CN109613106B - Multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and application method thereof - Google Patents
Multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and application method thereof Download PDFInfo
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Classifications
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- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
Abstract
The application relates to a multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and a use method thereof, wherein the device comprises four parts, namely a device bracket, an extraction module, a filtering module and a sample collection module, wherein: the device is characterized in that the device comprises a plurality of modules, a plug, a device support, a device and a device, wherein the device is characterized in that each module is connected through the connecting channel and controls the circulation of a sample through the plug, the device support is used for supporting and fixing each module, the device can be used for simultaneously arranging and combining a plurality of modules to realize the pretreatment of a plurality of complex environments and biological samples, the required sample quantity is extremely small, the rapid, high-flux and high-sensitivity detection of small molecular compounds can be realized, the device is modularized in terms of sample pretreatment and mass spectrum detection, and the sample can be directly dripped on a commercial MALDI target plate after the pretreatment, so that the sample preparation time is saved, the operation steps are simplified, and the detection efficiency is improved.
Description
Technical Field
The application belongs to the field of pretreatment, and particularly relates to a multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and a use method thereof.
Background
The matrix assisted laser desorption tandem time of flight mass spectrometry (MALDI-TOF MS) technique was a rapid, simple, high throughput mass spectrometry technique developed from the 80 s of the 20 th century. The principle is that the object to be measured is mixed with the solution of the selected matrix compound, after the solvent volatilizes, the mixture and the matrix form co-crystallization, at the moment, a certain pulse of laser is used for irradiation, the matrix which is easy to absorb laser energy firstly absorbs the laser energy and then transmits the laser energy to the object to be measured through the co-crystallization, the ionization of the molecules of the object to be measured is excited, and the molecules are detected by a mass spectrometer after acceleration in a flight tube. The flight time and the mass-to-charge ratio are positively correlated, and the molecular weight of the object to be detected can be detected through a mass spectrogram. Because of its wide mass spectrum detection range, it does not need to separate by chromatography, and can analyze extremely complex samples.
In the analysis and detection of complex samples, the sample pretreatment technology is of great importance. In recent years, various sample pretreatment techniques have been vigorously developed, and are applied to pretreatment of various types of samples. Techniques such as solid phase extraction, solid phase microextraction, micro solid phase extraction, magnetic extraction, ultrasound-assisted extraction, ultrafiltration and dialysis are widely used in complex environmental and biological samples. However, due to the drawbacks of long pretreatment time, complex operation, difficulty in realizing high throughput and automation, researchers have also been looking for different methods to find new techniques to reduce pretreatment time and even avoid sample pretreatment.
Disclosure of Invention
Based on the above problems, a main object of the present application is to provide a multifunctional modular sample pretreatment device for high-throughput mass spectrometry detection and a method for using the same, which are used for solving at least one of the above technical problems.
In order to achieve the above object, the present application proposes a multifunctional modular sample pretreatment device to be used for high-throughput mass spectrometry detection and a method of using the same, comprising: the device comprises an extraction module, a filtering module, a sample collection module and a device bracket;
the extraction module is of a hollow column tube structure which is vertically arranged, a first opening is formed in the top end of the extraction module, a first sealing cover is arranged on the first opening, and an upward-inclined sample inlet is formed near the lower side of the opening;
the filter module comprises an upper filter module and a lower filter module, the upper filter module and the lower filter module are of hollow column tube structures which are vertically arranged, a second opening is formed in the top end of the upper filter module, a sealing cover is arranged on the second opening, an annular bulge is arranged at the bottom end of the upper filter module, an annular groove matched with the bulge is formed in the top end of the lower filter module, and a filter membrane is placed on the groove and is fixed with the bulge;
the bottom of the extraction module is communicated with the upper part of the upper filtering module through a first horizontal hollow column pipe, and a first plug is arranged on the first horizontal hollow column pipe;
the sample collection module is of a hollow column tube structure which is vertically arranged, a third opening is formed in the top end of the sample collection module, a third sealing cover is arranged on the third opening, a tube orifice is formed in one end of the third sealing cover and used for being connected with a vacuum pump, and a vertical conical pipeline is arranged at the bottom end of the sample collection module;
the bottom of the lower filtering module is communicated with the upper part of the sample collection module through a second horizontal hollow column pipe, and a second plug is arranged on the second horizontal hollow column pipe;
the device support is used for supporting the extraction module, the filtering module and the sample collection module, a clamping groove used for placing and fixing the MALDI target plate is formed in the middle of the device support, and the sample collection module is fixed above the clamping groove by the device support, so that a processed sample is dripped onto the MALDI target plate through the conical pipeline.
Further, the lengths of the hollow column tubes of the extraction module, the filtration module and the sample collection module are 1.5-5cm, the inner diameters are 0.5-3cm, and the outer diameters are 0.8-5cm.
Further, a clamping groove for placing the MALDI target plate in the middle of the device bracket is compatible with the MALDI target plate and is fixed through jackscrews.
Further, when the device is used, the third sealing cover is connected with the vacuum pump to provide auxiliary power for transferring the sample.
Further, in order to ensure accurate dripping of the sample onto the corresponding target spot of the MALDI target plate, the diameter of the outlet of the conical tube of the sample collection module is 0.5-1mm.
Further, a third plug for controlling the opening and closing of the modules is arranged on the conical pipeline at the bottom of the sample collection module.
Furthermore, the extraction module, the filtering module and the sample collection module are all arranged side by side, and the number of the simultaneous use can be adjusted according to the requirement of sample pretreatment so as to realize high-flux pretreatment and detection.
Further, the device holder may be sized according to the number of simultaneous uses of the extraction module, the filtration module, and the sample collection module.
Further, the first plug and the second plug are single taps or drain plugs depending on the number of simultaneous uses of the extraction module, the filtration module, and the sample collection module.
Further, when the extraction module, the filtration module and the sample collection module are arranged side by side, the plugs which are opened and closed among the control modules are arranged plugs, and at the moment, the circulation of the sample is controlled by pushing the arranged plugs.
Further, the modular device is formed by 3D printing rapid prototyping, precision prototyping, machining, cold and hot working or hand-forming techniques.
Further, the samples for processing by the modular device include macro and micro environmental samples and biological samples, preferably micro biological samples.
In another aspect, the application provides a method for performing matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry detection on a sample to be detected according to the above-described modular device, comprising the steps of:
step 1, adding a sample to be detected and an extraction material into the multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and the use method thereof according to any one of claims 1 to 12, and after the sample is treated by the extraction module, the filtration module and the sample collection module, dripping the finally collected sample on a target point corresponding to a commercial MALDI target plate by controlling the first plug and the second plug;
and 2, mass spectrum detection is carried out on the sample to be detected by using a matrix-assisted laser desorption-tandem flight time mass spectrometer.
Further, in step 1, adding a sample to be detected and an extraction material into an extraction module, controlling a first plug to introduce a mixed solution into a filtration module after extracting for 30min, keeping all plugs in an open state, connecting a pipe orifice on a third sealing cover with a vacuum pump, performing suction filtration for 30s, and discarding the filtered filtrate;
if the extraction material can be used as matrix-assisted laser desorption ionization analyte of MALDI, taking out filter residues, placing the filter residues into a sample collecting device, adding 10-200 mu L of ultrapure water, and dispersing into suspension under the assistance of ultrasound; if the extraction material can not be used as matrix-assisted laser desorption ionization analyte of MALDI, placing a filter membrane in the groove of the filter module, and buckling the upper filter module and the lower filter module together;
then adding 0.5-3mL of eluent into the filter module, carrying out ultrasonic assistance for 10min, and removing the filter membrane after filtering for 30s by a vacuum pump connected with the third sealing cover; finally, the second plug is controlled to introduce the sample into the sample collection module and drop it onto a commercial MALDI target plate mounted on the device holder.
The multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and the use method thereof have the following beneficial effects:
1. the multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and the use method thereof have flexible and controllable shapes and structures, can be assembled according to pretreatment requirements of different samples, thereby realizing different pretreatment operations, and are simple to operate, convenient and quick.
2. The multifunctional modularized sample pretreatment device for high-flux mass spectrometry detection and the application method thereof can simultaneously arrange and combine a plurality of modules to realize pretreatment of a plurality of complex environments and biological samples at the same time, thereby realizing rapid high-flux mass spectrometry detection.
3. The multifunctional modularized sample pretreatment device for high-flux mass spectrometry detection and the use method thereof have the advantages of extremely small volume, extremely small required sample quantity, high-sensitivity mass spectrometry detection of small molecular compounds can be realized through enrichment of an extraction module, separation of a filtering module and concentration of a sample collection module, and the detection limit can reach ppt or even be lower than ppt level.
4. The multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and the use method thereof modularize sample pretreatment and mass spectrometry detection, are compatible with a commercial MALDI target plate, can simplify operation steps, save sample preparation time and improve detection efficiency;
5. the sample pretreatment and high-flux mass spectrum detection modularized device is in modularized design, is convenient to disassemble and assemble, can be further added with other functional modules, and has good expansibility and compatibility.
6. The multifunctional modularized sample pretreatment device for high-flux mass spectrometry detection and the use method thereof have the characteristics of low cost, simple manufacturing process, stable signal, high flux and convenient use, and have the possibility of continuous optimization;
drawings
FIGS. 1 and 2 are printed examples of a multifunctional modular sample pretreatment device for high-throughput mass spectrometry and a method for using the same according to an embodiment of the present application, wherein FIG. 1 is a front view of ten modules used simultaneously, FIGS. 2 (a) and 2 (b) are front and top views of the device, FIGS. 2 (c) and 2 (d) are structures of a device holder, FIG. 2 (e) is a five-row plug for control when ten modules are used simultaneously, and FIG. 2 (f) is a commercial MALDI target plate;
FIG. 3 is a mass spectrum detection result of 1. Mu.L of a mixed standard sample by using a multifunctional modular sample pretreatment device for high-throughput mass spectrum detection and a use method thereof according to an embodiment of the present application, and FIGS. 3 (a) and 3 (b) are respectively detected in a negative ion mode and a positive ion mode;
fig. 4 shows the result of the actual mass spectrum detection of the lung perfusate by using the multifunctional modular sample pretreatment device for high throughput mass spectrum detection and the method for using the same according to an embodiment of the present application, wherein fig. 4 (a) and 4 (b) are respectively detected in a negative ion mode and a positive ion mode;
FIG. 5 shows the mass spectrum detection result of the actual serum labeled sample by using the multifunctional modularized sample pretreatment device for high-throughput mass spectrum detection and the use method thereof according to an embodiment of the present application, wherein FIGS. 5 (a) and 5 (b) are respectively detected in a negative ion mode and a positive ion mode;
fig. 6 shows a mass spectrum detection result of an actual single-drop whole blood labeled sample by using the multifunctional modularized sample pretreatment device for high-throughput mass spectrum detection and the use method thereof according to an embodiment of the present application, wherein fig. 6 (a) and 6 (b) are respectively detected in a negative ion mode and a positive ion mode;
fig. 7 shows the result of mass spectrum screening of a perfluorocompound of an actual single drop whole blood sample by using the multifunctional modular sample pretreatment device for high-throughput mass spectrometry according to an embodiment of the present application.
Detailed Description
The present application will be further described in detail below with reference to specific embodiments and with reference to the accompanying drawings, in order to make the objects, technical solutions and advantages of the present application more apparent.
The high-flux mass spectrometry technology applied to complex sample analysis plays an extremely important role in many fields such as food safety, histology research, environment and health. The pretreatment technology is the most important component, wherein the steps of pretreatment are simplified, high-throughput and modularization are realized, and the pretreatment technology is a barrier in the pretreatment technology of complex samples and is also an urgent problem to be solved. Therefore, the application provides a multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and a use method thereof, wherein the multifunctional modularized sample pretreatment device comprises an extraction module 2, filtering modules 3 and 4, a sample collection module 5 and a device bracket 8;
the extraction module 2 is of a hollow column tube structure which is vertically arranged, a first opening is formed in the top end of the extraction module 2, a first sealing cover is arranged on the first opening, and a sample inlet 1 which is inclined upwards is formed near the lower side of the opening;
the filtering modules 3 and 4 comprise an upper filtering module 3 and a lower filtering module 4, the upper filtering module 3 and the lower filtering module 4 are of hollow column structures which are vertically arranged, a second opening is formed in the top end of the upper filtering module 3, a sealing cover is arranged on the second opening, an annular bulge is arranged at the bottom end of the upper filtering module 3, an annular groove matched with the bulge is formed in the top end of the lower filtering module 4, and a filtering membrane 6 is placed on the groove and is fixed with the bulge;
the bottom of the extraction module 2 is communicated with the upper part of the upper filter module 3 through a first horizontal hollow column pipe, and a first plug 7 is arranged on the first horizontal hollow column pipe;
the sample collection module 5 is of a hollow column tube structure which is vertically arranged, a third opening is formed in the top end of the sample collection module 5, a third sealing cover is arranged on the third opening, a tube orifice is formed in one end of the third sealing cover and used for being connected with a vacuum pump, and a vertical conical pipeline is arranged at the bottom end of the sample collection module 5;
the bottom of the lower filtering module 4 is communicated with the upper part of the sample collection module 5 through a second horizontal hollow column pipe, and a second plug 9 is arranged on the second horizontal hollow column pipe;
the device support 8 is used for supporting the extraction module 2, the filtering modules 3 and 4 and the sample collection module 5, a clamping groove for placing and fixing a MALDI target plate is formed in the middle of the device support 8, and the sample collection module 5 is fixed above the clamping groove by the device support 8, so that a processed sample is dripped onto the MALDI target plate through the conical pipeline.
Further, the lengths of the hollow column tubes of the extraction module 2, the filtering modules 3 and 4 and the sample collection module 5 are 1.5-5cm, the inner diameters are 0.5-3cm, and the outer diameters are 0.8-5cm; in some embodiments of the application, the hollow column tubes of the extraction module 2, the filtration modules 3,4 and the sample collection module 5 have a length of 4cm, a tube inside radius of 0.74cm and a tube outside diameter of 1cm.
The clamping groove in the middle of the device bracket 8 for placing the MALDI target plate is compatible with the MALDI target plate and is fixed by jackscrews.
When the device is used, the third sealing cover is connected with the vacuum pump to provide auxiliary power for transferring the sample.
In order to ensure accurate dripping of the sample onto the corresponding target spot of the MALDI target plate, the diameter of the sample collection module 5 at the outlet of the tapered tube is 0.5-1mm. Preferably, the diameter of the bottom end of the sample collection module 5 at the outlet of the vertical conical tube is 0.8mm.
And a third plug for controlling the opening and closing of the modules is arranged on the conical pipeline at the bottom of the sample collection module 5.
The extraction module 2, the filtering modules 3 and 4 and the sample collection module 5 are all arranged side by side, and the number of the extraction modules, the filtering modules and the sample collection module 5 can be adjusted according to the requirement of sample pretreatment so as to realize high-throughput pretreatment and detection. In some embodiments of the application, 10 modules are used simultaneously, namely: the extraction module 2, the filtration modules 3,4 and the sample collection module 5 are all 10 arranged side by side, and the device support is also adjusted to support 10 modules used simultaneously.
The device holder 8 is sized according to the number of simultaneous use of the extraction module 2, the filtration modules 3,4 and the sample collection module 5.
The first and second plugs are single taps or drain plugs depending on the number of simultaneous use of the extraction module 2, the filtration modules 3,4 and the sample collection module 5. In some embodiments of the application, the plugs that control the opening and closing of the modules are 5-row plugs, and the circulation of the sample is controlled by pushing the row plugs.
When the extraction module 2, the filtering modules 3 and 4 and the sample collection module 5 are arranged side by side, the plugs which are opened and closed among the control modules are arranged plugs, and at the moment, the circulation of the samples is controlled by pushing the arranged plugs.
The modular device is formed by machining, cold and hot machining, precision forming, 3D printing rapid prototyping or hand forming techniques. Preferably, the device is manufactured by 3D printing.
The samples for processing by the modular device include macro and micro environmental samples and biological samples, preferably micro biological samples.
In another embodiment of the present application, the present application provides a method for performing matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry on a sample to be tested according to the aforementioned modular device, comprising the steps of:
step 1, adding a sample to be detected and an extraction material into the multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and the use method thereof according to any one of claims 1 to 12, and after the sample is treated by the extraction module 2, the filtration modules 3,4 and the sample collection module 5, dripping the finally collected sample on a target point corresponding to a commercial MALDI target plate by controlling the third plug;
and 2, mass spectrum detection is carried out on the sample to be detected by using a matrix-assisted laser desorption-tandem flight time mass spectrometer.
Further, in step 1, adding a sample to be detected and an extraction material into the extraction module 2, controlling the first plug 7 to introduce the mixed solution into the filtration modules 3 and 4 after extracting for 30min, keeping all plugs in an open state, connecting the plugs with a vacuum pump through a pipe orifice on the third sealing cover, and filtering for 30s, and then discarding the filtered filtrate;
if the extraction material can be used as matrix-assisted laser desorption ionization analyte of MALDI, taking out filter residues, placing the filter residues into a sample collecting device, adding 10-200 mu L of ultrapure water, and dispersing into suspension under the assistance of ultrasound; if the extraction material can not be used as matrix-assisted laser desorption ionization analyte of MALDI, placing filter membranes in the grooves of the filter modules 3,4, and fastening the upper filter module 3 and the lower filter module 4 together;
then adding 0.5-3mL of eluent into the filtering modules 3 and 4, carrying out ultrasonic assistance for 10min, and removing the filtering membrane after filtering for 30s by a vacuum pump connected with the third sealing cover; finally, the second plug 9 is controlled to introduce the sample into the sample collection module 5 and drop it onto a commercial MALDI target plate mounted on the device holder.
The modular device of the present application can handle samples including single drop whole blood samples, and small molecules detected can reach ppt levels or even below the detection limit of ppt levels.
The multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and the use method thereof are described in detail below through specific embodiments.
Examples
As shown in fig. 1, the embodiment provides a multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection and a use method thereof, and in the embodiment, the device is manufactured by 3D printing, and the main materials are photosensitive resin and rubber; the device is integrally assembled by nine parts, as shown in the abstract drawing, wherein 1 is a sample inlet, 2 is an extraction module, 3,4 are the upper half part and the lower half part of a filter module, 5 is a sample collection module, 6 is a filter membrane, 7 is a plug, and 8 is a device bracket. The target plate is used for containing a sample to be detected, which needs to be detected by a MALDI-TOF mass spectrometer. As shown in fig. 1 and 2, fig. 1 is a front view of ten modules used simultaneously, fig. 2 (a) and 2 (b) are a front view and a top view of the device, fig. 2 (c) and 2 (d) are structures of a device holder, fig. 2 (e) is a five-row plug for control when ten modules are used simultaneously, and fig. 2 (f) is a commercial MALDI target plate.
The multifunctional modularized sample pretreatment device for high-flux mass spectrometry detection, the application method thereof and the MALDI-TOF mass spectrometer are used for carrying out bisphenol S with the concentration of 0.9 mug/mL; 2,2', 4' -tetrabromodiphenyl ether at a concentration of 4.5 μg/mL; pentachlorophenol with concentration of 0.9 mug/mL; estradiol at a concentration of 4.5 μg/mL; perfluorooctyl sulfonic acid at a concentration of 0.09 μg/mL; tetrabromobisphenol A at a concentration of 0.9. Mu.g/mL; trivalent arsenic at a concentration of 1.4 μg/mL; N-hexadecyl-N, N-dimethylbenzyl ammonium chloride at a concentration of 0.4 μg/mL; tetradecyldimethyl ammonium chloride at a concentration of 0.4 μg/mL; dodecyl dimethyl benzyl ammonium chloride at a concentration of 0.4 μg/mL; cetyl trimethylammonium bromide at a concentration of 0.09 μg/mL; n, N, N-trimethyl-1-tetradecylammonium bromide at a concentration of 0.09. Mu.g/mL was subjected to detection analysis. First, 20. Mu.L of the sample was pipetted with a pipette to dilute into the device of this example with 980. Mu.L of water, and 100. Mu.L of 10mg/mL of extraction material was added; the processed sample is dripped on a connected target plate, the target plate is put into a MALDI-TOF mass spectrometer for detection after the sample is dried, the model of the used instrument is Bruker DaltonicsAutoflex III Smartbean MALDI-TOF mass spectrometer, the frequency of the used instrument is 355nm Nd: YAG with 100Hz, the laser power is set to 40%, and negative ions and positive ions are respectively adopted for detection. The measurement results are shown in fig. 3, fig. 4, fig. 5, fig. 6, and fig. 7, wherein fig. 3 is a mass spectrum detection result of 1 μl of the mixed standard sample, fig. 4 is an actual lung perfusate mass spectrum detection result, fig. 5 is an actual serum labeled sample mass spectrum detection result, fig. 6 is an actual single-drop whole blood labeled sample mass spectrum detection result, and fig. 7 is an actual single-drop whole blood sample mass spectrum detection result. From fig. 3 to 6, it can be seen that all 12 organic matters to be detected have stronger detection signals, which proves that the device of the embodiment can modularize sample pretreatment and high flux. As can be seen from fig. 7, 3 perfluorinated compounds can be screened from a single drop of whole blood sample, which proves that the device of the embodiment not only can modularize sample pretreatment and high-throughput detection, but also has the function of screening non-target compounds by mass spectrometry, and can further extend the application of the device.
According to the embodiment, the multifunctional modularized sample pretreatment device for high-flux mass spectrometry detection and the use method thereof can be used for simultaneously arranging and combining a plurality of modules to realize pretreatment of a plurality of complex environments and biological samples, realizing rapid high-flux mass spectrometry detection, realizing high-sensitivity detection of small molecular compounds, and realizing the detection limit of ppt or even lower than ppt. The device is convenient to disassemble and assemble, is convenient to use, has continuous optimization possibility, and has wide application prospect.
The foregoing description of the embodiments has been provided for the purpose of illustrating the general principles of the application, and is not meant to limit the application thereto, but to limit the application thereto, and any modifications, equivalents, improvements and equivalents thereof may be made without departing from the spirit and principles of the application.
Claims (15)
1. A multifunctional modular sample pretreatment device to be used for high throughput mass spectrometry detection, comprising: an extraction module (2), a filtration module (3, 4), a sample collection module (5) and a device bracket (8);
the extraction module (2) is of a hollow column tube structure which is vertically arranged, a first opening is formed in the top end of the extraction module (2), a first sealing cover is arranged on the first opening, and a sample inlet (1) which is inclined upwards is formed near the lower side of the opening;
the filter modules (3, 4) comprise an upper filter module (3) and a lower filter module (4), the upper filter module (3) and the lower filter module (4) are of hollow column tube structures which are vertically arranged, a second opening is formed in the top end of the upper filter module (3), a sealing cover is arranged on the second opening, an annular bulge is arranged at the bottom end of the upper filter module (3), an annular groove matched with the bulge is formed in the top end of the lower filter module (4), and a filter membrane (6) is placed on the groove and is fixed with the bulge;
the bottom of the extraction module (2) is communicated with the upper part of the upper filtering module (3) through a first horizontal hollow column pipe, and a first plug (7) is arranged on the first horizontal hollow column pipe;
the sample collection module (5) is of a hollow column tube structure which is vertically arranged, a third opening is formed in the top end of the sample collection module (5), a third sealing cover is arranged on the third opening, a tube orifice is formed in one end of the third sealing cover and used for being connected with a vacuum pump, and a vertical conical pipeline is arranged at the bottom end of the sample collection module (5);
the bottom of the lower filtering module (4) is communicated with the upper part of the sample collecting module (5) through a second horizontal hollow column pipe, and a second plug (9) is arranged on the second horizontal hollow column pipe;
the device support (8) is used for supporting the extraction module (2), the filtering modules (3, 4) and the sample collection module (5), a clamping groove used for placing and fixing a MALDI target plate is formed in the middle of the device support (8), and the sample collection module (5) is fixed above the clamping groove by the device support (8) so that a processed sample can be dripped onto the MALDI target plate through the conical pipeline.
2. The multifunctional modular sample pretreatment device for high-throughput mass spectrometry according to claim 1, wherein the hollow column tubes of the extraction module (2), the filtration modules (3, 4) and the sample collection module (5) are each 1.5-5cm in length, 0.5-3cm in inner diameter and 0.8-5cm in outer diameter.
3. A multifunctional modular sample pretreatment device for high-throughput mass spectrometry according to claim 1 or 2, characterized in that the clamping groove for placing MALDI target plate in the middle of the device holder (8) is compatible with MALDI target plate and fixed by jackscrews.
4. The multifunctional modular sample pretreatment apparatus for high-throughput mass spectrometry of claim 1, wherein in use, the third seal cap is coupled to a vacuum pump to provide auxiliary power for sample transfer.
5. Multifunctional modular sample pretreatment device for high-throughput mass spectrometry according to claim 1 or 2, characterized in that the diameter of the outlet of the conical tube of the sample collection module (5) is 0.5-1mm in order to ensure accurate dripping of the sample onto the corresponding target spot of the MALDI target plate.
6. The multifunctional modular sample pretreatment apparatus for high-throughput mass spectrometry according to claim 1 or 2, wherein a third plug for controlling the opening and closing of the modules is provided on the tapered pipe at the bottom of the sample collection module (5).
7. The multifunctional modular sample pretreatment device for high-throughput mass spectrometry according to claim 1 or 2, wherein the extraction module (2), the filtration modules (3, 4) and the sample collection module (5) are arranged side by side, and the number of simultaneous uses can be adjusted according to the need of sample pretreatment to realize high-throughput pretreatment and detection.
8. Multifunctional modular sample pretreatment device for high-throughput mass spectrometry according to claim 1 or 2, characterized in that the device holder (8) is adjustable in size according to the number of simultaneous uses of the extraction module (2), the filtration modules (3, 4) and the sample collection module (5).
9. Multifunctional modular sample pretreatment device for high-throughput mass spectrometry according to claim 1 or 2, characterized in that the number of simultaneous use of the first plug and the second plug according to the extraction module (2), the filtration module (3, 4) and the sample collection module (5) is single tap or drain plug.
10. The multifunctional modular sample pretreatment apparatus for high-throughput mass spectrometry according to claim 9, wherein when the extraction module (2), the filtration modules (3, 4) and the sample collection module (5) are provided in a plurality of side-by-side arrangement, the plugs controlling the opening and closing between the modules are drain plugs, and the circulation of the sample is controlled by pushing the drain plugs.
11. The multifunctional modular sample pretreatment apparatus for high-throughput mass spectrometry according to claim 1 or 2, wherein the modular sample pretreatment apparatus is formed by 3D printing rapid prototyping, precision prototyping, machining, cold and hot working or hand-forming techniques.
12. The multifunctional modular sample pretreatment apparatus for high-throughput mass spectrometry according to claim 1 or 2, wherein the samples for treatment by the modular sample pretreatment apparatus include both constant and micro-sized environmental samples and biological samples.
13. The multifunctional modular sample pretreatment apparatus for high-throughput mass spectrometry of claim 12, wherein the sample for treatment by the modular sample pretreatment apparatus is a trace amount of biological sample.
14. A method for performing matrix-assisted laser Desorption ionization tandem time-of-flight mass spectrometry on a sample to be tested by a multifunctional modular sample pretreatment device for high throughput mass spectrometry according to claim 6, comprising the steps of:
step 1, adding a sample to be detected and an extraction material into the multifunctional modularized sample pretreatment device for high-throughput mass spectrometry detection according to claim 6, and dripping the finally collected sample on a target point corresponding to a commercial MALDI target plate by controlling the third plug after the extraction module (2), the filtering modules (3, 4) and the sample collection module (5) are processed;
and 2, mass spectrum detection is carried out on the sample to be detected by using a matrix-assisted laser desorption-tandem flight time mass spectrometer.
15. Method according to claim 14, characterized in that in step 1, the sample to be tested and the extraction material are added to the extraction module (2), after 30min of extraction the first plug (7) is controlled to introduce the mixed liquor into the filtration module (3, 4), all plugs are kept in an open state, after connection with the vacuum pump through the nozzle on the third sealing cap and suction filtration for 30s, the suction filtered filtrate is discarded;
if the extraction material can be used as matrix-assisted laser desorption ionization analyte of MALDI, taking out filter residues, placing the filter residues into a sample collecting device, adding 10-200 mu L of ultrapure water, and dispersing into suspension under the assistance of ultrasound; if the extraction material can not be used as matrix-assisted laser desorption ionization analyte of MALDI, placing a filter membrane in the grooves of the filter modules (3, 4) and buckling the upper filter module (3) and the lower filter module (4) together;
then adding 0.5-3mL of eluent into the filtering modules (3, 4), carrying out ultrasonic assistance for 10min, and discarding the filtering membrane after filtering for 30s by a vacuum pump connected with the third sealing cover; finally, a second plug (9) is controlled to introduce the sample into the sample collection module (5) and drop it onto a commercial MALDI target plate mounted on the device holder.
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