Microorganism capture liquid, preparation method and application in low-viscosity fuel

Abstract

The present invention provides a highly stable microorganism capture liquid, a preparation method and an application in low-viscosity fuel. The microorganism capture liquid comprises, by mass percentage, ≥90% water, 20mM-200mM 4-hydroxyethylpiperazineethanesulfonic acid, 0.08%-0.12% Tween, 0.12%-0.25% quaternary ammonium surfactant, 0.25%-0.4% Triton, 0.02%-0.10% benzalkonium bromide, 0.2%-0.4% polyether polyol, totaling 100%, wherein the microorganism capture liquid is used to capture microorganisms from an oil phase with high stability.

Description

Technical Field

[0001] The present invention relates to the field of microbial detection, and in particular to a high-stability microbial capture liquid, a preparation method and an application thereof in low-viscosity fuel. Background Art

[0002] When low kinematic viscosity oil is contaminated by microorganisms, it will cause filter blockage, oil gauge failure and equipment corrosion, which seriously threatens transportation safety. Therefore, it is of great significance to detect low kinematic viscosity oil. Common methods for detecting microorganisms in low kinematic viscosity oil include indirect detection method, culture method and equipment rapid detection method. The indirect detection method indirectly judges whether the oil system is contaminated by microorganisms by changes in indicators such as oil appearance, surface tension and chemical composition. Since its indicators cannot be quantified, it has not been widely used; the culture method requires a strict sterile operating environment, skilled operating skills and cumbersome operating procedures, which usually takes 3 to 7 days and cannot meet the requirements of on-site testing; the equipment rapid detection method has a short detection time and can quickly detect whether the sample is contaminated by microorganisms. Among them, the adenosine triphosphate (ATP) bioluminescence method is widely used due to its faster response speed and higher sensitivity.

[0003] In the process of realizing the concept of the present invention, the inventors found that when the capture solution used to capture microorganisms in the oil phase is used for ATP bioluminescence detection, the relative light unit (RLU) value of the test is greatly affected by time. Summary of the invention

[0004] In view of this, the present invention provides a highly stable microorganism capture liquid, a preparation method and an application of the microorganism capture liquid in low-viscosity fuel.

[0005] The first aspect of the present invention provides a microorganism capture liquid, which includes, by mass percentage: ≥90% water, 4-hydroxyethylpiperazineethanesulfonic acid with a concentration of 20mM~200mM, 0.08%~0.12% Tween, 0.12%~0.25% quaternary ammonium salt surfactant, 0.25%~0.4% Triton, 0.02%~0.10% benzalkonium bromide, 0.2%~0.4% polyether polyol, totaling 100%, wherein the microorganism capture liquid is used to capture microorganisms from an oil phase with high stability.

[0006] According to an embodiment of the present invention, the polyether polyol includes at least one of the following: poloxamer 188, poloxamer 407, and a block copolymer of propylene oxide and polyethylene oxide.

[0007] According to an embodiment of the present invention, the concentration of 4-hydroxyethylpiperazineethanesulfonic acid is 200 mM, and the water includes sterile water.

[0008] According to an embodiment of the present invention, Tween includes at least one of the following: Tween 20, Tween 40, Tween 60, Tween 80, and Tween 85.

[0009] According to an embodiment of the present invention, the quaternary ammonium salt surfactant includes at least one of the following: hexadecyltrimethylammonium bromide and hexadecyltrimethylammonium chloride.

[0010] According to an embodiment of the present invention, the pH value of the microorganism capture liquid is 7.0-9.0.

[0011] The second aspect of the present invention provides a method for preparing a microorganism capture liquid, comprising: preparing a 4-hydroxyethylpiperazineethanesulfonic acid solution with a concentration of 20mM~200mM; dissolving 0.08%~0.12% by mass of Tween, 0.12%~0.25% by mass of a quaternary ammonium salt surfactant, 0.25%~0.4% by mass of Triton, and 0.2%~0.4% by mass of a polyether polyol in the 4-hydroxyethylpiperazineethanesulfonic acid solution in order to obtain a mixed solution; and sterilizing the mixed solution to obtain a microorganism capture liquid.

[0012] According to an embodiment of the present invention, the mixed solution is sterilized to obtain the microorganism-capturing solution, including: heating the mixed solution in a 60° C. water bath for 30 minutes to obtain the microorganism-capturing solution.

[0013] The third aspect of the present invention provides an application of a microorganism capture liquid in fuel, wherein the microorganism capture liquid is used to capture microorganisms in the fuel so as to detect the microorganisms in the fuel using the adenosine triphosphate bioluminescence method; the microorganism capture liquid is the above-mentioned microorganism capture liquid or the microorganism capture liquid prepared by the above-mentioned preparation method.

[0014] According to an embodiment of the present invention, the kinematic viscosity of the fuel is ≥1.3 mm 2 ·s -1 , and ≤24.0mm 2 ·s -1 .

[0015] According to an embodiment of the present invention, cationic surfactants such as quaternary ammonium surfactants and benzalkonium bromide in the microorganism capture liquid can act on the negative charge of the microorganism cell wall to enhance the capture efficiency of the microorganism, and nonionic surfactants such as Tween, Triton and polyether polyols can reduce the oil-water interfacial tension, promote oil-water mixing, and are conducive to the contact of components such as quaternary ammonium surfactants and benzalkonium bromide with the microorganism in the microorganism capture liquid, further improving the capture efficiency. In addition, 4-hydroxyethylpiperazineethanesulfonic acid is a biological buffer that can maintain a constant pH value in a wide pH range, provides a suitable living environment for microorganisms, and helps to maintain the activity and stability of microorganisms. In addition, polyether polyols can also improve the hydrolysis resistance of the capture liquid, etc. Due to the synergistic effect between 4-hydroxyethylpiperazineethanesulfonic acid, Tween, quaternary ammonium surfactants, Triton, benzalkonium bromide and polyether polyol components, that is, through the synergistic effect of multiple action mechanisms such as maintaining a suitable pH environment, reducing oil-water interfacial tension, interacting with the negative charge of the microbial cell wall, and resisting hydrolysis, it is possible to maintain the microbial capture efficiency of the microbial capture liquid while providing longer-lasting microbial capture performance, thereby improving the stability of the capture system. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The above and other objects, features and advantages of the present invention will become more apparent through the following description of the embodiments of the present invention with reference to the accompanying drawings, in which:

[0017] Figure 1 The diagram is a diagram showing the effect of the microorganism capture liquid obtained according to Example 1 of the present invention and the commercial capture liquid on the detection of microorganisms in fuel. DETAILED DESCRIPTION

[0018] Below, embodiments of the present invention will be described with reference to the accompanying drawings. However, it should be understood that these descriptions are exemplary only and are not intended to limit the scope of the present invention. In the following detailed description, for ease of explanation, many specific details are set forth to provide a comprehensive understanding of embodiments of the present invention. However, it is apparent that one or more embodiments may also be implemented without these specific details. In addition, in the following description, descriptions of known structures and technologies are omitted to avoid unnecessary confusion of concepts of the present invention.

[0019] The terms used herein are only for describing specific embodiments and are not intended to limit the present invention. The terms "comprise", "include", etc. used herein indicate the existence of features, steps, operations and / or components, but do not exclude the existence or addition of one or more other features, steps, operations or components.

[0020] All terms (including technical and scientific terms) used herein have the meanings commonly understood by those skilled in the art unless otherwise defined. It should be noted that the terms used herein should be interpreted as having a meaning consistent with the context of this specification and should not be interpreted in an idealized or overly rigid manner.

[0021] When using expressions such as "at least one of A, B, and C, etc.", they should generally be interpreted according to the meaning of the expression commonly understood by those skilled in the art (for example, "a system having at least one of A, B, and C" should include but is not limited to a system having A alone, B alone, C alone, A and B, A and C, B and C, and / or A, B, C, etc.).

[0022] In the related technology, the ATP bioluminescence method for low-viscosity oil microbial detection generally has the following three steps: oil microbial extraction, microbial ATP extraction, and ATP detection. Before measuring the ATP of microorganisms, ATP needs to be extracted from the microorganisms. The extraction efficiency of ATP directly affects the biomass determination results. The method of extracting microbial ATP has gone through the process from heating and boiling to using ATP extractant. The principle of ATP detection is: luciferin, as an oxidizable substrate, combines with luciferase and ATP in the presence of magnesium ions to form a luciferase-luciferin-adenosine monophosphate (AMP) complex, which then combines with oxygen to emit light. The intensity of the light emitted is positively correlated with the ATP content at a certain concentration. For living microorganisms within a certain growth period, there is a relatively constant level of ATP content, so that there is a good linear relationship between ATP concentration and living microbial content. Therefore, the microbial content in the sample can be indirectly determined by detecting the fluorescence intensity generated by ATP during the reaction.

[0023] In the process of realizing the concept of the present invention, the inventors found that when the capture solution used to capture microorganisms in the oil phase is used for ATP bioluminescence detection, the relative light unit (RLU) value of the test is greatly affected by time.

[0024] Based on this, the first aspect of the present invention provides a highly stable microorganism capture liquid, a preparation method and an application in low-viscosity fuel. The microorganism capture liquid comprises, by mass percentage: ≥90% water, 20mM~200mM 4-hydroxyethylpiperazineethanesulfonic acid, 0.08%~0.12% Tween, 0.12%~0.25% quaternary ammonium salt surfactant, 0.25%~0.4% Triton, 0.02%~0.10% benzalkonium bromide, 0.2%~0.4% polyether polyol, totaling 100%, and the microorganism capture liquid is used to capture microorganisms from the oil phase.

[0025] According to an embodiment of the present invention, the concentration of 4-hydroxyethylpiperazineethanesulfonic acid may be, but is not limited to, 20 mM, 30 mM, 40 mM, 50 mM, 100 mM, 150 mM, 200 mM. Preferably, the concentration of 4-hydroxyethylpiperazineethanesulfonic acid may be 200 mM.

[0026] According to an embodiment of the present invention, the mass percentage of Tween may be, for example, but not limited to: 0.08%, 0.09%, 0.10%, 0.11%, 0.12%.

[0027] According to an embodiment of the present invention, the mass percentage of the quaternary ammonium salt surfactant may be, for example, but not limited to, 0.12%, 0.14%, 0.16%, 0.18%, 0.20%, 0.22%, 0.24%, and 0.25%.

[0028] According to an embodiment of the present invention, the mass percentage of Triton may be, for example, but not limited to, 0.25%, 0.275%, 0.30%, 0.325%, 0.35%, 0.375%, and 0.4%.

[0029] According to an embodiment of the present invention, the mass percentage of benzalkonium bromide may be, for example, but not limited to, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or 0.10%.

[0030] According to an embodiment of the present invention, the mass percentage of the polyether polyol may be, for example, but not limited to, 0.2%, 0.225%, 0.25%, 0.275%, 0.30%, 0.325%, 0.35%, 0.375%, and 0.4%.

[0031] According to an embodiment of the present invention, since it is considered that the presence of microorganisms in water without sterilization may interfere with the microorganisms in the oil phase, as a preferred embodiment, the water may include sterile water.

[0032] According to the embodiments of the present invention, it is found that the content of surfactant not only affects the capture ability of microorganisms in the oil phase, but also affects the subsequent detection of microorganisms in the oil phase. Therefore, in the microorganism capture solution provided by the present invention, surfactants, such as quaternary ammonium salt surfactants, Triton, benzalkonium bromide and polyether polyols, when their respective contents are lower than the above-mentioned respective mass percentages, cannot quickly and as far as possible capture the microorganisms in the oil phase. When their respective contents are higher than the above-mentioned respective mass percentages, the detection of microorganism content by bioluminescence method will inhibit the luciferase activity in the bioluminescent reagent and reduce the luminous intensity, thereby reducing the detection efficiency.

[0033] According to an embodiment of the present invention, cationic surfactants such as quaternary ammonium surfactants and benzalkonium bromide in the microorganism capture liquid can act on the negative charge of the microorganism cell wall to enhance the capture efficiency of the microorganism, and nonionic surfactants such as Tween, Triton and polyether polyols can reduce the oil-water interfacial tension, promote oil-water mixing, and are conducive to the contact of components such as quaternary ammonium surfactants and benzalkonium bromide with the microorganism in the microorganism capture liquid, further improving the capture efficiency. In addition, 4-hydroxyethylpiperazineethanesulfonic acid is a biological buffer that can maintain a constant pH value in a wide pH range, provides a suitable living environment for microorganisms, and helps to maintain the activity and stability of microorganisms. In addition, polyether polyols can also improve the hydrolysis resistance of the capture liquid, etc. Due to the synergistic effect between 4-hydroxyethylpiperazineethanesulfonic acid, Tween, quaternary ammonium surfactants, Triton, benzalkonium bromide and polyether polyol components, that is, through the synergistic effect of multiple action mechanisms such as maintaining a suitable pH environment, reducing oil-water interfacial tension, interacting with the negative charge of the microbial cell wall, and resisting hydrolysis, it is possible to maintain the microbial capture efficiency of the microbial capture liquid while providing longer-lasting microbial capture performance, thereby improving the stability of the capture system.

[0034] According to an embodiment of the present invention, the polyether polyol may include at least one of the following: poloxamer 188, poloxamer 407, and a block copolymer of propylene oxide and polyethylene oxide.

[0035] According to the embodiments of the present invention, polyether polyol, as an organic polymer, has good water resistance and synergistically acts with other components in the microorganism capture solution to provide a wider testing time when testing the RLU value, thereby improving the stability of the testing system.

[0036] According to an embodiment of the present invention, the Tween may include at least one of the following: Tween 20, Tween 40, Tween 60, Tween 80, and Tween 85.

[0037] According to the embodiment of the present invention, Tween, as a nonionic surfactant, will not ionize in water, and thus will not affect the pH value of the microorganism capture liquid. In addition, Tween can reduce the surface tension between water and oil, help immiscible liquids mix, and form a stable emulsion, which is beneficial for the microorganism capture liquid to capture microorganisms from the oil phase.

[0038] According to an embodiment of the present invention, the quaternary ammonium salt surfactant may include at least one of the following: hexadecyltrimethylammonium bromide and hexadecyltrimethylammonium chloride.

[0039] According to the embodiments of the present invention, the quaternary ammonium salt surfactant has good stability.

[0040] According to an embodiment of the present invention, the pH value of the microorganism capture liquid is 7.0-9.0.

[0041] According to an embodiment of the present invention, the pH value of the microorganism capture liquid is 7.0-9.0, which is conducive to the caching effect of 4-hydroxyethylpiperazineethanesulfonic acid. In addition, when detecting the microorganisms captured by the microorganism capture liquid, the microorganism capture liquid that is too acidic or too alkaline will inhibit the activity of luciferase in the ATP bioluminescence reagent, thereby reducing the detection efficiency.

[0042] Based on the microorganism capture liquid provided in the above embodiment of the present invention, the present invention further provides a method for preparing the microorganism capture liquid, including operations S1 to S3.

[0043] In operation S1, prepare a 4-hydroxyethylpiperazineethanesulfonic acid solution with a concentration of 20 mM to 200 mM.

[0044] In operation S2, 0.08% to 0.12% of Tween, 0.12% to 0.25% of quaternary ammonium surfactant, 0.25% to 0.4% of Triton, and 0.2% to 0.4% of polyether polyol in percentage by weight are dissolved in 4-hydroxyethylpiperazineethanesulfonic acid solution in sequence to obtain a mixed solution.

[0045] In operation S3, the mixed solution is sterilized to obtain a microorganism capture solution.

[0046] According to an embodiment of the present invention, the 4-hydroxyethylpiperazineethanesulfonic acid solution can be prepared according to a conventional solution preparation method, which is not specifically limited in the present invention.

[0047] According to an embodiment of the present invention, after preparing a 4-hydroxyethylpiperazineethanesulfonic acid solution with a concentration of 20 mM to 200 mM, 1 mol / L NaOH and 1 mol / L HCl may be used to adjust the pH of the 4-hydroxyethylpiperazineethanesulfonic acid solution.

[0048] According to the embodiments of the present invention, the preparation method provided by the present invention has simple preparation procedures and can reduce the process cost of preparing the microorganism capture liquid.

[0049] According to an embodiment of the present invention, the above operation S3, sterilizing the mixed solution to obtain the microorganism capturing solution, may include the operation of: heating the mixed solution in a 60° C. water bath for 30 minutes to obtain the microorganism capturing solution.

[0050] According to an embodiment of the present invention, the prepared microorganism capture liquid can be stored in a 4° C. refrigerator for later use.

[0051] Based on the microorganism capture liquid and the preparation method of the microorganism capture liquid provided in the above embodiments of the present invention, the present invention also provides an application of the microorganism capture liquid in fuel, wherein the microorganism capture liquid is used to capture microorganisms in the fuel so as to detect the microorganisms in the fuel using the adenosine triphosphate bioluminescence method; the microorganism capture liquid is the above-mentioned microorganism capture liquid or the microorganism capture liquid prepared by the above-mentioned preparation method.

[0052] According to an embodiment of the present invention, the kinematic viscosity of the fuel is ≥1.3 mm 2 ·s -1 , and ≤24.0mm 2 ·s -1 .

[0053] According to the embodiments of the present invention, the safety of transportation can be improved by performing microbial detection on oil with low kinematic viscosity.

[0054] It should be noted that the part on the preparation method of the microorganism capture liquid and the part on the application of the microorganism capture liquid in fuel in the embodiments of the present invention correspond to the part on the microorganism capture liquid in the embodiments of the present invention. The description of the part on the preparation method of the microorganism capture liquid and the part on the application of the microorganism capture liquid in fuel specifically refers to the part on the microorganism capture liquid and will not be repeated here.

[0055] The following is a more specific example to illustrate the high stability microorganism capture liquid, its preparation method and its application in low viscosity fuel. It should be noted that the example is only a specific embodiment of the present invention and cannot limit the protection scope of the present invention.

[0056] Embodiment 1:

[0057] The microorganism capture solution includes the following components: 4-hydroxyethylpiperazineethanesulfonic acid: 200mM, pH=7.8, Tween 20: 0.02%, Tween 40: 0.02%, Tween 60: 0.02%, Tween 80: 0.02%, hexadecyltrimethylammonium bromide: 0.10%, benzalkonium bromide: 0.02%, Triton: 0.25%, poloxamer 188: 0.1%, poloxamer 407: 0.1%, propylene oxide and polyethylene oxide: 0.1%.

[0058] The preparation method of the above-mentioned microorganism capture liquid is as follows:

[0059] Step 1: Weigh 4-hydroxyethylpiperazineethanesulfonic acid and dissolve it in 50 mL of water. The pH value is 7.8. Use 1 mol / L NaOH and 1 mol / L HCl to adjust the pH value to 7.0.

[0060] Step 2: Weigh Tween 20: 0.01g, Tween 40: 0.01g, Tween 60: 0.01g, Tween 80: 0.01g, hexadecyltrimethylammonium bromide: 0.05g, benzalkonium bromide: 0.01g, Triton: 0.125g, Poloxamer 188: 0.05g, Poloxamer 407: 0.05g, propylene oxide and polyethylene oxide: 0.05g, ultrasonicate for 10min, and dissolve in 4-hydroxyethylpiperazineethanesulfonic acid solution;

[0061] Step 3: Sterilize the prepared solution: After preparation, heat it in a 60℃ water bath for 30 minutes, cool it to room temperature and store it in a 4℃ refrigerator.

[0062] The microorganism capture liquid is used to detect the microorganisms in the fuel. The detection method is to add the microorganism capture liquid to the fuel sample, mix it evenly and let it stand to obtain a stratified sample of the water phase and the oil phase. Obtain an aqueous phase sample from the stratified sample as the sample to be tested. Add adenosine triphosphate lysis liquid to the sample to be tested, shake it thoroughly, and obtain a lysis sample. Add the lysis sample to the bioluminescent reagent, mix it thoroughly, and use a chemiluminescence detector to collect the relative light unit value, wherein the bioluminescent reagent includes luciferin and luciferase. Based on the relative light unit value, determine the degree of microbial contamination of the fuel sample. The kinematic viscosity of the fuel sample is ≥1.3mm 2 ·s -1 , and ≤24.0mm 2 ·s -1 The detection efficiency was about 86.3%. After 20 minutes of storage, the RLU value decay rate was about 2.4%.

[0063] Example 2

[0064] The microorganism capture solution includes the following components: 4-hydroxyethylpiperazineethanesulfonic acid: 200mM, pH=7.0; Tween 20: 0.06%, Tween 60: 0.06%, hexadecyltrimethylammonium bromide: 0.15%, benzalkonium bromide: 0.02%, Triton: 0.4%, poloxamer 188: 0.1%, poloxamer 407: 0.1%, propylene oxide and polyethylene oxide: 0.1%.

[0065] The preparation method of the above-mentioned microorganism capture liquid is:

[0066] Step 1: Weigh 4-hydroxyethylpiperazineethanesulfonic acid and dissolve it in 50 mL of water. Use 1 mol / L NaOH and 1 mol / L HCl to adjust the pH value to 7.0.

[0067] Step 2: Weigh Tween 20: 0.03g, Tween 60: 0.03g, cetyltrimethylammonium bromide: 0.075g, benzalkonium bromide: 0.01g, Triton: 0.2g, Poloxamer 188: 0.05g, Poloxamer 407: 0.05g, propylene oxide and polyethylene oxide: 0.05g, ultrasonically for 10min, and dissolve in 4-hydroxyethylpiperazineethanesulfonic acid solution;

[0068] Step 3: Sterilize the prepared solution: After preparation, heat it in a 60℃ water bath for 30 minutes, cool it to room temperature and store it in a 4℃ refrigerator.

[0069] The microorganism capture liquid was used to detect microorganisms in the fuel. The detection method was similar to that in the above-mentioned Example 1 and will not be described in detail here. The detection efficiency was about 87.8%. After being placed for 20 minutes, the RLU value decay rate was about 11.6%.

[0070] Example 3

[0071] The microorganism capture solution includes the following components: 4-hydroxyethylpiperazineethanesulfonic acid: 200mM, pH=9.0, Tween 20: 0.02%, Tween 40: 0.02%, Tween 60: 0.02%, Tween 80: 0.02%, hexadecyltrimethylammonium bromide: 0.10%, benzalkonium bromide: 0.02%, Triton: 0.25%, Poloxamer 188: 0.2%, propylene oxide and polyethylene oxide: 0.2%.

[0072] The preparation method of the above-mentioned microorganism capture liquid is as follows:

[0073] Step 1: Weigh 4-hydroxyethylpiperazineethanesulfonic acid and dissolve it in 50 mL of water. Use 1 mol / L NaOH and 1 mol / L HCl to adjust the pH value to 7.0.

[0074] Step 2: Weigh Tween 20: 0.01g, Tween 40: 0.01g, Tween 60: 0.01g, Tween 80: 0.01g, hexadecyltrimethylammonium bromide: 0.05g, benzalkonium bromide: 0.01g, Triton: 0.125g, Poloxamer 188: 0.1g, propylene oxide and polyethylene oxide: 0.1g, ultrasonically for 10min, and dissolve in 4-hydroxyethylpiperazineethanesulfonic acid solution;

[0075] Step 3: Sterilize the prepared solution: After preparation, heat it in a 60℃ water bath for 30 minutes, cool it to room temperature and store it in a 4℃ refrigerator.

[0076] The microorganism capture liquid was used to detect microorganisms in the fuel. The detection method was similar to that in the above-mentioned Example 1 and will not be described in detail here. The detection efficiency was about 85.6%. After being placed for 20 minutes, the RLU value decay rate was about 7.5%.

[0077] Comparative Example 1:

[0078] The microorganism capture solution includes the following components: 4-hydroxyethylpiperazineethanesulfonic acid: 200mM, pH=7.8, Tween 20: 0.02%, Tween 40: 0.02%, Tween 60: 0.02%, Tween 80: 0.02%, hexadecyltrimethylammonium bromide: 0.10%, benzalkonium bromide: 0.02%, and Triton: 0.25%.

[0079] The preparation method of the above-mentioned microorganism capture liquid is:

[0080] Step 1: Weigh 4-hydroxyethylpiperazineethanesulfonic acid and dissolve it in 50 mL of water. The pH value is 7.8. Use 1 mol / L NaOH and 1 mol / L HCl to adjust the pH value to 7.0.

[0081] Step 2: Weigh Tween 20: 0.01g, Tween 40: 0.01g, Tween 60: 0.01g, Tween 80: 0.01g, hexadecyltrimethylammonium bromide: 0.05g, benzalkonium bromide: 0.01g, Triton: 0.125g in sequence, sonicate for 10min, and dissolve in 4-hydroxyethylpiperazineethanesulfonic acid solution;

[0082] Step 3: Sterilize the prepared solution: After preparation, heat it in a 60℃ water bath for 30 minutes, cool it to room temperature and store it in a 4℃ refrigerator.

[0083] The microorganism capture liquid was used to detect microorganisms in the fuel. The detection method was similar to that in the above-mentioned Example 1 and will not be described in detail here. The detection efficiency was about 79.4%. After being placed for 20 minutes, the RLU value decay rate was about 31.4%.

[0084] Comparing Example 1 with Comparative Example 1, it can be seen that, compared with the components of the microorganism capture liquid of Example 1 of the present invention, Comparative Example 1 lacks polyether polyols, such as: 0.1% poloxamer 188, 0.1% poloxamer 407, 0.1% propylene oxide and polyoxyethylene. The detection efficiency of the microorganism capture liquid of Comparative Example 1 for detecting microorganisms in fuel is lower than the detection efficiency of the microorganism capture liquid of Example 1 for detecting microorganisms in fuel. The RLU value decay rate of the microorganism capture liquid of Comparative Example 1 after being placed for 20 minutes is higher than the RLU value decay rate of the microorganism capture liquid of Example 1 after being placed for 20 minutes. Comparative Example 2

[0085] The microorganism capture solution includes the following components: 4-hydroxyethylpiperazineethanesulfonic acid: 200mM, pH=7.8, Tween 20: 0.02%, Tween 40: 0.02%, Tween 60: 0.02%, Tween 80: 0.02%, hexadecyltrimethylammonium bromide: 0.10%, benzalkonium bromide: 0.02%, poloxamer 188: 0.1%, poloxamer 407: 0.1%, propylene oxide and polyethylene oxide: 0.1%.

[0086] The preparation method of the above-mentioned microorganism capture liquid is:

[0087] Step 1: Weigh 4-hydroxyethylpiperazineethanesulfonic acid and dissolve it in 50 mL of water. Use 1 mol / L NaOH and 1 mol / L HCl to adjust the pH value to 7.0.

[0088] Step 2: Weigh Tween 20: 0.01g, Tween 40: 0.01g, Tween 60: 0.01g, Tween 80: 0.01g, hexadecyltrimethylammonium bromide: 0.05g, benzalkonium bromide: 0.01g, poloxamer 188: 0.05g, poloxamer 407: 0.05g, propylene oxide and polyethylene oxide: 0.05g in sequence, sonicate for 10min, and dissolve in 4-hydroxyethylpiperazineethanesulfonic acid solution;

[0089] Step 3: Sterilize the prepared solution: After preparation, heat it in a 60℃ water bath for 30 minutes, cool it to room temperature and store it in a 4℃ refrigerator.

[0090] The microorganism capture liquid was used to detect microorganisms in the fuel. The detection method was similar to that in the above-mentioned Example 1 and will not be described in detail here. The detection efficiency was about 75.4%. After being placed for 20 minutes, the RLU value decay rate was about 37.4%.

[0091] Comparing Example 1 with Comparative Example 2, it can be seen that the composition of the microorganism capture solution of Example 1 of the present invention lacks 0.25% of Triton. The detection efficiency of the microorganism capture solution of Comparative Example 2 for detecting microorganisms in fuel is lower than the detection efficiency of the microorganism capture solution of Example 1 for detecting microorganisms in fuel. The RLU value decay rate of the microorganism capture solution of Comparative Example 2 after being placed for 20 minutes is higher than the RLU value decay rate of the microorganism capture solution of Example 1 after being placed for 20 minutes.

[0092] Comparative Example 3

[0093] The microorganism capture solution includes the following components: 4-hydroxyethylpiperazineethanesulfonic acid: 200mM, pH=7.8, Tween 20: 0.02%, Tween 40: 0.02%, Tween 60: 0.02%, Tween 80: 0.02%, hexadecyltrimethylammonium bromide: 0.10%, Triton: 0.25%, Poloxamer 188: 0.1%, Poloxamer 407: 0.1%, propylene oxide and polyethylene oxide: 0.1%.

[0094] The preparation method of the above-mentioned microorganism capture liquid is as follows:

[0095] Step 1: Weigh 4-hydroxyethylpiperazineethanesulfonic acid and dissolve it in 50 mL of water. Use 1 mol / L NaOH and 1 mol / L HCl to adjust the pH value to 7.0.

[0096] Step 2: Weigh Tween 20: 0.01g, Tween 40: 0.01g, Tween 60: 0.01g, Tween 80: 0.01g, hexadecyltrimethylammonium bromide: 0.05g, Triton: 0.125g, Poloxamer 188: 0.05g, Poloxamer 407: 0.05g, propylene oxide and polyethylene oxide: 0.05g, ultrasonicate for 10min, and dissolve in 4-hydroxyethylpiperazineethanesulfonic acid solution;

[0097] Step 3: Sterilize the prepared solution: After preparation, heat it in a 60℃ water bath for 30 minutes, cool it to room temperature and store it in a 4℃ refrigerator.

[0098] The microorganism capture liquid was used to detect microorganisms in the fuel. The detection method was similar to that in the above-mentioned Example 1 and will not be described in detail here. The detection efficiency was about 78.1%. After being placed for 20 minutes, the RLU value decay rate was about 36.7%.

[0099] By comparing Example 1 with Comparative Example 3, it can be seen that the composition of the microorganism capture liquid of Comparative Example 3 is less than that of the microorganism capture liquid of Example 1 of the present invention, and 0.02% benzalkonium bromide is lacking. The detection efficiency of the microorganism capture liquid of Comparative Example 3 for detecting microorganisms in fuel is lower than the detection efficiency of the microorganism capture liquid of Example 1 for detecting microorganisms in fuel. The RLU value decay rate of the microorganism capture liquid of Comparative Example 3 after being placed for 20 minutes is higher than the RLU value decay rate of the microorganism capture liquid of Example 1 after being placed for 20 minutes.

[0100] According to the embodiments of the present invention, through comparative analysis of comparative examples 1 to 3 and embodiment 1, it can be seen that only when the microorganism capture liquid component is the microorganism capture liquid provided by the embodiments of the present invention, the microorganism capture liquid has a high detection efficiency for microorganisms in fuel, and the RLU value decay rate of the test is low after the microorganism capture liquid is placed for 20 minutes. It can be proved that the microorganism capture liquid provided by the present invention can provide a longer-lasting microorganism capture performance while maintaining the microorganism capture efficiency of the microorganism capture liquid due to the synergistic effect between 4-hydroxyethylpiperazineethanesulfonic acid, Tween, quaternary ammonium salt surfactant, Triton, benzalkonium bromide and polyether polyol, thereby improving the stability of the capture system.

[0101] Figure 1 The diagram is a diagram showing the effect of the microorganism capture liquid obtained according to Example 1 of the present invention and the commercial capture liquid on the detection of microorganisms in fuel.

[0102] The commercial capture liquid can be used to detect microorganisms in the fuel. The detection method is similar to that in the above-mentioned Example 1 and will not be described in detail here. It should be noted that the commercial capture liquid, ATP lysis liquid, and bioluminescent reagent (including luciferin and luciferase) involved in the examples and comparative examples of the present invention are all from HY-LiTE aviation fuel pen. Figure 1 As shown, the microbial capture liquid provided by the present invention (i.e., the capture liquid shown in the figure) has a detection efficiency for detecting microorganisms in fuel that is comparable to that of the commercial capture liquid, and has a better stability than the commercial capture liquid, i.e., the attenuation degree of the RLU value tested after being placed for 20 minutes is lower.

[0103] According to the embodiments of the present invention, comparative analysis with commercial capture liquids can also prove that the microorganism capture liquid provided by the present invention can provide longer-lasting microorganism capture performance while maintaining the microorganism capture efficiency of the microorganism capture liquid through the synergistic effect between 4-hydroxyethylpiperazineethanesulfonic acid, Tween, quaternary ammonium surfactants, Triton, benzalkonium bromide and polyether polyols, thereby improving the stability of the capture system.

[0104] It will be appreciated by those skilled in the art that the features described in the various embodiments of the present invention may be combined and / or combined in various ways, even if such combinations or combinations are not explicitly described in the present invention. In particular, without departing from the spirit and teachings of the present invention, the features described in the various embodiments of the present invention may be combined and / or combined in various ways. All of these combinations and / or combinations fall within the scope of the present invention.

[0105] The embodiments of the present invention are described above. However, these embodiments are only for the purpose of illustration, and are not intended to limit the scope of the present invention. Although each embodiment is described above, it does not mean that the measures in each embodiment cannot be used in combination. The scope of the present invention is defined by the attached claims and their equivalents. Without departing from the scope of the present invention, those skilled in the art may make various substitutions and modifications, which should all fall within the scope of the present invention.

Claims

1. A microorganism capture solution, characterized in that: In terms of mass percentage, the microorganism capture liquid comprises: ≥90% water, 4-hydroxyethylpiperazineethanesulfonic acid with a concentration of 20mM~200mM, 0.08%~0.12% Tween, 0.12%~0.25% quaternary ammonium salt surfactant, 0.25%~0.4% Triton, 0.02%~0.10% benzalkonium bromide, and 0.2%~0.4% polyether polyol, totaling 100%, wherein the polyether polyol is at least one of the following: poloxamer 188, poloxamer 407, and a block copolymer of propylene oxide and polyethylene oxide; the quaternary ammonium salt surfactant is at least one of the following: hexadecyltrimethylammonium bromide and hexadecyltrimethylammonium chloride.

2. The microorganism-capturing liquid according to claim 1, characterized in that: The concentration of the 4-hydroxyethylpiperazineethanesulfonic acid is 200 mM, and the water is sterile water.

3. The microorganism-capturing liquid according to claim 1, characterized in that: The Tween is at least one of the following: Tween 20, Tween 40, Tween 60, Tween 80, and Tween 85.

4. The microorganism-capturing liquid according to claim 1, characterized in that: The pH value of the capture solution is 7.0-9.

0.

5. A method for preparing a microorganism capture liquid, characterized in that: The method comprises: Prepare a 4-hydroxyethylpiperazineethanesulfonic acid solution with a concentration of 20 mM to 200 mM; Dissolve 0.08% to 0.12% of Tween, 0.12% to 0.25% of a quaternary ammonium salt surfactant, 0.25% to 0.4% of Triton, 0.02% to 0.10% of benzalkonium bromide, and 0.2% to 0.4% of a polyether polyol in the 4-hydroxyethylpiperazineethanesulfonic acid solution in order to obtain a mixed solution; Sterilizing the mixed solution to obtain the microorganism capturing solution; Wherein, the polyether polyol is at least one of the following: poloxamer 188, poloxamer 407, block copolymer of propylene oxide and polyethylene oxide; the quaternary ammonium salt surfactant is at least one of the following: hexadecyltrimethylammonium bromide, hexadecyltrimethylammonium chloride.

6. The method according to claim 5, characterized in that The step of sterilizing the mixed solution to obtain the microorganism capturing solution comprises: heating the mixed solution in a 60° C. water bath for 30 minutes to obtain the microorganism capturing solution.

7. Application of a microorganism capture liquid in fuel, characterized in that: The microorganism capture liquid is used to capture the microorganisms in the fuel so as to detect the microorganisms in the fuel using the adenosine triphosphate bioluminescence method; the microorganism capture liquid is the microorganism capture liquid according to any one of claims 1 to 4 or the microorganism capture liquid prepared by the preparation method according to any one of claims 5 to 6, wherein the kinematic viscosity of the fuel is ≥1.3 mm 2 ·s -1 , and ≤24.0mm 2 ·s -1 .

Images