CN109423294A - Soil remediation composition and application and geobiont repair method - Google Patents

Soil remediation composition and application and geobiont repair method Download PDF

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Publication number
CN109423294A
CN109423294A CN201710758724.6A CN201710758724A CN109423294A CN 109423294 A CN109423294 A CN 109423294A CN 201710758724 A CN201710758724 A CN 201710758724A CN 109423294 A CN109423294 A CN 109423294A
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China
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soil
weight
content
remediation composition
parts
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CN201710758724.6A
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Chinese (zh)
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王亭
郦和生
胡艳华
王岽
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中国石油化工股份有限公司
中国石油化工股份有限公司北京化工研究院
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Priority to CN201710758724.6A priority Critical patent/CN109423294A/en
Publication of CN109423294A publication Critical patent/CN109423294A/en

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    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K17/00Soil-conditioning materials or soil-stabilising materials
    • C09K17/40Soil-conditioning materials or soil-stabilising materials containing mixtures of inorganic and organic compounds
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B09DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09CRECLAMATION OF CONTAMINATED SOIL
    • B09C1/00Reclamation of contaminated soil
    • B09C1/08Reclamation of contaminated soil chemically
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B09DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09CRECLAMATION OF CONTAMINATED SOIL
    • B09C1/00Reclamation of contaminated soil
    • B09C1/10Reclamation of contaminated soil microbiologically, biologically or by using enzymes
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K17/00Soil-conditioning materials or soil-stabilising materials
    • C09K17/14Soil-conditioning materials or soil-stabilising materials containing organic compounds only
    • C09K17/18Prepolymers; Macromolecular compounds
    • C09K17/20Vinyl polymers
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K17/00Soil-conditioning materials or soil-stabilising materials
    • C09K17/14Soil-conditioning materials or soil-stabilising materials containing organic compounds only
    • C09K17/18Prepolymers; Macromolecular compounds
    • C09K17/20Vinyl polymers
    • C09K17/22Polyacrylates; Polymethacrylates
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K17/00Soil-conditioning materials or soil-stabilising materials
    • C09K17/14Soil-conditioning materials or soil-stabilising materials containing organic compounds only
    • C09K17/18Prepolymers; Macromolecular compounds
    • C09K17/32Prepolymers; Macromolecular compounds of natural origin, e.g. cellulosic materials
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20Bacteria; Culture media therefor

Abstract

The present invention relates to environment protection fields, and in particular to soil remediation composition and application and geobiont repair method.The soil remediation composition contains water-retaining agent, soil property regulator, nutriment, beta-cyclodextrin and single bacterium agent.The application is application of the soil remediation composition in rehabilitating soil.The method includes using soil remediation composition as described above to repair soil.By the microbial bacterial agent for introducing degradable petroleum hydrocarbon into soil, and introduce has the component (such as soil property regulator, nutriment, beta-cyclodextrin and water-retaining agent) of improving effect to promote microbial bacterial agent to breed in the soil soil soil property simultaneously, the pollutant in soil is promoted to be degraded, to obtain preferable soil remediation effect.Soil remediation composition of the invention not will cause soil secondary pollution, will not destroy soil environment, and at low cost, easy to operate.

Description

Soil remediation composition and application and geobiont repair method

Technical field

The present invention relates to environment protection fields, and in particular, to soil remediation composition and application and geobiont repair side Method.

Background technique

Durings oil exploration, exploitation, transport, refinement etc., due to misoperation or accident leakage, part petroleum is spilt It falls in soil, causes the petroleum hydrocarbon contaminated of soil.The whole world passes through the approach such as ground crude oil and accident leakage every year and is drained into Petroleum pollution in environment is about 8 × 106T, it is petroleum hydrocarbon contaminated to have become the focus of world attention.In the 1980s, beautiful There is serious petroleum hydrocarbon soil contamination problem in state, has half or so in 450,000 blocks of brown soils there are petroleum hydrocarbon contaminated, The petroleum hydrocarbon contaminated place of same period Canada is up to 60%, and there is also serious oil pollution, morals with factory in Britain many areas The country such as state, Holland, Britain has formulated relevant recovery project plan, strengthens the reparation and improvement of petroleum hydrocarbon contaminated soil.

Since China ranked among big oil-producing country, the world ten from 1978, more and more oil-gas reservoirs and oil gas field be explored with Exploitation, oil play increase, and about exploiting 700,000 tons of crude oil every year wherein about 1/10th enter soil environment causes China Newly-increased polluted soil has reached 100,000 tons or so every year.Soil is petroleum hydrocarbon contaminated, and the situation is tense, and hindering China can hold The process of supervention exhibition.

After petroleum hydrocarbon contaminant enters soil, the composition of soil texture and organic matter can be changed, petroleum hydrocarbon is easily viscous with grogs Even, soil aperture is blocked, soil water permeability, gas permeability are influenced, nutrient is limited from soil particle and enters the soil liquid, make soil The decline of earth fertility;And a large amount of organic carbons contained in petroleum hydrocarbon can make the carbon, nitrogen, phosphorus ratio numerous imbalances of soil.In addition, stone The metabolic intermediate of petroleum hydrocarbon in the soil has some characteristic groups, can absorb and complexation heavy metal ion, to influence Transport And Transformation of the heavy metal in soil/botanical system.Meanwhile it certain cannot can be permeated the ground by the hydrocarbon component of adsorption by soil Water causes the pollution of underground water, brings potential hazard for mankind's domestic water.

Soil has certain self-purification capacity to petroleum hydrocarbon contaminated in nature, but when the petroleum hydrocarbon into soil is more than environment Self-purification capacity when, petroleum hydrocarbon will be caused to accumulate and pollute in the soil.The existing method for repairing petroleum hydrocarbon contaminated soil Mainly there are physical restoration, chemical restoration, biological restoration and the combination of a variety of recovery techniques etc..

Microorganism is the important member in soil ecosystem, the substance circulation and energy flow in soil and biosphere In serve it is key.Many microorganisms can be degraded using petroleum hydrocarbon as sole carbon source and the energy by metabolism in nature Petroleum hydrocarbon contaminant in environment finally converts petroleum pollution to harmless carbon dioxide and water.Under aerobic conditions:

Petroleum hydrocarbon+microorganism+O2+ nutriment → CO+H2O+ by-product+microbial cell biomass.

The microbial profile of energy decomposing petroleum hydrocarbon is extensive, but is primarily present in contaminated area, because of natural microorganisms group It falls structure or population is not appropriate for oil polluted environment, after environment is by oil pollution, the survival of the fittest, so that adapting to petroleum wastewater The population of dye is developed, and the growth of other microbial populations is suppressed or is eliminated.Microorganism remediation generally refers to utilize soil Indigenous microorganism in earth adds under the environmental condition of optimization to efficient degradation microorganism of the pollution environment supplement through taming Fast decomposing organic pollutant repairs contaminated soil, and can be divided into biology in situ reparation (will be by petroleum hydrocarbon contaminated soil The method that is handled in situ) and Ex situ bioremediation (petroleum hydrocarbon contaminated soil is dug out, other than contaminated site or is transported The method handled to special reparation place).

Compared to physics and chemical method, repairing method of microorganism have at low cost, high treating effect, it is without secondary pollution, answer With range it is wide, do not destroy the advantages that soil environment, be suitable for being widely used in handling petroleum hydrocarbon contaminated soil.China is in Pollution Field Ground soil restoring technology, engineering application and equipment research and development etc. are started late, at present still in the low-level stage.

In terms of petroleum hydrocarbon contaminated soil recovery technique, research work is often based on laboratory or pilot-scale, live stone Petroleum hydrocarbon pollution amelioration case is rarely seen.In terms of petroleum hydrocarbon contaminated soil prosthetic appliance, rest on Scientific research equipment exploitation or experiment sample more Machine pilot scale stage, the industrial application of prosthetic appliance are very few.Therefore, filtering out has efficient degradation effect to petroleum hydrocarbon contaminant Bacterial strain, and cooperate suitable facilitated technique, there is highly important researching value to the biological prosthetic of petroleum hydrocarbon contaminated soil.

Summary of the invention

The purpose of the invention is to overcome the prior art to repair using biological prosthetic method to petroleum hydrocarbon contaminated soil The defect of multiple effect difference provides a kind of soil remediation composition and geobiont repair method, according to the method described in the present invention Preferable soil remediation effect, and the compositions and methods of the invention repairing especially suitable for petroleum hydrocarbon contaminated soil can be obtained It is multiple, it is particularly suitable for the reparation of the micro- contaminated soil of petroleum hydrocarbon and volatility, half volatile petroleum hydrocarbon contaminated soil.

To achieve the goals above, one aspect of the present invention provides a kind of soil remediation composition, soil remediation combination Object contains water-retaining agent, soil property regulator, nutriment, beta-cyclodextrin and single bacterium agent.

Preferably, the single bacterium agent is Stenotrophomonas (Stenotrophomonas), enlightening thatch Salmonella (Dietzia), rattan Yellow micrococcus luteus (Micrococcus luteus) or acinetobacter calcoaceticus (Acinetobacter).

Another aspect of the present invention provides soil remediation composition as described above, and in rehabilitating soil, (especially petroleum hydrocarbon is dirty The soil of dye) in application.

Another aspect of the invention provides a kind of geobiont repair method, and this method includes being repaired using soil of the invention Compound composition repairs soil.

In geobiont repair method of the present invention, by introducing degradable pollutant in soil into soil The microbial bacterial agent of (such as petroleum hydrocarbon), and by introducing component (such as soil property regulator, battalion that there is improving effect to soil soil property Support substance, beta-cyclodextrin and water-retaining agent etc.) to promote microbial bacterial agent to breed in the soil, promote the pollutant in soil to be dropped Solution, to obtain preferable soil remediation effect.

In addition, carrying out soil remediation using soil remediation composition of the present invention, secondary pollution not will cause, also not Soil environment can be destroyed, and at low cost, it is easy to operate.And soil remediation composition of the invention is especially suitable for petroleum hydrocarbon dirt The reparation for contaminating soil, is particularly suitable for the reparation of volatility, half volatile petroleum hydrocarbon contaminated soil.

Specific embodiment

The endpoint of disclosed range and any value are not limited to the accurate range or value herein, these ranges or Value should be understood as comprising the value close to these ranges or value.For numberical range, between the endpoint value of each range, respectively It can be combined with each other between the endpoint value of a range and individual point value, and individually between point value and obtain one or more New numberical range, these numberical ranges should be considered as specific open herein.

On the one hand, the present invention provides a kind of soil remediation composition, which contains water-retaining agent, soil property Regulator, nutriment, beta-cyclodextrin and single bacterium agent.

According to the present invention, the soil be meaning known in this field on soil, be minerals, organic matter, work it is organic The mixture of body and moisture and air etc..Preferably, by weight, minerals account for the 90- of solid fraction (soil dry weight) 95% or more, organic matter accounts for about 1-10%, it is seen that soil constituent is based on minerals.The soil organism is exactly in soil with each Organic compound existing for kind form.In addition to this there are also the soil liquid, it be soil moisture and its contained dissolution substance and The general name of suspended material.The soil liquid is the medium that plant and microorganism absorb nutrients from soil and pollutant in soil The main path migrated in earth.

In soil remediation composition of the present invention, without containing the component that can destroy soil environment, therefore, repairing It not will cause secondary pollution while soil.

According to the present invention, the single bacterium agent can be that can be used in the strain of petroleum hydrocarbon degradation, but this hair in the prior art Bright inventor has found in the course of the study, Stenotrophomonas (Stenotrophomonas), enlightening thatch Salmonella (Dietzia), Micrococcus luteus (Micrococcus luteus), acinetobacter calcoaceticus (Acinetobacter) are to crude oil, n-alkane, branch The ingredients such as alkane and aromatic hydrocarbons have preferable degrading activity.

Wherein, Stenotrophomonas (Stenotrophomonas) belongs to the aerobic Gram-negative bar of stringent non-fermented type Bacterium, nutritional requirement be not high.The present inventor it has been investigated that, Stenotrophomonas have unique biochemical activity and generation Thank characteristic, degradable benzene, toluene, ethylbenzene and aniline.

Wherein, Hai Dici Salmonella (Dietzia) is a kind of strain with degrading crude oil and long chain alkane class ability, and is had There is the ability of the decomposing petroleum hydrocarbon under hypersaline environment.In addition, the present inventor also found in the course of the study, which exists During decomposing petroleum hydrocarbon, surface reactive material can be generated, is conducive to microorganism and the direct contact of petroleum hydrocarbon is utilized.

Wherein, micrococcus luteus (Micrococcus luteus) belongs to obligate aerobic gram-positive bacteria, life condition It is required that loose.The present inventor has found that the bacterium is to n-alkane, the isoparaffin in petroleum component in the course of the study And the degradation effect of cycloalkane is preferable.

Wherein, acinetobacter calcoaceticus (Acinetobacter) is non-zymocyte, which passes through terminal oxidized way under aerobic conditions Diameter degradation saturated hydrocarbons, growth ability is strong, and breeding is rapid.

According to the present invention, the grain size distribution of solid particle or combinations of particle sizes are known as the mechanical composition of soil in soil, also known as The soil texture.It can be classified to soil according to the mechanical composition of soil.The quality of soil is to influence soil fertility height, can plough Property quality and one of Fundamentals of contaminant capacity size.Inventor is the study found that the adjustable soil of soil property regulator Solid particle granularity, making soil grades, more evenly oxygen content is higher, and it is saturating can to break soil hardening, chesson, raising soil Gas reduces the soil weight, promote soil microbial activities, reinforce contact of the microorganism with pollutant, enhancing soil rich water seeps Saturating power.Therefore, soil property regulator of the invention can be in the prior art with the substance of function as above, it is preferred that the soil Matter regulator can be selected from one of sawdust, stalk and zeolite powder or a variety of.

According to the present invention, the water-retaining agent is also known as high water absorbency polymer, can absorb and be equivalent to own wt hundred times extremely Thousands of times of deionized water and decades of times forms gelatinous mass after water swelling, in dried strip to hundred times or more of saliferous moisture It again can be by moisture slow release under part.Inventor the study found that water-retaining agent using proper, can significantly improve soil water content, Cubical expansivity, liquid phase percentage, total porosity and capillary porosity percentage improve soil texture, mitigate ground temperature variation, promote Into microbial activities, increase soil fertility to stimulate and microorganism is maintained to grow.Therefore, water-retaining agent of the invention can be existing With the substance of function as above in technology, it is preferred that the water-retaining agent is selected from acrylate copolymer, acrylamide polymer and shallow lake One of powder-acrylic copolymer is a variety of.

In the present invention, the nutriment can be the soil nutrient containing nitrogen and P elements.Preferably, The nutriment is urea and/or corn pulp.

In the present invention, beta-cyclodextrin is the derivative of cyclodextrin, and hydrophobic cavity can form with organic matter and generate envelope Object is stablized, fixed organic pollutant, reduces the volatility of organic pollutant.Beta-cyclodextrin can also increase hydrophobicity organic contamination The dissolution of object, enters it in the soil liquid from soil surface, promotes microorganism to its preferably degradation and utilization.

It will group of the above several substances (water-retaining agent, soil property regulator, nutriment, beta-cyclodextrin and single bacterium agent) to be suitable for At being added in petroleum hydrocarbon contaminated soil, the effect for forming an entirety to play collaboration compatibility is organically combined between each component, So as to effectively repair petroleum hydrocarbon contaminated soil.

According to the present invention, the content of each component can be become in a wider scope in the soil remediation composition Change, under preferable case, in the soil remediation composition, the water-retaining agent based on 100 parts by weight, the soil property regulator Content be 100-1500 parts by weight, more preferably 300-1250 parts by weight are (for example, 300 parts by weight, 500 parts by weight, 700 weights In the range of any two in amount part, 900 parts by weight, 1100 parts by weight, 1250 parts by weight and these point values are constituted Arbitrary value);The content of the nutriment be 10-500 parts by weight, more preferably 10-360 parts by weight (for example, 10 parts by weight, 20 parts by weight, 30 parts by weight, 40 parts by weight, 50 parts by weight, 80 parts by weight, 120 parts by weight, 150 parts by weight, 180 parts by weight, In 200 parts by weight, 250 parts by weight, 280 parts by weight, 300 parts by weight, 330 parts by weight, 360 parts by weight and these point values Any two constitute in the range of arbitrary value);The content of the beta-cyclodextrin is 500-3000 parts by weight, more preferably 600-2500 parts by weight are (for example, 600 parts by weight, 800 parts by weight, 1000 parts by weight, 1200 parts by weight, 1500 parts by weight, 1800 The range that any two in parts by weight, 2000 parts by weight, 2300 parts by weight, 2500 parts by weight and these point values are constituted Interior arbitrary value);The content of the single bacterium agent is 107-1012CFU, preferably 108-1011CFU, such as (108CFU、109CFU、 1010CFU、1011Any two in CFU and these point values constitute in the range of arbitrary value).

Wherein, CFU is Colony Forming Unit, is defined as a certain amount of bacterium solution after diluting and passes through the side of casting or coating Method allows the microbial single-cell in it to be dispersed on culture medium flat plate one by one, and after cultivating, each living cells just forms a bacterium It falls.

Second aspect, the present invention provides application of the soil remediation composition as described above in rehabilitating soil.

According to description as above, soil remediation composition of the invention is especially suitable for petroleum hydrocarbon contaminated soil progress It repairs.

Wherein, petroleum hydrocarbon refers to the mixture containing hydrocarbon found in crude oil.Petroleum hydrocarbon is current environment In one of the organic pollutant, including gasoline, kerosene, diesel oil, lubricating oil, paraffin and pitch etc. that are widely present, be a variety of hydro carbons (normal alkane, branched paraffin, cycloalkane, aromatic hydrocarbons) and a small amount of other organic matters, such as sulfide, nitride, naphthenic are mixed Close object.

The third aspect, the present invention provides a kind of biological renovation methods of soil, this method comprises: using as described above Soil remediation composition repairs soil.

The present inventor with soil it should be noted that contacting soil remediation composition as above to soil When earth is repaired, each component in the soil remediation composition can be added separately in soil, can also be matched one It rises and is added in soil together.If each component as above is in what manner added in soil by opinion, be accordingly to be regarded as be using soil Remediation composition repairs soil.

According to the present invention, based on the soil to be repaired of 100 parts by weight, the dosage of the water-retaining agent can be 0.01-1 weight Measure part, preferably 0.1-0.3 parts by weight, specifically, for example, can for 0.1,0.15,0.2,0.25,0.3 parts by weight and these Any two in point value constitute in the range of arbitrary value.

According to the present invention, based on the soil to be repaired of 100 parts by weight, the dosage of the soil property regulator can be 0.1- 5 parts by weight, preferably 0.5-2.5 parts by weight specifically, such as can be 0.5,0.8,1.1,1.4,1.7,2.0,2.3,2.5 weights Measure the arbitrary value in the range of any two in part and these point values are constituted.

According to the present invention, the chemical composition of microbial cell and nutrient type determine the nutritional need of cell, petroleum drop Solving bacterium is mainly chemoheterotrophic bacteria, and when carrying out microbiological treatment to petroleum hydrocarbon contaminated soil, petroleum hydrocarbon is that microorganism mentions in system For available a large amount of carbon substrates, but mainly carbon, protium, the shortage of other elements that petroleum hydrocarbon provides may make microorganism The petroleum hydrocarbon for being difficult to degrade in soil.Inventor the study found that add the nutriments such as nitrogen phosphorus, make in soil C N the ratio between P connect The ratio between nearly microbial body interior element, can effectively facilitate the mass propagation of microorganism.Due to ammoniacal nitrogen (NH4 +- N) class nutriment exists Ammonia can be converted under weak basic condition and is diffused into air, and the loss of nitrogen class nutriment is caused.And Phosphorus nutrients Matter is relatively stable in the soil, not easily runs off.Therefore present invention primarily contemplates the additive amounts of nitrogen class nutriment imitates degradation The influence of fruit.In method of the present invention, it is preferable that the dosage of the nutriment makes carbon in soil nitrogen mass ratio C: N is (5-15): 1, specifically, such as can be 5:1,6:1,7:1,8:1,9:1,10:1,11:1,12:1,13:1,14:1,15: 1 and these ratios in the ratio range that is constituted of any two in other any ratios.Wherein, the content of C is according to soil The 90% of petroleum hydrocarbon gross mass calculates in earth.When the nutriment is urea and/or corn pulp, the content of N is according to urea The 3.5% of 46.67% or corn pulp gross weight of gross mass calculates.

According to the present invention, based on the soil to be repaired of 100 parts by weight, the dosage of the beta-cyclodextrin can be 1-5 weight Part is measured, specifically, such as can be any two in 1,1.5,2,2.5,3,3.5,4,4.5,5 parts by weight and these point values Arbitrary value in the range of constituting.

In accordance with the present invention it is preferred that the soil to be repaired based on 100 parts by weight, the dosage of the microbial inoculum can be with 109- 1011CFU specifically, such as can be 1 × 109、5×109、1×1010、5×1010、1×1011、5×1011CFU and these Any two in point value constitute in the range of arbitrary value.

In accordance with the present invention it is preferred that keeping Soil Water Content in repair process is 15-25%.

According to the present invention, the soil is preferably petroleum hydrocarbon contaminated soil, especially the micro- contaminated soil of petroleum hydrocarbon and is waved Hair property, half volatile petroleum hydrocarbon contaminated soil.

Embodiment

The present invention will be described in detail by way of examples below.

1, material therefor in following embodiment and comparative example

Polyacrylic acid is purchased from company, Changzhou Chun Jiang Chemical Co., Ltd., article No. SL-324;

Polyacrylamide is purchased from company, Shenyang An Ruixiang chemical industry Hardwear Pty Ltd., and article No. is polyacrylamide -03;

Starch with Acrylic Acid polymer is purchased from company, Tuo Zhihong Science and Technology Ltd.;

Zeolite powder is purchased from company, Lingshou County Hua Wei mineral products processing factory;

Urea is purchased from company, Jinan Xin Hong Chemical Co., Ltd.;

Corn pulp is purchased from company, Shandong Hong Xing corn development corporation, Ltd.;

Acinetobacter calcoaceticus is purchased from China General Microbiological culture presevation administrative center, deposit number CGMCC1.10395T

Enlightening thatch Salmonella is purchased from China General Microbiological culture presevation administrative center, deposit number CGMCC1.6332T

Micrococcus luteus is purchased from China General Microbiological culture presevation administrative center, deposit number CGMCC1.10554;

Stenotrophomonas is purchased from China General Microbiological culture presevation administrative center, deposit number CGMCC1.6393;

N-dodecane is purchased from Aladdin company, article No. D108175.

Soil is the dry soil to sterilize at 250 DEG C for 24 hours.

2, the petroleum hydrocarbon contaminated biological prosthetic effect assessment of soil

1. microbial inoculum is enriched in minimal medium of the target contaminant as sole carbon source;2. being centrifugated, obtain It is enriched with microbial inoculum;3. microbial inoculum is put into soil together with water-retaining agent, soil conditioner, beta-cyclodextrin, nutriment;4. in soil Target contaminant is added in earth;5. periodic detection soil petroleum hydrocarbon content and soil bacteria sum calculate the degradation effect of petroleum hydrocarbon Fruit.

Concrete operation step are as follows:

(1) it takes 1.5mL bacterium solution to be added in the triangular flask equipped with 150mL minimal medium, adds target contaminant, 150r/min vibrates 7 days, obtains enrichment bacterium solution.

(2) obtained enrichment culture liquid 8000r/min centrifugal solid-liquid is separated, discards upper layer culture medium solution, nothing is added 8000r/min centrifugal solid-liquid separation again after machine salt culture medium 100mL sufficiently vibrates, discards upper solution, layer solid of keeping on file is standby With.

(3) water-retaining agent, soil conditioner, beta-cyclodextrin, nutriment and bottom solid are added in soil, are sufficiently stirred It mixes, while deionized water is added, soil moisture content is made to maintain a certain range.

(4) target contaminant is added into soil, stirs evenly.

(5) soil petroleum hydrocarbon content and soil bacteria sum are detected weekly.

(6) water of every two days benefits keeps soil moisture content in a certain range, digs weekly 3 times.

Minimal medium: NH4Cl is 0.67g, Na2CO3For 1.06g, MgSO4·7H2O is 0.1g, CaCl2For 0.1g, K2HPO4·3H2O is 1.5g, KH2PO4For 0.5g, FeCl3·6H2O is 0.1g, deionized water 1L, pH value 7.0-7.5.

3, petroleum hydrocarbon assay in soil

Using the dry fresh pedotheque of anhydrous sodium sulfate, using carbon tetrachloride as extracting solution, mechanical shaking extraction pedotheque In petroleum substance, measure the concentration of petroleum-type.The content of petroleum-type is respectively 2930cm-1 (CH by wave number2C-H in group The stretching vibration of key), (c h bond stretches in aromatic rings by 2960cm-1 (stretching vibration of c h bond in CH3 group) and 3030cm-1 Contracting vibration) absorbance at bands of a spectrum calculated.

Specific determination step are as follows:

(1) pedotheque being collected is removed into the sundries such as gravel, root system of plant, be sufficiently mixed uniformly.

(2) 10.000g pedotheque is accurately weighed, 10g anhydrous sodium sulfate is added, mixes well, places 30min, solidification After crush.

(3) pedotheque that will have been dried is fully transferred in 50ml centrifuge tube, and 25ml carbon tetrachloride is added, sufficiently mixed It closes, is placed in ultrasonator, shake 10min.

(4) the centrifuge tube 8000r/min centrifugal solid-liquid after concussion is separated, collects upper layer carbon tetrachloride.

(5) 25ml carbon tetrachloride is added again into the remaining subsoil solid of centrifuge tube, is sufficiently mixed, is placed in ultrasound In oscillator, 10min is shaken.

(6) the centrifuge tube 8000r/min centrifugal solid-liquid after concussion is separated, collects upper layer carbon tetrachloride, and collects before Upper layer carbon tetrachloride mixing, it is to be measured.

(7) according to the relevant regulations of GB 7172, the content w of moisture in pedotheque is measuredw

The content ω (mg/kg) of Hydrocarbon in Soil, is calculated according to following formula.

In formula:

The content of ω --- Hydrocarbon in Soil, mg/kg;

ρ1--- the sample petroleum-type concentration checked on standard curve, mg/L;

ρ0--- the blank sample petroleum-type concentration checked on standard curve, mg/L;

The extension rate of D --- extracting solution;

V --- extracting solution constant volume, 50ml;

ms--- sampling amount, g;

ww--- soil moisture content, %.

The measurement range of this method is 50-500mg/kg, and such as soil petroleum hydrocarbon content to be measured exceeds this range, after should diluting Measurement.

Said determination is carried out to preprosthetic soil sample, after starting reparation, said determination is carried out to soil sample weekly, counts petroleum Hydrocarbon content, wherein the unit of petroleum hydrocarbon content is mg/kg.

4, soil bacteria sum measures

Measuring method: plate culture colony counting method

Specific determination step are as follows:

Weigh 10g pedotheque, be put into the triangular flask for filling 90mL aseptic deionized water, be placed on shaking table with 160 times/ The speed shaken at room temperature 4h of min, is sufficiently mixed soil sample with water, and the microbial cell in soil is fully dispersed, from soil It separates.This is 10-1Soil supension draws this soil supension of 1mL in the sterile water of 9mL NaCl containing 8.5g/L, and use is sterile Suction pipe pressure-vaccum 3 times mixings, are made 10-2Soil supension.And so on be successively made 10-3、10-4、10-5、10-6、10-7、10-8It is different The soil supension of dilution.It selects after dilution appropriate by combining colony counting method to measure the total number of bacteria of soil supension.

Total number of bacteria x (CFU/g), is calculated according to following formula in soil.

In formula:

X --- soil bacteria sum, CFU/g;

The total number of bacteria of e --- soil supension, CFU/ml;

The volume of V --- soil supension, ml;

M --- sampling amount, g;

W --- soil moisture content, %.

Meat extract peptone culture medium: NaCl 10g, peptone 10g, beef extract 5g, agar 18g, deionized water For 1L, pH value 7.2-7.4.

Said determination is carried out to preprosthetic soil sample, is started after being repaired according to the method for embodiment or experimental example, often Week carries out said determination to soil sample, counts soil sample total number of bacteria (unit 106CFU/g)。

Following embodiment and comparative example are all made of following tests method.Testing soil used is the drying of 250 DEG C of sterilizings for 24 hours Soil.Bacterium solution is centrifugated, takes solid portion that deionized water is added and is sufficiently mixed, be added in soil.

Embodiment 1

It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1;It weighs 0.96g urea and is dissolved in 100g In ionized water, aqueous solution 2 is obtained;It takes Stenotrophomonas to be mixed in 300g deionized water, obtains aqueous solution 3;Weigh 45g sawdust, 90g beta-cyclodextrin is mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein Stenotrophomonas Dosage makes the amount of single bacterium agent in soil be shown in Table 2.Then 19.75g n-dodecane is added into uniformly mixed soil, mixing is equal It is even.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Soil petroleum hydrocarbon is detected weekly Content and soil bacteria sum, are as a result shown in Tables 1 and 2 respectively.

Embodiment 2

It weighs 3.6g polyacrylamide to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 12.86g corn pulp and be dissolved in In 100g deionized water, aqueous solution 2 is obtained, takes enlightening thatch Salmonella to be mixed in 300g deionized water, obtains aqueous solution 3, weigh 45g straw Stalk, 90g beta-cyclodextrin are mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, are uniformly mixed, wherein Di Ci The dosage of Salmonella makes the amount of single bacterium agent in soil be shown in Table 2.19.75g n-dodecane, mixing are added into uniformly mixed soil Uniformly.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Soil petroleum is detected weekly Hydrocarbon content and soil bacteria sum, are as a result shown in Tables 1 and 2 respectively.

Embodiment 3

It weighs 8.4g Starch with Acrylic Acid polymer to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 0.96g urea It is dissolved in 100g deionized water, obtains aqueous solution 2, micrococcus luteus is taken to be mixed in 300g deionized water, obtain aqueous solution 3, claim It takes 45g zeolite powder, 90g beta-cyclodextrin to be mixed in 2400g soil, aqueous solution 1,2,3 is added in soil, be uniformly mixed, wherein The dosage of micrococcus luteus makes the amount of single bacterium agent in soil be shown in Table 2.19.75g positive 12 is added into uniformly mixed soil Alkane is uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Detection soil weekly Earth petroleum hydrocarbon content and soil bacteria sum, are as a result shown in Tables 1 and 2 respectively.

Embodiment 4

It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 0.96g urea and be dissolved in 100g In ionized water, aqueous solution 2 is obtained, acinetobacter calcoaceticus is taken to be mixed in 300g deionized water, obtains aqueous solution 3, weighs 18g sawdust, 90g Beta-cyclodextrin is mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein the dosage of acinetobacter calcoaceticus So that the amount of single bacterium agent is shown in Table 2 in soil.19.75g n-dodecane is added into uniformly mixed soil, is uniformly mixed.Every two It mends a water, keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Weekly detect soil petroleum hydrocarbon content and Soil bacteria sum, is as a result shown in Tables 1 and 2 respectively.

Embodiment 5

It weighs 6g polyacrylamide to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 12.86g corn pulp and be dissolved in In 100g deionized water, aqueous solution 2 is obtained, takes enlightening thatch Salmonella to be mixed in 300g deionized water, obtains aqueous solution 3, weigh 72g straw Stalk, 90g beta-cyclodextrin are mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, are uniformly mixed, wherein enlightening thatch Salmonella Dosage make the amount of single bacterium agent in soil be shown in Table 2.19.75g n-dodecane is added into uniformly mixed soil, mixing is equal It is even.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Soil petroleum hydrocarbon is detected weekly Content and soil bacteria sum, are as a result shown in Tables 1 and 2 respectively.

Embodiment 6

It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 1.92g urea and be dissolved in 100g In ionized water, aqueous solution 2 is obtained, Stenotrophomonas is taken to be mixed in 300g deionized water, obtains aqueous solution 3, weighs 45g zeolite Powder, 90g beta-cyclodextrin are mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, are uniformly mixed, wherein oligotrophy unit cell The dosage of bacterium makes the amount of single bacterium agent in soil be shown in Table 2.19.75g n-dodecane is added into uniformly mixed soil, mixing is equal It is even.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Soil petroleum hydrocarbon is detected weekly Content and soil bacteria sum, are as a result shown in Tables 1 and 2 respectively.

Embodiment 7

It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 0.29g urea and be dissolved in 100g In ionized water, obtain aqueous solution 2, micrococcus luteus taken to be mixed in 300g deionized water, obtain aqueous solution 3, weigh 45g sawdust, 90g beta-cyclodextrin is mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein micrococcus luteus Dosage makes the amount of single bacterium agent in soil be shown in Table 2.19.75g n-dodecane is added into uniformly mixed soil, is uniformly mixed. Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Soil petroleum hydrocarbon content is detected weekly And soil bacteria sum, Tables 1 and 2 is as a result seen respectively.

Embodiment 8

Weigh 6g polyacrylic acid to be dissolved in 200g deionized water, obtain aqueous solution 1, weigh 0.96g urea be dissolved in 80g go from In sub- water, aqueous solution 2 is obtained, acinetobacter calcoaceticus is taken to be mixed in 200g deionized water, obtains aqueous solution 3, weighs 45g sawdust, 90g β- Cyclodextrin is mixed in 2520g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein the dosage of acinetobacter calcoaceticus makes The amount for obtaining single bacterium agent in soil is shown in Table 2.19.75g n-dodecane is added into uniformly mixed soil, is uniformly mixed.Every two days A water is mended, keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Soil petroleum hydrocarbon content and soil are detected weekly Earth total number of bacteria, is as a result shown in Tables 1 and 2 respectively.

Embodiment 9

It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 0.96g urea and be dissolved in 200g In ionized water, obtain aqueous solution 2, take enlightening thatch Salmonella to be mixed in 320g deionized water, obtain aqueous solution 3, weigh 45g sawdust, 90g beta-cyclodextrin is mixed in 2280g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein take enlightening thatch Salmonella Dosage makes the amount of single bacterium agent in soil be shown in Table 2.19.75g n-dodecane is added into uniformly mixed soil, is uniformly mixed. Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Soil petroleum hydrocarbon content is detected weekly And soil bacteria sum, Tables 1 and 2 is as a result seen respectively.

Embodiment 10

It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1;It weighs 0.96g urea and is dissolved in 100g In ionized water, aqueous solution 2 is obtained;It takes Stenotrophomonas to be mixed in 300g deionized water, obtains aqueous solution 3;Weigh 45g sawdust, 144g beta-cyclodextrin is mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein Stenotrophomonas Dosage make the amount of single bacterium agent in soil be shown in Table 2.Then 19.75g n-dodecane, mixing are added into uniformly mixed soil Uniformly.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Soil petroleum is detected weekly Hydrocarbon content and soil bacteria sum, are as a result shown in Tables 1 and 2 respectively.

Embodiment 11

It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1;It weighs 0.96g urea and is dissolved in 100g In ionized water, aqueous solution 2 is obtained;It takes Stenotrophomonas to be mixed in 300g deionized water, obtains aqueous solution 3;Weigh 45g sawdust, 36g beta-cyclodextrin is mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein Stenotrophomonas Dosage makes the amount of single bacterium agent in soil be shown in Table 2.Then 19.75g n-dodecane is added into uniformly mixed soil, mixing is equal It is even.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Soil petroleum hydrocarbon is detected weekly Content and soil bacteria sum, are as a result shown in Tables 1 and 2 respectively.

Comparative example 1

It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, any microbial inoculum is not added.

Comparative example 2

It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, do not add water-retaining agent.

Comparative example 3

It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, do not add nutriment.

Comparative example 4

It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, soil property regulator is not added.

Comparative example 5

It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, do not add beta-cyclodextrin.

Table 1

Initially 7d 14d 21d 28d 35d 42d 49d Embodiment 1 6583 6023 5174 3713 1949 1218 913 578 Embodiment 2 6583 6174 5570 4292 2983 2070 1766 1644 Embodiment 3 6583 5962 4931 3288 1583 1036 791 517 Embodiment 4 6583 6328 5783 4626 3653 2740 2496 2314 Embodiment 5 6583 5718 4566 2922 1218 791 487 365 Embodiment 6 6583 5900 4810 3166 1462 973 731 457 Embodiment 7 6583 6205 5600 4321 3044 2131 1826 1704 Embodiment 8 6583 6144 5418 4579 3217 2604 2062 2000 Embodiment 9 6583 5839 4748 3044 1400 913 608 426 Embodiment 10 6583 5992 4992 3348 1704 1066 822 548 Embodiment 11 6583 6083 5357 3956 2435 1583 1400 1218 Comparative example 1 6583 6122 5939 5071 4342 3656 3474 3337 Comparative example 2 6583 6303 6108 5682 4204 3473 3278 3132 Comparative example 3 6583 6073 5858 4840 3984 3180 2965 2804 Comparative example 4 6583 6180 6032 5339 4754 4207 4061 3952 Comparative example 5 6583 6118 5296 4018 3330 2574 1852 1461

Note: petroleum hydrocarbon content unit is mg/kg soil.

Table 2

Note: total number of bacteria unit is 106CFU/g soil.

By above table as can be seen that microorganism can be quickly when carrying out soil remediation using the solution of the present invention Reach the peak of growth curve, and degrades to pollutant.And when the scheme in comparative example 2-5 being used to be repaired, soil Condition is relatively poor, and the growth of microorganism determines the degradation situation of pollutant, is repaired using the scheme in comparative example 1 When, due to not adding additional petroleum hydrocarbon degradation bacterium, petroleum hydrocarbon incipient degradation speed is slower.

The preferred embodiment of the present invention has been described above in detail, and still, the present invention is not limited thereto.In skill of the invention In art conception range, can with various simple variants of the technical solution of the present invention are made, including each technical characteristic with it is any its Its suitable method is combined, and it should also be regarded as the disclosure of the present invention for these simple variants and combination, is belonged to Protection scope of the present invention.

Claims (10)

1. a kind of soil remediation composition, which is characterized in that the soil remediation composition contains water-retaining agent, soil property regulator, battalion Support substance, beta-cyclodextrin and single bacterium agent.
2. soil remediation composition according to claim 1, wherein the single bacterium agent is Stenotrophomonas (Stenotrophomonas), enlightening thatch Salmonella (Dietzia), micrococcus luteus (Micrococcus luteus) or acinetobacter calcoaceticus (Acinetobacter)。
3. soil remediation composition according to claim 1 or 2, wherein the water-retaining agent is selected from acrylate copolymer, third One of acrylamide polymer and Starch with Acrylic Acid copolymer are a variety of.
4. soil remediation composition described in any one of -3 according to claim 1, wherein the soil property regulator is selected from saw One of end, stalk, zeolite powder are a variety of.
5. soil remediation composition described in any one of -4 according to claim 1, wherein the nutriment is to contain carbon The soil nutrient of element, nitrogen and P elements;
Preferably, the nutriment is urea and/or corn pulp.
6. any soil remediation composition in -5 according to claim 1, wherein the water conservation based on 100 parts by weight Agent, the content of the soil property regulator are 100-1500 parts by weight, and the content of the nutriment is 10-500 parts by weight, described The content of beta-cyclodextrin is 500-3000 parts by weight, and the content of the single bacterium agent is 1010-1013CFU。
7. application of the soil remediation composition in rehabilitating soil described in any one of claim 1-6.
8. a kind of biological renovation method of soil, which is characterized in that this method comprises: using any one of claim 1-6 The soil remediation composition repairs soil.
9. restorative procedure according to claim 8, wherein the soil to be repaired based on 100 parts by weight, the soil are repaired The dosage of compound composition makes the content of the water-retaining agent be 0.01-1 parts by weight, and the content of the soil property regulator is 0.1-5 Parts by weight, the content of the nutriment make carbon in soil nitrogen mass ratio C:N be (5-15): 1, the content of the beta-cyclodextrin For 1-5 parts by weight, the content of the single bacterium agent is 108-1012CFU。
10. restorative procedure according to claim 8 or claim 9, wherein keep Soil Water Content for 15-25 weight in repair process Measure %;
Preferably, the soil is petroleum hydrocarbon contaminated soil.
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