CN114592498A - Method for inducing calcium carbonate precipitation by combining plant urease with urease inhibitor - Google Patents
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Abstract
The invention discloses a method for inducing calcium carbonate precipitation by using urease inhibitor and plant urease, belonging to the field of geotechnical engineering and rock mass reinforcement and adopting the following steps: 1. urease is extracted from soybean. 2. And adding a urease inhibitor into the mixed solution of the calcium chloride and the urea to prepare the cementing solution. 3. And sequentially pouring the urease solution and the cementing solution into the rock-soil body to be reinforced, and standing and maintaining to finish the rock-soil body reinforcement. According to the method, a long bacterial culture period is not needed, and a small amount of urease inhibitor is added into the cementing liquid, so that the reaction rate is slowed down, the blockage near a grouting opening is prevented, smooth grouting can be ensured, the cementing liquid is fully infiltrated before the pores are filled with calcium carbonate crystals, the rock mass reinforcement uniformity is improved, and the method is suitable for reinforcing large-volume complex fractured rock masses; has good mechanical property and can achieve the expected reinforcing effect.
Description
Technical Field
The invention belongs to the field of geotechnical engineering and rock mass reinforcement, and particularly relates to a method for inducing calcium carbonate precipitation by combining plant urease with a urease inhibitor.
Background
MICP (microbial induced carbonate precipitation) is a new technology, and is mainly used for reinforcing sandy soil and rock masses so as to achieve the purpose of improving the mechanical properties of the rock-mass. Compared with the traditional cement paste reinforcement, the MICP technology has the characteristics of environmental protection and environmental friendliness, and becomes a popular research field. However, the MICP technology also has considerable drawbacks: the development of the field is severely restricted by complicated and fussy bacterial culture process, overlong culture period, large activity instability of bacteria under the influence of environment, difficulty in mass culture and the like. In addition, the MICP technology faces another difficult problem, namely, the enzymatic reaction speed is difficult to control, the porosity of rock-soil mass is often reduced faster, the blockage of the pores near grouting holes is caused, and the slurry permeability is reduced continuously.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a method for inducing calcium carbonate precipitation by combining plant urease with a urease inhibitor, so as to solve the problems that the MICP technology in the prior art is complex and tedious in bacterial culture process, too long in culture period, large in environmental influence, unstable in activity and difficult to culture in large quantities; and the enzymatic reaction speed is difficult to control, so that the porosity of the rock-soil body is reduced quickly, and the problems of blockage of pores near grouting holes and continuous reduction of the slurry permeability are caused.
In order to achieve the purpose, the invention adopts the following technical scheme to realize the purpose:
the invention discloses a method for inducing calcium carbonate precipitation by combining plant urease with a urease inhibitor, which comprises the following steps:
the method comprises the following steps: drying soybean to obtain soybean powder, mixing with deionized water to obtain soybean powder solution, centrifuging the soybean powder solution, and removing supernatant to obtain urease solution;
step two: mixing calcium chloride and urea to prepare basic cementing liquid;
step three: adding a urease inhibitor into the basic cementing liquid prepared in the step two to prepare cementing liquid;
step four: and (4) injecting the urease solution prepared in the step one and the cementing solution prepared in the step three into the rock mass to be reinforced.
Preferably, in the first step, the soybeans are dried in an oven at 40 ℃ for 6 hours, and the soybean flour is sieved out by a 100-mesh steel sieve after being crushed.
Preferably, in the step one, the mass concentration of the bean flour solution is 20 g/L-100 g/L.
Preferably, in the step one, the soybean powder solution is uniformly stirred, the warp cloth is filtered to a beaker, the beaker is kept still for 2 to 3 hours, then supernatant liquid of the beaker is taken and placed in a centrifuge tube to be centrifuged for 15min at 3000r/min and 4 ℃, and the supernatant liquid in the centrifuge tube is taken and stored in an environment at 4 ℃.
Preferably, in step two, the calcium chloride and urea are mixed with analytically pure reagents in a ratio of 1: 1, and the mass concentration of the solution is 0.25 mol/L-1.5 mol/L.
Preferably, in the third step, the content of the urease inhibitor is 0.1% -1% of the cementing liquid.
Preferably, in step three, the urease inhibitor adopts n-butyl thiophosphoryl triamide, namely NBPT.
Preferably, in the fourth step, the volume ratio of the plant urease solution to the cementing solution is 1: 1.
compared with the prior art, the invention has the following beneficial effects:
according to the method for inducing calcium carbonate precipitation by combining plant urease with urease inhibitor, the urease is extracted from plants, and a large amount of industrially produced finished urease can be directly purchased, so that a long bacterial culture period is bypassed, the requirements of engineering practice on construction period and materials are met, and the method is suitable for reinforcing large-volume rock masses; a small amount of urease inhibitor is added into the cementing liquid, so that the reaction rate is slowed down on the premise of not influencing the occurrence of enzymatic reaction, the blockage near a grouting opening is prevented, the smooth grouting can be ensured, the cementing liquid is fully infiltrated before the pores are filled with calcium carbonate crystals, the rock mass reinforcing uniformity is improved, and the method is suitable for reinforcing large-volume complex fractured rock masses; the method for reinforcing the rock mass by combining the plant urease with the urease inhibitor can generate calcite-shaped calcium carbonate crystals in the cracks of the rock mass, has good mechanical properties and can achieve the expected reinforcing effect.
Furthermore, in the third step, the content of the urease inhibitor is 0.1% -1% of the cementing liquid. Urease inhibitors can significantly reduce urease activity resulting in a reduction in the rate of calcium carbonate production. However, in rock mass grouting experiments, calcium carbonate can be rapidly precipitated due to too high urease activity, so that a grouting hole is blocked, and the generation speed of the calcium carbonate can be slowed down by adding a proper amount of urease inhibitor. According to experimental research, in a rock mass grouting test, the generation speed of calcium carbonate is reduced by a urease inhibitor, and the generation rate of calcium carbonate in a rock mass can be increased. With the increase of the urease inhibitor from 0 to 1 percent, the calcium carbonate generation rate in the grouting rock body shows the phenomenon that the calcium carbonate generation rate is increased and then decreased.
Furthermore, the urease inhibitor n-butyl thiophosphoric triamide, namely NBPT, adopted by the invention is a nontoxic and harmless additive widely applied to agriculture and forestry, and is energy-saving, emission-reducing and environment-friendly.
Drawings
FIG. 1 is a flow chart of the operation of the present invention.
Detailed Description
In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It should be noted that the terms "first," "second," and the like in the description and claims of the present invention and in the drawings described above are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used is interchangeable under appropriate circumstances such that the embodiments of the invention described herein are capable of operation in sequences other than those illustrated or described herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
The invention is described in further detail below with reference to the accompanying drawings:
referring to fig. 1, which is an operational flow diagram of the present invention, a method for inducing calcium carbonate precipitation by plant urease in combination with a urease inhibitor comprises:
the method comprises the following steps: and (4) extracting the plant urease. Drying and crushing soybeans into soybean powder, mixing deionized water to prepare a soybean powder solution, centrifuging the soybean powder solution, and removing supernatant to obtain the urease solution.
Step two: mixing calcium chloride and urea to prepare basic cementing liquid.
Step three: and (4) adding the urease inhibitor into the basic cementing liquid in the step two to prepare the cementing liquid.
Step four: and (3) injecting the urease solution in the step one and the cementing solution in the step three into the rock mass to be reinforced.
Further, in the first step, the soybeans are dried in an oven for 6 hours at 40 ℃, and the soybean powder is sieved out by a 100-mesh steel sieve after being crushed.
Further, in the step one, the mass concentration of the bean flour solution is about 20 g/L-100 g/L.
Further, in the first step, the soybean powder solution is uniformly stirred, the warp cloth is filtered to a beaker, the beaker is kept still for 2 to 3 hours, then supernatant liquid of the beaker is taken and placed in a centrifuge tube to be centrifuged for 15 minutes at 3000r/min and 4 ℃, and the supernatant liquid in the centrifuge tube is taken and stored in an environment at 4 ℃.
Further, in step two, calcium chloride and urea are mixed with analytical reagent in a ratio of 1: 1, and the mass concentration of the solution is 0.25 mol/L-1.5 mol/L.
Furthermore, in the third step, the content of the urease inhibitor is 0.1% -1% of the cementing liquid.
Further, in the third step, n-butyl thiophosphoryl triamide, namely NBPT is used as the urease inhibitor.
Further, in the third step, the volume ratio of the plant urease solution to the cementing solution is 1: 1.
further, in the fourth step, the rock mass after grouting is maintained under normal temperature environment until the expected strength is achieved.
Existing studies have shown that sources of urease can be extracted from plant species, especially legumes, in addition to being obtained by culturing urease-producing bacteria. Plants such as soybean contain abundant urease, and can be used as a large amount of stable urease sources. In addition, how to control the enzymatic reaction leaves the engineer with sufficient working time. The prior research shows that the urease inhibitor can reduce the activity of urease and lead the catalytic reaction of the urease to be slowly carried out. The invention provides a method for inducing calcium carbonate precipitation by combining plant urease with a urease inhibitor. The mixed solution is poured into the rock mass crack at a stable speed, urease catalyzes the generation of carbonate ions at a low catalytic speed in the rock mass crack, and calcium carbonate crystals are generated by combining with calcium ions, so that the aim of reinforcing the rock mass is fulfilled. The method disclosed by the invention is low in cost, green and environment-friendly, and simple in process, and the technical problem of MICP is basically solved.
The invention relates to a method for inducing calcium carbonate precipitation by combining plant urease with a urease inhibitor, which is described in the following more specific embodiment:
example 1
1. And (4) extracting the plant urease. Drying the soybeans in an oven at 40 ℃ for 6 hours, crushing, screening the soybean powder by a 100-mesh steel screen, and mixing the soybean powder with deionized water to prepare a soybean powder solution with the mass concentration of 50 g/L. Stirring the bean flour solution uniformly, filtering the solution to a beaker by using a warp cloth, standing for 2 to 3 hours, taking supernatant of the beaker, placing the supernatant in a centrifuge tube, centrifuging for 15min at the temperature of 4 ℃ at 3000r/min, and taking the supernatant in the centrifuge tube to store in the environment of 4 ℃.
2. And (5) preparing cementing liquid. Calcium chloride and urea were mixed with analytical reagents in a ratio of 1: 1, the mass concentration of the solution is 0.25mol/L, and then, the urease inhibitor n-butyl thiophosphoric triamide, namely NBPT, is added, and the solution accounts for 1 percent of the total volume of the solution.
3. Grouting and reinforcing. And (3) sequentially filling the urease solution and the cementing solution into the pores of the rock body by using a peristaltic pump, and maintaining at normal environmental temperature until the expected strength is reached.
Example 2
1. And (4) extracting the plant urease. Drying the soybeans in an oven at 40 ℃ for 6 hours, crushing, screening the soybean powder by a 100-mesh steel screen, and mixing the soybean powder with deionized water to prepare a soybean powder solution with the mass concentration of 60 g/L. Stirring the bean flour solution uniformly, filtering the solution to a beaker by using a warp cloth, standing for 2 to 3 hours, taking supernatant of the beaker, placing the supernatant in a centrifuge tube, centrifuging for 15min at the temperature of 4 ℃ at 3000r/min, and taking the supernatant in the centrifuge tube to store in the environment of 4 ℃.
2. And (5) preparing cementing liquid. Calcium chloride and urea were mixed with analytical reagents in a ratio of 1: 1, the mass concentration of the solution is 0.25mol/L, and then, the urease inhibitor n-butyl thiophosphoric triamide, namely NBPT, is added, and the solution accounts for 1 percent of the total volume of the solution.
3. Grouting and reinforcing. And (3) sequentially pouring the urease solution and the cementing solution into the sandy soil by using a peristaltic pump, and maintaining at normal environmental temperature until the expected strength is reached.
Example 3
1. And (4) extracting the plant urease. Drying the soybeans in an oven at 40 ℃ for 6 hours, crushing, screening the soybean powder by a 100-mesh steel screen, and mixing the soybean powder with deionized water to prepare a soybean powder solution with the mass concentration of 50 g/L. Stirring the bean flour solution uniformly, filtering the solution to a beaker by using a warp cloth, standing for 2 to 3 hours, taking supernatant of the beaker, placing the supernatant in a centrifuge tube, centrifuging for 15min at the temperature of 4 ℃ at 3000r/min, and taking the supernatant in the centrifuge tube to store in the environment of 4 ℃.
2. And (5) preparing cementing liquid. Calcium chloride and urea were mixed with analytical reagents in a ratio of 1: 1, the mass concentration of the solution is 0.7mol/L, and then, urease inhibitor n-butyl thiophosphoric triamide, namely NBPT, is added, and the mass concentration accounts for 0.1 percent of the total volume of the solution.
3. Grouting and reinforcing. The urease solution and the cementing solution are sequentially filled into the pores of the rock mass by using a peristaltic pump, and the rock mass is maintained at normal environmental temperature until the expected strength is reached
Example 4
1. And (4) extracting the plant urease. Drying the soybeans in an oven at 40 ℃ for 6 hours, crushing, screening the soybean powder by a 100-mesh steel screen, and mixing the soybean powder with deionized water to prepare a soybean powder solution with the mass concentration of 20 g/L. Stirring the bean flour solution uniformly, filtering the solution to a beaker by using a warp cloth, standing for 2 to 3 hours, taking supernatant of the beaker, placing the supernatant in a centrifuge tube, centrifuging for 15min at the temperature of 4 ℃ at 3000r/min, and taking the supernatant in the centrifuge tube to store in the environment of 4 ℃.
2. And (5) preparing cementing liquid. Calcium chloride and urea were mixed with analytical reagents in a ratio of 1: 1, the mass concentration of the solution is 0.25mol/L, and then, the urease inhibitor n-butyl thiophosphoric triamide, namely NBPT, is added, and the solution accounts for 1 percent of the total volume of the solution.
3. Grouting and reinforcing. The urease solution and the cementing solution are sequentially filled into the pores of the rock mass by using a peristaltic pump, and the rock mass is maintained at normal environmental temperature until the expected strength is reached
Example 5
1. And (4) extracting the plant urease. Drying the soybeans in an oven at 40 ℃ for 6 hours, crushing, screening the soybean powder by a 100-mesh steel screen, and mixing the soybean powder with deionized water to prepare a soybean powder solution with the mass concentration of 100 g/L. Stirring the bean flour solution uniformly, filtering the solution to a beaker by using a warp cloth, standing for 2 to 3 hours, taking supernatant of the beaker, placing the supernatant in a centrifuge tube, centrifuging for 15min at the temperature of 4 ℃ at 3000r/min, and taking the supernatant in the centrifuge tube to store in the environment of 4 ℃.
2. And (5) preparing cementing liquid. Calcium chloride and urea were mixed with analytical reagents in a ratio of 1: 1, the mass concentration of the solution is 0.25mol/L, and then, the urease inhibitor n-butyl thiophosphoric triamide, namely NBPT, is added, and the total volume of the solution is 0.5 percent.
3. Grouting and reinforcing. And (3) sequentially pouring the urease solution and the cementing solution into the sandy soil by using a peristaltic pump, and maintaining at normal environmental temperature until the expected strength is reached.
Example 6
1. And (4) extracting the plant urease. Drying the soybeans in an oven at 40 ℃ for 6 hours, crushing, screening the soybean powder by a 100-mesh steel screen, and mixing the soybean powder with deionized water to prepare a soybean powder solution with the mass concentration of 50 g/L. Stirring the bean flour solution uniformly, filtering the solution to a beaker by using a warp cloth, standing for 2 to 3 hours, taking supernatant of the beaker, placing the supernatant in a centrifuge tube, centrifuging for 15min at the temperature of 4 ℃ at 3000r/min, and taking the supernatant in the centrifuge tube to store in the environment of 4 ℃.
2. And (5) preparing cementing liquid. Calcium chloride and urea were purified using analytical reagents in a 1: 1, the mass concentration of the solution is 1.5mol/L, and then, the urease inhibitor n-butyl thiophosphoric triamide, namely NBPT, is added, and the solution accounts for 1 percent of the total volume of the solution.
3. Grouting and reinforcing. And (3) sequentially pouring the urease solution and the cementing solution into the sandy soil by using a peristaltic pump, and maintaining at normal environmental temperature until the expected strength is reached.
Tests prove that the urease activity is higher when the pH value of the solution is more alkaline, so that the urease activity is better when the urea content in the mixed solution is slightly higher. Therefore, the proportion of urea and calcium chloride in the mixed cementing liquid is 1: 1, because the mixed solution after the reaction is completed has no residual urea; the reason for selecting the concentration of the urea-calcium chloride mixed solution to be 0.25mol/L to 1.5mol/L is that: the urease activity is different due to the difference of urease extraction processes and the difference of soybean raw materials, even if the urea amount which can be catalyzed by the urease solution with the same concentration is not consistent, the concentration of the mixed solution should be selected correspondingly according to the urease activity. The test result proves that the urease activity extracted from fresh soybeans in a laboratory can reach 2000U/L. And when the activity of the urease solution is more than 1000U/L, the calcium carbonate precipitation rate of 0.25mol/L urea calcium chloride solution is 100% (the calcium carbonate precipitation rate is the ratio of the actual calcium carbonate precipitation amount to the theoretical calcium carbonate precipitation amount), the urea calcium chloride concentration is increased, the calcium carbonate precipitation amount is increased, and the calcium carbonate precipitation rate is reduced. The concentration of urease inhibitor may be limited within a certain range to limit urease activity, i.e., the higher the concentration of urease inhibitor, the lower the urease activity and the lower the rate of calcium carbonate precipitation reaction. The concentration of the urease inhibitor is selected according to the urease activity, the urease concentration and the actual engineering requirements. The parameters mentioned in the urease extraction step of step one of the present invention need to be further explained. Drying for 6 hours in an environment of 40 ℃, screening bean flour by a 100-mesh steel screen after crushing, standing for 2 to 3 hours when the mass concentration of a bean flour solution is 20g/L to 100g/L, centrifuging for 15min at the temperature of 4 ℃ at 3000r/min, and storing supernatant in an environment of 4 ℃. The selection of parameters is common practice in laboratories, the soybean drying step needs to remove most of moisture in fresh soybeans, and cannot generate adverse effect on urease activity, and the temperature and the drying time which can meet the conditions can be selected. The bean flour needs to be screened out to prepare a fine powder solution, and in a certain concentration range, the higher the concentration of the urease is, and the preparation of the bean flour solution is selected according to the requirements. The extracted urease solution can maintain activity to the maximum extent under refrigeration.
The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical idea of the present invention falls within the protection scope of the claims of the present invention.
Claims (8)
1. A method for inducing calcium carbonate precipitation by combining plant urease with a urease inhibitor is characterized by comprising the following steps:
the method comprises the following steps: drying soybean to obtain soybean powder, mixing with deionized water to obtain soybean powder solution, centrifuging the soybean powder solution, and removing supernatant to obtain urease solution;
step two: mixing calcium chloride and urea to prepare basic cementing liquid;
step three: adding a urease inhibitor into the basic cementing liquid prepared in the step two to prepare cementing liquid;
step four: and (4) injecting the urease solution prepared in the step one and the cementing solution prepared in the step three into the rock mass to be reinforced.
2. The method of claim 1, wherein in step one, the soybeans are dried in an oven at 40 ℃ for 6 hours, and the soybean meal is sieved out by a 100-mesh steel sieve after being crushed.
3. The method for inducing calcium carbonate precipitation by combining plant urease with a urease inhibitor as claimed in claim 1, wherein in the first step, the mass concentration of the soybean flour solution is 20 g/L-100 g/L.
4. The method for inducing the precipitation of calcium carbonate by using the combination of plant urease and the urease inhibitor as claimed in claim 1, wherein in the first step, the soybean powder solution is stirred uniformly, the warp yarn cloth is filtered to a beaker, the beaker is kept still for 2 to 3 hours, then supernatant liquid of the beaker is taken and placed in a centrifuge tube to be centrifuged for 15 minutes at 3000r/min and 4 ℃, and the supernatant liquid in the centrifuge tube is taken and stored in an environment at 4 ℃.
5. The method for inducing calcium carbonate precipitation by combining plant urease with urease inhibitor according to claim 1, wherein in step two, the calcium chloride and urea are mixed by using analytically pure reagents in a ratio of 1: 1, and the mass concentration of the solution is 0.25 mol/L-1.5 mol/L.
6. The method for inducing calcium carbonate precipitation by combining plant urease with a urease inhibitor according to claim 1, wherein in the third step, the content of the urease inhibitor is 0.1% -1% of the cementing solution.
7. The method for inducing calcium carbonate precipitation by combining plant urease with urease inhibitor according to claim 1, wherein the urease inhibitor is n-butyl thiophosphoric triamide (NBPT) in step three.
8. The method for inducing calcium carbonate precipitation by combining plant urease with a urease inhibitor according to claim 1, wherein in the fourth step, the volume ratio of the plant urease solution to the cementing solution is 1: 1.
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