Cancer tissue mass spectrometry detection method and detection system thereof

Abstract

The invention relates to the technical field of mass spectrum detection tools, in particular to a cancer tissue mass spectrum analysis detection method and a detection system thereof. Comprising the steps of sample pretreatment and device preparation; selecting an extraction solution and generating a charged mixed solution; atomization and gas-liquid mixing; performing mass spectrum analysis and signal acquisition; and data processing and metabolite analysis. According to the cancer tissue mass spectrometry detection method and the detection system thereof, the cut cancer tissue sample is placed on the filter membrane infiltrated by the extract liquor, and the connecting device is sealed. 3 kV high voltage is applied to the ion source device, so that the methanol extraction liquid flowing through the tissue is electrified. And the charged mixed liquid enters the Y-shaped sprayer and is atomized by high-purity nitrogen provided by the gas conveying device. And the atomized charged liquid drops are directly introduced into a mass spectrometer for analysis. And finally, a mass spectrum signal analysis system processes data, metabolites in tissues are quickly and accurately recognized, the detection efficiency and sensitivity are remarkably improved, and sample requirements and dependence on experience are reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of mass spectrometry detection tools, and in particular to a cancer tissue mass spectrometry analysis and detection method and a detection system thereof. Background Art

[0002] Mass spectrometry has become an important tool in many analytical detection methods due to its advantages such as high sensitivity, strong specificity and rapid response. Despite this, traditional mass spectrometry methods often fail to perform to their full potential when faced with samples with complex matrices, which to some extent limits their application in a wider range of fields. In 2004, Professor Cooks RG proposed the desorption electrospray ionization mass spectrometry (DESI-MS) technology, which can directly perform mass spectrometry analysis on the sample surface without any sample pretreatment, greatly promoting the research and development of direct mass spectrometry technology. Direct mass spectrometry technology is an emerging mass spectrometry technology that can quickly perform qualitative and quantitative analysis of samples, especially in situ, online, real-time, non-destructive, high-throughput and low-loss analysis of complex matrix samples. It has shown great application potential in analytical research in many fields.

[0003] The current pathological diagnostic system relies on HE-stained tissue morphological analysis as its core method, identifying tumors through the microstructural characteristics of tissue samples. Although this technology is considered the "gold standard," it suffers from limitations such as long processing time (≥30 minutes), strong reliance on technician experience, and large sample volumes, making it difficult to meet the actual demand for "fast and accurate" testing solutions in society. Summary of the Invention

[0004] The present invention aims to at least improve one of the technical problems existing in the prior art. To this end, the present invention provides a cancer tissue mass spectrometry detection method and a detection system thereof.

[0005] The technical solutions of the present invention are as follows:

[0006] A method for mass spectrometry analysis of cancer tissue, comprising:

[0007] S1, build a cancer tissue mass spectrometry analysis and detection scenario;

[0008] S2, sample pretreatment and device preparation;

[0009] S3, extraction liquid selection and charged mixed liquid generation;

[0010] S4, atomization and gas-liquid mixing;

[0011] S5, mass spectrometry analysis and signal acquisition;

[0012] S6, Data processing and metabolite elucidation.

[0013] In a possible technical solution, further in S1, the detection scenario includes:

[0014] Tissue sampling and extraction mechanism, used for sampling and extraction of cancer tissues;

[0015] an ion source mechanism connected to the tissue sampling and extraction mechanism;

[0016] a gas delivery mechanism connected to the ion source mechanism;

[0017] The mass spectrometry signal analysis system is connected to the ion source mechanism and is used to perform mass spectrometry analysis on the sample.

[0018] In a possible technical solution, further, the tissue sampling and extraction mechanism includes:

[0019] Syringe pumps;

[0020] Extraction liquid injection needle, connected to the syringe pump;

[0021] A syringe head connected to the extraction liquid injection needle;

[0022] Tissue extraction mixture filter membrane, installed in the syringe head

[0023] An adapter, connected to the syringe head;

[0024] The capillary is connected to the adapter, and the other end is connected to the ion source mechanism.

[0025] In a possible technical solution, further, the ion source mechanism includes:

[0026] A Y-shaped gas-liquid mixing sprayer connected to the capillary tube;

[0027] Fixer, used to fix the Y-type gas-liquid mixing sprayer.

[0028] In a possible technical solution, further, the gas delivery mechanism includes:

[0029] High-purity nitrogen cylinders;

[0030] The voltage divider is connected to the high-purity nitrogen gas bottle and the ion source mechanism through a gas delivery copper tube.

[0031] In a possible technical solution, the tissue sampling and extraction mechanism further includes:

[0032] A voltage device has one end connected between the extraction liquid injection needle and the syringe head, and the other end connected to the mass spectrometry signal analysis system.

[0033] A cancer tissue mass spectrometry analysis and detection system, wherein the above method is used to perform mass spectrometry analysis and detection on cancer tissue.

[0034] According to the cancer tissue mass spectrometry detection method and detection system of the present invention, the excised cancer tissue sample is placed on a filter membrane soaked in an extract (methanol) and a sealed connection device is used. A high voltage of 3kV is applied to the ion source device to charge the methanol extract flowing through the tissue. The charged mixed liquid enters the Y-type sprayer and is atomized by the high-purity nitrogen provided by the gas delivery device. The atomized charged droplets are directly introduced into the mass spectrometer for analysis. Finally, the mass spectrometry signal analysis system processes the data, quickly and accurately identifies metabolites in the tissue, significantly improves detection efficiency and sensitivity, and reduces sample requirements and dependence on experience.

[0035] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0037] Figure 1 is a flow chart of a method for mass spectrometry analysis and detection of cancer tissue according to an embodiment of the present invention;

[0038] Figure 2 3 is a schematic diagram of a scenario structure of a method for mass spectrometry analysis and detection of cancer tissue according to an embodiment of the present invention.

[0039] Reference numerals:

[0040] The syringe pump 1, the extraction liquid injection needle 2, the syringe head 3, the tissue extraction mixture filter membrane 31, the adapter 4, the capillary 5, the Y-type gas-liquid mixing sprayer 6, the mass spectrometry signal analysis system 7, the high-purity nitrogen bottle 8, the voltage divider 9, and the voltage device 10 are connected. DETAILED DESCRIPTION

[0041] The embodiments of the present invention are described in detail below. The embodiments described with reference to the accompanying drawings are exemplary. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0042] It should be noted that when an element is referred to as being “fixed to” another element, it may be directly on the other element or there may be an intermediate element. When an element is referred to as being “connected to” another element, it may be directly connected to the other element or there may be an intermediate element.

[0043] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention pertains. The terms used herein in the specification of the present invention are for the purpose of describing specific embodiments only and are not intended to limit the present invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0044] In the specification, claims, and accompanying drawings of this application, the terms "first," "second," "third," and the like are used to distinguish different objects and are not used to describe a particular order. Furthermore, the terms "including," "comprising," "having," and any variations thereof are intended to cover non-exclusive inclusions. For example, a list of steps or elements may be included, or alternatively, steps or elements not listed may be included, or other steps or elements may be included that are inherent to the process, method, product, or apparatus.

[0045] Only part relevant to the present application is shown in the accompanying drawings, not all of it. Before discussing exemplary embodiments in more detail, it should be mentioned that some exemplary embodiments are described as processing or methods depicted as flow charts. Although flow charts describe various operations (or steps) as sequential processing, many operations therein can be implemented in parallel, concurrently or simultaneously. In addition, the order of various operations can be rearranged. When its operation is completed, the processing can be terminated, but can also have additional steps not included in the accompanying drawings. The processing can correspond to methods, functions, procedures, subroutines, subprograms, etc.

[0046] As used in this specification, the terms "component," "module," "system," "unit," and the like are used to refer to computer-related entities, hardware, firmware, a combination of hardware and software, software, or software in execution. For example, a unit can be, but is not limited to, a process running on a processor, a processor, an object, an executable file, an execution thread, a program, and / or distributed between two or more computers. In addition, these units can be executed from various computer-readable media having various data structures stored thereon. Units can communicate, for example, through local and / or remote processes based on signals having one or more data packets (e.g., data from a second unit interacting with another unit in a local system, a distributed system, and / or a network. For example, the Internet interacts with other systems via signals).

[0047] Example 1

[0048] A method for mass spectrometry analysis of cancer tissue, comprising:

[0049] S1, build a cancer tissue mass spectrometry analysis detection scenario, the detection scenario includes:

[0050] Tissue sampling and extraction mechanism, used for sampling and extraction of cancer tissues;

[0051] an ion source mechanism connected to the tissue sampling and extraction mechanism;

[0052] a gas delivery mechanism connected to the ion source mechanism;

[0053] The mass spectrometry signal analysis system is connected to the ion source mechanism and is used to perform mass spectrometry analysis on the sample.

[0054] S2, sample pretreatment and device preparation, specifically, begins by carefully placing a fresh or appropriately preserved cancer tissue sample (1 mm³), obtained during surgery or biopsy, onto a specially prepared tissue extraction mixture filter membrane. This membrane, typically made of an inert, porous material, serves the critical function of allowing fluid flow while effectively trapping tissue fragments, preventing them from entering the delicate analytical pipelines and causing clogging.

[0055] Subsequently, the tissue-carrying filter membrane assembly is carefully integrated into the entire customized analytical device. This device usually contains an extraction liquid reservoir, a high-voltage electrode interface, a Y-type gas-liquid mixing sprayer, and a transmission line connected to the mass spectrometer. Ensuring the absolute air-tightness and liquid-tightness of all connections (such as pipe joints, membrane assembly seals, sprayer interfaces, etc.) is the top priority of this step. Any tiny leak will not only lead to the loss of extraction liquid or charged droplets and reduce detection sensitivity, but may also cause safety hazards or interfere with the mass spectrometry vacuum system due to the presence of high voltage (3kV).

[0056] S3, extraction liquid selection and charged mixed liquid generation, specifically the selection of high-purity methanol as the tissue extraction liquid. Due to its good polarity and solubility, methanol can effectively extract a variety of small molecule metabolites (such as amino acids, organic acids, lipids, sugars, etc.) in tissues. At the same time, its low surface tension and high volatility make it very suitable for the subsequent electrospray process. Methanol flows through a specific pipeline through the filter membrane where the tissue sample is placed. In this process, it dissolves and carries out the metabolite components in the tissue to form a "tissue-methanol mixed extract."

[0057] A high-voltage DC voltage of 3000 volts (3kV) is applied at a critical point in the device. This voltage is typically applied to a metal capillary or electrode through which the extractant flows. This high voltage imparts a significant charge (usually positive, depending on the voltage polarity) to the methanol-tissue metabolite mixture flowing through this point. This process essentially pre-ionizes the mixture at low pressure, laying the foundation for the subsequent efficient formation of charged droplets.

[0058] S4, atomization and gas-liquid mixing, specifically, the charged methanol-tissue metabolite mixture is pressure-driven and delivered to a Y-shaped gas-liquid mixing nebulizer (also known as the ESI source needle). At the intersection of the nebulizers, high-purity, high-flow nitrogen (N2) is ejected at high speed from another channel.

[0059] The high-speed nitrogen flow exerts intense shear and impact forces on the charged liquid stream, instantly breaking it into extremely fine charged droplets (mist) with diameters typically in the micrometer range. Nitrogen plays multiple roles here: first, providing the atomization force; second, acting as a drying gas to promote rapid evaporation of the solvent in the subsequent droplets; and third, acting as a shielding gas to prevent oxygen ingress and potential oxidation side reactions, while maintaining spray stability. This atomization process significantly increases the liquid's surface area, accelerating solvent evaporation and ionization efficiency.

[0060] S5, mass spectrometry analysis and signal acquisition, specifically, the charged droplet cloud formed, rich in tissue metabolite ions, is guided into the vacuum interface (usually the atmospheric pressure interface) of the mass spectrometer (MS) under the combined action of the electric field (usually formed by the voltage difference between the nebulizer and the mass spectrometer inlet) and the pressure difference.

[0061] Inside the mass spectrometer, as the solvent (methanol) rapidly evaporates, the size of the charged droplets shrinks dramatically, and the surface charge density of the droplets increases dramatically, ultimately leading to a Coulomb explosion or ion evaporation, completely releasing the metabolite molecules (or their adducts and fragments) contained within them as gas-phase ions. These ions are effectively collected and focused in the ion source region of the mass spectrometer.

[0062] The ions are then separated according to their mass-to-charge ratio (m / z) in a mass analyzer (e.g., quadrupole, time-of-flight, ion trap, etc.), and the detector records the signal intensity generated by ions of different m / z. The resulting mass spectrum is a raw mass spectrum that reflects the individual components in the complex mixture.

[0063] S6, data processing and metabolite analysis, specifically involves importing the raw mass spectrometry signal stream into a tissue ionization extraction signal analysis system (usually referring to the accompanying data acquisition and processing software). This system performs complex signal processing tasks, including background noise subtraction, baseline correction, peak detection (identifying signal peaks), peak integration (calculating peak area or height to represent relative abundance), and precise calibration of the mass axis.

[0064] Most importantly, the system identifies metabolites by comparing the detected signal peaks with a built-in or custom metabolite database (containing the precise molecular weight, isotope distribution, possible adduct ion forms, and characteristic fragment ion information of known metabolites). Ultimately, the system outputs a detailed report listing all metabolite types successfully extracted and identified from the cancer tissue sample and their relative or absolute content information.

[0065] It should be noted that, in this embodiment, the tissue sampling and extraction mechanism includes:

[0066] Syringe pump 1;

[0067] Extraction liquid injection needle 2, connected to the injection pump 1;

[0068] The syringe head 3 is connected to the extraction liquid injection needle 2;

[0069] The tissue extraction mixture filter membrane 31 is installed in the syringe head 3

[0070] An adapter 4 connected to the syringe head 3;

[0071] The capillary 5 is connected to the adapter 4, and the other end is connected to the ion source mechanism.

[0072] It should be noted that, in this embodiment, the ion source mechanism includes:

[0073] A Y-shaped gas-liquid mixing sprayer 6 is connected to the capillary tube;

[0074] Fixer, used to fix the Y-type gas-liquid mixing sprayer.

[0075] It should be noted that, in this embodiment, the gas delivery mechanism includes:

[0076] High purity nitrogen cylinder 8;

[0077] The voltage divider 9 is connected to the high-purity nitrogen bottle and the ion source mechanism through a gas delivery copper tube.

[0078] It should be noted that, in this embodiment, the tissue sampling and extraction mechanism further includes:

[0079] One end of the voltage device 10 is connected between the extraction liquid injection needle and the syringe head, and the other end is connected to the mass spectrometry signal analysis system 7.

[0080] According to the cancer tissue mass spectrometry detection method and detection system of the present invention, the excised cancer tissue sample is placed on a filter membrane soaked in an extract (methanol) and a sealed connection device is used. A high voltage of 3kV is applied to the ion source device to charge the methanol extract flowing through the tissue. The charged mixed liquid enters the Y-type sprayer and is atomized by the high-purity nitrogen provided by the gas delivery device. The atomized charged droplets are directly introduced into the mass spectrometer for analysis. Finally, the mass spectrometry signal analysis system processes the data, quickly and accurately identifies metabolites in the tissue, significantly improves detection efficiency and sensitivity, and reduces sample requirements and dependence on experience.

[0081] Example 2

[0082] A cancer tissue mass spectrometry analysis and detection system, wherein the above method is used to perform mass spectrometry analysis and detection on cancer tissue.

[0083] A cancer tissue mass spectrometry analysis and detection system in the embodiments of the present application can be a device, or a component, integrated circuit, or chip in a terminal. The device can be a mobile electronic device or a non-mobile electronic device. For example, the mobile electronic device can be a mobile phone, a tablet computer, a laptop computer, a PDA, an ultra-mobile personal computer (UMPC), a netbook, or a personal digital assistant (PDA), etc., and the non-mobile electronic device can be a server, a network attached storage (NAS), a personal computer (PC), etc., which is not specifically limited in the embodiments of the present application.

[0084] A cancer tissue mass spectrometry analysis and detection system in an embodiment of the present application may be a device having an operating system. The operating system may be an Android operating system, an iOS operating system, or other possible operating systems, which are not specifically limited in the embodiment of the present application.

[0085] The cancer tissue mass spectrometry analysis and detection system provided in the embodiment of the present application can achieve Figure 1 The various processes implemented in the method embodiment of a cancer tissue mass spectrometry detection method will not be described here in detail to avoid repetition.

[0086] According to the cancer tissue mass spectrometry detection system of an embodiment of the present invention, the excised cancer tissue sample can be placed on a filter membrane soaked in an extract (methanol) and a sealed connection device. A high voltage of 3kV is applied to the ion source device to charge the methanol extract flowing through the tissue. The charged mixed liquid enters the Y-type sprayer and is atomized by the high-purity nitrogen provided by the gas delivery device. The atomized charged droplets are directly introduced into the mass spectrometer for analysis. Finally, the mass spectrometry signal analysis system processes the data, quickly and accurately identifies metabolites in the tissue, significantly improves detection efficiency and sensitivity, and reduces sample requirements and dependence on experience.

[0087] Optionally, an embodiment of the present application further provides an electronic device, including a processor, a memory, and a program or instruction stored in the memory and executable on the processor. When the program or instruction is executed by the processor, each process of the above-mentioned cancer tissue mass spectrometry analysis and detection method embodiment is implemented, and the same technical effect can be achieved. To avoid repetition, it will not be described here.

[0088] An embodiment of the present application also provides a readable storage medium, on which a program or instruction is stored. When the program or instruction is executed by a processor, each process of the above-mentioned cancer tissue mass spectrometry analysis and detection method embodiment is implemented, and the same technical effect can be achieved. To avoid repetition, it will not be repeated here.

[0089] The processor is the processor in the electronic device in the above embodiment. The readable storage medium includes a computer-readable storage medium, such as a computer read-only memory (ROM), a random access memory (RAM), a magnetic disk, or an optical disk.

[0090] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like to indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore should not be understood as a limitation to the invention.

[0091] Throughout this specification, reference to terms such as "one embodiment," "some embodiments," "illustrative embodiments," "example," "specific example," or "some examples" means that a specific feature, structure, material, or characteristic described in conjunction with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, illustrative expressions of the above terms do not necessarily refer to the same embodiment or example.

[0092] Obviously, the described embodiments are only some of the embodiments of the present application, rather than all of the embodiments. Mentioning "embodiment" in this article means that the specific features, structures or characteristics described in conjunction with the embodiment may be included in at least one embodiment of the present embodiment application. The appearance of this phrase in various positions in the specification does not necessarily mean that they are all the same embodiments, nor are they independent or alternative embodiments that are mutually exclusive with other embodiments. It can be understood explicitly and implicitly by those skilled in the art that the embodiments described herein can be combined with other embodiments. Based on the embodiments in this application, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection of this application.

[0093] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to the embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the claims and their equivalents.

Claims

1. A method for mass spectrometry analysis of cancer tissue, characterized in that: include: S1, build a cancer tissue mass spectrometry analysis and detection scenario; S2, sample pretreatment and device preparation; S3, extraction liquid selection and charged mixed liquid generation; S4, atomization and gas-liquid mixing; S5, mass spectrometry analysis and signal acquisition; S6, Data processing and metabolite elucidation.

2. The method for mass spectrometry analysis of cancer tissue according to claim 1, characterized in that: In S1, the detection scenarios include: Tissue sampling and extraction mechanism, used for sampling and extraction of cancer tissues; an ion source mechanism connected to the tissue sampling and extraction mechanism; a gas delivery mechanism connected to the ion source mechanism; The mass spectrometry signal analysis system is connected to the ion source mechanism and is used to perform mass spectrometry analysis on the sample.

3. The method for mass spectrometry analysis of cancer tissue according to claim 2, characterized in that: The tissue sampling and extraction mechanism comprises: Syringe pumps; Extraction liquid injection needle, connected to the syringe pump; A syringe head connected to the extraction liquid injection needle; Tissue extraction mixture filter membrane, installed in the syringe head An adapter, connected to the syringe head; The capillary is connected to the adapter, and the other end is connected to the ion source mechanism.

4. The method for mass spectrometry analysis of cancer tissue according to claim 2, wherein: The ion source mechanism comprises: A Y-shaped gas-liquid mixing sprayer connected to the capillary tube; Fixer, used to fix the Y-type gas-liquid mixing sprayer.

5. The method for mass spectrometry analysis of cancer tissue according to claim 2, wherein: The gas delivery mechanism includes: High-purity nitrogen cylinders; The voltage divider is connected to the high-purity nitrogen gas bottle and the ion source mechanism through a gas delivery copper tube.

6. The method for mass spectrometry analysis of cancer tissue according to claim 3, characterized in that: The tissue sampling and extraction mechanism further includes: A voltage device has one end connected between the extraction liquid injection needle and the syringe head, and the other end connected to the mass spectrometry signal analysis system.

7. A cancer tissue mass spectrometry analysis and detection system, characterized in that: The method according to any one of claims 1 to 7 is used to perform mass spectrometry analysis on cancer tissue.

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