotating shaft in the present invention;

[0034] Figure 6 is a cross-sectional schematic diagram of a fixing ring in the present invention;

[0035] Figure 7 It is a schematic diagram of the structure of the corrugated plate in the present invention;

[0036] Figure 8 It is a schematic diagram of the structure of the first fixing block and the second fixing block in the present invention.

[0037] In the figure: 1, mixing barrel; 101, motor; 102, fixing ring; 103, connecting groove; 2, first fixing block; 201, first grinding surface; 202, second grinding surface; 3, rotating shaft; 301, limiting groove; 302, sliding block; 303, spring; 304, sampling slot; 305, embedded cylinder; 306, magnetic ring; 307, connecting cavity; 308, through hole; 4, second fixing block; 401, third grinding surface; 402, fourth grinding surface; 403, sealing plate; 404, corrugated plate; 405, first pressure surface; 406, guide surface; 407, second pressure surface; 408, connecting plate; 409, connecting ring; 410, first gap; 5, fixing rod; 501, elastic rod; 502, elastic sheet. DETAILED DESCRIPTION

[0038] In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0039] Example 1: An analytical method for detecting acid phosphatase activity in soil, mainly comprising the following steps:

[0040] Step 1, prepare a soil raw material, an acetate buffer solution with a pH value of 5, a disodium nitrobenzene phosphate solution and a NaOH solution, then spray the soil with water, and fully stir the sprayed soil to make its humidity basically consistent, and record the weight of the sprayed water, and measure the actual specific gravity of the soil raw material, wherein the disodium nitrobenzene phosphate solution is used as a substrate, the concentration of the acetate buffer solution is 5.0umol / ml, and the concentration of NaOH is 40mg / ml;

[0041] Step 2: Take 10 grams of soil, dry it, and calculate the moisture content of the soil;

[0042] Step 3: Take 30 grams of soil raw material and add it into a mixing device, add 80 ml of acetate buffer to the soil and stir evenly to form a crude enzyme solution, and keep stirring it continuously so that the soil and acetate buffer are always in a stirring state;

[0043] Step 4: Take 48-well ELISA plates G and H, set 8 groups of samples and 16 groups of controls in ELISA plate G, use a sampler to take samples, and each group of samples uses 100ul crude enzyme solution and 150ul substrate as culture medium; 16 groups of controls include 8 groups of substrate controls and 8 groups of crude enzyme solution controls, the substrate controls use 150ul substrate and 100ul acetate buffer as culture medium, and the crude enzyme solution controls use 100ul crude enzyme solution and 150ul acetate buffer as culture medium;

[0044] Step 5: Place the ELISA plate G filled with the culture solution in step 4 in a 30°C incubator and culture for 60 minutes;

[0045] Step 6. Then take out the ELISA plate G after the culture is completed, and transfer 100ul of the culture solution in the corresponding wells of the ELISA plate H to the corresponding wells of the ELISA plate G, and then add 10ul of NaOH solution to each well to terminate the reaction. Finally, place it in the ELISA reader for colorimetry at a wavelength of 405nm to obtain the sample Abs, substrate control Abs and crude enzyme solution control Abs. The blank Abs of the ELISA plate H is first obtained at a wavelength of 405nm;

[0046] Step 7: Calculate the acid phosphatase activity using the p-nitrophenol standard curve.

[0047] Example 2: Reference Figure 1-Figure 8 , an analytical device used in an analytical method for detecting acid phosphatase activity in soil, comprising a mixing device, an ELISA plate, an ELISA instrument, a sampler and an incubator, wherein the mixing device is used for mixing the soil with the acetate buffer in step three, the stirring device comprises a mixing barrel 1, a fixing ring 102 is fixedly connected in the mixing barrel 1, a plurality of connecting grooves 103 are circumferentially arranged on the fixing ring 102, a rotating shaft 3 is rotatably arranged in the mixing barrel 1, and the fixing ring 102 is sealingly rotatably connected to the lower end of the rotating shaft 3, a sampling groove 304 is arranged in the rotating shaft 3, a plurality of through holes 308 are circumferentially arranged on the rotating shaft 3, a plurality of first fixing blocks 2 are circumferentially fixedly connected in the mixing barrel 1, and a plurality of connecting grooves 103 are circumferentially arranged on the first fixing block 2 A first grinding surface 201 and a second grinding surface 202 are provided, and the first fixed block 2 matches the connecting groove 103. A plurality of second fixed blocks 4 are fixedly connected to the rotating shaft 3 in a circumferential manner. A third grinding surface 401 and a fourth grinding surface 402 are provided on the second fixed block 4. The third grinding surface 401 matches the first grinding surface 201, and the fourth grinding surface 402 matches the second grinding surface 202. A sealing plate 403 matching the first fixed block 2 is fixedly connected to the upper side of the second fixed block 4. A motor 101 is fixedly connected to the lower side of the mixing barrel 1. The output end of the motor 101 is fixedly connected to the rotating shaft 3. A plurality of buffer components matching the second fixed block 4 are provided on the rotating shaft 3.

[0048] The buffer assembly includes a plurality of limiting grooves 301 circumferentially arranged on the rotating shaft 3 , a slider 302 is slidably arranged in the limiting groove 301 , a spring 303 is symmetrically fixedly connected between the slider 302 and the limiting groove 301 , and a connecting plate 408 is fixedly connected between the slider 302 and the sealing plate 403 .

[0049] When the soil and the acetate buffer are mixed, the soil and the acetate buffer are added into the mixing barrel 1, and the motor 101 drives the plurality of second fixed blocks 4 to rotate forwardly. The rotating second fixed blocks 4 mix the soil and the acetate buffer in the mixing barrel 1, and when the fourth grinding surface 402 contacts the second grinding surface 202, the soil between the fourth grinding surface 402 and the second grinding surface 202 is ground to prevent the soil from sticking together and affecting the mixing effect with the acetate buffer, thereby effectively preventing the acetate buffer from being unevenly mixed with the soil and affecting the accuracy of its analysis.

[0050] When it is necessary to sample the crude enzyme solution mixed in the mixing barrel 1, the control motor 101 is reversed, and the rotating shaft 3 drives the second fixed block 4 to reverse. When the sealing plate 403 is rotated to contact and seal with the upper end of the first fixed block 2, the connecting groove 103 connects the two through holes 308, and the crude enzyme solution located between the first fixed block 2 and the second fixed block 4 is squeezed into the sampling groove 304 under the action of pressure. When the second fixed block 4 contacts the first fixed block 2, the connecting groove 103 is staggered with the through hole 308, which can prevent the crude enzyme solution in the sampling groove 304 from flowing back. After multiple squeezing, the crude enzyme solution level in the sampling groove 304 rises. , and each time the crude enzyme liquid enters the sampling tank 304 through the through hole 308, it will impact the crude enzyme liquid in the sampling tank 304 to make it fully mixed, thereby effectively preventing the crude enzyme liquid in the sampling tank 304 from precipitating. When the liquid level in the mixing barrel 1 is lower than the first fixed block 2, part of the gas is squeezed into the sampling tank 304, thereby further improving the mixing effect of the crude enzyme liquid in the sampling tank 304. Finally, the motor 101 is turned off, and the rotating shaft 3 stops when the second fixed block 4 is in contact with the first fixed block 2. At this time, the liquid level in the sampling tank 304 rises, which is convenient for sampling with a sampler, effectively solving the problem of sampling difficulties in large-caliber mixing devices.

[0051] When the first grinding surface 201 contacts the third grinding surface 401 or the second grinding surface 202 contacts the fourth grinding surface 402, the squeezed second fixed block 4 moves upward, causing the slider 302 to slide upward in the limiting groove 301. When the second fixed block 4 moves away from the first fixed block 2, the slider 302 is reset under the action of the spring 303.

[0052] After sampling for a certain period of time, the motor 101 is controlled again to drive the rotating shaft 3 to rotate forward. At this time, when the third grinding surface 401 gradually moves away from the first grinding surface 201, negative pressure is generated between the first fixed block 2 and the second fixed block 4. Then, the connecting groove 103 connects the through hole 308, and the crude enzyme liquid in the sampling groove 304 is quickly back-drawn. The back-drawing speed is fast, and the impurities remaining at the bottom of the sampling groove 304 can be extracted, effectively preventing excessive soil impurities from remaining at the bottom of the sampling groove 304. At the same time, the crude enzyme liquid quickly sprayed out from the through hole 308 has a certain impact force, which effectively improves its mixing effect.

[0053] Example 3: Reference Figure 1-Figure 8 , an analytical device used in an analytical method for detecting acid phosphatase activity in soil, is basically the same as Example 2, further, a sealing plate 403 is fixedly connected to a wave plate 404 on the side away from the second fixed block 4, and the wave plate 404 is provided with a first pressure surface 405, a guide surface 406 and a second pressure surface 407, and the guide surface 406 matches the first fixed block 2.

[0054] An end of the first pressure-bearing surface 405 close to the rotating shaft 3 is inclined toward an end close to the mixing barrel 1 .

[0055] When sampling is required, the counter-rotating shaft 3 drives the second fixed block 4 to rotate. At this time, the guide surface 406 of the wave plate 404 first contacts the first fixed block 2 to guide the wave plate 404. When the first fixed block 2 contacts the sealing plate 403, the first pressure surface 405 has a certain downward pressure under the impact of the crude enzyme solution and the gas, thereby effectively preventing the pressure between the first fixed block 2 and the second fixed block 4 from being too high, causing the crude enzyme solution to leak out from between the sealing plate 403 and the first fixed block 2.

[0056] Example 4: Reference Figure 1-Figure 8 , an analytical device used in an analytical method for detecting acid phosphatase activity in soil, is basically the same as Example 2, and further, multiple connecting plates 408 are fixedly connected by connecting rings 409.

[0057] The multiple connecting plates 408 are connected by the connecting ring 409, so that the multiple second fixing blocks 4 can be guaranteed to move synchronously and provide more uniform downward pressure.

[0058] A plurality of fixed rods 5 are fixedly connected to the rotating shaft 3 in a circular shape, and an elastic rod 501 is fixedly connected to the lower end of the fixed rod 5, and the elastic rod 501 is arranged between the wave plate 404 and the second fixed block 4. When the elastic rod 501 is squeezed and deformed by the first fixed block 2 and shakes, the shaking elastic rod 501 hits the wave plate 404.

[0059] An elastic sheet 502 is fixedly connected to the end of the elastic rod 501 .

[0060] When the crude enzyme liquid in the mixing barrel 1 is mixed, the elastic rod 501 drives the elastic sheet 502 to stir the crude enzyme liquid in the mixing barrel 1, thereby effectively improving the mixing effect of the crude enzyme liquid. When the elastic rod 501 contacts the first fixed block 2, it is squeezed and tilted to one side, and when the elastic rod 501 moves away from the first fixed block 2, the elastic rod 501 shakes under the action of elastic force. At this time, the shaking elastic rod 501 drives the elastic sheet 502 to stir the crude enzyme liquid in the mixing barrel 1 in multiple dimensions, thereby effectively improving the mixing effect. The shaking elastic rod 501 hits the wave plate 404, effectively preventing residual soil on the wave plate 404, and the second fixed block 4 rotating forwardly stirs in the crude enzyme liquid. When the liquid level is greater than the sealing plate 403, part of the crude enzyme liquid splashes upward along the second pressure surface 407, thereby further improving its mixing effect.

[0061] A first gap 410 is defined between the through hole 308 and the second fixing block 4 .

[0062] An embedded cylinder 305 is sealingly and slidably connected in the sampling groove 304 , a magnetic ring 306 is fixedly connected to the top of the embedded cylinder 305 , and a connecting cavity 307 is provided in the embedded cylinder 305 .

[0063] By setting the first gap 410 between the through hole 308 and the second fixed block 4, a small gap is formed between the first grinding surface 201 and the third grinding surface 401 after the through hole 308 is completely fitted with one side of the first fixed block 2, thereby effectively preventing soil particles from being squeezed into the sampling groove 304. At the same time, the soil can be ground, thereby effectively improving the mixing effect of the crude enzyme solution.

[0064] An embedded cylinder 305 is arranged in the sampling groove 304, and a connecting cavity 307 with a smaller diameter is provided in the embedded cylinder 305, so as to provide a better liquid surface for less raw materials and facilitate sampling thereof.

[0065] It will be apparent to those skilled in the art that the invention is not limited to the details of the exemplary embodiments described above and that the invention can be implemented in other specific forms without departing from the spirit or essential features of the invention. Therefore, the embodiments should be considered exemplary and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations falling within the meaning and scope of the equivalent elements of the claims be included in the invention. Any reference numeral in a claim should not be considered as limiting the claim to which it relates.

[0066] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. An analytical method for detecting acid phosphatase activity in soil, characterized in that: The main steps include: Step 1, prepare a soil raw material, an acetate buffer solution with a pH value of 5, a disodium nitrobenzene phosphate solution and a NaOH solution, then spray the soil with water, and fully stir the sprayed soil to make its humidity basically consistent, and record the weight of the sprayed water to measure the actual specific gravity of the soil raw material, wherein the disodium nitrobenzene phosphate solution is used as a substrate; Step 2: Take 10 grams of soil, dry it, and calculate the moisture content of the soil; Step 3: Take 30 grams of soil raw material and add it into a mixing device, add 80 ml of acetate buffer to the soil and stir evenly to form a crude enzyme solution, and keep stirring it continuously so that the soil and acetate buffer are always in a stirring state; Step 4, take ELISA plate G, set 8 groups of samples and 16 groups of controls in ELISA plate G, use a sampler to take samples, each group of samples uses 100ul crude enzyme solution and 150ul substrate as culture medium; 16 groups of controls include 8 groups of substrate controls and 8 groups of crude enzyme solution controls, the substrate controls use 150ul substrate and 100ul acetate buffer as culture medium, and the crude enzyme solution controls use 100ul crude enzyme solution and 150ul acetate buffer as culture medium; Step 5: Place the ELISA plate G filled with the culture solution in step 4 in a 30°C incubator and culture for 60 minutes; Step 6: Take out the ELISA plate G after the culture is completed, transfer 100ul of the culture solution in the corresponding well of the ELISA plate H to the corresponding well of the ELISA plate G, and then add 10ul of NaOH solution to each well to terminate the reaction. Finally, place the plate in an ELISA reader for colorimetry at a wavelength of 405nm to obtain the sample Abs, substrate control Abs and crude enzyme solution control Abs; Step 7: Calculate the acid phosphatase activity using the p-nitrophenol standard curve.

2. An analytical device used in the analytical method for detecting acid phosphatase activity in soil as claimed in claim 1, characterized in that: The invention comprises a mixing device, an ELISA plate, an ELISA instrument, a sampler and an incubator, wherein the mixing device is used for mixing the soil with the acetate buffer in step three, the stirring device comprises a mixing barrel, a fixing ring is fixedly connected in the mixing barrel, a plurality of connecting grooves are circumferentially arranged on the fixing ring, a rotating shaft is rotatably arranged in the mixing barrel, and the fixing ring is sealingly rotatably connected to the lower end of the rotating shaft, a sampling groove is arranged in the rotating shaft, a plurality of through holes are circumferentially arranged on the rotating shaft, a plurality of first fixing blocks are fixedly connected in the mixing barrel, a first fixing block is provided with a first connecting groove, and a plurality of connecting grooves are circumferentially arranged on the first connecting block. A grinding surface and a second grinding surface, and the first fixed block matches the connecting groove, a plurality of second fixed blocks are fixedly connected to the rotating shaft in a circumferential manner, a third grinding surface and a fourth grinding surface are arranged on the second fixed block, the third grinding surface matches the first grinding surface, and the fourth grinding surface matches the second grinding surface, a sealing plate matching the first fixed block is fixedly connected to the upper side of the second fixed block, a motor is fixedly connected to the lower side of the mixing barrel, the output end of the motor is fixedly connected to the rotating shaft, and a plurality of buffer components matching the second fixed blocks are arranged on the rotating shaft.

3. The analytical method for detecting acid phosphatase activity in soil according to claim 2, characterized in that: The buffer assembly includes a plurality of limiting grooves circumferentially arranged on the rotating shaft, a slider is slidably arranged in the limiting groove, a spring is symmetrically fixedly connected between the slider and the limiting groove, and a connecting plate is fixedly connected between the slider and the sealing plate.

4. The analytical method for detecting acid phosphatase activity in soil according to claim 2, characterized in that: A wave plate is fixedly connected to one side of the sealing plate away from the second fixed block. The wave plate is provided with a first pressure-bearing surface, a guide surface and a second pressure-bearing surface. The guide surface matches the first fixed block.

5. The analytical method for detecting acid phosphatase activity in soil according to claim 4, characterized in that: The end of the first pressure-bearing surface close to the rotating shaft is inclined toward the end close to the mixing barrel.

6. The analytical method for detecting acid phosphatase activity in soil according to claim 3, characterized in that: The plurality of connecting plates are fixedly connected via connecting rings.

7. The analytical method for detecting acid phosphatase activity in soil according to claim 4, characterized in that: A plurality of fixing rods are fixedly connected to the rotating shaft in a circle, and an elastic rod is fixedly connected to the lower end of the fixing rod, and the elastic rod is arranged between the wave plate and the second fixing block. When the elastic rod is squeezed and deformed by the first fixing block and shakes, the shaking elastic rod hits the wave plate.

8. The analytical method for detecting acid phosphatase activity in soil according to claim 7, characterized in that: The end of the elastic rod is fixedly connected with an elastic sheet.

9. The analytical method for detecting acid phosphatase activity in soil according to claim 2, characterized in that: A first gap is provided between the through hole and the second fixing block.

10. The analytical method for detecting acid phosphatase activity in soil according to claim 2, characterized in that: An embedded cylinder is sealingly and slidably connected in the sampling groove, a magnetic ring is fixedly connected to the top of the embedded cylinder, and a connecting cavity is arranged in the embedded cylinder.

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