CN114958820B - Water-soluble natural wax-dissolving agent and preparation method and application thereof - Google Patents

Water-soluble natural wax-dissolving agent and preparation method and application thereof Download PDF

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CN114958820B
CN114958820B CN202210510298.5A CN202210510298A CN114958820B CN 114958820 B CN114958820 B CN 114958820B CN 202210510298 A CN202210510298 A CN 202210510298A CN 114958820 B CN114958820 B CN 114958820B
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dissolving agent
wax
water
soluble natural
natural wax
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CN114958820A (en
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王书亮
张晓燕
周禄江
付春艳
刘永兵
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Southwest Petroleum University
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    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
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    • C09K8/00Compositions for drilling of boreholes or wells; Compositions for treating boreholes or wells, e.g. for completion or for remedial operations
    • C09K8/52Compositions for preventing, limiting or eliminating depositions, e.g. for cleaning
    • C09K8/524Compositions for preventing, limiting or eliminating depositions, e.g. for cleaning organic depositions, e.g. paraffins or asphaltenes
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    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
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    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/08Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
    • C12N11/082Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds
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    • C12R2001/00Microorganisms ; Processes using microorganisms
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    • C12R2001/38Pseudomonas
    • C12R2001/385Pseudomonas aeruginosa
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02WCLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00Technologies for solid waste management
    • Y02W30/50Reuse, recycling or recovery technologies
    • Y02W30/74Recovery of fats, fatty oils, fatty acids or other fatty substances, e.g. lanolin or waxes

Abstract

The invention discloses a water-soluble natural wax-dissolving agent and a preparation method and application thereof, wherein the preparation steps of the wax-dissolving agent are as follows: (1) culturing pseudomonas aeruginosa in seed solution; (2) Preparing bacterial suspension from bacterial mud obtained after centrifuging the seed liquid; (3) The bacterial suspension reacts with a mixed solution composed of polyvinyl alcohol and calcium chloride and a sodium alginate solution to prepare PA/SA/PVA gel balls; (4) Solidifying the PA/SA/PVA gel ball to obtain PA/SA/PVA liquid core microcapsule; (5) preparing a culture solution; (6) Adding the liquid core microcapsule into a culture solution for fermentation to prepare a fermentation solution containing rhamnolipid; (7) passing the fermentation broth through a ceramic filter membrane to obtain a micro-filtrate; and (8) adding alkali and inorganic salt into the micro-filtrate. The water-soluble natural wax-dissolving agent provided by the invention has excellent wax-preventing performance, and the main raw materials of the wax-dissolving agent can be prepared by using the waste cooking oil as a fermentation carbon source, so that the production cost of the wax-dissolving agent is effectively reduced, and the wax-dissolving agent has the advantage of environmental protection.

Description

Water-soluble natural wax-dissolving agent and preparation method and application thereof
Technical Field
The invention belongs to the technical field of oilfield development, and particularly relates to a water-soluble natural wax-dissolving agent, and a preparation method and application thereof.
Background
In the process of crude oil exploitation, the temperature and the pressure of crude oil liquid are continuously reduced in the process of conveying the crude oil liquid from an oil well to the ground, wax contained in the crude oil liquid is usually crystallized and separated out, and the separated wax is continuously deposited on the sucker rod and the inner wall of an oil pipe; serious wax precipitation affects normal production operations of oil wells, and therefore, a great deal of effort is required to perform frequent well flushing and pump inspection operations to remove precipitated wax. The methods currently used for preventing and removing paraffin deposits in crude oil production mainly comprise the following three types: 1. a mechanical paraffin removal method, a thermal paraffin removal and chemical paraffin removal and prevention method, and a chemical paraffin removal and prevention method; the mechanical paraffin removal method and the thermal paraffin removal method are rarely adopted because the oil well has to be stopped when in operation and the energy consumption is larger; the chemical paraffin removal and prevention method has the advantages of convenient use, good application effect, no need of stopping production in the operation process and the like, and becomes the most common technology of paraffin removal and prevention of oil wells (Gao Wenyuan. Research on paraffin removal and prevention agents of oil wells [ D ]. Daqing petroleum institute, 2009.).
Chemical paraffin removal and inhibition generally refers to a method of adding paraffin solvents from an annular space to an oil pipe to prevent and remove paraffin deposits, and the method has the effects of reducing congealing, viscosity and blocking in addition to paraffin removal and inhibition (Chen Fu, qu Jinming, wang Fuxiang, etc. the current state of research and development of paraffin removal and inhibition agents for oil wells [ J ] oil and gas chemical industry, 2003, 32 (4): 243-245.). In recent years, with the increasing use of chemical paraffin removal and prevention methods, paraffin removal and prevention agents (also called paraffin dissolving agents) have been developed correspondingly. However, most of the main raw materials of the conventional paraffin removal and prevention agents are nondegradable surfactants artificially synthesized by petrochemical products, and the surfactants are required to be used in the use process by aromatic hydrocarbon solvents with strong toxicity such as benzene, dimethylbenzene and the like, so that the environment is damaged and the physical health of workers is damaged after long-term use (Dejian, and the like; the development and application of water-soluble and oil-soluble paraffin removal and prevention agents [ D ] Daqing petroleum institute, 2003.). Therefore, the design and synthesis of the green degradable nontoxic water-soluble wax dissolving method is very necessary and has a wide application prospect.
Disclosure of Invention
The invention aims to provide a water-soluble natural wax-dissolving agent, and a preparation method and application thereof.
In order to achieve the above object, the solution of the present invention is:
the preparation method of the water-soluble natural wax-dissolving agent is characterized by comprising the following steps:
(1) Inoculating pseudomonas aeruginosa on a nutrient gravy agar medium, performing constant-temperature activation, transferring the strain subjected to the constant-temperature activation into seed liquid, and performing shake flask culture to obtain seed liquid containing the pseudomonas aeruginosa;
(2) Centrifuging the seed solution prepared in the step (1) to remove supernatant, and adding the obtained bacterial mud into sterile physiological saline to prepare bacterial suspension;
(3) Adding the bacterial suspension prepared in the step (2) into a mixed solution composed of polyvinyl alcohol and calcium chloride, uniformly stirring, dropwise adding the obtained mixed solution into a sodium alginate solution, and filtering to obtain spherical sodium alginate/polyvinyl alcohol gel balls;
(4) Washing the sodium alginate/polyvinyl alcohol gel ball prepared in the step (3), then placing the washed gel ball into a calcium chloride solution for solidification, removing water on the surface of the solidified gel ball, and then freezing to prepare the sodium alginate/polyvinyl alcohol liquid core microcapsule;
(5) Adding a carbon source into an inorganic culture medium to prepare an inorganic salt culture solution containing the carbon source;
(6) Thawing the sodium alginate/polyvinyl alcohol liquid core microcapsule prepared in the step (4), adding the thawed sodium alginate/polyvinyl alcohol liquid core microcapsule into the inorganic salt culture solution containing carbon sources prepared in the step (5), and placing the inorganic salt culture solution into a fermentation tank for constant-temperature culture to prepare a rhamnolipid-containing fermentation liquor;
(7) Passing the rhamnolipid-containing fermentation liquor prepared in the step (6) through a ceramic filter membrane to remove larger proteins and insoluble impurities in the fermentation liquor, so as to prepare micro-filtrate;
(8) Adding alkali and inorganic salt into the micro-filtrate prepared in the step (7) to prepare the water-soluble natural wax-dissolving agent.
Preferably, the constant temperature activation temperature in step (1) is 37 ℃; the constant temperature activation time is 24 hours.
Preferably, the mass ratio of polyvinyl alcohol to calcium chloride sodium alginate in the mixed solution of polyvinyl alcohol and calcium chloride in the step (3) is 1:1:2.
Preferably, the freezing temperature in the step (4) is-18 ℃ and the freezing time is 4-6h.
Preferably, the carbon source in step (5) is selected from one of waste cooking oil, glycerol or vegetable oil.
Preferably, the constant temperature culture temperature in the step (6) is 37 ℃ and the constant temperature culture time is 24 hours.
Preferably, the pore size of the ceramic filter membrane in the step (7) is 50 nm-1000 nm.
Preferably, the base in step (8) is one of ethanolamine or triethylamine; the inorganic salt is one of sodium sulfate or sodium chloride.
The water-soluble natural wax-dissolving agent prepared by the preparation method of the water-soluble natural wax-dissolving agent is characterized in that the mass ratio of rhamnolipid is 0.1-1.0%; the mass ratio of the alkali is 0.5 to 2.0 percent, the mass ratio of the inorganic salt is 0.1 to 1.0 percent, and the balance is water.
The water-soluble natural wax-dissolving agent prepared by the preparation method of the water-soluble natural wax-dissolving agent is applied to wax prevention and paraffin removal of petroleum pipelines.
Compared with the existing preparation method of wax-dissolving agent, the principle and gain effect of the invention are as follows:
1. according to the preparation method of the water-soluble natural wax-dissolving agent, in the process of preparing main raw materials of rhamnolipid, polyvinyl alcohol (PVA) and Sodium Alginate (SA) with good biocompatibility are selected as immobilization materials, and then an embedding method is used for immobilizing pseudomonas aeruginosa cells so as to prepare SA/PVA liquid core microencapsulated cells. The prepared liquid core microencapsulated cells can encapsulate microorganisms (such as pseudomonas aeruginosa) in an internal liquid environment through a layer of external capsule membrane so as to block the cells from entering the external environment, and the formed microcapsule membrane allows nutrients and microorganism products (such as rhamnolipid) to pass through bidirectionally; this not only facilitates the microorganisms to extract nutrients and release extracellular products, but also reduces the effects of product inhibition, thereby reducing the fermentation cycle and increasing the yield of extracellular products (Sun Z, lv G, li s.probing the role of microenvironment for microencapsulated sacchromyces cerevisiae under osmotic stress [ J ] Applied Microbial and Cell Physiology,2007, 75:1419-1427.).
2. The water-soluble natural wax-dissolving agent provided by the preparation method provided by the invention uses natural surfactant rhamnolipid as a main raw material and small amount of alkali and inorganic salt as auxiliary materials. The rhamnolipid in the wax-dissolving agent is used as a surfactant to enable the paraffin surface on the wall of the oil well to be reversed into a hydrophilic surface, so that the wax cannot be deposited on the surface continuously, and the effect of preventing the wax is achieved; in addition, the rhamnolipid is adsorbed on the surface of the oil pipe, so that the paraffin is beneficial to falling off from the surface of the pipe wall, and the effect of dissolving the paraffin is achieved. The addition of alkali and inorganic salts can further reduce the surface tension of rhamnolipids; therefore, when the wax-dissolving agent permeates into the gaps of the wax crystals of the loose structure, the binding force between wax molecules can be weakened, and the wax crystals are dispersed in the oil flow due to the disassembly.
3. The main raw material of the water-soluble natural wax-dissolving agent provided by the invention is degradable natural product rhamnolipid which is harmless to the environment, so that the sulfonate chemical synthesis surfactant which is high in toxicity and poor in biodegradability is successfully avoided.
4. The preparation method of the water-soluble natural wax-dissolving agent provided by the invention takes cooking waste oil or kitchen waste grease and the like as fermentation carbon sources to prepare and produce rhamnolipid, so that the production cost of the wax-dissolving agent is effectively reduced.
Drawings
FIG. 1 is a graph showing the recycling of PA/SA/PVA liquid core microcapsules.
Detailed Description
The present invention will be described in further detail with reference to examples. It is also to be understood that the following examples are given solely for the purpose of illustration and are not to be construed as limitations on the scope of the invention, and that the particular amounts of materials, reaction times and temperatures, process parameters, etc. shown are but one example of a suitable range, and that some insubstantial modifications and adaptations of the invention to those skilled in the art are within the scope of the invention.
The reagents and instrumentation used in the present invention require strict sterility before use.
Pseudomonas aeruginosa used in the invention is purchased from Beijing deposit biotechnology Co., ltd, and the strain deposit number is as follows:
ATCC 9027=dsm1128=ifo 13275 =nrrl-B-800; the strain was stored in a refrigerator at-80℃during the experiment.
The composition of the culture medium used in the invention is as follows:
nutrient gravy agar (g/L): 15.0 parts of agar powder; 3.0 parts of beef extract; peptone 10.0; 10.0% of NaCl; pH:7.0 to 7.2;
seed liquid (g/L): 3.0 parts of beef extract; peptone 10.0; 10.0% of NaCl; pH:7.0 to 7.2;
inorganic salt medium (g/L): naNO 3 6.0;NaCl 1.0;KCl 1.0;Na 2 HPO 4 0.3;KH 2 PO 4 3.0;MgSO 4 2.5;FeCl 3 0.16;CaCl 2 0.1;ZnSO 4 1.5;MnSO 4 1.5;CuSO 4 0.15、H 3 BO 3 1.5。
Example 1
The preparation method of the water-soluble natural wax-dissolving agent provided by the invention comprises the following steps:
(1) Inoculating pseudomonas aeruginosa (Pseudomonas aeruginosa, PA) on nutrient gravy agar medium, activating at constant temperature in a constant temperature box at 37 ℃ for 24 hours, transferring the strain subjected to constant temperature activation into 100mL of seed liquid (the inoculation amount is 5%), and culturing for 18 hours in shaking flask culture at 200rpm at 37 ℃ to obtain seed liquid containing pseudomonas aeruginosa;
(2) Centrifuging (12000 r/min,10 min) the seed solution prepared in the step (1) to remove supernatant, and washing the obtained bacterial sludge three times by using 2mL of 0.9% sterile physiological saline; adding the washed bacterial mud into 0.9% sterile physiological saline to prepare 5 g.L -1 Is a bacterial suspension of (a);
(3) Adding 25ml of the bacterial suspension prepared in the step (2) into a mixed solution consisting of polyvinyl alcohol (PVA) and calcium chloride under aseptic conditions, and uniformly stirring; wherein the concentration of PVA in the mixed solution is 6.0% w/v, caCl 2 The concentration of the mixture solution is 2.0% w/v, the mixture solution is sterilized for 30min at the high pressure of 121 ℃ before being used, then the mixture solution is cooled to room temperature for standby, and the concentration of the bacterial suspension after the mixture solution is added is 3.0% v/v; then the obtained mixed solution is added into 50ml of 1.0% w/v Sodium Alginate (SA) solution drop by drop under intense stirring, at the moment, the solution becomes turbid and rapidly forms gel balls, and spherical PA/SA/PVA gel balls are prepared after filtration;
(4) Washing the PA/SA/PVA gel beads prepared in the step (3) with 20ml of sterile water three times, and then adding the washed PA/SA/PVA gel beads into 50ml of 2.0% CaCl 2 Solidifying in the solution for 2.5h, absorbing the surface moisture, and freezing in a refrigerator at-18 ℃ for 4h to obtain the PA/SA/PVA liquid core microcapsule;
(5) Adding 1ml of cooking waste oil (soybean oil left after frying the deep-fried dough sticks, which is subjected to aseptic treatment after filtering before use) into 99ml of inorganic culture medium to prepare inorganic salt culture solution containing the cooking waste oil;
(6) Adding the PA/SA/PVA liquid core microcapsule prepared in the step (4) into the inorganic salt culture solution containing the waste cooking oil prepared in the step (5) according to the mass ratio of 5% after thawing, and then placing the inorganic salt culture solution into a fermentation tank to perform shaking culture at the constant temperature of 150r/min for 72 hours at the temperature of 37 ℃ to prepare a rhamnolipid-containing fermentation liquor;
(7) Passing the rhamnolipid-containing fermentation broth prepared in step (6) through a ceramic filter membrane (30 cm in length, 3cm in diameter, and 0.08m in filtration area) with a pore size of 500nm 2 ) Removing larger proteins and insoluble impurities in the fermentation liquor to obtain micro-filtrate; 2ml of the micro-filtrate was obtained by the anthrone sulfuric acid method (Neissa M, pinzon L, kwang J. Analysis of rhamnolipid biosurfactants by methylene blue complexation [ J)]Applied Microbiology and Biotechnology,2009, 82:975-981.) the total concentration of rhamnolipids contained was determined and the results are shown in table 1;
(8) Adding NaOH into the micro-filtrate prepared in the step (7) to adjust the pH value to 9.0, and then passing the obtained alkaline micro-filtrate through an ultrafiltration membrane element (PES polyethersulfone material, length of 30cm, diameter of 5cm and filtration area of 0.4 m) 2 ) Washing the filter membrane with 5ml water for 3 times after the filtration is finished, and collecting the filtrate to prepare rhamnolipid solution; taking 1ml of rhamnolipid solution for LC-MS analysis to determine the types and the contents of the rhamnolipids; the results of LC-MS analysis indicate that the rhamnolipid prepared by the method provided by the invention comprises the rhamnolipid Rha-C 10 -C 10 Rhamnolipid Rha-C 10 -C 10 Rhamnolipid Rha-C 10 -C 12 (or Rha-Rha-C) 12 -C 10 ) These three rhamnolipids. The structural formula of Rhamnolipid (Rhamnolipid) is shown as follows:
in the above formula: r in double rhamnose double lipid 1 Is L-alpha-rhamnopyranosyl, R 2 Beta-hydroxydecanoic acid; r in mono-rhamnose double lipid 1 Is H, R 2 Beta-hydroxydecanoic acid; r in double rhamnose monolipid 1 Is L-alpha-rhamnopyranosyl, R 2 Is H; r in monostearyl monoesters 1 Is H, R 2 H.
LC-MS analysis is to determine that the main product in the fermented solution is rhamnolipid, but the total rhamnolipid content is the most significant for the method provided by the invention; therefore, this step can be omitted in the following embodiments;
(9) And (3) adding 0.05g of triethylamine and 0.02g of NaCl into the rhamnolipid solution prepared in the step (8), and finally adding water to prepare 250ml of water-soluble natural wax-dissolving agent.
Referring to the Chinese oil and gas industry standard SY/T5370-1999) surface and interfacial tension determination method, using Texas-500C ultra-low interfacial tension meter, setting the rotation speed to 8000r/min and the temperature to 60 ℃ of the oil reservoir to determine the interfacial tension between the prepared water-soluble natural wax-dissolving agent and crude oil; the results are shown in Table 3.
Wax-proofing performance test reference (Xu Yanli, et al, study of water-soluble wax-proofing agent FL-3 and field test [ J ]. Petrochemical applications, 2013, 32 (4): 37-40.) for wax-soluble agents was conducted in indoor experiments using the "reverse bottle wax-proofing assay" to evaluate water-soluble wax-proofing agents; the method comprises the following specific steps: adding 250ml of crude oil with 30% of water content into a weighed brine bottle, adding 25mg of the wax-dissolving agent prepared in the step (9), putting the brine bottle in a water bath, keeping the temperature at 75 ℃ for 30min, taking out the brine bottle by hand, keeping the temperature at 60 ℃ for 2.5h, taking out the brine bottle, reversely buckling the brine bottle on the wide-mouth bottle at the temperature, keeping the temperature until the brine bottle does not drip oil, weighing and calculating, and finally comparing the sample with a blank sample without the wax-dissolving agent to calculate the wax-preventing rate of the sample, wherein the result is shown in Table 3.
Example 2
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: omitting the step (8), and directly fixing the volume of the micro-filtrate prepared in the step (7) to prepare the water-soluble natural wax-dissolving agent; the rest of the procedure is the same as in example 1. The interfacial tension characterization procedure between the wax-dissolving agent and crude oil was the same as in example 1, and the results are shown in Table 3. The wax-dissolving agent obtained was subjected to the same wax-preventing performance test as in example 1, and the results are shown in Table 3.
Example 3
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: in the step (5), the cooking waste oil is 2ml, the inorganic culture medium is 98ml, the step (8) is omitted, and the micro-filtrate prepared in the step (7) is directly subjected to constant volume to prepare the water-soluble natural wax-dissolving agent; the rest of the procedure is the same as in example 1. The results of the total concentration of rhamnolipid contained in step (7) are shown in table 1. The interfacial tension characterization procedure between the wax-dissolving agent and crude oil was the same as in example 1, and the results are shown in Table 3. The wax-dissolving agent obtained was subjected to the same wax-preventing performance test as in example 1, and the results are shown in Table 3.
Example 4
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: in the step (5), the cooking waste oil is 4ml, the inorganic culture medium is 96ml, the step (8) is omitted, and the micro-filtrate prepared in the step (7) is directly subjected to constant volume to prepare the water-soluble natural wax-dissolving agent; the rest of the procedure is the same as in example 1. The results of the total concentration of rhamnolipid contained in step (7) are shown in table 1.
Example 5
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: in the step (5), the cooking waste oil is 6ml, the inorganic culture medium is 94ml, the step (8) is omitted, and the micro-filtrate prepared in the step (7) is directly subjected to constant volume to prepare the water-soluble natural wax-dissolving agent; the rest of the procedure is the same as in example 1. The results of the total concentration of rhamnolipid contained in step (7) are shown in table 1.
TABLE 1 comparison of total rhamnolipid concentration at different waste cooking oil concentrations
Waste cooking oil concentration (v/v)%) Total concentration of rhamnolipid (g/L)
Example 1 1 7.1
Example 3 2 15.3
Example 4 4 16.6
Example 5 6 17.8
It can be seen from Table 1 that the total concentration of rhamnolipids increases with increasing concentration of waste cooking oil, taking into account that the comprehensive utilization of the carbon source is optimal when the concentration of waste cooking oil is 2 v/v%.
Example 6
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: in the step (5), the cooking waste oil is changed into waste glycerol (waste oil bath in a laboratory is subjected to filtration and aseptic treatment before use), the consumption of the waste glycerol is 2ml, and the consumption of the inorganic culture medium is 98ml; omitting the step (8), and directly fixing the volume of the micro-filtrate prepared in the step (7) to prepare the water-soluble natural wax-dissolving agent; the rest of the procedure is the same as in example 1. The results of the total concentration of rhamnolipid contained in step (7) are shown in table 2.
Example 7
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: in the step (5), the cooking waste oil is changed into kitchen waste grease (sold in the market, and filtration and aseptic treatment are needed before use), the amount of the kitchen waste grease is 2ml, and the amount of the inorganic culture medium is 98ml; omitting the step (8), and directly fixing the volume of the micro-filtrate prepared in the step (7) to prepare the water-soluble natural wax-dissolving agent; the rest of the procedure is the same as in example 1. The results of the total concentration of rhamnolipid contained in step (7) are shown in table 2.
Example 8
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: in the step (5), the cooking waste oil is changed into vegetable oil (commercial soybean oil which needs to be subjected to aseptic treatment before use), the vegetable oil dosage is 2ml, and the inorganic culture medium dosage is 98ml; omitting the step (8), and directly fixing the volume of the micro-filtrate prepared in the step (7) to prepare the water-soluble natural wax-dissolving agent; the rest of the procedure is the same as in example 1. The results of the total concentration of rhamnolipid contained in step (7) are shown in table 2.
Example 9
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: in the step (5), the cooking waste oil is changed into waste gasoline (commercially available, which is subjected to aseptic treatment before use), wherein the consumption of the waste gasoline is 2ml, and the consumption of the inorganic culture medium is 98ml; omitting the step (8), and directly fixing the volume of the micro-filtrate prepared in the step (7) to prepare the water-soluble natural wax-dissolving agent; the rest of the procedure is the same as in example 1. The results of the total concentration of rhamnolipid contained in step (7) are shown in table 2.
TABLE 2 comparison of total rhamnolipid concentrations using different carbon sources
Carbon source Total concentration of rhamnolipid (g/L)
Example 3 Waste cooking oil 15.3
Example 6 Waste glycerol 8.2
Example 7 Kitchen waste grease 14.6
Example 8 Vegetable oil 15.8
Example 9 Waste gasoline 12.5
It can be seen from table 1 that the different carbon sources have a great influence on the total concentration of rhamnolipids, but the total concentration of rhamnolipids produced by fermentation of waste cooking oil, kitchen waste oil and vegetable oil as carbon sources is not greatly different. Therefore, the method provided by the invention can completely utilize the cooking waste oil and the kitchen waste grease to prepare the water-soluble wax-dissolving agent, thereby changing waste into valuable and reducing the production cost of the wax-dissolving agent.
Example 10
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: in the step (5), the cooking waste oil is 2ml, the inorganic culture medium is 98ml, the step (8) is omitted, and in the step (9), triethylamine is replaced by ethanolamine; the rest of the procedure is the same as in example 1. The interfacial tension characterization procedure between the wax-dissolving agent and crude oil was the same as in example 1, and the results are shown in Table 3. The wax-dissolving agent obtained was subjected to the same wax-preventing performance test as in example 1, and the results are shown in Table 3.
Example 11
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: in the step (5), the cooking waste oil is 2ml, the inorganic culture medium is 98ml, the step (8) is omitted, and in the step (9), triethylamine is replaced by ammonia water; the rest of the procedure is the same as in example 1. The interfacial tension characterization procedure between the wax-dissolving agent and crude oil was the same as in example 1, and the results are shown in Table 3. The wax-dissolving agent obtained was subjected to the same wax-preventing performance test as in example 1, and the results are shown in Table 3.
Example 12
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: in the step (5), the cooking waste oil is 2ml, the inorganic culture medium is 98ml, the step (8) is omitted, and in the step (9), triethylamine is replaced by sodium hydroxide solution; the rest of the procedure is the same as in example 1. The interfacial tension characterization procedure between the wax-dissolving agent and crude oil was the same as in example 1, and the results are shown in Table 3. The wax-dissolving agent obtained was subjected to the same wax-preventing performance test as in example 1, and the results are shown in Table 3.
Example 13
The procedure for the preparation of the wax-dissolving agent is described in reference to example 1, with the difference that: in the step (5), the cooking waste oil is 2ml, the inorganic culture medium is 98ml, the step (8) is omitted, and no alkali is added in the step (9); the rest of the procedure is the same as in example 1. The interfacial tension characterization procedure between the wax-dissolving agent and crude oil was the same as in example 1, and the results are shown in Table 3. The wax-dissolving agent obtained was subjected to the same wax-preventing performance test as in example 1, and the results are shown in Table 3.
TABLE 3 comparison of Water-soluble Natural wax-dissolving Agents Using different bases
Alkali Interfacial tension (10) -3 mN/m) Wax control rate (%)
Example 1 Triethylamine 20.1 67
Example 2 Triethylamine 19.6 65
Example 10 Ethanolamine 2.2 93
Example 11 Ammonia water 97.8 46
Example 12 Sodium hydroxide 90.5 49
Example 13 N/A 324.7 38
It can be seen from table 3 that the interfacial tension and wax-preventing rate of the water-soluble natural wax-dissolving agent provided by the invention are both affected by the type of the added alkali, wherein the effect of the used organic alkali is obviously better than that of inorganic alkali, and the wax-preventing rate can be as high as 93% when the ethanolamine is used as alkali. It should be noted that the results in example 2 show that the rhamnolipid fermentation broth prepared by the PA/SA/PVA liquid core microcapsule fermentation method provided by the present invention can be used for preparing the water-soluble natural wax dissolving agent by simply filtering with a ceramic filter membrane.
Example 14
Step for preparing wax-dissolving agent reference example 10. The wax-dissolving agent obtained was tested for wax-inhibiting properties by referring to example 1, except that: the test was performed using crude oil having a water content of 50%, and the results are shown in table 4.
Example 15
Step for preparing wax-dissolving agent reference example 10. The wax-dissolving agent obtained was tested for wax-inhibiting properties by referring to example 1, except that: the test was performed using crude oil having a water content of 60%, and the results are shown in table 4.
Example 16
Step for preparing wax-dissolving agent reference example 10. The wax-dissolving agent obtained was tested for wax-inhibiting properties by referring to example 1, except that: the test was performed using crude oil having a water content of 70%, and the results are shown in table 4.
TABLE 4 wax control of water soluble natural wax solubles in crude oils of varying water content
Crude oil moisture content (%) Wax control rate (%)
Example 10 30 93
Example 14 50 95
Example 15 60 94
Example 16 70 94
From table 4, it can be seen that the interfacial wax control rate of the water-soluble natural wax-dissolving agent provided by the invention slightly increases to 94% or more with the increase of the water content of crude oil.
Example 17
The PA/SA/PVA core microcapsules used in example 3 were recovered and reused, and each use was cleaned with sterile water. The procedure for the preparation of rhamnolipids is as in example 1, repeated 9 times with the total concentration of rhamnolipids results shown in figure 1. As can be seen from fig. 1: after the PA/SA/PVA liquid core microcapsule provided by the invention is continuously cultured for 10 times, the total concentration of rhamnolipid in the obtained fermentation broth can still reach 8.3g/L, which shows that the microcapsule breakage number is small in the repeated experiment process.

Claims (7)

1. The preparation method of the water-soluble natural wax-dissolving agent comprises the steps of (by mass ratio) obtaining rhamnolipid in the water-soluble natural wax-dissolving agent of 0.1-1.0%; the mass ratio of the alkali is 0.5-2.0%, the mass ratio of the inorganic salt is 0.1-1.0%, and the balance is water; the method is characterized by comprising the following steps of:
(1) Inoculating pseudomonas aeruginosa on a nutrient gravy agar medium, performing constant-temperature activation, and transferring the strain subjected to the constant-temperature activation into a seed solution for culture to obtain a seed solution containing the pseudomonas aeruginosa;
(2) Adding the bacterial sludge obtained after removing the supernatant of the seed liquid prepared in the step (1) into sterile physiological saline to prepare bacterial suspension;
(3) Adding the bacterial suspension prepared in the step (2) into a mixed solution composed of polyvinyl alcohol and calcium chloride, stirring uniformly, then adding the obtained mixed solution into a sodium alginate solution dropwise, and filtering to obtain spherical sodium alginate/polyvinyl alcohol gel balls, wherein the mass ratio of polyvinyl alcohol, calcium chloride and sodium alginate in the mixed solution composed of polyvinyl alcohol and calcium chloride is 1:1:2;
(4) Washing the sodium alginate/polyvinyl alcohol gel ball prepared in the step (3), then placing the washed gel ball into a calcium chloride solution for solidification, removing water on the surface of the solidified gel ball, and then freezing to prepare the sodium alginate/polyvinyl alcohol liquid core microcapsule;
(5) Adding a carbon source into an inorganic culture medium to prepare an inorganic salt culture solution containing the carbon source;
(6) Adding the sodium alginate/polyvinyl alcohol liquid core microcapsule prepared in the step (4) into the inorganic salt culture solution containing carbon sources prepared in the step (5) for constant temperature culture to prepare fermentation liquor containing rhamnolipid;
(7) Passing the rhamnolipid-containing fermentation liquor prepared in the step (6) through a ceramic filter membrane to remove larger proteins and insoluble impurities in the fermentation liquor, so as to prepare micro-filtrate;
(8) Adding alkali and inorganic salt into the micro-filtrate prepared in the step (7) to prepare the water-soluble natural wax-dissolving agent; the alkali is one of ethanolamine or triethylamine; the inorganic salt is one of sodium sulfate or sodium chloride.
2. The method for preparing a water-soluble natural wax-dissolving agent according to claim 1, wherein the constant temperature activation temperature in the step (1) is 37 ℃; the constant temperature activation time was 24h.
3. The method for preparing a water-soluble natural wax-dissolving agent according to claim 1, wherein the freezing temperature in the step (4) is-18 ℃ and the freezing time is 4-6h.
4. The method for preparing a water-soluble natural wax-dissolving agent according to claim 1, wherein the carbon source in the step (5) is one selected from the group consisting of waste cooking oil, glycerin and vegetable oil.
5. The method for preparing a water-soluble natural wax-dissolving agent according to claim 1, wherein the constant temperature culture temperature in the step (6) is 37 ℃ and the constant temperature culture time is 24 hours.
6. The method for preparing a water-soluble natural wax-dissolving agent according to claim 1, wherein the pore diameter of the ceramic filter membrane in the step (7) is 50 nm-1000 nm.
7. Use of the water-soluble natural wax-dissolving agent prepared by the preparation method of the water-soluble natural wax-dissolving agent according to any one of claims 1 to 6 in wax control and paraffin removal of petroleum pipelines.
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