Disinfection cleaning solution for mass spectrometry flow cytometer and preparation method thereof

Abstract

The embodiment of the invention discloses a disinfection cleaning solution for a mass spectrometry flow cytometer and a preparation method of the disinfection cleaning solution. The disinfection cleaning solution is prepared from the following raw materials in percentage by weight: 0.1%-0.5% of trypsin, 0.1%-0.5% of IV type collagenase, 0.1%-0.5% of I type collagenase, 0.1%-1% of DNA enzyme, 0.1%-0.5% of protease, 0.01%-0.05% of EDTA (Ethylene Diamine Tetraacetic Acid), 0.1%-2% of a surfactant disinfectant and the balance of deionized water. When the disinfection cleaning solution is used for cleaning a key component glass atomizer of a mass spectrometry flow cytometry, residual components, including collagen, extracellular matrix, histone, DNA and the like, possibly existing in the using process can be effectively digested and enzymolyzed, and the disinfection cleaning solution has an excellent cleaning effect.

Description

Technical Field

[0001] This invention relates to the field of equipment cleaning technology, specifically to a disinfection and cleaning solution for mass spectrometry flow cytometers and its preparation method. Background Technology

[0002] Mass cytometry is a novel single-cell analysis technique that combines flow cytometry with mass spectrometry to enable multi-parameter analysis of individual cells. The core of mass cytometry is the use of rare-earth metal-labeled antibodies to simultaneously detect multiple biomarkers within and on the surface of a single cell through mass spectrometry. Mass cytometry is a powerful single-cell analysis tool that offers new possibilities for understanding the complexity of life and the mechanisms of disease. It has wide applications in biomedical research, including studying cellular functional states, signal transduction pathways, and gene expression; it can also be used for early disease diagnosis and efficacy evaluation. Furthermore, mass cytometry can be used for drug screening and optimization, contributing to the development of new drugs.

[0003] Mass cytometry involves complex and delicate components, which are prone to accumulating residues such as protein and DNA during use. Therefore, cleaning and disinfecting the nebulizer is crucial for experimental accuracy and equipment maintenance. First, cleaning and disinfection eliminate potential contamination, ensuring accurate experimental results. Bacterial, viral, or other microbial contamination of the nebulizer can interfere with experimental results. Second, regular cleaning and disinfection can extend the lifespan of the nebulizer. Accumulation of large amounts of protein and nucleic acid dirt or deposits on the nebulizer can lead to equipment damage or performance degradation. Therefore, regular cleaning and disinfection of the nebulizer is essential. Furthermore, cleaning and disinfection are part of laboratory safety protocols, preventing laboratory personnel from coming into contact with potentially hazardous substances. Summary of the Invention

[0004] Therefore, embodiments of the present invention provide a disinfection and cleaning solution for mass flow cytometers and a method for preparing the same.

[0005] To achieve the above objectives, the embodiments of the present invention provide the following technical solutions:

[0006] According to a first aspect of the present invention, the present invention provides a disinfection and cleaning solution for a mass spectrometry flow cytometer, the disinfection and cleaning solution comprising, by weight percentage, the following raw materials: 0.1%~0.5% trypsin, 0.1%~0.5% type IV collagenase, 0.1%~0.5% type I collagenase, 0.1%~1% DNase, 0.1%~0.5% protease, 0.01%~0.05% EDTA, 0.1%~2% surfactant disinfectant, and the balance being deionized water.

[0007] The samples tested may originate from single-cell suspensions obtained after digesting different tissues, but the solutions inevitably carry some small tissue components. Studies have found that trypsin digests substances between tissue cells, dispersing tissue cell clumps into individual cells; type IV collagenase degrades aged or abnormal collagen fibers in tissue cell clumps; type I collagenase digests collagen in tissue cell clumps, breaking it down into individual cells; DNase hydrolyzes DNA in tissue cell clumps; proteases digest proteins in tissue cells, causing protein breakdown; EDTA chelates calcium and magnesium ions, aiding in cleaning; and surfactant disinfectants have broad-spectrum bactericidal effects, used for microbial killing and disinfection. A mixture of trypsin, type IV collagenase, type I collagenase, DNase, proteases, EDTA, and surfactant disinfectants at the above concentrations can effectively digest and enzymatically dissolve residual components that may exist during use, including collagen, extracellular matrix, histones, and DNA, exhibiting excellent cleaning effects.

[0008] Furthermore, the disinfectant cleaning solution comprises the following ingredients by weight percentage: 0.25% trypsin, 0.2% type IV collagenase, 0.2% type I collagenase, 0.45% DNase, 0.3% protease, 0.02% EDTA, 1% surfactant disinfectant, and the balance being deionized water.

[0009] Furthermore, the surfactant disinfectant is selected from benzalkonium bromide, dichlorobenzylguanidine hexane, or polyhexamethylene guanidine hydrochloride.

[0010] Furthermore, the surfactant disinfectant is polyhexamethylene guanidine hydrochloride.

[0011] According to a second aspect of the present invention, the present invention provides a method for preparing the disinfection and cleaning solution for mass spectrometry flow cytometer as described above, the method comprising:

[0012] Weigh out each ingredient according to the formula;

[0013] Trypsin, type IV collagenase, type I collagenase, DNase, protease, EDTA, and surfactant disinfectant are added to deionized water and stirred until dissolved to obtain the disinfectant cleaning solution.

[0014] The embodiments of the present invention have the following advantages:

[0015] The disinfectant cleaning solution of this invention is used to clean the glass nebulizer, a key component of a mass flow cytometer. This effectively digests and enzymatically dissolves residual components that may exist during use, including collagen, extracellular matrix, histones, DNA, etc., and has excellent cleaning effect. Attached Figure Description

[0016] To more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely exemplary, and those skilled in the art can derive other embodiments based on the provided drawings without creative effort.

[0017] Figure 1 This is a schematic diagram of the glass nebulizer, a key component in the mass spectrometry flow cytometer provided by the present invention that requires cleaning.

[0018] Figure 2 This is a schematic diagram illustrating the use of a syringe to clean a glass atomizer, as provided by the present invention. Detailed Implementation

[0019] The following specific embodiments illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0020] Example 1

[0021] This embodiment provides a disinfection and cleaning solution for a mass spectrometry flow cytometer, the raw materials of which are: 0.25% trypsin, 0.2% type IV collagenase, 0.2% type I collagenase, 0.45% DNase, 0.3% protease, 0.02% EDTA, 1% polyhexamethylene guanidine hydrochloride, and the balance being deionized water.

[0022] The preparation method of the above cleaning solution is as follows: weigh each raw material according to the formula; add trypsin, type IV collagenase, type I collagenase, DNase, protease, EDTA and polyhexamethylene guanidine hydrochloride to deionized water, and stir to dissolve evenly.

[0023] Example 2

[0024] This embodiment provides a disinfection and cleaning solution for a mass flow cytometer, the raw materials of which are: 0.4% trypsin, 0.1% type IV collagenase, 0.3% type I collagenase, 0.2% DNase, 0.4% protease, 0.02% EDTA, 1.5% dichlorophenoxyguanidine, and the balance being deionized water.

[0025] The preparation method of the above disinfectant cleaning solution is the same as in Example 1.

[0026] Example 3

[0027] This embodiment provides a disinfection and cleaning solution for a mass spectrometry flow cytometer, the raw materials of which are: 0.3% trypsin, 0.2% type IV collagenase, 0.2% type I collagenase, 0.8% DNase, 0.3% protease, 0.04% EDTA, 0.3% benzalkonium bromide, and the balance being deionized water.

[0028] The preparation method of the above disinfectant cleaning solution is the same as in Example 1.

[0029] Example 4

[0030] This embodiment provides a disinfection and cleaning solution for a mass flow cytometer, the raw materials of which are: 0.1% trypsin, 0.4% type IV collagenase, 0.2% type I collagenase, 0.6% DNase, 0.5% protease, 0.01% EDTA, 0.5% polyhexamethylene guanidine hydrochloride, and the balance being deionized water.

[0031] Example 5

[0032] This embodiment provides a disinfection and cleaning solution for a mass spectrometry flow cytometer, the raw materials of which are: 0.5% trypsin, 0.2% type IV collagenase, 0.5% type I collagenase, 0.8% DNase, 0.2% protease, 0.05% EDTA, 2% polyhexamethylene guanidine hydrochloride, and the balance being deionized water.

[0033] The preparation method of the above disinfectant cleaning solution is the same as in Example 1.

[0034] Comparative Example 1

[0035] This comparative example provides a disinfection and cleaning solution for a mass flow cytometer, which differs from Example 1 only in that it does not use trypsin.

[0036] Comparative Example 2

[0037] This comparative example provides a disinfection and cleaning solution for a mass flow cytometer, which differs from Example 1 only in that it does not use type IV collagenase.

[0038] Comparative Example 3

[0039] This comparative example provides a disinfection and cleaning solution for a mass flow cytometer, which differs from Example 1 only in that it does not use type I collagenase.

[0040] Comparative Example 4

[0041] This comparative example provides a disinfection and cleaning solution for a mass flow cytometer, which differs from Example 1 only in that it does not use DNase.

[0042] Comparative Example 5

[0043] This comparative example provides a disinfection and cleaning solution for a mass flow cytometer, which differs from Example 1 only in that it does not use protease.

[0044] Comparative Example 6

[0045] This comparative example provides a disinfection and cleaning solution for a mass flow cytometer, which differs from Example 1 only in that the trypsin content is adjusted to 0.8%.

[0046] Comparative Example 7

[0047] This comparative example provides a disinfection and cleaning solution for mass flow cytometers, which differs from Example 1 only in that the content of type IV collagenase is adjusted to 0.6%.

[0048] Comparative Example 8

[0049] This comparative example provides a disinfection and cleaning solution for mass flow cytometers, which differs from Example 1 only in that the content of type I collagenase is adjusted to 0.8%.

[0050] Comparative Example 9

[0051] This comparative example provides a disinfection and cleaning solution for a mass flow cytometer, which differs from Example 1 only in that the DNase content is adjusted to 1.5%.

[0052] Comparative Example 10

[0053] This comparative example provides a disinfection and cleaning solution for a mass flow cytometer, which differs from Example 1 only in that the content of the protease is adjusted to 0.8%.

[0054] Comparative Example 11

[0055] This comparative example provides a disinfection and cleaning solution for mass spectrometry flow cytometers, which differs from Example 1 only in that the content of polyhexamethylene guanidine hydrochloride is adjusted to 2.5%.

[0056] Test Example 1

[0057] 1. Microbial testing

[0058] Before cleaning, microbial smears or cultures were performed on the flow nebulizer to obtain baseline data. The microbial smear or culture method was as follows: the nebulizer was placed in a self-sealing bag with 20 mL of PBS buffer, sealed, and then placed in an ultrasonic oscillator for 10 min of ultrasonic oscillation. The cleaning solution was then inoculated onto a petri dish and incubated at 37°C for 48 h. The culture was collected for later use. The cleaning solutions from Examples 1-5 and Comparative Examples 1-11 were used for cleaning, maintaining consistent cleaning time and temperature. The cleaning method was as follows: 20 μL of the collected culture was pipetted into sterile EP tubes, preparing 17 tubes in total. One tube was used as a control with 0.5 mL of PBS, and the remaining 16 tubes were incubated at room temperature for 1 h with 0.5 mL of the cleaning solutions from Examples 1-5 and Comparative Examples 1-11. After cleaning, microbial smears or cultures were performed again, and the data before and after cleaning were compared. If the number of microorganisms significantly decreased or disappeared after cleaning, the cleaning solution was effective. If the number of microorganisms does not change significantly after cleaning, it may be necessary to change the cleaning solution or adjust the cleaning method.

[0059] The microbial test results of the disinfectant cleaning solutions of Examples 1-5 and Comparative Examples 1-11 are shown in Table 1.

[0060] Table 1

[0061]

[0062] 2. The effectiveness of the cleaning fluid can also be further verified by observing the appearance of the flow nebulizer and checking its function. The most direct method is to check the pressure value displayed on the instrument when using the machine. Generally, a pressure of 9-13 indicates good flow of the glass nebulizer, while a pressure greater than 14 indicates poor flow and suggests residual blockage in the tubing.

[0063] Table 2 shows the pressure values ​​of the instruments after cleaning with the disinfectant cleaning solutions of Examples 1-5 and Comparative Examples 1-11.

[0064] Table 2

[0065]

[0066] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A disinfection and cleaning solution for a mass spectrometry flow cytometer, characterized in that, The disinfectant cleaning solution comprises the following ingredients by weight percentage: trypsin 0.1%~0.5%, type IV collagenase 0.1%~0.5%, type I collagenase 0.1%~0.5%, DNase 0.1%~1%, protease 0.1%~0.5%, EDTA 0.01%~0.05%, surfactant disinfectant 0.1%~2%, and the balance being deionized water.

2. The disinfection and cleaning solution for mass spectrometry flow cytometers according to claim 1, characterized in that, The disinfectant cleaning solution comprises the following ingredients by weight percentage: 0.25% trypsin, 0.2% type IV collagenase, 0.2% type I collagenase, 0.45% DNase, 0.3% protease, 0.02% EDTA, 1% surfactant disinfectant, and the balance being deionized water.

3. The disinfection and cleaning solution for mass spectrometry flow cytometers according to claim 1, characterized in that, The surfactant disinfectant is selected from benzalkonium bromide, dichlorobenzylguanidine hexane, or polyhexamethylene guanidine hydrochloride.

4. The disinfection and cleaning solution for mass spectrometry flow cytometers according to claim 1, characterized in that, The surfactant disinfectant is polyhexamethylene guanidine hydrochloride.

5. The method for preparing the disinfection and cleaning solution for mass spectrometry flow cytometer according to claim 1, characterized in that, The method includes: Weigh out each ingredient according to the formula; Trypsin, type IV collagenase, type I collagenase, DNase, protease, EDTA, and surfactant disinfectant are added to deionized water and stirred until dissolved to obtain the disinfectant cleaning solution.

6. A method for cleaning a mass flow cytometer, characterized in that, The method includes: Connect the syringe containing the disinfectant cleaning solution as described in claim 1 to the glass atomizer, and then gently aspirate the syringe.

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