Metabolin detection method and system based on liquid chromatography-tandem mass spectrometry

Abstract

The invention relates to the technical field of metabolite electrochemical analysis, in particular to a metabolite detection method and system based on liquid chromatography-tandem mass spectrometry, and the method comprises the steps: collecting an original metabolite sample from a metabolite pore plate by using an automatic sampler, and carrying out intelligent elution on the original metabolite sample to obtain a target eluent; separating the target eluent by using the original mobile phase set, a degassing box, a liquid chromatography solution conveying box and a chromatographic column to obtain a target metabolite component set, transmitting the target metabolite component set to a mass spectrum unit to obtain a target mass spectrum unit, and performing signal detection on the target metabolite component set by using the target mass spectrum unit to obtain a component mass spectrum, and analyzing the component mass spectrum based on a liquid chromatography-tandem mass spectrometry analysis unit to obtain a metabolite detection report. According to the invention, the automation degree of the whole process of metabolite detection can be improved, and the accuracy of metabolite detection results is enhanced.

Description

Technical Field

[0001] This invention relates to the field of electrochemical analysis technology of metabolites, and in particular to a method and system for metabolite detection based on liquid chromatography-tandem mass spectrometry. Background Technology

[0002] Metabolites, as direct products of biochemical reactions in organisms, play an irreplaceable role in the accurate detection of disease biomarkers, drug metabolism kinetics, and systems biology. Liquid chromatography-tandem mass spectrometry (LC-MS / MS), with its high efficiency in chromatographic separation and the high sensitivity and specificity of mass spectrometry, has become a key technology for the qualitative and quantitative analysis of metabolites in complex biological samples, providing a core tool for a deeper understanding of the molecular mechanisms of life processes.

[0003] Traditional methods typically rely on manual visual positioning of sample wells, manual sampling, and offline pretreatment procedures such as dilution, derivatization, and solid-phase extraction, followed by detection using liquid chromatography-tandem mass spectrometry. The drawbacks of this approach are that manual operation makes it difficult to guarantee the accuracy and repeatability of sampling locations, and offline pretreatment steps are easily affected by factors such as operator experience and environmental fluctuations, leading to inconsistent sample processing conditions, which in turn introduces operational errors and batch-to-batch variations, thus affecting the accuracy and repeatability of the final detection results. Summary of the Invention

[0004] This invention provides a method for metabolite detection based on liquid chromatography-tandem mass spectrometry and a computer-readable storage medium. Its main purpose is to improve the automation level of the entire metabolite detection process and enhance the accuracy of metabolite detection results.

[0005] To achieve the above objectives, the present invention provides a method for metabolite detection based on liquid chromatography-tandem mass spectrometry, comprising:

[0006] A liquid chromatography-tandem mass spectrometry (LC-MS / MS) device is constructed, comprising: a high-performance liquid chromatography (HPLC) unit, a mass spectrometry (MS) unit, and an HPLC-MS / MS analysis unit. The HPLC unit includes: a degassing chamber, a liquid chromatography solution delivery chamber, an autosampler, a chromatographic column, and a column oven, with the chromatographic column housed within the column oven. The MS unit is a triple quadrupole mass spectrometer, and the HPLC unit and the MS unit are connected via a capillary tube.

[0007] Receive metabolite detection instructions, and obtain the original mobile phase set based on the metabolite detection instructions, wherein the original mobile phase set includes multiple original mobile phases;

[0008] The original metabolite samples were collected from the pre-constructed metabolite well plate using the autosampler in the high performance liquid chromatography unit, and the original metabolite samples were intelligently eluted to obtain the target eluent.

[0009] The target eluent was separated using the original mobile phase set, degassing box, liquid chromatography solution delivery box and chromatographic column to obtain the target metabolite component set;

[0010] The target metabolite components are transported to the mass spectrometry unit via capillary tubes to obtain the target mass spectrometry unit, which includes: a mass spectrometry ion source, a mass analyzer, and a particle detector;

[0011] The target metabolite component set is detected by the target mass spectrometry unit to obtain the component mass spectrum. The component mass spectrum is then analyzed by the liquid chromatography-tandem mass spectrometry analysis unit to obtain the metabolite detection report.

[0012] Optionally, the step of collecting raw metabolite samples from a pre-constructed metabolite plate using an autosampler in a high-performance liquid chromatography unit includes:

[0013] The well plate imaging device and intelligent injection needle in the autosampler were identified. The autosampler also includes: online concentration detection device, metabolite dilution device, internal standard addition device and online solid phase extraction column.

[0014] The well plate imaging device is activated based on the metabolite detection command. The well plate imaging device then acquires images of the metabolite well plate to obtain the target well plate image.

[0015] Metabolite location is identified based on the target plate image to obtain the current metabolite location;

[0016] The system automatically samples metabolites from well plates using the current metabolite location, preset metabolite sampling volume, and intelligent injection to obtain raw metabolite samples.

[0017] Optionally, the step of identifying the metabolite location based on the target well plate image to obtain the current metabolite location includes:

[0018] Edge extraction is performed on the target well plate image to obtain the well plate grid edge image, which includes multiple well plate metabolite grids;

[0019] A grid arrangement map is generated based on multiple well plate metabolite grids in the well plate grid edge image;

[0020] The target metabolite grid position is obtained based on the metabolite detection command, and the target metabolite grid is identified in the grid arrangement diagram according to the target metabolite grid position;

[0021] Metabolite features are extracted from the target metabolite grid to obtain metabolite features;

[0022] Determine whether the metabolite characteristics fall within the preset range of space characteristics;

[0023] If the metabolite features are not within the range of the space features, then the target metabolite grid is recorded as a valid metabolite grid;

[0024] Obtain the image center coordinates of the effective metabolite grid, transform the image center coordinates into the real coordinate system, and obtain the current metabolite position.

[0025] Optionally, the intelligent elution of the original metabolite sample to obtain the target eluent includes:

[0026] The concentration of the original metabolite sample is estimated based on the online concentration detection device in the autosampler to obtain the current metabolite concentration.

[0027] The original metabolite sample was diluted using the metabolite dilution equipment and the current metabolite concentration to obtain a diluted metabolite sample.

[0028] Internal standard solution was added to diluted metabolite samples using an internal standard addition device to obtain a sample-internal standard mixture.

[0029] The sample internal standard mixture was derivatized to obtain a derivatized mixture;

[0030] The derivatized mixture was loaded into an online solid-phase extraction column to obtain a loaded solid-phase extraction column;

[0031] The loaded solid-phase extraction column is cleaned to obtain a cleaned solid-phase extraction column. The cleaned solid-phase extraction column is then eluted with a preset strong elution solvent to obtain the target eluent.

[0032] Optionally, the online concentration detection device in the autosampler estimates the concentration of the original metabolite sample to obtain the current metabolite concentration, including:

[0033] The original metabolite sample was subjected to ultrasonic detection using an online concentration detection device to obtain the current ultrasonic detection signal;

[0034] Feature extraction is performed on the current ultrasonic detection signal to obtain the current ultrasonic feature vector;

[0035] Identify the image color feature vector of the original metabolite sample in the target well plate image;

[0036] Based on the current ultrasonic feature vector and image color feature vector, neighboring data indexing is performed in the pre-constructed concentration prior database to obtain the ultrasonic neighboring concentration and the image color neighboring concentration. The concentration prior database includes: ultrasonic prior database and color prior database.

[0037] The current metabolite concentration is obtained by weighted summation of adjacent concentrations in ultrasound and adjacent concentrations in image color.

[0038] Optionally, the step of performing adjacent data indexing in a pre-constructed concentration prior database based on the current ultrasonic feature vector and image color feature vector to obtain adjacent ultrasonic concentrations and adjacent image color concentrations includes:

[0039] Multiple ultrasound prior data in the ultrasound prior database of the concentration prior database were identified. Each ultrasound prior data includes: a prior ultrasound feature vector and a prior metabolite concentration.

[0040] Based on the current ultrasonic feature vector, vector similarity is calculated for multiple ultrasonic prior data to obtain multiple ultrasonic vector similarities.

[0041] Identify the maximum ultrasonic similarity among multiple ultrasonic vector similarities, and record the ultrasonic prior data corresponding to the maximum ultrasonic similarity as the adjacent ultrasonic prior data;

[0042] The prior metabolite concentrations in adjacent ultrasound prior data are denoted as adjacent ultrasound concentrations.

[0043] Identify adjacent color intensities in an image based on its color feature vector in a color prior database.

[0044] Optionally, the step of weighted summing of adjacent concentrations of ultrasound and adjacent concentrations of image colors to obtain the current metabolite concentration includes:

[0045] The ultrasonic feature index set in the current ultrasonic feature vector and the color feature index set in the image color feature vector are statistically analyzed separately.

[0046] Ultrasonic feature indicators are extracted sequentially from the set of ultrasonic feature indicators. Based on the extracted ultrasonic feature indicators, multiple ultrasonic data with the same name are extracted from the ultrasonic prior database. Each ultrasonic data with the same name includes: prior metabolite concentration and the corresponding value of ultrasonic feature indicator.

[0047] The correlation of ultrasonic indicators is obtained by calculating the concentration correlation of multiple ultrasonic data with the same name.

[0048] By summarizing the correlations of each ultrasonic feature index, we obtain the ultrasonic index correlation set.

[0049] Obtain the color index correlation set corresponding to the color feature index set from the color prior database;

[0050] The mean values ​​of the ultrasonic index correlation set and the color index correlation set are calculated respectively to obtain the average ultrasonic correlation and the average color correlation.

[0051] The current metabolite concentration is obtained by weighting the adjacent concentrations of ultrasound and adjacent concentrations of image color based on the average ultrasound correlation and average color correlation.

[0052] Optionally, the step of weighting the adjacent concentrations of ultrasound and adjacent concentrations of image colors based on average ultrasound correlation and average color correlation to obtain the current metabolite concentration includes:

[0053] The current metabolite concentration can be calculated using the following formula:

[0054]

[0055] in, Indicates the current concentration of metabolites. This represents the average ultrasound correlation. Indicates average color correlation. Indicates the adjacent concentrations of ultrasound waves. This indicates the density of adjacent colors in an image.

[0056] Optionally, the separation of the target eluent using the original mobile phase set, degassing chamber, liquid chromatography solution delivery box, and chromatographic column to obtain a target metabolite component set includes:

[0057] The original mobile phase set is introduced into the degassing box to obtain the degassed mobile phase set, and the degassed mobile phase set is transported to the liquid chromatography solution delivery box to obtain the target solution delivery box;

[0058] The degassed mobile phase is mixed under high pressure using a target solution delivery box and a preset delivery ratio to obtain a mixed mobile phase.

[0059] The target eluent is delivered to the chromatographic column using an autosampler and a mixed mobile phase to obtain the target chromatographic column;

[0060] The target eluent is separated into components using a target chromatographic column to obtain a target metabolite component set, which includes multiple target metabolite components.

[0061] To achieve the above objectives, the present invention also provides a metabolite detection system based on liquid chromatography-tandem mass spectrometry, comprising:

[0062] A mass spectrometry device construction module is used to construct a liquid chromatography-tandem mass spectrometry device. The liquid chromatography-tandem mass spectrometry device includes: a high-performance liquid chromatography unit, a mass spectrometry unit, and a liquid chromatography-tandem mass spectrometry analysis unit. The high-performance liquid chromatography unit includes: a degassing chamber, a liquid chromatography solution delivery chamber, an autosampler, a chromatographic column, and a column oven. The chromatographic column is built into the column oven. The mass spectrometry unit is a triple quadrupole mass spectrometer. The high-performance liquid chromatography unit and the mass spectrometry unit are connected through a capillary tube.

[0063] The metabolite elution module is used to receive metabolite detection commands and obtain the original mobile phase set based on the metabolite detection commands. The original mobile phase set includes multiple original mobile phases. The original metabolite samples are collected from the pre-constructed metabolite well plate using the autosampler in the high performance liquid chromatography unit. The original metabolite samples are intelligently eluted to obtain the target eluent.

[0064] The metabolite separation module is used to separate the target eluent using the original mobile phase set, degassing box, liquid chromatography solution delivery box and chromatographic column to obtain a target metabolite component set. The target metabolite component set is then transferred to the mass spectrometry unit via capillary to obtain the target mass spectrometry unit, which includes a mass spectrometry ion source, a mass analyzer and a particle detector.

[0065] The detection report generation module is used to detect the target metabolite component set by using the target mass spectrometry unit to obtain the component mass spectrum, and analyze the component mass spectrum based on the liquid chromatography-tandem mass spectrometry analysis unit to obtain the metabolite detection report.

[0066] To address the above problems, the present invention also provides an electronic device, the electronic device comprising:

[0067] Memory, storing at least one instruction;

[0068] The processor executes the instructions stored in the memory to implement the metabolite detection method based on liquid chromatography-tandem mass spectrometry described above.

[0069] To address the aforementioned problems, the present invention also provides a computer-readable storage medium storing at least one instruction, which is executed by a processor in an electronic device to implement the above-described method for metabolite detection based on liquid chromatography-tandem mass spectrometry.

[0070] To address the problems described in the background section, this invention first constructs a liquid chromatography-tandem mass spectrometry (LC-MS / MS) device. The autosampler in this step possesses online pretreatment and intelligent positioning functions, and the use of a triple quadrupole mass spectrometer provides crucial hardware support for subsequent high-sensitivity and high-selectivity metabolite detection processes that require no manual intervention. Compared to traditional devices relying on manual operation and simple detectors, this step offers significant advantages in automation and detection performance. Furthermore, this scheme utilizes the autosampler within the high-performance liquid chromatography (HPLC) unit to collect raw metabolite samples from pre-constructed metabolite well plates. The raw metabolite samples undergo intelligent elution to obtain the target eluent. This step, by integrating modules such as well plate visual positioning, intelligent injection needles, online concentration detection and automated derivatization, and solid-phase extraction, replaces the traditional technology that relies entirely on manual intervention. This method, which eliminates the need for manual visual positioning, sampling, offline preparation, and processing, not only improves the accuracy and repeatability of sampling locations but also ensures consistency of pretreatment conditions across different samples through built-in automated steps. This fundamentally reduces errors and variability introduced by manual operation. Finally, the target eluent is separated using the original mobile phase set, degassing chamber, liquid chromatography solution delivery box, and chromatographic column to obtain the target metabolite component set. In this step, online degassing and high-pressure gradient mixing of the mobile phase are automatically completed by the system. Combined with a temperature-controlled chromatographic column, this provides stable and precisely controllable chromatographic conditions for the separation of metabolite components, overcoming the inconsistent separation results caused by manual mobile phase preparation, insufficient degassing, or temperature fluctuations in traditional methods. This provides a stable and reliable component input for subsequent mass spectrometry detection. Therefore, this invention improves the automation level of the entire metabolite detection process and enhances the accuracy of metabolite detection results. Attached Figure Description

[0071] Figure 1 This is a schematic flowchart of a metabolite detection method based on liquid chromatography-tandem mass spectrometry provided in an embodiment of the present invention;

[0072] Figure 2 This is a functional block diagram of a metabolite detection system based on liquid chromatography-tandem mass spectrometry provided in an embodiment of the present invention;

[0073] Figure 3 This is a schematic diagram of the structure of an electronic device for implementing the metabolite detection method based on liquid chromatography-tandem mass spectrometry, according to an embodiment of the present invention.

[0074] Explanation of reference numerals in the attached figures:

[0075] 10. Electronic device; 11. Processor; 12. Memory; 13. Bus.

[0076] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0077] It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0078] This application provides a method for metabolite detection based on liquid chromatography-tandem mass spectrometry (LC-MS / MS). The execution entity of this method includes, but is not limited to, at least one electronic device configured to execute the method provided in this application, such as a server or a terminal. In other words, the method can be executed by software or hardware installed on a terminal or server device, and the software may be a blockchain platform. The server includes, but is not limited to, a single server, a server cluster, a cloud server, or a cloud server cluster.

[0079] Reference Figure 1 The diagram shown is a schematic flowchart of a metabolite detection method based on liquid chromatography-tandem mass spectrometry (LC-MS / MS) according to an embodiment of the present invention. In this embodiment, the metabolite detection method based on LC-MS / MS includes:

[0080] S1. Construct a liquid chromatography-tandem mass spectrometry (LC-MS / MS) device, wherein the LC-MS / MS device includes: a high-performance liquid chromatography (HPLC) unit, a mass spectrometry (MS) unit, and an HPLC-MS / MS analysis unit. The HPLC unit includes: a degassing chamber, a liquid chromatography solution delivery chamber, an autosampler, a chromatographic column, and a column oven, with the chromatographic column housed within the column oven. The MS unit is a triple quadrupole mass spectrometer, and the HPLC unit and the MS unit are connected via a capillary tube.

[0081] As is clear, the liquid chromatography-tandem mass spectrometry (LC-MS / MS) device refers to an instrument system used for detecting and analyzing metabolites. This LC-MS / MS device consists of a high-performance liquid chromatography (HPLC) unit, a mass spectrometry (MS / MS) unit, and an HPLC-MS / MS analysis unit. The HPLC unit refers to a subsystem including a degassing chamber, a liquid chromatography solution delivery chamber, an autosampler, a chromatographic column, and a column oven. This HPLC unit is used for the automatic acquisition, pretreatment, separation, and elution of the raw metabolite sample. The degassing chamber is a device used to remove dissolved gases from the subsequent raw mobile phase. The liquid chromatography solution delivery chamber is a device used to receive and deliver the degassed raw mobile phase; for example, a binary or quaternary high-pressure gradient pump can be used as the liquid chromatography solution delivery chamber. The autosampler is a device that converts the raw metabolite sample into the target eluent; the specific configuration of the autosampler will be described in detail in subsequent embodiments. The chromatographic column refers to a device built into the column oven used to separate the various target metabolite components in the target eluent; for example, a reversed-phase column (such as a C18 column) or a hydrophilic interaction column. The column oven refers to the device used to contain and control the temperature of the chromatographic column. The mass spectrometry unit refers to a triple quadrupole mass spectrometer, which is used for ionization, mass analysis, and signal detection of the target metabolite fractions separated by chromatography. The liquid chromatography-tandem mass spectrometry analysis unit refers to the software system used to analyze the component mass spectra generated by the mass spectrometry unit and generate a final metabolite detection report; for example, Analyst or MassHunter software can be used as this liquid chromatography-tandem mass spectrometry analysis unit. The capillary refers to the connecting component used to transfer the target metabolite fractions separated by the high-performance liquid chromatography unit to the mass spectrometry unit.

[0082] S2. Receive metabolite detection instructions and obtain the original mobile phase set based on the metabolite detection instructions. The original mobile phase set includes multiple original mobile phases.

[0083] Understandably, the metabolite detection instruction refers to a manually initiated instruction to detect metabolites. The original mobile phase set refers to a collection of multiple original mobile phases, wherein the original mobile phase refers to a single solvent or solution used to constitute the mobile phase required for chromatographic separation. This original mobile phase is used as a carrier for subsequent chromatographic column separation, carrying the target eluent through the chromatographic column under the transport of the liquid chromatography solution delivery box to achieve the separation of target metabolite components. Examples include: aqueous phase (such as ultrapure water, aqueous solution containing volatile buffer salts or acid-base additives), organic phase (such as methanol, acetonitrile, isopropanol), etc.

[0084] S3. Use the autosampler in the high performance liquid chromatography unit to collect the original metabolite sample from the pre-constructed metabolite well plate, and perform intelligent elution on the original metabolite sample to obtain the target eluent.

[0085] As is understood, the metabolite plate refers to a standard laboratory container used to hold samples of metabolites to be tested. This metabolite plate has a regularly arranged array of wells to facilitate positioning and sampling by an automated sampler, such as a 96-well plate, a 384-well plate, or other standard multi-well plates. The raw metabolite sample refers to the metabolite sample to be tested collected from the metabolite plate, such as blood, urine, cell culture supernatant, tissue homogenate, or other biological samples after preliminary processing. The target eluent refers to the liquid obtained after intelligent elution of the raw metabolite sample.

[0086] Specifically, the method of collecting raw metabolite samples from pre-constructed metabolite well plates using an autosampler in a high-performance liquid chromatography unit includes:

[0087] The well plate imaging device and intelligent injection needle in the autosampler were identified. The autosampler also includes: online concentration detection device, metabolite dilution device, internal standard addition device and online solid phase extraction column.

[0088] The well plate imaging device is activated based on the metabolite detection command. The well plate imaging device then acquires images of the metabolite well plate to obtain the target well plate image.

[0089] Metabolite location is identified based on the target plate image to obtain the current metabolite location;

[0090] The system automatically samples metabolites from well plates using the current metabolite location, preset metabolite sampling volume, and intelligent injection to obtain raw metabolite samples.

[0091] It should be explained that the well plate imaging device refers to a camera device used to photograph metabolite well plates, such as a CCD camera or a CMOS camera. This well plate imaging device is mounted on the robotic arm or fixed bracket of the autosampler to acquire a complete top-view image of the metabolite well plate. The intelligent injection needle refers to an injection needle that can accurately locate and automatically sample based on the current metabolite position, such as a three-dimensional motion robotic arm equipped with a high-precision servo motor. The above-mentioned online concentration detection device, metabolite dilution device, internal standard addition device, and online solid-phase extraction column will be described in detail in subsequent embodiments. The target well plate image refers to the top-view image of the metabolite well plate acquired by the well plate imaging device. When the well plate imaging device receives a metabolite detection command, it will photograph the metabolite well plate to obtain the target well plate image. The current metabolite position refers to the specific location of the original metabolite sample to be collected in the metabolite well plate. The metabolite sampling volume refers to the volume of the original metabolite sample to be extracted. This metabolite sampling volume is set by the relevant testing personnel or according to the relevant device operation manual. The original metabolite sample refers to a volume S of metabolite extracted from the current metabolite location by the intelligent injection needle, where S represents the metabolite sampling volume. This solution, by introducing this well plate imaging device and intelligent injection needle, improves the accuracy and repeatability of sampling location compared to traditional techniques relying on manual positioning and sampling, and achieves fully automated operation, reducing errors and variability caused by manual operation.

[0092] Specifically, the step of identifying the metabolite location based on the target well plate image to obtain the current metabolite location includes:

[0093] Edge extraction is performed on the target well plate image to obtain the well plate grid edge image, which includes multiple well plate metabolite grids;

[0094] A grid arrangement map is generated based on multiple well plate metabolite grids in the well plate grid edge image;

[0095] The target metabolite grid position is obtained based on the metabolite detection command, and the target metabolite grid is identified in the grid arrangement diagram according to the target metabolite grid position;

[0096] Metabolite features are extracted from the target metabolite grid to obtain metabolite features;

[0097] Determine whether the metabolite characteristics fall within the preset range of space characteristics;

[0098] If the metabolite features are not within the range of the space features, then the target metabolite grid is recorded as a valid metabolite grid;

[0099] Obtain the image center coordinates of the effective metabolite grid, transform the image center coordinates into the real coordinate system, and obtain the current metabolite position.

[0100] It should be explained that the well plate grid edge image refers to a binary image obtained after edge extraction, retaining only the outline of the metabolite well plate and the edge information of each well. Edge extraction of the target well plate image involves using image processing algorithms such as the Canny operator and the Sobel operator to detect areas with abrupt changes in grayscale values, thereby outlining the boundaries of the metabolite well plate and each well (i.e., the well plate metabolite grid). The well plate metabolite grid refers to the rectangular or circular region identified on the well plate grid edge image, corresponding to the physical well position of the metabolite well plate. Each well plate metabolite grid represents an independent sample well in the metabolite well plate. The grid arrangement diagram refers to a position index diagram composed of multiple well plate metabolite grids, in which each well plate metabolite has a corresponding arrangement position. The target metabolite grid position refers to the specific grid position where the metabolite to be detected, as included in the metabolite detection instruction, is placed. For example, it could be the first row and first column. The origin is the lower left vertex of the metabolite well plate, the longer side starting from this lower left vertex is the first row, and the shorter side starting from this lower left vertex is the first column. This lower left vertex is specially marked to allow the origin to be located in the grid arrangement diagram. The target metabolite grid refers to the well plate metabolic grid in the grid arrangement diagram corresponding to the target metabolite grid position.

[0101] Furthermore, the metabolite features refer to image features such as color histogram, average brightness, and texture at the target metabolite grid. These metabolite features are used to determine whether the target metabolite grid is empty. If the metabolite features are not within the empty feature range, it indicates that the target metabolite grid contains metabolites, and subsequent operations can proceed. If the metabolite features are within the empty feature range, it indicates that the target metabolite grid is empty, possibly due to operator error, such as misremembering the target metabolite grid position. In this case, an error message needs to be generated to remind the operator to correct the error. The empty feature range refers to a set of multiple numerical intervals determined by statistically analyzing multiple image features of a large number of known empty well plate grids. The numerical intervals included in the empty feature range correspond one-to-one with the image features in the metabolite features. The above-mentioned metabolite features not being within the empty feature range means that each image feature in the metabolite features is not within the corresponding numerical interval in the empty feature range. The image center coordinates refer to the coordinates of the geometric center of the effective metabolite grid in the well plate grid edge image. The aforementioned real-world coordinate system transformation refers to establishing a mapping matrix between the pixel coordinate system of the well plate grid edge image and the physical world coordinate system of the autosampler robotic arm through a calibration process. This mapping matrix is ​​constructed as follows: using a calibration plate of known precise dimensions (such as the metabolite well plate in this scheme), multiple images of the calibration plate are captured from different angles by a camera in the well plate imaging device. The pixel coordinates of feature points (such as corner points and geometric center points) in each image are detected and paired with the known physical world coordinates of these feature points, thus obtaining pixel pairing groups composed of pixel coordinates and physical world coordinates. Based on these pixel pairing groups, algorithms such as the Zhang Zhengyou calibration method are used to solve for the intrinsic parameter matrix (including parameters such as focal length and principal point) and extrinsic parameter matrix of the camera in the well plate imaging device. Next, the hand-eye calibration method is used to confirm the hand-eye matrix between the camera and the end of the autosampler robotic arm. Finally, the mapping matrix is ​​constructed using the hand-eye matrix, intrinsic parameter matrix, and extrinsic parameter matrix. Using this mapping matrix, the image center coordinates of the identified effective metabolite grid can be converted into a real coordinate system that controls the movement of the smart injection needle, thereby determining the current metabolite position in this real coordinate system.

[0102] Specifically, the intelligent elution of the original metabolite sample to obtain the target eluent includes:

[0103] The concentration of the original metabolite sample is estimated based on the online concentration detection device in the autosampler to obtain the current metabolite concentration.

[0104] The original metabolite sample was diluted using the metabolite dilution equipment and the current metabolite concentration to obtain a diluted metabolite sample.

[0105] Internal standard solution was added to diluted metabolite samples using an internal standard addition device to obtain a sample-internal standard mixture.

[0106] The sample internal standard mixture was derivatized to obtain a derivatized mixture;

[0107] The derivatized mixture was loaded into an online solid-phase extraction column to obtain a loaded solid-phase extraction column;

[0108] The loaded solid-phase extraction column is cleaned to obtain a cleaned solid-phase extraction column. The cleaned solid-phase extraction column is then eluted with a preset strong elution solvent to obtain the target eluent.

[0109] It should be explained that the online concentration detection device refers to a device used to estimate the concentration of the original metabolite sample in real time, such as an ultrasonic sensor. The current metabolite concentration refers to the solute concentration of the original metabolite sample. The metabolite dilution device refers to a device used to automatically add diluent to the original metabolite sample, such as a high-precision syringe pump or a volumetric dilution pump. The diluted metabolite sample refers to the original metabolite sample after dilution. The internal standard addition device refers to a device used to quantitatively add internal standard solution to the diluted metabolite sample, such as a high-precision syringe pump or a nano-syringe. The internal standard solution refers to a compound solution with a known concentration and chemical properties similar to the target metabolite component but not present in the original metabolite sample, such as a stable isotope-labeled metabolite of the same type (e.g., 13C- or 2H-labeled amino acids). The sample internal standard mixture refers to the diluted metabolite sample after the addition of the internal standard solution. The derivatized mixture refers to the sample internal standard mixture after the derivatization reaction has been completed. Derivatization of the sample internal standard mixture involves adding a derivatizing reagent to the mixture, thereby chemically modifying the target metabolite component. The purpose of this derivatization is to improve the detection sensitivity of the target metabolite component and enhance chromatographic separation behavior. For example, it can imbue the target metabolite component, which is normally difficult to ionize or volatilize, with easily detectable functional groups.

[0110] Furthermore, the online solid-phase extraction column refers to a micro-extraction column filled with a mixed-mode adsorbent or ion-exchange packing material. The loaded solid-phase extraction column refers to an online solid-phase extraction column after the derivatization mixture has been loaded. The impurity-removing solid-phase extraction column refers to a loaded solid-phase extraction column after the cleaning step has been completed. Cleaning the loaded solid-phase extraction column involves passing a weak elution solvent (such as an aqueous solution containing a small amount of organic phase) through the column to wash away impurities that are non-specifically adsorbed, while the target metabolite component remains on the column. The strong elution solvent refers to a solvent with strong elution capacity that can efficiently elute the target metabolite component retained on the impurity-removing solid-phase extraction column, such as an 80% methanol / acetonitrile aqueous solution. Eluting the impurity-removing solid-phase extraction column based on the preset strong elution solvent involves loading the strong elution solvent into the impurity-removing solid-phase extraction column, eluting the purified target metabolite component and the component in the internal standard solution together from the loaded impurity-removing solid-phase extraction column, and obtaining the eluted liquid as the target eluent.

[0111] In detail, the online concentration detection device in the autosampler estimates the concentration of the original metabolite sample to obtain the current metabolite concentration, including:

[0112] The original metabolite sample was subjected to ultrasonic detection using an online concentration detection device to obtain the current ultrasonic detection signal;

[0113] Feature extraction is performed on the current ultrasonic detection signal to obtain the current ultrasonic feature vector;

[0114] Identify the image color feature vector of the original metabolite sample in the target well plate image;

[0115] Based on the current ultrasonic feature vector and image color feature vector, neighboring data indexing is performed in the pre-constructed concentration prior database to obtain the ultrasonic neighboring concentration and the image color neighboring concentration. The concentration prior database includes: ultrasonic prior database and color prior database.

[0116] The current metabolite concentration is obtained by weighted summation of adjacent concentrations in ultrasound and adjacent concentrations in image color.

[0117] It should be explained that the current ultrasonic detection signal refers to the original electrical signal received after the original metabolite sample is ultrasonically detected by an online concentration detection device, reflecting the acoustic characteristics of the sample (such as sound velocity, attenuation, impedance, etc.). Ultrasonic detection of the original metabolite sample using an online concentration detection device means: immersing the ultrasonic probe of the online concentration detection device into a container containing the original metabolite sample, then emitting ultrasonic waves of a specific frequency and receiving the ultrasonic signals reflected back from the interface of the original metabolite sample, thereby obtaining acoustic information related to the physicochemical properties of the original metabolite sample. The current ultrasonic feature vector refers to a numerical vector composed of multiple ultrasonic features obtained through feature extraction. Optionally, the ultrasonic features are sound velocity, ultrasonic signal amplitude attenuation coefficient, acoustic impedance, etc. Feature extraction of the current ultrasonic detection signal means: quantizing and calculating one or more of the above-mentioned ultrasonic features from the current ultrasonic detection signal using signal processing algorithms (such as calculating time of flight, measuring peak amplitude attenuation, performing fast Fourier transform, etc.), and combining these ultrasonic features into a numerical vector. The image color feature vector refers to the vector composed of the color features of the target metabolite grid where the original metabolite sample is located. Optionally, the color features may include color channel statistics, color histograms, color moments, etc. The concentration prior database refers to the total database including the ultrasound prior database and the color prior database. The ultrasound adjacent concentration refers to the metabolite concentration in the ultrasound prior data that is most similar to the current ultrasound feature vector in the ultrasound prior database (i.e., the subsequent prior metabolite concentration). The image color adjacent concentration refers to the metabolite concentration in the color prior data that is most similar to the image color feature vector in the color prior database.

[0118] In detail, the step of performing adjacent data indexing in a pre-constructed concentration prior database based on the current ultrasonic feature vector and image color feature vector to obtain adjacent ultrasonic concentrations and adjacent image color concentrations includes:

[0119] Multiple ultrasound prior data in the ultrasound prior database of the concentration prior database were identified. Each ultrasound prior data includes: a prior ultrasound feature vector and a prior metabolite concentration.

[0120] Based on the current ultrasonic feature vector, vector similarity is calculated for multiple ultrasonic prior data to obtain multiple ultrasonic vector similarities.

[0121] Identify the maximum ultrasonic similarity among multiple ultrasonic vector similarities, and record the ultrasonic prior data corresponding to the maximum ultrasonic similarity as the adjacent ultrasonic prior data;

[0122] The prior metabolite concentrations in adjacent ultrasound prior data are denoted as adjacent ultrasound concentrations.

[0123] Identify adjacent color intensities in an image based on its color feature vector in a color prior database.

[0124] It is clear that the aforementioned ultrasound prior data refers to a data combination composed of prior ultrasound feature vectors and prior metabolite concentrations. The construction method of the aforementioned ultrasound prior database is as follows: multiple metabolite samples with known concentrations (i.e., prior metabolite concentrations) are obtained; ultrasound detection is performed on each metabolite sample with a known concentration, thereby obtaining multiple prior ultrasound feature vectors; these multiple prior ultrasound feature vectors are combined with the known concentrations of the corresponding metabolite samples; the combined data is the ultrasound prior data, and the ultrasound prior data corresponding to each metabolite sample with a known concentration constitutes the concentration prior database. The ultrasound vector similarity refers to the vector cosine value between the current ultrasound feature vector and a prior ultrasound feature vector in a certain ultrasound prior data. The maximum ultrasound similarity refers to the ultrasound vector similarity with the largest value among multiple ultrasound vector similarities. The aforementioned color prior database contains multiple color prior data, and the construction method of this color prior database and the color prior data is the same as that of the aforementioned ultrasound prior database and the ultrasound prior data. Furthermore, the steps for identifying adjacent concentrations of image colors in the color prior database based on image color feature vectors are the same as the steps for obtaining adjacent ultrasound concentrations, and will not be repeated here.

[0125] Specifically, the weighted summation of adjacent concentrations of ultrasound and adjacent concentrations of image color to obtain the current metabolite concentration includes:

[0126] The ultrasonic feature index set in the current ultrasonic feature vector and the color feature index set in the image color feature vector are statistically analyzed separately.

[0127] Ultrasonic feature indicators are extracted sequentially from the set of ultrasonic feature indicators. Based on the extracted ultrasonic feature indicators, multiple ultrasonic data with the same name are extracted from the ultrasonic prior database. Each ultrasonic data with the same name includes: prior metabolite concentration and the corresponding value of ultrasonic feature indicator.

[0128] The correlation of ultrasonic indicators is obtained by calculating the concentration correlation of multiple ultrasonic data with the same name.

[0129] By summarizing the correlations of each ultrasonic feature index, we obtain the ultrasonic index correlation set.

[0130] Obtain the color index correlation set corresponding to the color feature index set from the color prior database;

[0131] The mean values ​​of the ultrasonic index correlation set and the color index correlation set are calculated respectively to obtain the average ultrasonic correlation and the average color correlation.

[0132] The current metabolite concentration is obtained by weighting the adjacent concentrations of ultrasound and adjacent concentrations of image color based on the average ultrasound correlation and average color correlation.

[0133] It should be explained that the ultrasonic feature index set refers to the set of ultrasonic feature types corresponding to each vector value in the current ultrasonic feature vector, such as: sound velocity index, attenuation index, impedance index, etc. The color feature index set refers to the set of color feature types corresponding to each vector value in the image color feature vector. The homologous ultrasonic data refers to the ultrasonic prior data corresponding to the extracted ultrasonic feature index. The ultrasonic index correlation refers to the correlation between the ultrasonic feature index and the prior metabolite concentration. Specifically, calculating the concentration correlation based on multiple homologous ultrasonic data means calculating the Pearson correlation coefficient between all prior metabolite concentrations and all corresponding ultrasonic feature index values ​​in the multiple homologous ultrasonic data sets; this Pearson correlation coefficient is the ultrasonic index correlation. The color index correlation set refers to the set of multiple color index correlations. Specifically, color index correlation refers to the correlation between a certain color feature index in the color prior database and the metabolite concentration; the calculation method for color index correlation is the same as that for ultrasonic index correlation. The average ultrasonic correlation refers to the average value of all ultrasonic index correlations in the ultrasonic index correlation set, and the average color correlation refers to the average value of all color index correlations in the color index correlation set.

[0134] In detail, the weighted calculation of adjacent concentrations of ultrasound and adjacent concentrations of image color based on average ultrasound correlation and average color correlation to obtain the current metabolite concentration includes:

[0135] The current metabolite concentration can be calculated using the following formula:

[0136]

[0137] in, Indicates the current concentration of metabolites. This represents the average ultrasound correlation. Indicates average color correlation. Indicates the adjacent concentrations of ultrasound waves. This indicates the density of adjacent colors in an image.

[0138] It is understandable that, since the correlations (i.e., average ultrasound correlation or average color correlation) between ultrasound features and color features and metabolite concentrations differ, adjacent concentrations corresponding to features with higher correlations can be assigned higher weights. This means that the correlation can be introduced into the current metabolite concentration calculation formula. Item and item.

[0139] S4. The target eluent is separated using the original mobile phase set, degassing box, liquid chromatography solution delivery box and chromatographic column to obtain the target metabolite component set.

[0140] Understandably, the target metabolite component set refers to a collection of multiple target metabolite components, wherein the target metabolite component refers to the compound corresponding to a single metabolite (or internal standard) peak with a specific retention time in the subsequent component mass spectrum obtained after separation.

[0141] In detail, the separation of the target eluent using the original mobile phase set, degassing chamber, liquid chromatography solution delivery box, and chromatographic column to obtain a target metabolite component set includes:

[0142] The original mobile phase set is introduced into the degassing box to obtain the degassed mobile phase set, and the degassed mobile phase set is transported to the liquid chromatography solution delivery box to obtain the target solution delivery box;

[0143] The degassed mobile phase is mixed under high pressure using a target solution delivery box and a preset delivery ratio to obtain a mixed mobile phase.

[0144] The target eluent is delivered to the chromatographic column using an autosampler and a mixed mobile phase to obtain the target chromatographic column;

[0145] The target eluent is separated into components using a target chromatographic column to obtain a target metabolite component set, which includes multiple target metabolite components.

[0146] It is clear that the degassed mobile phase set refers to the collection of each original mobile phase in the original mobile phase set after degasing in the degasing chamber. Degasing refers to removing dissolved gases (such as oxygen and nitrogen) from the original mobile phase using online vacuum degasing or helium bubbling. The target solution delivery tank refers to the liquid chromatography solution delivery tank that receives the degassed mobile phase set. The delivery ratio refers to a manually set volume ratio for mixing different degassed mobile phases, which can be set according to relevant industry regulations or literature. The mixed mobile phase is the mobile phase obtained by mixing all the degassed mobile phases in the degassed mobile phase set. High-pressure delivery mixing of the degassed mobile phase set using the target solution delivery tank and the preset delivery ratio refers to: multiple degassed mobile phases of corresponding proportions being extracted and delivered by different channels of the target solution delivery tank according to the delivery ratio. These extracted degassed mobile phases are thoroughly mixed in the high-pressure mixer within the target solution delivery tank, thereby forming a mixed mobile phase. The target chromatographic column refers to the chromatographic column through which the mixed mobile phase and target eluent flow. The separation of components in the target eluent using the target chromatographic column refers to the following: the target eluent is pumped into the target chromatographic column under the carrying of a mixed mobile phase. Due to the differences in the interaction forces (such as partitioning, adsorption, ion exchange, etc.) between different metabolite components in the target eluent and the stationary phase of the chromatographic column, these metabolite components pass through the target chromatographic column at different speeds under the impetus of the mixed mobile phase, thus being separated in terms of the time it takes to elute from the target chromatographic column, forming a time-discrete set of target metabolite components that are sequentially detected by the mass spectrometry unit.

[0147] S5. Based on the capillary, the target metabolite components are aggregated and transported to the mass spectrometry unit to obtain the target mass spectrometry unit, which includes: a mass spectrometry ion source, a mass analyzer and a particle detector.

[0148] It should be explained that the target mass spectrometry unit refers to the mass spectrometry unit that receives the target metabolite component set. The mass spectrometry ion source refers to the device that converts the target metabolite components, after chromatographic separation and capillary transmission, into gaseous ions, such as an electrospray ionization (ESI) source. The mass analyzer refers to the component that separates the gaseous ions according to their mass-to-charge ratio (m / z), and this mass analyzer is a triple quadrupole mass analyzer. The particle detector refers to the device used to convert the gaseous ions screened by the mass analyzer into a measurable electrical signal, such as an electron multiplier (EM).

[0149] S6. Detect the target metabolite component set using the target mass spectrometry unit to obtain the component mass spectrum. Analyze the component mass spectrum using the liquid chromatography-tandem mass spectrometry analysis unit to obtain a metabolite detection report.

[0150] Understandably, the component mass spectrum refers to a data spectrum reflecting the signal of the target metabolite component obtained by the target mass spectrometry unit. The specific method of using the target mass spectrometry unit to detect the signal of the target metabolite component set is as follows: each target metabolite component in the target metabolite component set is subjected to the following operation: the target metabolite component enters the mass spectrometry ion source in the target mass spectrometry unit, and the mass spectrometry ion source ionizes the target metabolite component after entering, thereby generating charged phase ions. The charged phase ions are introduced into the mass analyzer under vacuum. After being screened and fragmented by the mass analyzer, the signal intensity of the screened charged phase ions is detected by the particle detector, thereby obtaining the component mass spectrum. The above method of obtaining the component mass spectrum is existing technology and will not be described in detail here. The metabolite detection report refers to an output document containing the content of each target metabolite component in the target metabolite component set. The analysis of the component mass spectrum based on a liquid chromatography-tandem mass spectrometry (LC-MS / MS) unit involves: the LC-MS / MS unit automatically identifying chromatographic peaks in the component mass spectrum, calculating the peak area or peak height by integration, and then calculating the content of each target metabolite component according to a pre-established standard curve or internal standard method. Finally, the retention time and mass spectrometry fragment information of each target metabolite component are compared with a standard spectral library to confirm the identity of each target metabolite component. Based on the identity and corresponding content of all target metabolite components, a structured metabolite detection report is generated.

[0151] To address the problems described in the background section, this invention first constructs a liquid chromatography-tandem mass spectrometry (LC-MS / MS) device. The autosampler in this step possesses online pretreatment and intelligent positioning functions, and the use of a triple quadrupole mass spectrometer provides crucial hardware support for subsequent high-sensitivity and high-selectivity metabolite detection processes that require no manual intervention. Compared to traditional devices relying on manual operation and simple detectors, this step offers significant advantages in automation and detection performance. Furthermore, this scheme utilizes the autosampler within the high-performance liquid chromatography (HPLC) unit to collect raw metabolite samples from pre-constructed metabolite well plates. The raw metabolite samples undergo intelligent elution to obtain the target eluent. This step, by integrating modules such as well plate visual positioning, intelligent injection needles, online concentration detection and automated derivatization, and solid-phase extraction, replaces the traditional technology that relies entirely on manual intervention. This method, which eliminates the need for manual visual positioning, sampling, offline preparation, and processing, not only improves the accuracy and repeatability of sampling locations but also ensures consistency of pretreatment conditions across different samples through built-in automated steps. This fundamentally reduces errors and variability introduced by manual operation. Finally, the target eluent is separated using the original mobile phase set, degassing chamber, liquid chromatography solution delivery box, and chromatographic column to obtain the target metabolite component set. In this step, online degassing and high-pressure gradient mixing of the mobile phase are automatically completed by the system. Combined with a temperature-controlled chromatographic column, this provides stable and precisely controllable chromatographic conditions for the separation of metabolite components, overcoming the inconsistent separation results caused by manual mobile phase preparation, insufficient degassing, or temperature fluctuations in traditional methods. This provides a stable and reliable component input for subsequent mass spectrometry detection. Therefore, this invention improves the automation level of the entire metabolite detection process and enhances the accuracy of metabolite detection results.

[0152] like Figure 2 The diagram shown is a functional block diagram of a metabolite detection system based on liquid chromatography-tandem mass spectrometry provided in an embodiment of the present invention.

[0153] The metabolite detection system 100 based on liquid chromatography-tandem mass spectrometry (LC-MS / MS) of the present invention can be installed in an electronic device. Depending on the functions implemented, the metabolite detection system 100 may include a mass spectrometry device construction module 101, a metabolite elution module 102, a metabolite separation module 103, and a detection report generation module 104. The module described in this invention can also be referred to as a unit, which refers to a series of computer program segments that can be executed by the processor of an electronic device and can perform a fixed function, and which are stored in the memory of the electronic device.

[0154] The mass spectrometry device construction module 101 is used to construct a liquid chromatography-tandem mass spectrometry device, wherein the liquid chromatography-tandem mass spectrometry device includes: a high performance liquid chromatography unit, a mass spectrometry unit, and a liquid chromatography-tandem mass spectrometry analysis unit. The high performance liquid chromatography unit includes: a degassing chamber, a liquid chromatography solution delivery chamber, an autosampler, a chromatographic column, and a column oven, and the chromatographic column is built into the column oven. The mass spectrometry unit is a triple quadrupole mass spectrometer, and the high performance liquid chromatography unit and the mass spectrometry unit are connected through a capillary tube.

[0155] The metabolite elution module 102 is used to receive metabolite detection instructions, obtain the original mobile phase set based on the metabolite detection instructions, wherein the original mobile phase set includes multiple original mobile phases, and collects original metabolite samples from the pre-constructed metabolite well plate using the autosampler in the high performance liquid chromatography unit, and performs intelligent elution on the original metabolite samples to obtain the target eluent.

[0156] The metabolite separation module 103 is used to separate the target eluent using the original mobile phase set, degassing box, liquid chromatography solution delivery box and chromatographic column to obtain a target metabolite component set, and to transfer the target metabolite component set to the mass spectrometry unit based on the capillary to obtain the target mass spectrometry unit, wherein the target mass spectrometry unit includes: a mass spectrometry ion source, a mass analyzer and a particle detector.

[0157] The detection report generation module 104 is used to perform signal detection on the target metabolite component set using the target mass spectrometry unit to obtain the component mass spectrum, and analyze the component mass spectrum based on the liquid chromatography-tandem mass spectrometry analysis unit to obtain a metabolite detection report.

[0158] In detail, the modules in the metabolite detection system 100 based on liquid chromatography-tandem mass spectrometry described in this embodiment of the invention employ the same methods as described above. Figure 1 The method used is the same as the metabolite detection method based on liquid chromatography-tandem mass spectrometry described above, and can produce the same technical effect, so it will not be repeated here.

[0159] like Figure 3 The diagram shown is a schematic representation of an electronic device for implementing a metabolite detection method based on liquid chromatography-tandem mass spectrometry, according to an embodiment of the present invention.

[0160] The electronic device 1 may include a processor 10, a memory 11 and a bus 12, and may also include a computer program stored in the memory 11 and executable on the processor 10, such as a metabolite detection method program based on liquid chromatography-tandem mass spectrometry.

[0161] The memory 11 includes at least one type of readable storage medium, such as flash memory, portable hard drive, multimedia card, card-type memory (e.g., SD or DX memory), magnetic memory, disk, optical disk, etc. In some embodiments, the memory 11 can be an internal storage unit of the electronic device 1, such as a portable hard drive. In other embodiments, the memory 11 can be an external storage device of the electronic device 1, such as a plug-in portable hard drive, smart media card (SMC), secure digital card (SD), flash card, etc., equipped on the electronic device 1. Furthermore, the memory 11 includes both internal storage units and external storage devices of the electronic device 1. The memory 11 can be used not only to store application software and various types of data installed on the electronic device 1, such as the code of a metabolite detection method program based on liquid chromatography-tandem mass spectrometry, but also to temporarily store data that has been output or will be output.

[0162] In some embodiments, the processor 10 may be composed of integrated circuits, such as a single packaged integrated circuit or multiple integrated circuits with the same or different functions, including combinations of one or more central processing units (CPUs), microprocessors, digital processing chips, graphics processors, and various control chips. The processor 10 is the control unit of the electronic device, connecting various components of the entire electronic device via various interfaces and lines. It executes programs or modules stored in the memory 11 (e.g., a metabolite detection method program based on liquid chromatography-tandem mass spectrometry) and calls data stored in the memory 11 to perform various functions of the electronic device 1 and process data.

[0163] The bus 12 can be a peripheral component interconnect (PCI) bus or an extended industry standard architecture (EISA) bus, etc. The bus 12 can be divided into an address bus, a data bus, a control bus, etc. The bus 12 is configured to realize the connection and communication between the memory 11 and at least one processor 10, etc.

[0164] Figure 3 Only electronic devices with components are shown; it will be understood by those skilled in the art that... Figure 3The structure shown does not constitute a limitation on the electronic device 1, and may include fewer or more components than shown, or combine certain components, or have different component arrangements.

[0165] For example, although not shown, the electronic device 1 may also include a power supply (such as a battery) to power the various components. Preferably, the power supply can be logically connected to the at least one processor 10 through a power management device, thereby enabling functions such as charging management, discharging management, and power consumption management. The power supply may also include one or more DC or AC power supplies, recharging devices, power fault detection circuits, power converters or inverters, power status indicators, and other arbitrary components. The electronic device 1 may also include various sensors, Bluetooth modules, Wi-Fi modules, etc., which will not be described in detail here.

[0166] Furthermore, the electronic device 1 may also include a network interface. Optionally, the network interface may include a wired interface and / or a wireless interface (such as a Wi-Fi interface, a Bluetooth interface, etc.), which is typically used to establish communication connections between the electronic device 1 and other electronic devices.

[0167] Optionally, the electronic device 1 may further include a user interface, which may be a display, an input unit (such as a keyboard), and optionally, a standard wired interface or a wireless interface. Optionally, in some embodiments, the display may be an LED display, a liquid crystal display, a touch-sensitive liquid crystal display, or an OLED (Organic Light-Emitting Diode) touchscreen, etc. The display may also be appropriately referred to as a screen or display unit, used to display information processed in the electronic device 1 and to display a visual user interface.

[0168] The metabolite detection method program based on liquid chromatography-tandem mass spectrometry stored in the memory 11 of the electronic device 1 is a combination of multiple instructions. When run in the processor 10, it can achieve the following:

[0169] A liquid chromatography-tandem mass spectrometry (LC-MS / MS) device is constructed, comprising: a high-performance liquid chromatography (HPLC) unit, a mass spectrometry (MS) unit, and an HPLC-MS / MS analysis unit. The HPLC unit includes: a degassing chamber, a liquid chromatography solution delivery chamber, an autosampler, a chromatographic column, and a column oven, with the chromatographic column housed within the column oven. The MS unit is a triple quadrupole mass spectrometer, and the HPLC unit and the MS unit are connected via a capillary tube.

[0170] Receive metabolite detection instructions, and obtain the original mobile phase set based on the metabolite detection instructions, wherein the original mobile phase set includes multiple original mobile phases;

[0171] The original metabolite samples were collected from the pre-constructed metabolite well plate using the autosampler in the high performance liquid chromatography unit, and the original metabolite samples were intelligently eluted to obtain the target eluent.

[0172] The target eluent was separated using the original mobile phase set, degassing box, liquid chromatography solution delivery box and chromatographic column to obtain the target metabolite component set;

[0173] The target metabolite components are transported to the mass spectrometry unit via capillary tubes to obtain the target mass spectrometry unit, which includes: a mass spectrometry ion source, a mass analyzer, and a particle detector;

[0174] The target metabolite component set is detected by the target mass spectrometry unit to obtain the component mass spectrum. The component mass spectrum is then analyzed by the liquid chromatography-tandem mass spectrometry analysis unit to obtain the metabolite detection report.

[0175] Specifically, the processor 10's implementation method for the above instructions can be found in [reference needed]. Figures 1 to 3 The descriptions of the relevant steps in the corresponding embodiments are not repeated here.

[0176] Furthermore, if the modules / units integrated in the electronic device 1 are implemented as software functional units and sold or used as independent products, they can be stored in a computer-readable storage medium. The computer-readable storage medium can be volatile or non-volatile. For example, the computer-readable medium may include: any entity or device capable of carrying the computer program code, a recording medium, a USB flash drive, a portable hard drive, a magnetic disk, an optical disk, a computer memory, or a read-only memory (ROM).

[0177] The present invention also provides a computer-readable storage medium storing a computer program, which, when executed by a processor of an electronic device, can perform the following:

[0178] A liquid chromatography-tandem mass spectrometry (LC-MS / MS) device is constructed, comprising: a high-performance liquid chromatography (HPLC) unit, a mass spectrometry (MS) unit, and an HPLC-MS / MS analysis unit. The HPLC unit includes: a degassing chamber, a liquid chromatography solution delivery chamber, an autosampler, a chromatographic column, and a column oven, with the chromatographic column housed within the column oven. The MS unit is a triple quadrupole mass spectrometer, and the HPLC unit and the MS unit are connected via a capillary tube.

[0179] Receive metabolite detection instructions, and obtain the original mobile phase set based on the metabolite detection instructions, wherein the original mobile phase set includes multiple original mobile phases;

[0180] The original metabolite samples were collected from the pre-constructed metabolite well plate using the autosampler in the high performance liquid chromatography unit, and the original metabolite samples were intelligently eluted to obtain the target eluent.

[0181] The target eluent was separated using the original mobile phase set, degassing box, liquid chromatography solution delivery box and chromatographic column to obtain the target metabolite component set;

[0182] The target metabolite components are transported to the mass spectrometry unit via capillary tubes to obtain the target mass spectrometry unit, which includes: a mass spectrometry ion source, a mass analyzer, and a particle detector;

[0183] The target metabolite component set is detected by the target mass spectrometry unit to obtain the component mass spectrum. The component mass spectrum is then analyzed by the liquid chromatography-tandem mass spectrometry analysis unit to obtain the metabolite detection report.

[0184] In the embodiments provided by this invention, it should be understood that the disclosed devices, systems, and methods can be implemented in other ways. For example, the system embodiments described above are merely illustrative, and actual implementations may have other classification methods.

[0185] The modules described as separate components may or may not be physically separate. The components shown as modules may or may not be physical units; that is, they may be located in one place or distributed across multiple network units. Some or all of the modules can be selected to achieve the purpose of this embodiment according to actual needs.

[0186] Furthermore, the functional modules in the various embodiments of the present invention can be integrated into one processing unit, or each unit can exist physically separately, or two or more units can be integrated into one unit. The integrated unit can be implemented in hardware or in the form of hardware plus software functional modules.

[0187] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the present invention.

[0188] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A method for detecting metabolites based on liquid chromatography-tandem mass spectrometry, characterized in that, The method includes: A liquid chromatography-tandem mass spectrometry (LC-MS / MS) device is constructed, comprising: a high-performance liquid chromatography (HPLC) unit, a mass spectrometry (MS) unit, and an HPLC-MS / MS analysis unit. The HPLC unit includes: a degassing chamber, a liquid chromatography solution delivery chamber, an autosampler, a chromatographic column, and a column oven, with the chromatographic column housed within the column oven. The MS unit is a triple quadrupole mass spectrometer, and the HPLC unit and the MS unit are connected via a capillary tube. Receive metabolite detection instructions, and obtain the original mobile phase set based on the metabolite detection instructions, wherein the original mobile phase set includes multiple original mobile phases; The original metabolite samples were collected from the pre-constructed metabolite well plate using the autosampler in the high performance liquid chromatography unit, and the original metabolite samples were intelligently eluted to obtain the target eluent. The target eluent was separated using the original mobile phase set, degassing box, liquid chromatography solution delivery box and chromatographic column to obtain the target metabolite component set; The target metabolite components are transported to the mass spectrometry unit via capillary tubes to obtain the target mass spectrometry unit, which includes: a mass spectrometry ion source, a mass analyzer, and a particle detector; The target metabolite component set is detected by the target mass spectrometry unit to obtain the component mass spectrum. The component mass spectrum is then analyzed by the liquid chromatography-tandem mass spectrometry analysis unit to obtain the metabolite detection report.

2. The method for metabolite detection based on liquid chromatography-tandem mass spectrometry as described in claim 1, characterized in that, The method of collecting raw metabolite samples from pre-constructed metabolite plates using an autosampler in a high-performance liquid chromatography unit includes: The well plate imaging device and intelligent injection needle in the autosampler were identified. The autosampler also includes: online concentration detection device, metabolite dilution device, internal standard addition device and online solid phase extraction column. The well plate imaging device is activated based on the metabolite detection command. The well plate imaging device then acquires images of the metabolite well plate to obtain the target well plate image. Metabolite location is identified based on the target plate image to obtain the current metabolite location; The system automatically samples metabolites from well plates using the current metabolite location, preset metabolite sampling volume, and intelligent injection to obtain raw metabolite samples.

3. The method for metabolite detection based on liquid chromatography-tandem mass spectrometry as described in claim 2, characterized in that, The step of identifying metabolite locations based on the target plate image to obtain the current metabolite location includes: Edge extraction is performed on the target well plate image to obtain the well plate grid edge image, which includes multiple well plate metabolite grids; A grid arrangement map is generated based on multiple well plate metabolite grids in the well plate grid edge image; The target metabolite grid position is obtained based on the metabolite detection command, and the target metabolite grid is identified in the grid arrangement diagram according to the target metabolite grid position; Metabolite features are extracted from the target metabolite grid to obtain metabolite features; Determine whether the metabolite characteristics fall within the preset range of space characteristics; If the metabolite features are not within the range of the space features, then the target metabolite grid is recorded as a valid metabolite grid; Obtain the image center coordinates of the effective metabolite grid, transform the image center coordinates into the real coordinate system, and obtain the current metabolite position.

4. The method for metabolite detection based on liquid chromatography-tandem mass spectrometry as described in claim 3, characterized in that, The intelligent elution of the original metabolite sample to obtain the target eluent includes: The concentration of the original metabolite sample is estimated based on the online concentration detection device in the autosampler to obtain the current metabolite concentration. The original metabolite sample was diluted using the metabolite dilution equipment and the current metabolite concentration to obtain a diluted metabolite sample. Internal standard solution was added to diluted metabolite samples using an internal standard addition device to obtain a sample-internal standard mixture. The sample internal standard mixture was derivatized to obtain a derivatized mixture; The derivatized mixture was loaded into an online solid-phase extraction column to obtain a loaded solid-phase extraction column; The loaded solid-phase extraction column is cleaned to obtain a cleaned solid-phase extraction column. The cleaned solid-phase extraction column is then eluted with a preset strong elution solvent to obtain the target eluent.

5. The method for metabolite detection based on liquid chromatography-tandem mass spectrometry as described in claim 4, characterized in that, The online concentration detection device in the autosampler estimates the concentration of the original metabolite sample to obtain the current metabolite concentration, including: The original metabolite sample was subjected to ultrasonic detection using an online concentration detection device to obtain the current ultrasonic detection signal; Feature extraction is performed on the current ultrasonic detection signal to obtain the current ultrasonic feature vector; Identify the image color feature vector of the original metabolite sample in the target well plate image; Based on the current ultrasonic feature vector and image color feature vector, neighboring data indexing is performed in the pre-constructed concentration prior database to obtain the ultrasonic neighboring concentration and the image color neighboring concentration. The concentration prior database includes: ultrasonic prior database and color prior database. The current metabolite concentration is obtained by weighted summation of adjacent concentrations in ultrasound and adjacent concentrations in image color.

6. The method for metabolite detection based on liquid chromatography-tandem mass spectrometry as described in claim 5, characterized in that, The step of indexing adjacent data in a pre-constructed concentration prior database based on the current ultrasonic feature vector and image color feature vector to obtain adjacent ultrasonic concentrations and adjacent image color concentrations includes: Multiple ultrasound prior data in the ultrasound prior database of the concentration prior database were identified. Each ultrasound prior data includes: a prior ultrasound feature vector and a prior metabolite concentration. Based on the current ultrasonic feature vector, vector similarity is calculated for multiple ultrasonic prior data to obtain multiple ultrasonic vector similarities. Identify the maximum ultrasonic similarity among multiple ultrasonic vector similarities, and record the ultrasonic prior data corresponding to the maximum ultrasonic similarity as the adjacent ultrasonic prior data; The prior metabolite concentrations in adjacent ultrasound prior data are denoted as adjacent ultrasound concentrations. Identify adjacent color intensities in an image based on its color feature vector in a color prior database.

7. The method for metabolite detection based on liquid chromatography-tandem mass spectrometry as described in claim 6, characterized in that, The weighted summation of adjacent concentrations of ultrasound and adjacent concentrations of image color to obtain the current metabolite concentration includes: The ultrasonic feature index set in the current ultrasonic feature vector and the color feature index set in the image color feature vector are statistically analyzed separately. Ultrasonic feature indicators are extracted sequentially from the set of ultrasonic feature indicators. Based on the extracted ultrasonic feature indicators, multiple ultrasonic data with the same name are extracted from the ultrasonic prior database. Each ultrasonic data with the same name includes: prior metabolite concentration and the corresponding value of ultrasonic feature indicator. The correlation of ultrasonic indicators is obtained by calculating the concentration correlation of multiple ultrasonic data with the same name. By summarizing the correlations of each ultrasonic feature index, we obtain the ultrasonic index correlation set. Obtain the color index correlation set corresponding to the color feature index set from the color prior database; The mean values ​​of the ultrasonic index correlation set and the color index correlation set are calculated respectively to obtain the average ultrasonic correlation and the average color correlation. The current metabolite concentration is obtained by weighting the adjacent concentrations of ultrasound and adjacent concentrations of image color based on the average ultrasound correlation and average color correlation.

8. The method for metabolite detection based on liquid chromatography-tandem mass spectrometry as described in claim 7, characterized in that, The method of weighting adjacent concentrations of ultrasound and adjacent concentrations of image color based on average ultrasound correlation and average color correlation to obtain the current metabolite concentration includes: The current metabolite concentration can be calculated using the following formula: in, Indicates the current concentration of metabolites. This represents the average ultrasound correlation. Indicates average color correlation. Indicates the adjacent concentrations of ultrasound waves. This indicates the density of adjacent colors in an image.

9. The method for metabolite detection based on liquid chromatography-tandem mass spectrometry as described in claim 8, characterized in that, The target eluent is separated using the original mobile phase set, degassing chamber, liquid chromatography solution delivery box, and chromatographic column to obtain a set of target metabolite components, including: The original mobile phase set is introduced into the degassing box to obtain the degassed mobile phase set, and the degassed mobile phase set is transported to the liquid chromatography solution delivery box to obtain the target solution delivery box; The degassed mobile phase is mixed under high pressure using a target solution delivery box and a preset delivery ratio to obtain a mixed mobile phase. The target eluent is delivered to the chromatographic column using an autosampler and a mixed mobile phase to obtain the target chromatographic column; The target eluent is separated into components using a target chromatographic column to obtain a target metabolite component set, which includes multiple target metabolite components.

10. A metabolite detection system based on liquid chromatography-tandem mass spectrometry, characterized in that, The system includes: A mass spectrometry device construction module is used to construct a liquid chromatography-tandem mass spectrometry device. The liquid chromatography-tandem mass spectrometry device includes: a high-performance liquid chromatography unit, a mass spectrometry unit, and a liquid chromatography-tandem mass spectrometry analysis unit. The high-performance liquid chromatography unit includes: a degassing chamber, a liquid chromatography solution delivery chamber, an autosampler, a chromatographic column, and a column oven. The chromatographic column is built into the column oven. The mass spectrometry unit is a triple quadrupole mass spectrometer. The high-performance liquid chromatography unit and the mass spectrometry unit are connected through a capillary tube. The metabolite elution module is used to receive metabolite detection commands and obtain the original mobile phase set based on the metabolite detection commands. The original mobile phase set includes multiple original mobile phases. The original metabolite samples are collected from the pre-constructed metabolite well plate using the autosampler in the high performance liquid chromatography unit. The original metabolite samples are intelligently eluted to obtain the target eluent. The metabolite separation module is used to separate the target eluent using the original mobile phase set, degassing box, liquid chromatography solution delivery box and chromatographic column to obtain a target metabolite component set. The target metabolite component set is then transferred to the mass spectrometry unit via capillary to obtain the target mass spectrometry unit, which includes a mass spectrometry ion source, a mass analyzer and a particle detector. The detection report generation module is used to detect the target metabolite component set by using the target mass spectrometry unit to obtain the component mass spectrum, and analyze the component mass spectrum based on the liquid chromatography-tandem mass spectrometry analysis unit to obtain the metabolite detection report.

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