An extraction method and application of biomass surfactant for dispersing oil spills
By using a freeze-thaw-ultrasonic disruption method to break down diatoms, surfactants are extracted from diatom biomass, solving the problems of poor biodegradability of chemically synthesized agents and high energy consumption in cell disruption in existing oil spill treatments, thus achieving efficient and environmentally friendly oil spill dispersion treatment.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- DALIAN MARITIME UNIVERSITY
- Filing Date
- 2026-01-13
- Publication Date
- 2026-06-09
Smart Images
Figure CN122168303A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of surfactant technology, specifically relating to an extraction method and application of a biomass-based surfactant for dispersing oil spills. Background Technology
[0002] With the continuous development of the petrochemical industry, oil spills caused by oil exploration, extraction, and transportation activities occur frequently, posing a serious threat to the marine ecological environment. Traditional oil spill treatment methods mostly use chemical dispersants, which are sprayed onto the water surface at the site of the oil spill to quickly disperse the floating oil into small droplets, thereby accelerating the migration and transformation of the droplets under the influence of waves and other forces. One type of nonionic dispersant, GM-2, primarily uses fatty alcohol polyoxyethylene ethers as its surfactant, which is often difficult to completely degrade and has long-term toxic effects on marine aquatic plants and animals.
[0003] Most existing oil spill dispersants rely on chemically synthesized surfactants, which suffer from poor biodegradability, significant potential toxicity to marine ecosystems, and the risk of secondary pollution during oil spill response. While microalgal surfactants possess excellent biodegradability and environmental friendliness, current preparation methods are limited by low efficiency and high cost. Existing cell disruption methods, such as high-pressure homogenization or solvent extraction, often suffer from high energy consumption, high cost, or the potential to denature the target material. In recent years, biosurfactants have attracted attention due to their superior biodegradability and environmental compatibility. Bioremediation offers advantages such as low cost, no secondary pollution, and short remediation time. Furthermore, applying marine phytoplankton such as microalgae to bioremediation can help control harmful ecological phenomena like red tides and improve the availability of microalgae.
[0004] Therefore, developing a physical, gentle, and efficient cell disruption method is key to improving the extraction efficiency of surfactant biomass. For example, utilizing the synergistic effect of freeze-thaw cycles and ultrasound to disrupt microorganisms with hard siliceous cell walls, such as diatoms, is considered a highly promising technological approach. Summary of the Invention
[0005] The purpose of this invention is to provide an extraction method and application of biomass-based surfactants for dispersing oil spills. This extraction method is low in cost, simple in process, environmentally friendly and safe. Furthermore, the biomass-based surfactant has the advantages of thorough degradation and being non-toxic and environmentally friendly.
[0006] To address the shortcomings mentioned in the background art, this invention provides a method for extracting biomass-based surfactants for dispersing oil spills. The method involves extracting surfactant biomass from diatom biomass through freeze-thaw and ultrasonic disruption, which is then used for dispersing and treating oil spills.
[0007] According to one aspect of the present invention, a method for extracting biomass-based surfactants for dispersing oil spills is provided, comprising the following steps: Step 1, Diatom Cultivation: Inoculate *Phaeodactylum tricornutum* into artificial seawater with a salinity of 30‰, add F / 2 nutrient solution, and cultivate in a constant temperature and light incubator to obtain *Phaeodactylum tricornutum* in the stable concentration period; Step 2, freeze-thaw-ultrasonic disruption: Place the stable-concentration *Phaeodactylum tricornutum* obtained in Step 1 in a freezer and then thaw it in a water bath. Repeat the freeze-thaw process 1-3 times to break up the diatom cells and obtain the freeze-thawed diatoms. Then, use an ultrasonic disruptor to disrupt the diatoms and obtain diatoms containing biomass surfactants.
[0008] Based on the above technical solution, the volume mixing ratio of the triangular brown finger algae and the artificial seawater with a salinity of 30‰ in step 1 is 500~1000ml:3000ml, wherein the artificial seawater with a salinity of 30‰ is sterilized at 100℃ for 20~30min.
[0009] Based on the above technical solution, the addition of F / 2 nutrient solution in step 1 is specifically as follows: F / 2 nutrient solution is added at a volume ratio of 1:1000 every four days.
[0010] Based on the above technical solution, the cultivation conditions in step 1 are as follows: The culture temperature is 19~21℃; The light intensity for cultivation was 3000 lx, simulating sunlight; The light-dark cycle of the culture was 14h:10h.
[0011] Based on the above technical solution, the concentration of *Phaeodactylum tricornutum* during the concentration stabilization period in step 1 is 3~4.5×10⁻⁶. 6 cells / mL.
[0012] Based on the above technical solution, the freezing conditions in step 2 are as follows: The freezing temperature is -21 to -19°C; The freezing time is 11-13 hours.
[0013] Based on the technical solution, the thawing conditions in step 2 are as follows: The thawing temperature is 29~31℃; The thawing time is 20-30 minutes.
[0014] Based on the above technical solution, the specific crushing process in step 2 is as follows: place the ultrasonic crusher below the surface of the diatomaceous earth liquid so that it is located in the middle of the diatomaceous earth liquid, and crush it with ultrasonic power of 80~320W for 14~16 minutes under ice bath conditions.
[0015] Based on the above technical solution, the lipid content of the diatom containing biomass-based surfactant in step 2 is 62.5~99.1 mg / L, and the protein content is 81.4~119.4 mg / L.
[0016] According to another aspect of the present invention, the application of a biomass-based surfactant extracted using a biomass-based surfactant extraction method in dispersing oil spills is provided, comprising the following steps: Diatoms containing biomass-based surfactants were mixed with seawater with a salinity of 30‰, and Iranian heavy oil was added. The mixture was shaken, and the oil-water interfacial tension was changed through adsorption, thus dispersing the spilled oil into oil droplets. Based on the above technical solution, the volume mixing ratio of the diatom containing biomass-type surfactant to seawater with a salinity of 30‰ is 90~130ml:270~310ml.
[0017] The mass ratio of the Iranian heavy oil to seawater with a salinity of 30‰ is 0.19~0.21g: 276750~312625g.
[0018] The conditions for the oscillation are as follows: The rotational speed of the oscillation is 155~165 rpm; The oscillation temperature is 20~21℃; The oscillation time is 115~125 min.
[0019] Based on the above technical solution, the oil-water interfacial tension is 15.96~17.77 mN / m; The dispersion rate of the spilled oil was 45.19%~56.95%; The average volumetric size of the oil droplets formed by the oil spill dispersion is 71.36~84.45 μm.
[0020] Beneficial effects (1) The technical solution disclosed in this invention combines freeze-thaw cycles with ultrasonic disruption, which synergistically enhances the efficient release of intracellular substances from *Phaeodactylum tricornutum*. During freeze-thaw, water inside and outside the cells freezes to form ice crystals. These expanding ice crystals physically pierce or break through the cell wall and cell membrane structure, forming initial damage channels. Subsequent ultrasonic disruption utilizes the high-pressure shock waves and microjets generated by its cavitation effect to further tear and break the already damaged cell wall, thereby fully releasing intracellular lipids, proteins, and other surface-active substances into the algal solution. Compared with a single disruption method, this combined approach has higher disruption efficiency and better maintains the activity of surface-active biomass under ice bath conditions, improving the extraction efficiency of surface-active biomass and reducing production costs.
[0021] (2) The technical solution disclosed in this invention utilizes diatoms, a rich biomass resource with wide availability and renewability. The resulting microalgal surface-active biomass has good biodegradability and environmental friendliness, is non-toxic to marine ecosystems, and can effectively avoid the secondary pollution problems caused by traditional chemical oil dispersants. Attached Figure Description
[0022] Figure 1 This is the result of the oil spill dispersion rate test in an application example of the present invention; Figure 2 The results of oil-water interfacial tension testing are shown in an application example of this invention. Figure 3 This shows the volume-average particle size of the dispersed oil droplets in an application example of the present invention; Figure 4 This shows the particle size of the dispersed oil droplets in an application example of the present invention. Detailed Implementation
[0023] To make the objectives and technical solutions of this invention clearer, the following embodiments are provided for further explanation. However, the scope of protection of this invention is not limited to these embodiments; the embodiments are merely for illustrative purposes. Those skilled in the art should understand that any changes or equivalent substitutions that do not depart from the concept of this invention are included within the scope of protection of this invention.
[0024] Unless otherwise specified, all reagents and raw materials used in this invention are obtained through purchase.
[0025] The selection and cultivation of diatoms in this embodiment of the invention are as follows: Before cultivation, 3000 ml of artificial seawater with a salinity of 30‰ is sterilized at high temperature. After cooling to room temperature, 0.5-1 L of diatoms (Phaeodactylum tricornutum, obtained from the Liaoning Provincial Marine and Fisheries Research Institute) are inoculated. Then, every four days, F / 2 nutrient solution (purchased from Dalian Zhenyang Biotechnology Co., Ltd.) is added to the diatom solution at a volume ratio of 1:1000 (F / 2 nutrient solution to inoculated diatoms). The algal species name and cultivation time are labeled, and the solution is placed in a constant temperature and light incubator at a temperature of 20(±1)℃ and a light intensity of 3000 lx to simulate sunlight, with a light-dark cycle of 14h:10h. After self-propagation of diatoms, the conical flask is shaken three times daily to prevent diatom cells from adhering to the bottom of the flask and growing or even dying in large numbers, thus obtaining an algal solution with a stable concentration.
[0026] Example 1: Take 300 ml of *Phaeodactylum tricornutum* culture that has reached a stable concentration (concentration 4 × 10⁻⁶). 6Cell disruption was performed using cells / mL. The diatoms were frozen at -20°C for 12 hours. The completely frozen diatom solution was then removed and thawed in a 30°C water bath for 20-30 minutes. The frozen solution was then placed in an ultrasonic disruptor (80W, probe in the middle of the liquid) and sonicated for 15 minutes in an ice bath. The treated diatoms were then mixed with seawater (salinity 30‰) at a volume ratio of 100ml:300ml to obtain 1×10⁻⁶ cells / mL. 6 cells / mL diatomaceous earth solution (lipid content 62.5 mg / L, protein content 81.4 mg / L of surfactant biomass).
[0027] Example 2: Take 300 ml of *Phaeodactylum tricornutum* culture that has reached a stable concentration (concentration 4 × 10⁻⁶). 6 Cell disruption was performed using a method involving cell / mL ratio. The cell disruption step included: freezing the diatomaceous liquid at -20°C for 12 hours, followed by thawing in a 30°C water bath for 20-30 minutes; repeating this freeze-thaw process three times; then ultrasonically disrupting the liquid after the three freeze-thaw treatments under ice bath conditions at an ultrasonic power of 80W for 15 minutes; finally, mixing the treated diatoms with seawater of salinity 30‰ at a volume ratio of 100ml:300ml to obtain 1×10⁻⁶ cells / mL. 6 cells / mL diatomaceous earth solution (surface active biomass with lipid content of 82.6 mg / L and protein content of 101.6 mg / L).
[0028] Example 3: The difference from Example 1 is that the ultrasonic power was 320W. The treated diatoms were mixed with seawater with a salinity of 30‰ at a volume ratio of 100ml:300ml to obtain 1×10⁻⁶ diatoms. 6 cells / mL diatomaceous earth solution (lipid content 92.3 mg / L, protein content 115.2 mg / L of surfactant biomass).
[0029] Example 4: The difference from Example 2 is that the ultrasonic power was 320W. The treated diatoms were mixed with seawater with a salinity of 30‰ at a volume ratio of 100ml:300ml to obtain 1×10⁻⁶ diatoms. 6 cells / mL diatomaceous earth solution (surface active biomass with lipid content of 99.1 mg / L and protein content of 119.4 mg / L).
[0030] Comparative Example 1 The difference from Example 1 is that the diatomaceous earth solution was only subjected to ultrasonic treatment at a power of 80W. The treated diatoms were then mixed with seawater of salinity 30‰ at a volume ratio of 100 ml:300 ml to obtain 1×10⁻⁶ diatoms. 6 cells / mL diatomaceous earth solution (lipid content 47.8 mg / L, protein content 68.3 mg / L of surfactant biomass).
[0031] Comparative Example 2 The difference from Example 1 is that only a freeze-thaw process was performed, and the number of freeze-thaw cycles was one. The treated diatoms were mixed with seawater with a salinity of 30‰ at a volume ratio of 100ml:300ml to obtain 1×10⁻⁶ diatoms. 6 cells / mL diatomaceous earth solution (lipid content 31.1 mg / L, protein content 57.0 mg / L of surfactant biomass).
[0032] Application examples The 1×10 prepared in Examples 1-4 6 A mixture of diatoms (cells / mL) and seawater was placed in a baffled conical flask, and 0.2 g of Iranian heavy oil (API specific gravity 24.01, sulfur content 3.51 W%, purchased from Hengli Petrochemical Co., Ltd.) was added sequentially. The mixture was then oscillated for 120 min at 20℃ and 160 rpm to simulate a marine environment. The dispersion rate of the biomass-based surfactant in the diatom mixture on the Iranian heavy oil was tested (see...). Figure 1 Oil-water interfacial tension (see...) Figure 2 ), the average volumetric size of the dispersed oil droplets (see Figure 3 ) and the size of the dispersed oil droplets (see Figure 4 ),Depend on Figures 1-4 It is known that, under the same number of freeze-thaw cycles, increasing the ultrasonic power helps to more fully break down the cell wall, thereby releasing more intracellular lipids and proteins and increasing the yield of surface-active biomass.
[0033] The above description is merely a few embodiments of this application and is not intended to limit this application in any way. Although this application discloses preferred embodiments as described above, it is not intended to limit this application. Any changes or modifications made by those skilled in the art without departing from the scope of the technical solution of this application using the disclosed technical content are equivalent to equivalent implementation cases and fall within the scope of the technical solution.
Claims
1. A method for extracting biomass-based surfactants for dispersing oil spills, characterized in that, Includes the following steps: Step 1, Diatom Cultivation: Inoculate *Phaeodactylum tricornutum* into artificial seawater with a salinity of 30‰, add F / 2 nutrient solution, and cultivate in a constant temperature and light incubator to obtain *Phaeodactylum tricornutum* in the stable concentration period; Step 2, Freeze-thaw-ultrasonic disruption: The *Phaeodactylum tricornutum* obtained in Step 1, during its concentration stabilization period, was frozen in a refrigerator and then thawed in a water bath. This freeze-thaw cycle was repeated 1-3 times to rupture the diatom cells, yielding freeze-thawed diatoms. These diatoms were then disrupted using an ultrasonic disruptor to obtain diatoms containing biomass-based surfactants. The treated diatoms were mixed with seawater with a salinity of 30‰ at a volume ratio of 90-130 ml: 270-310 ml to obtain 1×10⁻⁶ diatoms. 6 cells / mL diatomaceous earth solution.
2. The extraction method according to claim 1, characterized in that, The volume ratio of the *Phaeodactylum tricornutum* and the artificial seawater with a salinity of 30‰ in step 1 is 500-1000 ml: 3000 ml, wherein the artificial seawater with a salinity of 30‰ is sterilized at 100℃ for 20-30 min. The addition of F / 2 nutrient solution in step 1 is specifically as follows: F / 2 nutrient solution is added at a volume ratio of 1:1000 every four days. The cultivation conditions described in step 1 are as follows: The culture temperature is 19~21℃; The light intensity for cultivation was 3000 lx; The light-dark cycle of the culture was 14h:10h; The concentration of *Phaeodactylum tricornutum* during the stable concentration period described in step 1 is 3~4.5 × 10⁻⁶. 6 cells / mL.
3. The extraction method according to claim 1, characterized in that, The freezing conditions in step 2 are as follows: The freezing temperature is -21 to -19°C; The freezing time is 11-13 hours.
4. The extraction method according to claim 1, characterized in that, The thawing conditions described in step 2 are as follows: The thawing temperature is 29~31℃; The thawing time is 20-30 minutes.
5. The extraction method according to claim 1, characterized in that, The specific crushing process in step 2 is as follows: place the ultrasonic crusher below the surface of the diatomaceous earth solution so that it is in the middle of the diatomaceous earth solution, and crush it with ultrasonic power of 80~320W for 14~16 minutes under ice bath conditions.
6. The extraction method according to claim 1, characterized in that, The step 2 involves a biomass-based surfactant at a concentration of 1×10⁻⁶. 6 The lipid content in diatoms with a cell / mL concentration ranges from 62.5 to 99.1 mg / L, and the protein content ranges from 81.4 to 119.4 mg / L.
7. The application of a biomass-based surfactant extracted using the extraction method according to any one of claims 1 to 6 in dispersing oil spills, characterized in that, Includes the following steps: Diatoms containing biomass-based surfactants were mixed with seawater with a salinity of 30‰, and Iranian heavy oil was added. The mixture was shaken, and the oil-water interfacial tension was changed through adsorption, thus dispersing the spilled oil into oil droplets.
8. The application according to claim 7, characterized in that, The volume mixing ratio of the diatom containing biomass-based surfactants to seawater with a salinity of 30‰ is 90~130ml: 270~310ml. The mass ratio of the Iranian heavy oil to seawater with a salinity of 30‰ is 0.19~0.21g: 276750~312625g.
9. The application according to claim 7, characterized in that, The conditions for the oscillation are as follows: The rotational speed of the oscillation is 155~165 rpm; The oscillation temperature is 20~21℃; The oscillation time is 115~125 min.
10. The application according to claim 7, characterized in that, The oil-water interfacial tension is 15.96~17.77 mN / m; The dispersion rate of the oil spill was 45.19%~56.95%; The average volumetric size of the oil droplets formed by the oil spill dispersion is 71.36~84.45 μm.