CN116626207B

CN116626207B - Full-automatic liquid-phase mass spectrum detection system and detection method special for fat-soluble vitamins

Info

Publication number: CN116626207B
Application number: CN202310857698.8A
Authority: CN
Inventors: 刘小平; 徐杰; 王亚博; 杜建兵; 刘华芬
Original assignee: Beijing Kailaipu Biotechnology Co ltd
Current assignee: Beijing Kailaipu Biotechnology Co ltd
Priority date: 2023-07-13
Filing date: 2023-07-13
Publication date: 2023-10-20
Anticipated expiration: 2043-07-13
Also published as: CN116626207A

Abstract

The invention provides a full-automatic liquid phase mass spectrum detection system and method special for fat-soluble vitamins, wherein the system comprises a sample pretreatment module, a sample injection module, a column temperature box module, a mass spectrum module, a pump module, a solvent bottle module and a computer module; the sample pretreatment module receives a blood sampling pipe frame filled with a sample, carries out pretreatment on the sample in the blood sampling pipe frame, and transfers the sample to the sample injection module after the pretreatment is finished, and the sample injection module sends the sample to the column temperature box module through the pump module; the separation of fat-soluble vitamins is completed in the column temperature box module; the separated substances enter a mass spectrum module, the mass spectrum module transmits the acquired signals to a computer module, and the computer module feeds back detection results according to the acquired data. The invention designs a full-automatic liquid phase mass spectrometer special for fat-soluble vitamins, which has a full-automatic one-stop detection and analysis function, reduces the investment of personnel, improves the working efficiency and reduces the error probability.

Description

Full-automatic liquid-phase mass spectrum detection system and detection method special for fat-soluble vitamins

Technical Field

The invention relates to a detection technology used in the medical field, in particular to a full-automatic mass spectrometer technology special for fat-soluble vitamins.

Background

Fat-soluble vitamins refer to vitamins which are insoluble in water and soluble in fats and organic solvents, and mainly comprise vitamin A, vitamin D, vitamin E, vitamin K and the like. The detection of the content of fat-soluble vitamins is very important in the field of biomedical detection, and can provide intermediate result data for further diagnosis of a plurality of health indexes. In the prior art, lipid-soluble vitamins are detected by adopting a liquid chromatography method, according to the information fed back by the current market, in the detection process, the steps of pretreatment, compound separation, detection and the like are usually needed for a sample by a tester, the personnel and time cost are high, in addition, the steps are independently carried out in the industry, such as pretreatment, separation and mass spectrum processes, which are respectively carried out on different instruments, or different personnel are carried out at different time/place, finally mass spectrum is carried out uniformly, so that an analysis result is obtained, the pretreatment and sample injection process method is not fixed, and the stability and reliability of the result are difficult to ensure.

In general, the current market lacks high-sensitivity, high-specificity, high-accuracy and high-efficiency detection equipment which can finish one-stop from the start of detecting a sample to the output of a detection result, and the current liquid phase mass spectrum is general detection equipment, lacks special instruments and matched methods for analyzing fat-soluble vitamins, and further lacks special instruments and matched methods for simultaneously analyzing multiple fat-soluble vitamins under a complex matrix. Therefore, how to realize the full-automatic analysis of fat-soluble vitamins and ensure high sensitivity, high specificity and accuracy is a bottleneck for improving the analysis and detection efficiency.

Disclosure of Invention

The invention aims to solve the technical problem of providing a full-automatic liquid-phase mass spectrum detection system and a full-automatic liquid-phase mass spectrum detection method for fat-soluble vitamins, which can automatically finish the processing from a sample to an analysis result of the fat-soluble vitamins, have excellent specificity, sensitivity and accuracy, and can detect the fat-soluble vitamins with very low concentration.

For this purpose, the invention provides the following technical scheme:

the invention provides a full-automatic liquid-phase mass spectrum detection system special for fat-soluble vitamins, which comprises a sample pretreatment module, a sample injection module, a column temperature box module, a mass spectrum module, a pump module, a solvent bottle module and a computer module;

the sample pretreatment module is arranged in the middle of the mass spectrum detection system and is configured to receive a blood sampling pipe frame filled with a sample, and carry out pretreatment on the sample in the blood sampling pipe frame, and after the pretreatment is completed, the sample pretreatment module transfers the blood sampling pipe frame to the lower part of the sample injection module;

the sample injection module is arranged at the upper part of the mass spectrum detection system and is configured to absorb the pre-processed sample from the sample pre-processing module and send the sample to the column temperature box module through the pump module;

the column incubator module is arranged on one side of the sample injection module and is configured to temporarily contain the pretreated sample and control the temperature of the sample so as to separate the fat-soluble vitamins;

The mass spectrum module is arranged at the lower part of the mass spectrum detection system, substances separated by the column temperature box module enter the mass spectrum module through a pipeline, the mass spectrum module transmits acquired signals to the computer module, and the computer module feeds back detection results according to the acquired data.

Further, the mass spectrum module comprises a mass spectrum shell, wherein the bottom of the mass spectrum shell is provided with universal casters, a vacuum module is arranged in the mass spectrum shell, the vacuum module comprises a vacuum cavity, one end of the vacuum cavity is arranged on one surface of the mass spectrum shell, an ion channel interface plate extending out of the mass spectrum shell is arranged at the end of the vacuum cavity, the ion channel interface plate is configured to be connected with the ion source module, one side of the vacuum cavity is provided with a molecular pump, the molecular pump is communicated with an external space through a vacuum tube and is configured to be used for vacuumizing, and a triple quadrupole rod module is arranged in the vacuum cavity;

the ion source module comprises an ion source cavity, an ion source fixing device is arranged at the upper part of the ion source cavity, an ion source probe which can extend into the cavity is arranged on the ion source fixing device, the ion source probe is configured to be used for ionization of a compound, a calibration structure is also arranged on the ion source fixing device and is configured to be used for calibrating the space position of the ion source probe, a heating rod is arranged in the ion source cavity, and an observation window is arranged on the surface of the ion source cavity;

The triple quadrupole rod module comprises a quadrupole rod support, wherein a second quadrupole rod is fixedly connected to the middle of the quadrupole rod support, two ends of the middle quadrupole rod are respectively connected with a first quadrupole rod and a third quadrupole rod, the first quadrupole rod is configured to be used for screening and focusing of parent ions, the second quadrupole rod is an ion reaction tank and is configured to break the parent ions into child ions, the third quadrupole rod is configured to be used for screening and focusing of the child ions, one side of the quadrupole rod support is provided with a quadrupole rod fixing plate, and a detector is arranged on the quadrupole rod fixing plate and is used for counting the number of the child ions and converting the number of the child ions into an electric signal;

the triple quadrupole rod module further comprises a communication control module configured to transmit the detector-converted electrical signal to the computer module.

Further, the sample pretreatment module comprises a pretreatment chamber, wherein an access blood collection tube rack module is arranged in the pretreatment chamber, is configured to be used for placing a blood collection tube sample to be treated and analyzed and a blood collection tube sample which is already treated, and is configured to be capable of adding a new blood collection tube sample and taking out the blood collection tube sample which is already treated at any time;

The pretreatment chamber is also internally provided with a centrifugal module, a pipetting gun module, a mixing module, a deep pore plate module, a solvent bottle module, a tip head and a reagent tank module, wherein the centrifugal module is arranged near the blood collection tube rack module and is configured to be used for centrifuging a blood collection tube and the deep pore plate to separate serum and plasma in a sample and/or separate precipitated protein from the sample; the pipette gun module is configured to draw serum and protein precipitants and internal standard reagents from the centrifugal module and the solvent bottle module respectively and put the reagents into the deep hole plate, the deep hole plate module is configured to move the deep hole plate to a sample injection position and a mixing position, the mixing module is configured to mix the serum and protein precipitants and internal standard reagents in the deep hole plate, the solvent bottle module is configured to store mobile phase reagents and sample injection needle cleaning reagents, the tip head and reagent tank module is configured to store tip head, precipitants, internal standard reagents and waste tanks required by pipetting, and the mechanical arm module is configured to move the blood sampling pipe frame and/or the deep hole plate, so that the blood sampling pipe frame module, the centrifugal module and the hole plate module can be moved in and out, and the scanning procedure of the blood sampling pipe is completed.

A damping foot pad is arranged below the pretreatment chamber and is configured to reduce vibration caused by the centrifugal module, so that the influence of the centrifugal vibration on the mass spectrum module is reduced;

the pretreatment chamber is also internally provided with a gun head and a cleaning pool module, the gun head and the cleaning pool module are arranged near the pipetting gun module and are configured to be used for placing and cleaning the gun head for sucking the serum sample, and in addition, the pretreatment chamber is also provided with a rack cabinet door structure such as an acrylic door and the like and is used for partially or temporarily closing the pretreatment chamber so as to avoid environmental pollution;

further, the centrifugal module comprises a centrifugal machine seat, a motor is arranged in the centrifugal machine seat, the motor is connected with a motor shaft through a coupler, the motor shaft is rotationally connected with a centrifugal carrier, a blood collection tube carrier placing position is arranged on the centrifugal carrier and used for loading blood collection tubes, the blood collection tube carrier used for loading the blood collection tubes can be placed on the blood collection tube carrier placing position, the centrifugal carrier comprises at least two symmetrical blood collection tube carrier placing positions, each blood collection tube carrier placing position can be loaded with two leveling reagent tubes, according to the weight measured by the blood collection tube rack module, the balancing amount and the balancing scheme are calculated according to a preset calculation method, the liquid transfer gun module is controlled to absorb reagents from the tip head and the reagent tank module to the leveling reagent tubes according to the calculated balancing scheme, so that the load on the centrifugal machine is balanced, and the centrifugal machine is ensured to work more stably.

The centrifugal carrier comprises a middle fixing plate and side fixing plates, wherein the motor is connected with the middle fixing plate in a rotating mode, the side fixing plates are U-shaped, two side portions of the U-shaped are arranged on two sides of a carrier placing position of a blood collection tube and are provided with mounting grooves and sliding rails, elastic elements are further arranged on the middle fixing plate corresponding to each side fixing plate respectively, the side fixing plates are connected with the elastic elements through carrier fixing plate connecting pieces, steps with different heights are arranged at the bottom positions of the U-shaped, carrier fixing plates and limiting fixing plates are arranged on the side fixing plates, the carrier fixing plates can be driven to move along the sliding rails through the limiting fixing plates, and when the carrier fixing plates move, the carrier fixing plates can be adjusted to stretch out and draw back and/or lock states by steps with different heights.

According to the invention, the blood collection tube carrier is locked or unlocked through the small range of the side fixing plate in the centrifugal carrier, the carrier fixing plate is in limiting rigid connection with the fixing plate, and the translation of the limiting fixing plate can be completed through the mechanical arm, so that the purpose of locking or unlocking the blood collection tube carrier is achieved. The principle that the carrier fixing plate stretches out and draws back to lock or unlock the blood collection tube carrier is as follows: when the carrier fixing plate connecting piece is at the initial position, the carrier fixing plate connecting piece is in a state that the elastic element is retracted, the corresponding blood collection tube carrier is in an unlocking state, when the mechanical arm moves the fixing plate limiting piece to drive the carrier fixing plate connecting piece to move rightwards (as seen in the drawing) to the highest position, the mechanical arm automatically slides to a first step, then automatically slides to a second step (a transfer position) and stays at the first step, the carrier fixing plate connecting piece is in an extending state, and the blood collection tube carrier is in a locking state; when the unlocking is needed, the mechanical arm moves the fixing plate limiting part again to drive the carrier fixing plate connecting part to move right (as seen in the drawing) to the third step, and the mechanical arm can return to the initial state at the moment, and the whole process cannot slide reversely, so that the reliability is ensured.

Further, the pipette gun module includes: an X-axis module, a Y-axis module and a pipette; the top of the pipette gun is slidably connected to the X-axis module, the X-axis module is slidably connected to the Y-axis module, and the X-axis module and the Y-axis module are configured to enable the pipette gun to move in two dimensions;

the pipette includes: a plurality of pipette assemblies and a spacing adjustment assembly, the pipette assemblies including a plurality of pipette guns including at least one fixed pipette gun and a plurality of movable pipette guns, the fixed pipette guns being fixed to the spacing adjustment assembly without participating in the spacing adjustment movement, the plurality of movable pipette guns being movably connected to the spacing adjustment assembly, respectively, and configured to be able to adjust the spacing therebetween by the spacing adjustment assembly;

the space adjusting assembly comprises space adjusting supporting plates, the space adjusting supporting plates are arranged in pairs, a plurality of screw rods which respectively correspond to the movable pipetting guns are arranged between the space adjusting supporting plates, space adjusting sliding blocks are arranged on each screw rod, the space adjusting sliding blocks are respectively connected with the movable pipetting guns through sliding block connecting pieces, space adjusting motors are arranged on the space adjusting supporting plates, the space adjusting motors are connected with the screw rods through gear sets, each gear set comprises a plurality of gears with different double speeds, the gear speed in the middle is low, the gear speed in the outer side is gradually increased, and each screw rod is connected with the gear through a coupling; according to the invention, one liquid-transferring gun is fixed and not moving, the liquid-transferring gun beside the liquid-transferring gun is far away from the fixed liquid-transferring gun along with the rotation of the interval adjusting motor, then the liquid-transferring gun beside the liquid-transferring gun is far away from the fixed liquid-transferring gun, and the liquid-transferring guns at different positions are adjusted and moved through gears with different times of speed, so that equidistant adjustment is realized, and the liquid-sucking requirement is met.

And a plurality of guide sliding blocks which respectively correspond to the movable liquid-transferring guns are arranged on the liquid-transferring gun guide sliding rail, and each guide sliding block is respectively connected with one movable liquid-transferring gun and used for guiding the movement of the movable liquid-transferring gun.

Further, the pipette includes a pipette fixture configured to position a lateral position of the pipette assembly, the pipette fixture having a Z-axis motor and a Z-axis slide rail set thereon configured to drive the pipette up and down;

the pipetting gun further comprises a pipetting motor, the pipetting motor is connected with a pipetting piston through a pipetting piston rod, the pipetting piston is configured to move in a piston cavity to finish pipetting action, the pipetting piston is connected with a pipetting head, a head removing piece is arranged between the pipetting piston and the pipetting head, one side of the head removing piece is fixedly connected with a head removing auxiliary mechanism, and the head removing auxiliary mechanism comprises a metal fixing piece, a spring, an electromagnet and a spring fixing piece. The Z-axis motor and the Z-axis sliding rail group complete the movement of the pipetting gun in the Z-axis direction, the pipetting motor, the pipetting piston and the piston cavity cooperate to complete the pipetting action, when the Z-axis motor moves downwards, the pipetting head extrudes into the tip head to grasp the tip head, when the tip head needs to be retracted, the electromagnet is electrified to attract the metal block right above, the spring is compressed, the head removing piece translates downwards, and the tip head is extruded out of the pipetting head.

The liquid suction head comprises a rubber sealing ring, a rubber fixing piece and a metal elastic piece. After the liquid suction head is extruded into the tip head, the rubber sealing ring can play a sealing role, so that the air tightness and the liquid suction precision are ensured, two metal elastic sheets are embedded into the inner side of the liquid suction head and are placed at an angle of 180 degrees, after the liquid suction head is extruded into the tip head, the metal elastic sheets are tightly contacted with the tip head, and whether the tip head contacts the liquid level or not and the amount of sucked liquid can be judged by detecting the resistance values of the two ends of the metal elastic sheets, so that the position of the liquid suction head can be well controlled, and the liquid suction head is prevented from entering the liquid level too deeply, so that collision or precipitation at the bottom of suction is caused, and damage or blockage of a pipeline to the liquid suction head is avoided; meanwhile, because the liquid level detection function is provided, the deep hole plate can be used for directly sampling, compared with the conventional equipment which needs to suck a very small amount of samples from the upper surface of the deep hole plate to the shallow hole plate, and then the shallow hole plate is used for sampling, the invention reduces the cost, the procedures and the time and improves the efficiency.

Further, the pump module includes: the two mobile phase switching valves are communicated with the degasser, two pipelines of the degasser are respectively connected with two mobile phase pumps, the two mobile phase pumps are connected to the primary mixer, one outlet of the primary mixer is connected to the waste liquid port, and the other outlet of the primary mixer is connected with the mixer.

The invention comprises an A mobile phase switching valve and a B mobile phase switching valve, wherein 4 paths of mobile phase switching valves are connected to four mobile phase bottles in a solvent bottle module, and strong washing and weak washing (required by a cleaning pipeline) are also connected to reagent bottles corresponding to the solvent bottle module; the left side of the degasser is an inlet end which is respectively connected with an A mobile phase switching valve, a B mobile phase switching valve and a strong washing pipeline and a weak washing pipeline, so that bubbles in the pipeline are removed, the pipeline is stable in hydraulic pressure, and the sample injection is accurate. The mobile phase A and the mobile phase B are respectively connected to the mobile phase A pump and the mobile phase B pump after being degassed by a degasser, the mobile phase pumps provide power for the two paths of mobile phases, the output end of the mobile phase A and the mobile phase B are connected to a primary mixer, the primary mixing of the mobile phases A and the mobile phases B can be completed in the primary mixer, the mobile phases A and the mobile phases B are connected to the mixer after being mixed, and the mobile phases A and the mobile phases B are connected to a sample injection module after being mixed. In addition, when the pipeline of the pump module needs to be flushed, the output end can be switched to the waste liquid port without entering the sample injection module.

Further, the sample injection module is communicated with the pump module, the pump module comprises a three-way switching valve, the three-way switching valve is respectively connected with a strong washing liquid inlet, a weak washing liquid inlet and an output end, the three-way switching valve is connected with the three-way switching valve through a switching motor and selects a communicated valve port through a piston, the piston is configured to suck cleaning liquid through a coupler and a sample injection liquid suction motor and then output the cleaning liquid from the output end, the output end is connected to a six-way valve module, and the six-way valve module is connected with the liquid suction needle module;

The strong and weak washing pipeline output by the pump module is respectively connected into the strong washing liquid inlet and the weak washing liquid inlet, the switching motor rotates to switch and select one of the three paths of the piston and the strong washing liquid inlet, the weak washing liquid inlet and the output end to be communicated, the piston sucks washing liquid and then outputs the washing liquid from the output end, the output end is connected to the six-way valve module, and the six-way valve module is connected with the liquid suction needle module to realize liquid suction and flushing functions.

Specifically, six-way valve includes 6 interfaces, connects mobile phase interface (No. 1 hole), chromatographic column interface (No. 6 holes), imbibition needle interface (No. 4 holes), sample loop (2, no. 5 holes), output pipeline (No. 3 holes) respectively, and six-way valve has two positions, bit A and bit B respectively:

a position: the holes 1-2,3-4 and 5-6 are conducted in pairs;

b bit: 1-6,2-3,4-5 holes are communicated;

and a sample injection ring is connected between the holes 2 and 5, and the holes 2 and 5 are in a conducting state. When the strong and weak needle washing is needed, the six-way valve is in the A position, strong and weak washing liquid enters from the hole 3, and enters the 4-10 liquid suction needle module through the hole 4, so that the needle washing is completed; when the sample is required to be sucked, the position B is switched, at the moment, the sample is sucked by the 4-10 liquid suction needle module and enters the 4 # hole to the 5 # hole, enters the sample injection ring, stays in the sample injection ring, and is timely switched to the position A, at the moment, the mobile phase enters the six-way valve from the 1 # hole and reaches the 2 # hole, the sample carrying the sample injection ring flows from the 5 # hole to the 6 # hole, and the 6 # hole is connected with the chromatographic column (in the column temperature box module), so that the sample injection is completed.

The sample injection module further comprises a sample injection mobile XY module, the sample injection mobile XY module comprises a Y-axis motor support, a Y-axis sliding block set is connected to the Y-axis motor support, an X-axis seat is connected to the Y-axis sliding block set, the X-axis seat can slide along the Y-axis sliding block set through driving of a Y-axis motor, an X-axis motor shaft and an X-axis motor are arranged on the X-axis seat, a liquid suction needle connecting sliding block is connected to the X-axis motor shaft, and the liquid suction needle connecting sliding block is connected with the liquid suction needle module;

the liquid suction needle module comprises a liquid suction needle motor seat, the liquid suction needle motor seat is connected with a liquid suction needle connecting sliding block, a liquid suction needle motor is arranged in the liquid suction needle motor seat, the liquid suction needle motor is connected with a screw rod sliding block through a screw rod, and the screw rod sliding block moves in a guide groove and drives the liquid suction needle to move up and down to suck liquid. The liquid suction needle motor seat is arranged on a liquid suction needle connecting sliding block of the sample injection moving XY module, so that X and Y axis direction movement is realized; the motor of the liquid suction needle rotates to drive the screw rod sliding block to slide up and down along the guide groove, and the liquid suction needle is arranged on the screw rod sliding block, so that the movement of the liquid suction needle in the Z-axis direction is realized, and the liquid suction action is completed.

Further, the column temperature control box module comprises a sample feeding pipe, the sample feeding pipe is led into the temperature control box module and is connected with a chromatographic column in the temperature control box module, the chromatographic column is fixedly installed on the temperature control module through a chromatographic column buckle, a ventilation fan is arranged at the rear end of the temperature control box module, a column temperature box door is arranged at the front end of the temperature control box module, and the periphery of the column temperature box door is sealed through a sealing ring;

The temperature control module comprises an inner cavity radiator, a rear side radiator, heat preservation cotton, an inner cavity radiator fan, a rear side fan and a semiconductor refrigerating sheet, wherein the semiconductor refrigerating sheet is arranged in a mounting groove of the rear side radiator and the inner cavity radiator, heat conduction silicone grease is coated on two sides of the semiconductor refrigerating sheet, heat preservation cotton is used for preserving heat all around, the rear side radiator and the inner cavity radiator are arranged on a fixing plate, the inner cavity radiator fan and the rear side fan are respectively arranged on the outer sides of the inner cavity radiator and the rear side radiator, and the fan is used for improving working efficiency of the semiconductor refrigerating sheet.

The sample injection pipe is connected with the chromatographic column interface of the six-way valve module of the sample injection module, a sample sucked by the sample injection module enters the chromatographic column to finish separation, and the separated compound (fat-soluble vitamin) enters the mass spectrum module. The temperature control module is used for creating proper temperature for the chromatographic column and ensuring the separation effect of the chromatographic column.

A full-automatic liquid phase mass spectrum detection method special for fat-soluble vitamins adopts the detection system and comprises the following steps:

(1) Sample pretreatment:

(1.1) centrifuging the blood collection tube in the carrier at 3000-8000rpm for 5-10 minutes;

(1.2) drawing 50-200. Mu.L of serum into a 96-well deep well plate;

(1.3) precisely removing 20-100 mu L of protein precipitant into a 96-well deep hole plate;

(1.4) precisely adding 100-400 mu L of internal standard solution;

(1.5) placing the 96 deep hole plate in a mixer, and swirling for 5-10 minutes at 600-3000 rpm;

(1.6) centrifuging at 2000-4000rpm for 5-10 minutes;

(2) Flow matching and sample injection: driving a sample to be sampled by a pump module according to a mixed proportion flowing sample, and carrying out gradient elution on a flowing phase; wherein, the mobile phase ratio is: one of the mobile phases is an aqueous solution containing 0.1% formic acid, and the other mobile phase is a methanol solution containing 0.1% formic acid; the flow rate of the pump is controlled to be 0.6-0.8 mL/min;

(3) Separating: the module carries the sample into a temperature control box module and finishes the separation of fat-soluble vitamins in the temperature control box module;

(4) Mass spectrometry: the separated fat-soluble vitamins enter a mass spectrum module for analysis, and the analysis result of the quality control is fed back to a computer module.

The invention provides a full-automatic liquid-phase mass spectrum detection system special for fat-soluble vitamins and a matching method, which are known by integrating the technical scheme adopted by the invention. In the structure and the method, the special mass spectrum process for the fat-soluble vitamins is realized through the following modules and operation processes:

1) After the blood collection tube is centrifuged, whole blood presents the upper serum, the lower plasma is layered, fat-soluble vitamins are extracted from the serum, the process is completed by matching a pipette gun module of a sample pretreatment module, a gun head and a cleaning pool module, the pipette gun module grabs the gun head and the cleaning pool module to be used for sucking the serum, and the blood collection tube is deeply sucked to complete the serum, so that the pipette gun module has the detection functions of liquid level and liquid suction amount, and has specificity on detection signals of fat-soluble vitamins in the serum because of different reagents or different physical characteristics of the solution.

2) In order to ensure the accuracy of analysis and quantification, when serum pretreatment is carried out, a tip head is grabbed by a pipette module to absorb an internal standard reagent and the internal standard reagent is added into a deep pore plate, a specific internal standard solution is matched according to fat-soluble vitamins, and at the moment, the pipette module also detects the liquid level and the liquid absorption amount and has specificity for detection signals of the specific internal standard; in addition, in order to reduce the clogging of the pipeline by the sample after serum pretreatment, the deep-hole plate is centrifuged by utilizing a centrifugal module, and impurities are further precipitated to the bottom.

3) The invention is provided with the column temperature box module, and can realize good separation of the fat-soluble vitamins in complex matrixes and high efficiency by matching specific chromatographic columns associated with the fat-soluble vitamins and matching mobile phases with specific concentration ratios by the solvent bottle module and washing the fat-soluble vitamins according to a set gradient.

4) The ion source module in the mass spectrum module is responsible for ionization of the compound, the effect of ionization directly influences the response and the signal to noise ratio of signals, the sensitivity is further influenced, specific ionization conditions are needed for fat-soluble vitamins, the specific ionization conditions comprise an ionization method, the flow rate of nitrogen of an ion source, the temperature of the environment during ionization and the like, and in addition, the flow rate of nitrogen in a Q2 quadrupole in the triple quadrupole module also has an important influence on the quantitative analysis of fat-soluble vitamins.

The beneficial effects of the invention are as follows:

1. according to the invention, a liquid phase mass spectrometer specially aiming at full-automatic fat-soluble vitamins is designed through a pipetting gun module, a gun head and cleaning pool module, a tip head and reagent tank module, a centrifugal module, a column temperature box module, an ion source module, a triple quadrupole rod module and a matched method (pretreatment method and sample injection method), so that the full-automatic one-stop detection and analysis function is realized, the investment of personnel is reduced, the working efficiency is improved, and the error probability is reduced.

2. The invention adopts LC-MS (liquid chromatography tandem mass spectrometry) technology, the LC-MS is more sensitive in the aspect of analyzing fat-soluble vitamins, and the quantity and the range of solvents which can be used by the LC-MS are wider, the advantage is reflected in complex matrixes, the ionization technology of the LC-MS can provide better ionization efficiency, and the ion inhibition effect is reduced, so that the detection sensitivity and the accuracy of the fat-soluble vitamins in the complex matrixes are improved. Compared with other analysis technologies such as GC-MS (gas chromatography tandem mass spectrometry) and electrochemiluminescence, the method has the advantages of specificity, sensitivity and accuracy.

3. In the mass spectrum module, the triple quadrupole mass spectrum technology is adopted, so that the fat-soluble vitamins can be distinguished through the mass-to-charge ratio, and compared with similar TOF or other mass spectrums, the triple quadrupole mass spectrum technology has unique advantages, and can simultaneously analyze a plurality of fat-soluble vitamins.

Drawings

Fig. 1 is a schematic view of the overall structure of the present invention.

Fig. 2 is a schematic diagram of the structure of a mass spectrometry module of the present invention.

Fig. 3 is a schematic structural view of the vacuum module of the present invention.

Fig. 4 is a schematic structural view of an ion source module of the present invention.

Fig. 5 is a schematic structural view of a triple four-bar module of the present invention.

Fig. 6 is a schematic structural view of a sample pretreatment module of the present invention.

Fig. 7 is a schematic structural view of the in-out blood collection tube rack module of the present invention.

Fig. 8 is a schematic structural view of the centrifugal module of the present invention.

Fig. 9 is a schematic structural view of a blood collection tube carrier of the present invention.

FIG. 10 is a schematic view showing a locking structure of the cartridge carrier of the present invention, in which the carrier fixing plate connector is in an initial state.

FIG. 11 is a second schematic view of the locking structure of the cartridge carrier of the present invention, in which the carrier plate connector is located at the highest step.

Fig. 12 is a third schematic view of the locking structure of the blood collection tube carrier of the present invention, in which the carrier fixing plate connector is located at the second step.

FIG. 13 is a schematic view showing a locking structure of a blood collection tube carrier of the present invention, in which a carrier fixing plate connector is located at a third step.

Fig. 14 is a fifth schematic view of the locking structure of the blood collection tube carrier of the present invention, wherein the carrier fixing plate connector is located at the fourth step.

Fig. 15 is a schematic structural view of the mixing module of the present invention.

FIG. 16 is a schematic view of the structure of the tip head and reagent tank module of the present invention.

Fig. 17 is a schematic view of a moving structure of the pipette module of the present invention.

Fig. 18 is a schematic diagram of a spacing adjustment structure of a pipette module of the present invention.

Fig. 19 is a schematic structural view of the pipette of the present invention.

Fig. 20 is a schematic view of the structure of the liquid suction head of the present invention.

Fig. 21 is a schematic structural view of the deep hole plate module of the present invention.

Fig. 22 is a schematic structural view of the deep well plate carrier of the present invention.

Fig. 23 is a schematic structural view of the deep hole plate carrier tray of the present invention.

FIG. 24 is a schematic view of the gun head and purge tank module of the present invention.

Fig. 25 is a schematic structural view of the robot arm module of the present invention.

Fig. 26 is a schematic structural view of the primary mechanical arm of the present invention.

Fig. 27 is a schematic structural view of a secondary mechanical arm of the present invention.

Fig. 28 is a schematic structural view of a tertiary mechanical arm of the present invention.

Fig. 29 is a schematic view of the structure of the gripper of the present invention.

Fig. 30 is a schematic structural view of the pump module of the present invention.

Fig. 31 is a schematic structural diagram of a sample injection module according to the present invention.

Fig. 32 is a schematic view of the a-position state of the six-way valve of the present invention.

Fig. 33 is a B-bit state schematic diagram of the six-way valve of the present invention.

Fig. 34 is a schematic structural diagram of the sample injection mobile XY module of the present invention.

Fig. 35 is a schematic view of the structure of the pipette needle module of the present invention.

Fig. 36 is a schematic structural view of the column oven module of the present invention.

Fig. 37 is a schematic structural view of a temperature control module of the present invention.

Fig. 38 is a flow chart of the detection method of the present invention.

Reference numerals in the drawings: the sample pretreatment module 100, the pretreatment chamber 101, the access rack module 102, the access rack area 1021, the access rack area 1022, the pressure sensor 1023, the scanner 1024, the centrifuge module 103, the centrifuge base 1031, the coupling 1032, the motor shaft 1033, the centrifuge carrier 1034, the blood collection rack placement position 1035, the blood collection rack carrier 1036, the leveling reagent tube 1037, the middle fixing plate 1038, the side fixing plate 1039, the elastic member 1130, the carrier fixing plate connector 1131, the locking member 1132, the carrier fixing plate 1133, the limit fixing plate 1134, the slide rail 1135, the mounting groove 1136, the shelf 1137, the first step 1138, the second step 1139, the fixing plate cover 1230, the third step 1231, the fourth step 1232, the pipette module 104, the x-axis module 1041, the y-axis module 1042, the pipette 1043, the fixing pipette 1044, the plurality of movable pipette 1045, the pitch adjusting support plate 1046, the screw 1047, the pitch adjusting slider 1048, slider connection 1049, pitch adjustment motor 1140, gear set 1141,1 gear 1142,2 gear 1143,3 gear 1144, pipette guide slide 1145, guide slider 1146, pipette fixture 1147, z-axis motor 1148, z-axis slide rail set 1149, pipette motor 1240, pipette piston rod 1241, pipette piston 1242, piston cavity 1243, pipette head 1244, rubber seal 12441, rubber mount 12442 and metal spring 12443, tip piece 1245, metal mount 1246, spring 1247, electromagnet 1248, spring mount 1249, mixing module 105, mixing motor 1051, mixing coupler 1052, eccentric 1053, mixing wheel 1054, deep hole plate module 106, deep hole plate 1061, deep hole plate carrier 1062, deep hole plate carrier 10621, deep hole plate fixture spring 10622, deep hole plate leveling agent tube 10623, refrigeration chamber 10624, deep hole plate carrier tray 1063, deep hole plate refrigeration module 10631, deep hole plate tray 10632 and buffer rubber pad 5633, deep-hole plate lifting module 1064, deep-hole plate carrier bottom hole 1065, solvent bottle module 107, tip head and reagent tank module 108, tip head carrier 1081, waste tank 1082, reagent tank 1083 for precipitants, etc., internal standard bottle 1084, reagent carrier 1085, arm module 109, primary arm 1091, primary arm support 10911, primary motor 10912, primary motor support 10913, primary coupling 10914, primary arm shaft 10915, secondary arm 1092, secondary end cap 10921, secondary motor mount 10922, secondary arm motor 10923, secondary arm housing 10924, secondary coupling 10925, rotary joint 10926, secondary arm shaft 10927, tertiary arm 1093, tertiary end cap 10931, tertiary arm housing 10932, tertiary arm motor mount 10933, tertiary arm motor 10934, tertiary arm motor support 10935, mechanical claw 1094, mechanical claw end cap 10941, mechanical claw connection block 42, a gripper rotating motor 10943, a gripper housing 10944, a gripper opening and closing motor 10945, a gripper bearing 10946, a gripper 10947, a lancet coupling groove 10948, an anti-slip groove 10949, a gun head and wash tank module 110, a sterilization wash tank 1101, a venom inlet 1102, a disinfectant liquid outlet 1103, a cleaning liquid inlet 1104, a cleaning liquid outlet 1105, a multiplexing type pipette head 1106, a partition 1107, an acrylic door 111, a sampling module 200, a three-way switching valve 201, a strong-wash inlet 202, a weak-wash inlet 203, an output end 204, a switching motor 205, a selection piston 206, a sampling coupler 207, a sampling pipetting motor 208, a six-way valve 209, a pipetting needle module 210, a pipetting needle motor seat 2101, a pipetting needle motor 2102, a pipetting needle screw 2103, a pipetting needle screw slider 2104, a guide groove 2105, a mobile phase interface 211, a chromatographic column interface 212, a pipetting needle interface 213, a sampling ring 214, an output end line 215, a sampling movement module 216, y-axis motor mount 2161, Y-axis slider group 2162, X-axis shaft mount 2163, Y-axis motor 2164, X-axis motor shaft 2165, X-axis motor 2166, pipette needle connection slider 2167, column oven module 300, sample tube 301, column 302, column buckle 303, temperature control module 304, ventilation fan 305, column oven door window 306, seal 307, cavity radiator 308, rear side radiator 309, cavity radiator fan 311, rear side fan 312, semiconductor refrigeration sheet 313, thermal insulation cotton 314, mounting plate 315, mass spectrometry module 400, mass spectrometry housing 401, vacuum module 402, vacuum chamber 4021, ion channel interface plate 4022, ion source module 403, ion source cavity 4031, ion source fixture 4032, the ion source probe 4033, the horizontal micrometer 4034, the vertical micrometer 4035, the heating rod 4036, the observation window 4037, the molecular pump 404, the vacuum tube 405, the vacuum flange 406, the triple quadrupole module 407, the quaternary support 4071, the second quaternary support 4072, the first quaternary support 4073, the third quaternary support 4074, the quaternary support plate 4075, the detector 4076, the pump module 500, the mobile phase switching valve 501, the degasser 502, the mobile phase pump 503, the primary mixer 504, the waste liquid port 505, the mixer 506, the three-way switching valve 507, the switching motor 508, the solvent chamber module 600, the computer module 700, the blood sampling tube rack 800, the rack structure 900, and the universal castor 901.

Detailed Description

The present application will be described and illustrated with reference to the accompanying drawings and examples in order to make the objects, technical solutions and advantages of the present application more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the application. All other embodiments, which can be made by a person of ordinary skill in the art based on the embodiments provided by the present application without making any inventive effort, are intended to fall within the scope of the present application.

Reference in the specification to "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment may be included in at least one embodiment of the application. The appearances of such phrases in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments. It is to be expressly and implicitly understood by those of ordinary skill in the art that the described embodiments of the application can be combined with other embodiments without conflict.

Unless defined otherwise, technical or scientific terms used herein should be given the ordinary meaning as understood by one of ordinary skill in the art to which this application belongs. The terms "a," "an," "the," and similar referents in the context of the application are not to be construed as limiting the quantity, but rather as singular or plural. The terms "comprising," "including," "having," and any variations thereof, are intended to cover a non-exclusive inclusion; the terms "connected," "coupled," and the like in connection with the present application are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. The term "plurality" as used herein means greater than or equal to two.

Example 1, refer to fig. 1-37.

As shown in fig. 1, the invention provides a full-automatic liquid phase mass spectrum detection system special for fat-soluble vitamins, which at least comprises a sample pretreatment module 100, a sample injection module 200, a column temperature box module 300, a mass spectrum module 400, a pump module 500, a solvent chamber module 600 and a computer module 700; the sample pretreatment module 100 is disposed in the middle of the mass spectrum detection system, and is configured to receive a blood sampling tube rack 800 containing samples, perform pretreatment on the samples therein, and after the pretreatment is completed, the sample pretreatment module 100 transfers the blood sampling tube rack 800 to the position below the sample injection module 200; the sample injection module 200 is arranged at the upper part of the mass spectrum detection system and is used for sucking the pre-processed sample and sending the sample into the column incubator module 300 through the pump module 500; the column incubator module 300 is disposed at one side of the sample introduction module 200, and is used for receiving the pretreated sample, controlling the temperature of the sample, and completing the separation of fat-soluble vitamins in the sample; the mass spectrum module 400 is arranged at the lower part of the mass spectrum detection system, the compound separated by the column temperature box module 300 enters the mass spectrum module 400 through a pipeline, the mass spectrum module analyzes the compound and transmits the acquired signal to the computer module 700, and the computer module feeds back the detection result and generates a detection report according to the acquired data and preset content. The whole system forms integrated equipment through the rack structure 900, and the rack lower part is equipped with the universal castor 901 for equipment has the mobility.

As shown in fig. 2-3, the mass spectrum module 400 includes a mass spectrum housing 401, the mass spectrum housing 401 is a part of the whole frame structure 900, the universal casters 901 are disposed at the bottom of the mass spectrum housing 401, a vacuum module 402 is disposed in the mass spectrum housing 401, the vacuum module 402 includes a vacuum cavity 4021, and the vacuum cavity 4021 is used for isolating an ion channel from an external environment; one end of the vacuum cavity 4021 is disposed on one surface of the mass spectrum housing 401, an ion channel interface board 4022 extending out of the mass spectrum housing 401 is disposed at the end of the vacuum cavity 4021, the ion channel interface board 4022 is used for removing ionized impurities, reducing noise signals and reducing ion clusters (ion aggregation effect), the ion channel interface board 4022 can be connected with an ion source module 403, a molecular pump 404 is disposed on one side of the vacuum cavity 4021, the molecular pump is an air extractor, the molecular pump drives gas to move through high-speed rotation of an impeller, the molecular pump 404 is communicated with an external space through a vacuum tube 405 for vacuumizing, specifically, a vacuum flange 406 is disposed at an end of the vacuum tube 405, and the vacuum flange 406 is fixed on the mass spectrum housing 401 and is communicated with the outside of the device (and can also be taken over in addition) for discharging the vacuumized gas.

As shown in fig. 4, the ion source module 403 includes an ion source chamber 4031, and the ion source chamber 4031 creates a relatively stable environment for ionizing isolated external environments; the upper part of the ion source cavity 4031 is provided with an ion source fixing device 4032, the ion source fixing device 4032 is provided with an ion source probe 4033 which can extend into the ion source cavity 4031, the ion source probe 4033 is used for providing a probe pipeline, and a sample separated from the temperature control box module enters from the probe pipeline and is ionized under the action of an electric field; the ion source fixing device 4032 is further provided with a calibration structure, and the calibration structure comprises a horizontal micrometer 4034 and a vertical micrometer 4035, which are used for calibrating the spatial position (horizontal position+vertical position) of the ion source probe, so that the ionized compound can smoothly enter the ion channel; a heating rod 4036 is further arranged in the ion source cavity 4031 and used for assisting ionization and volatilizing the mobile phase solvent; the surface of the ion source cavity 4031 is provided with an observation window 4037, and the observation window 4037 is made of transparent acrylic material and is used for observing the working state of the ion source.

As shown in fig. 5, the mass spectrometry module 400 further comprises a triple quadrupole module 407, the triple quadrupole module 407 being disposed within the vacuum chamber 4021; the triple quadrupole rod module 407 is installed in the vacuum cavity 4021, the triple quadrupole rod module 407 comprises a quaternary rod support 4071 for supporting the whole triple quadrupole rod module 407, a second quaternary rod 4072 is fixedly connected to the middle of the quaternary rod support 4071, two ends of the middle quaternary rod are respectively connected with a first quaternary rod 4073 and a third quaternary rod 4074, the first quaternary rod 4073 is located at the upstream end of the triple quadrupole rod module 407 and used for screening and focusing parent ions, and only parent ions meeting a set mass-to-charge ratio can pass through the first quaternary rod 4073; the second quaternary lever 4072 is an ion reaction tank, the energetic parent ions enter the reaction tank containing nitrogen, the parent ions are broken into child ions, the third quaternary lever 4074 is used for screening and focusing the child ions, one side of the quaternary lever bracket 4071 is provided with a quaternary lever fixing plate 4075, the quaternary lever fixing plate 4075 is provided with a detector 4076, and the detector 4076 is used for measuring the number of the child ions and converting the number of the child ions into an electric signal; the triple quadrupole rod module further comprises a communication control module, wherein the communication control module is connected with the detector 4076 and sends the electric signal converted by the detector to the computer module 700.

As shown in fig. 6, the sample pretreatment module includes a pretreatment chamber 101, an in-out blood collection tube rack module 102 is disposed in the pretreatment chamber 101, the in-out blood collection tube rack module 102 is configured to place a blood collection tube sample to be treated and a blood collection tube sample that has been treated, and can add a new blood collection tube sample and take a finished blood collection tube sample to the in-out blood collection tube rack module at any time, and the in-out blood collection tube rack module 102 can be kept in a non-stop state all the time, so that the device can be kept to operate efficiently. The pretreatment chamber 101 is also internally provided with a centrifugal module 103, a pipette module 104, a mixing module 105, a deep hole plate module 106, a solvent bottle module 107, a tip head and a reagent tank module 108. Wherein, the centrifugal module 103 is arranged near the in-out blood sampling pipe frame module 102 and is used for centrifuging the blood sampling pipe and the deep pore plate, separating serum and plasma in the sample and separating precipitated protein from the sample; the pipette gun module 104 is used for respectively sucking serum and protein precipitants and internal standard reagents from the solvent bottle module of the centrifugal module 103 and the tip head and reagent tank module 108 and placing the reagents into the deep hole plate, the deep hole plate module 106 is used for moving the deep hole plate to a sample feeding position and a mixing position, the mixing module 105 is used for mixing the serum and protein precipitants and internal standard reagents in the deep hole plate, the solvent bottle module 107 is used for storing mobile phase reagents and sample injection needle cleaning reagents, the tip head and reagent tank module 108 is configured for storing tip head, precipitants, internal standard reagents, waste tanks and the like required by pipetting, the sample pretreatment module 100 further comprises a mechanical arm module 109, and the mechanical arm module 109 is used for moving the blood sampling pipe frame and/or the deep hole plate, realizing the movement of the blood sampling pipe frame module, the centrifugal module and the hole plate module and finishing the code scanning procedure of the blood sampling pipe.

In addition, a gun head and cleaning pool module 110 is further arranged in the pretreatment chamber 101, and the gun head and cleaning pool module 110 is arranged near the pipetting gun module 104 and is used for placing and cleaning a gun head for sucking a serum sample; a damper foot pad 1011 is disposed below the pretreatment chamber 101, and the damper foot pad 1011 is used to reduce vibration from the centrifugal module from being transferred to the mass spectrum module 400. The pretreatment chamber 101 is used for partially or temporarily closing the pretreatment chamber through a rack cabinet door structure such as an acrylic door 111 and the like, so that environmental pollution is avoided.

As shown in fig. 7, the in-out sample rack module 102 includes an in-sample rack area 1021 and an out-sample rack area 1022, wherein the in-sample rack area 1021 is used for placing the blood collection tube rack 800 ready for processing, and the out-sample rack area 1022 is used for placing the processed blood collection tube rack 800; the bottom of each blood collection tube rack 800 is provided with a pressure sensor 1023, so that the weight of each blood collection tube rack can be measured, and the leveling operation can be conveniently performed after the blood collection tube racks are transferred to the centrifuge module 103; the code scanner 1024 is installed in the middle of the side face of the sample frame entering and exiting module 102, after the blood sampling tube rack 800 is transferred from the sample frame entering module 102 to the centrifugal module 103, the mechanical arm module 109 can draw out the blood sampling tube (4 blood sampling tubes each time), and the code on the blood sampling tube is scanned and recorded through the code scanner 1024 to acquire information, so that the information is used as a unique code corresponding to the whole sample processing, and the tracking is convenient.

As shown in fig. 8, the centrifugal module 103 includes a centrifugal machine base 1031, a motor is disposed in the centrifugal machine base 1031, the motor is connected to a motor shaft 1033 through a coupling 1032, the motor shaft 1033 is rotatably connected to a centrifugal carrier 1034, a blood collection tube carrier placement position 1035 is disposed on the centrifugal carrier 1034, the blood collection tube carrier 1036 for loading the blood collection tube 800 can be placed on the blood collection tube carrier placement position 1035, in order to facilitate the centrifugal operation, the centrifugal carrier 1034 includes at least two symmetrical blood collection tube carrier placement positions 1035, in this embodiment, the centrifugal carrier 1034 includes four blood collection tube carrier placement positions 1035, and the four blood collection tube carrier placement positions 1035 are uniformly distributed in the circumferential direction of the centrifugal carrier. Each blood collection tube carrier placement position 1035 can be loaded with two leveling reagent tubes 1037, the balancing amount and the balancing scheme are calculated according to the weight measured by the blood collection tube rack module, the pipette module is controlled to absorb reagents from the tip head and the reagent tank module to the leveling reagent tubes 1037 according to the calculated balancing scheme, the reagents are used for balancing, the load on a centrifugal machine is balanced, the centrifugal machine is ensured to work more stably, and in the invention, the balancing scheme can be carried out by using algorithms or software in the prior art.

The centrifugal carrier 103 comprises a middle fixing plate 1038 and side fixing plates 1039, the motor shaft 1033 is connected with the middle fixing plate 1038, the side fixing plates 1039 are U-shaped, two side parts of the U-shaped are arranged on two sides of the blood collection tube carrier placement position 1035, mounting grooves 1136 and sliding rails 1135 are arranged on the two sides of the blood collection tube carrier 1036, shelves 1137 are arranged on two sides of the blood collection tube carrier 1036, the blood collection tube carrier 1036 is placed on the centrifugal carrier 103 through combination of the shelves 1137 and the mounting grooves 1136, and a fixing plate cover plate 1230 can be covered on the middle fixing plate 1038 and used for locking the blood collection tube carrier 1036.

As shown in fig. 9, the middle fixing plate 1038 is further provided with an elastic element 1130 corresponding to each side fixing plate 1039, the side fixing plates 1039 are connected with the elastic element 1130 through a carrier fixing plate connector 1131, a locking element 1132 is disposed at the bottom of the U-shape, the side fixing plates 1039 are provided with a carrier fixing plate 1133 and a limiting fixing plate 1134, the limiting fixing plate 1134 is fixedly connected to the carrier fixing plate 1133, the carrier fixing plate 1133 can be driven by the limiting fixing plate 1134 to move along the sliding rail 1135, and when the carrier fixing plate 1133 moves, the carrier fixing plate 1133 can be locked or released by the locking element 1132 at different positions.

In the invention, the mechanical arm module 109 drives the carrier fixing plate 1133 in the centrifugal carrier 1034 to move, so as to lock or unlock the blood collection tube carrier 1036, the carrier fixing plate 1133 is rigidly connected with the limiting fixing plate 1134, the mechanical arm module 109 can complete the translation of the limiting fixing plate 1134, and the purpose of locking or unlocking the blood collection tube carrier 1036 is achieved by matching with the fixing plate cover plate 1230.

As shown in fig. 10-14, the principle of the carrier fixing plate stretching to lock or unlock the blood collection tube carrier is as follows: when the carriage fixation plate connector 1131 is in the initial position (the state shown in fig. 10), the elastic element 1130 is in the retracted state, the locking element 1132 is in the relaxed state, and the corresponding blood collection tube carriage 1036 is in the unlocked state, and can be removed from the centrifugal carriage 1034 at any time; when the mechanical arm module 109 moves the limit fixing plate 1134 to drive the carrier fixing plate connector 1131 to move to the right to the highest position (the state shown in fig. 11), the elastic element 1130 is stretched to store energy, the outer end of the elastic element 1130 automatically slides down to the first step 1138 due to the action of gravity, then the outer end of the carrier fixing plate connector 1131 automatically slides down to the second step 1139 (the transition position) due to the lateral V-shape of the locking element 1132 and the elastic force of the elastic element 1130 (the state shown in fig. 12), and the carrier fixing plate connector 1131 stays at the position without other external force due to the integral and step-shaped arrangement of the locking element 1132, the locking element 1132 is in the action of the combined fixing plate cover 1230, the blood collection tube carrier 1036 is in the locking state, and the position of the blood collection tube carrier 1036 is ensured not to change in the centrifugation process; when the centrifugal process is completed and unlocking is required, the mechanical arm module 109 moves the limiting fixing plate 1134 outwards again to drive the carrier fixing plate connector 1131 to move to the right to the third step 1231 (as shown in fig. 13), after the third step 1231 is rotated, the mechanical arm module returns to the initial state (the state shown in fig. 10) under the action of the elastic element 1130 after passing through the fourth step 1232 (as shown in fig. 14), and the whole unlocking and locking process cannot slide reversely through the structural design of the locking element 1132, so that reliability is ensured.

As shown in fig. 15 and 21, the mixing module 105 includes a mixing motor 1051, a mixing shaft 1052, an eccentric 1053, and a mixing wheel 1054; the deep hole plate module 106 comprises a deep hole plate 1061, a deep hole plate carrier 1062, a deep hole plate carrier tray 1063 and a deep hole plate lifting module 1064, the mixing module 105 is located at a position below the deep hole plate module 106 corresponding to the deep hole plate 1061, the deep hole plate carrier tray 1063 is slidably mounted on the deep hole plate lifting module 1064 and can move up and down under the driving of the deep hole plate carrier tray 1064, the deep hole plate carrier 1062 is arranged in the deep hole plate carrier tray 1063, the deep hole plate 1061 is arranged in the deep hole plate carrier 1062, a deep hole plate carrier bottom hole 1065 is formed in the bottom of the deep hole plate carrier 1062, when the deep hole plate 1061 is lowered to the lowest position, the deep hole plate carrier bottom hole 1065 is combined with a mixing wheel 1054 of the mixing module 105, and the mixing wheel 1054 is sequentially connected through an eccentric wheel 1053, a coupling 1052 and a mixing motor 1051, so that the deep hole plate 1061 is mixed.

As shown in fig. 22, the deep well plate carrier 1062 includes a deep well carrier 10621, a deep well plate fixing spring 10622, and a deep well plate leveling reagent tube 10623. The deep hole carrier bottom comprises deep hole plate carrier bottom holes 1065 (for matching and mixing) and a refrigerating chamber 10624 (for adjusting the temperature of the deep hole plate), and the deep hole plate leveling reagent pipes 10623 are used for balancing when the deep hole plate is centrifuged, so that the centrifugal operation is more stable.

As shown in fig. 23, the deep hole plate carrier tray 1063 includes a deep hole plate cooling module 10631, a deep hole plate tray 10632, and a buffer rubber pad 10633, and specifically, the deep hole plate cooling module 10631 includes a semiconductor cooling fin, a cooling fan, and a cooling module bracket. The deep pore plate refrigerating module 10631 is deep into the refrigerating chamber and is positioned at the bottom of the deep pore plate, so that the temperature control requirement of 5-10 ℃ can be provided for samples in the deep pore plate; the buffer rubber pad 10633 may play a role of buffering when the mixing operation is performed.

As shown in FIG. 16, tip head and reagent tank module 108 includes tip head carrier 1081, waste tank 1082, reagent tank 1083, such as precipitant, internal standard bottle 1084, and reagent carrier 1085. Wherein, tip head carrier 1081 can load different specification tip heads, waste tank 1082 can load waste tip heads and reagents, reagent tank 1083 such as precipitant, internal standard bottle 1084 are all installed in reagent carrier 1085, pipette module 104 and manipulator module 109 can sample from tip heads and reagent tank module 108.

As shown in fig. 17, the pipette module 104 includes an X-axis module 1041, a Y-axis module 1042, and a pipette 1043; the top of the pipette is slidably connected to the X-axis module 1041, the X-axis module 1041 is slidably connected to the Y-axis module 1042, and the X-axis module 1041 and the Y-axis module 1042 enable the pipette 1043 to move in two dimensions.

As shown in fig. 18, the pipette gun 1043 includes a plurality of pipette gun assemblies and a spacing adjustment assembly, the pipette gun assemblies include a plurality of pipette gun bodies including at least one fixed pipette gun 1044 and a plurality of movable pipette guns 1045, wherein the fixed pipette guns 1044 are fixed on the spacing adjustment assembly and do not participate in the spacing adjustment movement, and correspondingly, the plurality of movable pipette guns 1045 are movably connected with the spacing adjustment assembly, respectively, and are configured to be able to adjust the spacing therebetween by the spacing adjustment assembly.

Specifically, the distance adjusting assembly includes a distance adjusting support plate 1046, the distance adjusting support plates 1046 are arranged in pairs, a plurality of screw rods 1047 corresponding to the plurality of movable pipetting guns 1045 are arranged between the distance adjusting support plates 1046, a distance adjusting slide block 1048 is arranged on each screw rod 1047, the distance adjusting slide block 1048 is connected with the plurality of movable pipetting guns 1045 through a slide block connecting piece 1049, a distance adjusting motor 1140 is arranged on the distance adjusting support plates 1046, the distance adjusting motor 1140 is connected with the plurality of screw rods 1047 through a gear set 1141, the gear set 1141 includes a plurality of gears with different times of speed, the gear at the middle is low in times of speed, the gear at the outer side is gradually increased in times of speed, and each screw rod is connected with the gear through a coupling; for example, in the present invention, the fixed pipette 1044 is fixed without moving and connecting gears, the movable pipette 1045 closest to the fixed pipette 1044 is connected to the middle 1-fold gear 1142 by a screw, the next-closest movable pipette 1045 is connected to the 2-fold gear 1143 by a screw, the outermost movable pipette 1045 is connected to the 3-fold gear 1144 by a screw, and when the interval adjusting motor 1140 is started, the three movable pipettes 1045 are respectively far from or near the fixed pipette 1044 at 1-fold, 2-fold and 3-fold speeds, thereby achieving equidistant adjustment of the pipettes and meeting the pipetting demands.

A pipette guiding rail 1145 is further disposed between the spacing adjusting support plates 1046, and a plurality of guiding sliders 1146 corresponding to the movable pipette 1045 are disposed on the pipette guiding rail 1145, and each guiding slider 1146 is connected to one movable pipette 1045, for guiding movement of the movable pipette 1045.

As shown in fig. 19, the pipette (which may be either a stationary pipette or a movable pipette, both of which are identical in function of pipetting) includes a pipette fixture 1147, the pipette fixture 1147 being used to position the lateral position of the pipette assembly, i.e., the pipette fixture 1147 allows the pipette to be fixed to a spacing adjustment assembly, where it may be directly fixed to a constant position component, such as a spacing adjustment support plate 1046, for a stationary pipette, and directly fixed to a guide slide 1146, and may be spacing adjusted, for a movable pipette. In addition, the pipette fixing block 1147 is provided with a Z-axis motor 1148 and a Z-axis slide rail set 1149, and the Z-axis motor 1148 and the Z-axis slide rail set 1149 are used for driving the pipette to move up and down; the pipette further comprises a pipetting motor 1240, the pipetting motor 1240 is connected with a pipetting piston 1242 through a pipetting piston rod 1241, the pipetting piston 1242 can move in a piston cavity 1243 to complete pipetting action, the pipetting piston 1242 is connected with a pipetting head 1244, a head removing member 1245 is arranged between the pipetting piston 1242 and the pipetting head 1244, the head removing member 1245 is a sleeve sleeved on the pipetting head and can slide on the pipetting head, other structures (such as a tip head sleeved on the pipetting head) outside the pipetting head are pushed, so that head removing is achieved, one side of the head removing member 1245 is fixedly connected with a head removing auxiliary mechanism, and the head removing auxiliary mechanism comprises a metal fixing member 1246, a pipetting spring 1247, an electromagnet 1248 and a spring fixing member 1249. The movement of the pipette in the Z-axis direction is completed by the Z-axis motor 1148 and the Z-axis sliding rail set 1149, the action of pipetting is completed by the pipetting motor 1240, the pipetting piston 1242 and the piston cavity 1243 in a matched mode, when the Z-axis motor 1148 moves downwards, the pipetting head 1244 is squeezed into the tip head to grasp the tip head, when the tip head needs to be withdrawn, the electromagnet 1248 is electrified to attract the metal fixing part 1246 right above, the spring 1247 is compressed, the head removing part 1245 moves downwards in a translating mode, and the tip head is extruded out of the pipetting head 1244.

As shown in fig. 20, the pipette head 1244 includes a rubber gasket 12441, a rubber fixing part 12442, and a metal spring plate 12443. When the liquid suction head 1244 is extruded into the tip head, the rubber sealing ring 12441 can play a sealing role, so that the air tightness and the liquid suction precision are ensured, the inner side is embedded with two metal elastic sheets 12443, the two metal elastic sheets are placed at an angle of 180 degrees, when the liquid suction head 1244 is extruded into the tip head, the metal elastic sheets 12443 are tightly contacted with the tip head, whether the tip head contacts the liquid level or not can be judged by detecting the resistance values at the two ends of the metal elastic sheets 12443, and the liquid suction quantity can be well controlled, therefore, the position of the liquid suction head can be well controlled, and the phenomenon that the liquid suction head enters the liquid level too deeply to collide or suck the bottom sediment to cause the damage of the liquid suction head or the blockage of a pipeline is avoided; meanwhile, because the liquid level detection function is provided, the deep hole plate can be used for directly sampling, compared with the conventional equipment which needs to suck a very small amount of samples from the upper surface of the deep hole plate to the shallow hole plate, and then the shallow hole plate is used for sampling, the invention reduces the cost, the procedures and the time and improves the efficiency.

As shown in fig. 24, the gun head and wash basin module 110 includes a sanitizing cartridge 1101, a sanitizing liquid inlet 1102, a sanitizing liquid outlet 1103, a cleaning liquid inlet 1104, a cleaning liquid outlet 1105, and a reusable pipette head 1106. The disinfection cleaning box 1101 is divided into a disinfection tank and a disinfection liquid cleaning tank, the disinfection cleaning box is separated by a partition plate 1107, when the liquid suction head 1244 of the liquid suction gun module sucks the multiplexing liquid suction head, the butyl rubber plug in the liquid suction tube can be directly penetrated and the serum sample is sucked (the operations of uncovering and capping are reduced, the efficiency is higher), the cleaning box is transferred to the deep hole plate 1061, the multiplexing liquid suction head is deeply penetrated into the disinfection tank for disinfection, then is deeply penetrated into the disinfection liquid cleaning tank for cleaning, and then the liquid suction operation of the next sample is performed.

As shown in fig. 25, the robot module 109 includes a primary robot 1091, a secondary robot 1092, a tertiary robot 1093, and a gripper 1094. As shown in fig. 26, the primary mechanical arm 1091 includes a primary mechanical arm support 10911, a primary motor 10912, a primary motor support 10913, a primary coupler 10914, and a primary mechanical arm rotating shaft 10915, where the primary mechanical arm support 1091 is used for fixing the mechanical arm module 109, the primary motor 1092 provides rotational power, the primary motor support 1093 is used for fixing the motor, the coupler 1094 and the primary mechanical arm rotating shaft 1095 transmit the rotational power to the secondary mechanical arm, and the primary mechanical arm rotating direction is shown by an arrow in the figure. As shown in fig. 27, the secondary mechanical arm 1092 includes a secondary end cap 10921, a secondary motor base 10922, a secondary mechanical arm motor 10923, a secondary mechanical arm housing 10924, a secondary coupling 10925, a rotary connection block 10926 and a secondary mechanical arm rotating shaft 10927, and the secondary end cap 10921 is used for protecting the motor, so as to facilitate maintenance of the interface; a secondary mechanical arm motor 10923 provides power for rotation; the secondary coupler 10925, the rotary connection block 10926, and the secondary arm shaft 10927 transmit the motor rotation to the tertiary arm 1093, with the secondary arm rotation direction indicated by the arrow in the figure. As shown in fig. 28, the tertiary mechanical arm 1093 includes a tertiary end cap 10931, a tertiary mechanical arm housing 10932, a tertiary mechanical arm motor mount 10933, a tertiary mechanical arm motor 10934, and a tertiary mechanical arm motor support 10935, and the rotation direction of the tertiary mechanical arm is shown by an arrow in the figure. As shown in fig. 29, the gripper 1094 includes a gripper end cover 10941, a gripper connecting block 10942, a gripper rotating motor 10943, a gripper housing 10944, a gripper opening and closing motor 10945, a gripper bearing 10946, a gripper 10947, and the like, the gripper connecting block 10942 is connected with a secondary mechanical arm 1092, the gripper rotating motor 10943 is used for changing the orientation of the gripper 10947, and after the gripper opening and closing motor 10945 and the gripper bearing 10946 are assembled, the opening and closing of the gripper 10947 is controlled; the mechanical claw member 10947 comprises 4 parallel blood collection tube coupling grooves 10948, so that the mechanical claw is convenient to grasp the blood collection tubes for code scanning operation, and anti-slip patterns 10949 are arranged on the inner side of the blood collection tube coupling grooves 10948, so that the grabbing is more reliable.

As shown in fig. 30, the pump module 500 includes two mobile phase switching valves 501, the two mobile phase switching valves 501 are communicated with a degasser 502, two pipelines of the degasser 502 are respectively connected with two mobile phase pumps 503, the two mobile phase pumps 503 are connected with a primary mixer 504, one outlet of the primary mixer 504 is connected with a waste liquid port 505, and the other outlet is connected with a mixer 506; in this embodiment, the two mobile phase switching valves 501 are an a mobile phase switching valve and a B mobile phase switching valve respectively, and 4 paths of mobile phase switching valves are connected to four mobile phase bottles in the solvent bottle module 600, and strong washing and weak washing (required by a cleaning pipeline) are also connected to reagent bottles corresponding to the solvent bottle module; the left side of the degasser 502 is an inlet end, and is respectively connected with an A mobile phase switching valve, a B mobile phase switching valve, a strong washing pipeline and a weak washing pipeline, so that bubbles in the pipelines are removed, the pipeline is stable in hydraulic pressure, and the sample injection is accurate. The mobile phase A and the mobile phase B are respectively connected to the mobile phase A pump and the mobile phase B pump after being degassed by a degasser, the mobile phase pumps provide power for the two paths of mobile phases, the output end of the mobile phase A and the mobile phase B are connected to a primary mixer, the primary mixing of the mobile phases A and the mobile phases B can be completed in the primary mixer, the mobile phases A and the mobile phases B are connected to the mixer after being mixed, and the mobile phases A and the mobile phases B are connected to a sample injection module after being mixed. In addition, when the pipeline of the pump module needs to be flushed, the output end can be switched to the waste liquid port without entering the sample injection module.

As shown in fig. 31, the sample injection module 200 is connected to the pump module 500, the sample injection module 200 further includes a three-way switching valve 201, the three-way switching valve 201 is respectively connected to the strong wash liquid inlet 202, the weak wash liquid inlet 203 and the output end 204, the three-way switching valve 201 is connected to the three-way switching valve 201 through a switching motor 205, and the communicated valve ports are selected through a selection piston 206, the selection piston 206 sucks the cleaning liquid through a sample injection coupling 207 and a sample injection liquid suction motor 208 and then outputs the cleaning liquid from the output end 204, the output end 204 is connected to a six-way valve 209, and the six-way valve 209 is connected to a liquid suction needle 210 to realize liquid suction and flushing functions.

As shown in fig. 32-33, the six-way valve 209 includes 6 ports, which are respectively connected to a mobile phase port 211 (No. 1 hole), a chromatographic column port 212 (No. 6 hole), a liquid suction needle port 213 (No. 4 hole), a sample injection ring 214 (No. 2,5 holes), and an output end pipeline 215 (No. 3 hole), and the six-way valve 209 has two positions, namely a position (fig. 32) and B position (fig. 33): at position a: the holes 1-2,3-4 and 5-6 are conducted in pairs; b bit: 1-6,2-3,4-5 holes are communicated; the sample injection ring 214 is connected between the holes 2 and 5, and the holes 2 and 5 are in a conducting state. When the strong and weak needle washing is needed, the six-way valve is in the A position, strong and weak washing liquid enters from the hole 3 and enters the liquid suction needle 210 through the hole 4, and the needle washing is completed; when the sample is required to be sucked, the position is switched to the position B, at the moment, the sample is sucked by the liquid suction needle, enters the hole No. 4 to the hole No. 5, enters the sample injection ring, stays in the sample injection ring 214, is switched to the position A in time, at the moment, the mobile phase enters the six-way valve from the hole No. 1, reaches the hole No. 2, and flows to the hole No. 6 from the hole No. 5 carrying the sample injection ring, and the hole No. 6 is connected with the column incubator module 300, so that the sample injection is completed.

As shown in fig. 34, the sample injection module 200 further includes a sample injection moving XY module 216, the sample injection moving XY module 216 includes a Y-axis motor support 2161, a Y-axis slider group 2162 is connected to the Y-axis motor support 2161, an X-axis seat 2163 is connected to the Y-axis slider group 2162, the X-axis seat 2163 can be driven by a Y-axis motor 2164 to slide along the Y-axis slider group 2162, an X-axis motor shaft 2165 and an X-axis motor 2166 are disposed on the X-axis seat 2163, a liquid suction needle connecting slider 2167 is connected to the X-axis motor shaft 2165, and the liquid suction needle connecting slider 2167 is connected to the liquid suction needle module 210.

As shown in fig. 35, the liquid suction needle module 210 includes a liquid suction needle motor base 2101, the liquid suction needle motor base 2101 is connected with a liquid suction needle connecting slider 2167, a liquid suction needle motor 2102 is arranged in the liquid suction needle motor base 2101, the liquid suction needle motor 2102 is connected with a liquid suction needle screw rod slider 2104 through a liquid suction needle screw rod 2103, and the liquid suction needle screw rod slider 2104 moves in a guide groove 2105 and drives the liquid suction needle to move up and down to perform liquid suction. The liquid suction needle motor base 2101 is arranged on a liquid suction needle connecting sliding block 2167 of the sample injection moving XY module, so that X and Y axis direction movement is realized; the suction needle motor 2102 rotates to drive the suction needle screw rod sliding block 2104 to slide up and down along the guide groove 2105, and the suction needle is arranged on the suction needle screw rod sliding block 2104, so that the movement of the suction needle in the Z-axis direction is realized, and the suction action is completed.

As shown in fig. 36, the column incubator module 300 includes a sample tube 301, the sample tube 301 is led into the temperature control box module, and is connected to a chromatographic column 302 in the temperature control box module, the chromatographic column 302 is fixedly mounted on a temperature control module 304 through a chromatographic column buckle 303, a ventilation fan 305 is disposed at the rear end of the temperature control box module 300, a column oven door window 306 is disposed at the front end of the temperature control box module 300, and the periphery of the column oven door window 306 is sealed by a sealing ring 307.

As shown in fig. 37, the temperature control module 304 includes an inner cavity radiator 308, a rear side radiator 309, heat insulation cotton 314, an inner cavity cooling fan 311, a rear side fan 312 and a semiconductor cooling fin 313, the semiconductor cooling fin 313 is mounted in mounting grooves of the rear side radiator 309 and the inner cavity radiator 308, heat conductive silicone grease is coated on two sides of the semiconductor cooling fin 313, heat is insulated by the heat insulation cotton 314, the rear side radiator 309 and the inner cavity radiator 308 are mounted on a fixing plate 315, the inner cavity cooling fan 311 and the rear side fan 312 are mounted on the outer sides of the inner cavity radiator 308 and the rear side radiator 309 respectively, and the fan is used for improving the working efficiency of the semiconductor cooling fin.

The sample injection tube 301 is connected with the chromatographic column interface of the six-way valve module of the sample injection module 200, the sample sucked by the sample injection module enters the chromatographic column to complete separation, and the separated compound (fat-soluble vitamin) enters the mass spectrum module. The temperature control module is used for creating proper temperature for the chromatographic column and ensuring the separation effect of the chromatographic column.

After the mass spectrum is completed, the mass spectrum module sends the analysis result to the computer module 700, and the computer module generates a mass spectrum report according to a preset report template.

Example 2, refer to fig. 38.

The embodiment provides a full-automatic liquid-phase mass spectrometry detection method special for fat-soluble vitamins, which adopts the detection system described in the embodiment 1 and comprises the following steps:

s1, sample pretreatment:

1.1, placing the blood collection tube in a carrier, and centrifuging at 3000-8000rpm for 5-10 minutes;

1.2 sucking 50-200. Mu.L serum into 96-well deep well plate;

1.3 precisely removing 20-100 mu L of protein precipitant into a 96-well deep hole plate;

1.4 precisely adding 100-400 mu L of internal standard solution;

1.5 placing a 96 deep hole plate in a mixer, and swirling for 5-10 minutes at 600-3000 rpm;

1.6 centrifuging at 2000-4000rpm for 5-10 min;

s2, flow matching and sample injection: driving a sample to be sampled by a pump module according to a mixed proportion flowing sample, and carrying out gradient elution on a flowing phase; wherein, the mobile phase ratio is: one of the mobile phases is an aqueous solution containing 0.1% formic acid, and the other mobile phase is a methanol solution containing 0.1% formic acid; the flow rate of the pump is controlled to be 0.6-0.8 mL/min;

in the sample injection process, the temperature of the deep hole plate is controlled at 10-15 ℃, and the sample injection amount is controlled at 10-40 mu L.

With parameters fully in accordance with the invention, gradient elution can be carried out in the manner of Table 1.

Table 1: gradient elution

S3, separating: the module carries the sample into a temperature control box module and finishes the separation of fat-soluble vitamins in the temperature control box module;

the separation and temperature control conditions of the temperature control box module are as follows:

chromatographic column: C-18,2.1X 50mm, filler particle size 3 μm or equivalent performance;

column oven temperature: 30-40 ℃.

S4, mass spectrum: the separated fat-soluble vitamins enter a mass spectrum module for analysis, and the analysis result of the substances is fed back to a computer module, and the mass spectrum conditions are shown in table 2.

Table 2: mass spectrometry conditions

The stability, sensitivity and accuracy of the present invention will be demonstrated by the following examples using equipment tests, which are exemplified by VD2 and VD3, which are representative of fat-soluble vitamins.

Example 3 precision test.

Sample conditions: taking blank serum into a blood collection tube, adding the standard solution of VD2 and VD3 into the blank serum respectively to enable the concentration to be 6.2 ng/mL and 8.1 ng/mL respectively, and then enabling the blood collection tube to be connected to a blood collection tube frame, and carrying out pretreatment and analysis along with the blood collection tube frame.

Test results: as in table 3.

Table 3 test items: precision (n=10)

The invention has higher stability for measuring the fat-soluble vitamin according to the test data of a plurality of samples.

Example 4, accuracy (relative deviation%) test.

Sample conditions:

1) And (3) using a low-concentration VD2 and VD3 standard quality control substance to the blood collection tube, and carrying out pretreatment and analysis along with a blood collection tube rack, wherein the VD2 concentration target value is as follows: 6.98 ng/mL, VD3 concentration target: 10.5 ng/mL;

2) And (3) using a high-concentration VD2 and VD3 standard quality control substance to the blood collection tube, and carrying out pretreatment and analysis along with a blood collection tube rack, wherein the VD2 concentration target value is as follows: target VD3 concentration of 25.2 ng/mL: 34.4 ng/mL.

Test results: as in table 4.

Table 4 test items: accuracy (relative deviation%)

* Clinically, relative deviations within + -15% are generally required.

According to the accuracy test data, the invention has higher accuracy for testing the fat-soluble vitamins.

Example 5, sensitivity test.

Sample conditions:

taking blank serum into a blood collection tube, adding the blank serum into the blood collection tube by using a VD2 and VD3 standard solution respectively to ensure that the concentration is respectively 3.5 ng/mL and 3.0 ng/mL, and then, enabling the blood collection tube to be connected to a blood collection tube rack, and carrying out pretreatment and analysis along with the blood collection tube rack.

Test results: as in table 5.

Table 5 test items: sensitivity of

* The detection limit represents the lowest concentration value that can be measured.

The sensitivity test data show that the invention has higher sensitivity for testing fat-soluble vitamins.

The test in the embodiment shows that the invention has higher stability, accuracy and sensitivity in the aspect of testing the fat-soluble vitamins, and meanwhile, the invention is special for the detection equipment of the fat-soluble vitamins, thereby having specificity for the detection of the fat-soluble vitamins.

Claims

1. The full-automatic liquid phase mass spectrum detection system special for the fat-soluble vitamins is characterized by comprising a sample pretreatment module, a sample injection module, a column temperature box module, a mass spectrum module, a pump module, a solvent bottle module and a computer module;

the mass spectrum module is arranged at the lower part of the mass spectrum detection system, substances separated by the column temperature box module enter the mass spectrum module through a pipeline, the mass spectrum module transmits acquired signals to the computer module, and the computer module feeds back detection results according to the acquired data;

the sample pretreatment module comprises a pretreatment chamber, wherein an access blood sampling pipe rack module is arranged in the pretreatment chamber, is configured to be used for placing a blood sampling pipe sample to be treated and analyzed and a blood sampling pipe sample which is already treated, and is configured to be capable of adding a new blood sampling pipe sample and taking out the blood sampling pipe sample which is already treated at any time;

the pretreatment chamber is also internally provided with a centrifugal module, a pipetting gun module, a mixing module, a deep pore plate module, a solvent bottle module, a tip head and a reagent tank module, wherein the centrifugal module is arranged near the blood collection tube rack module and is configured to be used for centrifuging a blood collection tube and the deep pore plate, separating serum and plasma in a sample and separating precipitated protein and the sample; the pipette gun module is configured to draw serum and protein precipitants, internal standard reagents from the centrifugation module and tip head and reagent tank module respectively, and place the reagents into the deep well plate, the deep well plate module is configured to move the deep well plate to a sample injection position and a mixing position, the mixing module is configured to mix the serum and protein precipitants, internal standard reagents in the deep well plate, the solvent bottle module is configured to store mobile phase reagents and sample injection needle cleaning reagents, and the tip head and reagent tank module is configured to store tip head, precipitants, internal standard reagents and waste tanks required for pipetting; the sample pretreatment module further comprises a mechanical arm module, wherein the mechanical arm module is configured to move the blood sampling pipe rack and/or the deep hole plate, so that the blood sampling pipe rack and/or the deep hole plate can move among the blood sampling pipe rack module, the centrifugal module and the hole plate module, and the code scanning process of the blood sampling pipe is completed; the pretreatment chamber is also internally provided with a gun head and a cleaning pool module, and the gun head and the cleaning pool module are arranged near the pipetting gun module and are configured to be used for placing and cleaning the gun head for sucking the serum sample;

A damping foot pad is arranged below the pretreatment chamber and is configured to reduce vibration caused by the centrifugal module;

the centrifugal module comprises a centrifugal machine seat, a motor is arranged in the centrifugal machine seat, the motor is connected with a motor shaft through a coupler, the motor shaft is rotationally connected with a centrifugal carrier,

the centrifugal carrier is provided with a blood collection tube carrier placement position, the blood collection tube carrier can be placed on a blood collection tube carrier placement area, the centrifugal carrier comprises at least two symmetrical blood collection tube carrier placement position areas, the blood collection tube carrier is configured to be used for placing a plurality of blood collection tubes, and each blood collection tube carrier is internally provided with two leveling reagent tubes which are symmetrically distributed;

the centrifugal carrier comprises a middle fixing plate and side fixing plates, wherein the motor is connected with the middle fixing plate in a rotating mode, the side fixing plates are U-shaped, two side portions of the U-shaped are arranged on two sides of a carrier placing position of a blood collection tube and are provided with mounting grooves and sliding rails, elastic elements are further arranged on the middle fixing plate corresponding to each side fixing plate respectively, the side fixing plates and the elastic elements are connected through carrier fixing plate connecting pieces, steps with different heights are arranged at the bottom positions of the U-shaped, carrier fixing plates and limiting fixing plates are arranged on the side fixing plates, the carrier fixing plates can be driven to move along the sliding rails through the limiting fixing plates, and when the carrier fixing plates move, the carrier fixing plates can be adjusted to stretch out and draw back and/or lock states by steps with different heights.

2. The full-automatic liquid-phase mass spectrum detection system special for fat-soluble vitamins according to claim 1, wherein the mass spectrum module comprises a mass spectrum shell, universal casters are arranged at the bottom of the mass spectrum shell, a vacuum module is arranged in the mass spectrum shell, the vacuum module comprises a vacuum cavity, one end of the vacuum cavity is arranged on one surface of the mass spectrum shell, an ion channel interface plate extending out of the mass spectrum shell is arranged at the end of the vacuum cavity, the ion channel interface plate is configured to be connected with an ion source module, a molecular pump is arranged on one side of the vacuum cavity, the molecular pump is communicated with an external space through a vacuum tube and is configured to be used for vacuumizing, and a triple quadrupole rod module is arranged in the vacuum cavity;

The triple quadrupole rod module comprises a quadrupole rod support, wherein a second quadrupole rod is fixedly connected to the middle of the quadrupole rod support, two ends of the second quadrupole rod are respectively connected with a first quadrupole rod and a third quadrupole rod, the first quadrupole rod is configured to be used for screening and focusing of parent ions, the second quadrupole rod is an ion reaction tank and is configured to break the parent ions into child ions, the third quadrupole rod is configured to be used for screening and focusing of the child ions, one side of the quadrupole rod support is provided with a quadrupole rod fixing plate, and a detector is arranged on the quadrupole rod fixing plate and is used for counting the number of the child ions and converting the number of the child ions into electric signals;

3. The fully automated fat-soluble vitamin specific liquid phase mass spectrometry detection system of claim 1, wherein the pipette gun module comprises: an X-axis module, a Y-axis module and a pipette; the top of the pipette gun is slidably connected to the X-axis module, the X-axis module is slidably connected to the Y-axis module, and the X-axis module and the Y-axis module are configured to enable the pipette gun to move in two dimensions;

the space adjusting assembly comprises space adjusting supporting plates, the space adjusting supporting plates are arranged in pairs, a plurality of screw rods which respectively correspond to the movable pipetting guns are arranged between the space adjusting supporting plates, space adjusting sliding blocks are arranged on each screw rod, the space adjusting sliding blocks are respectively connected with the movable pipetting guns through sliding block connecting pieces, space adjusting motors are arranged on the space adjusting supporting plates, the space adjusting motors are connected with the screw rods through gear sets, each gear set comprises a plurality of gears with different double speeds, the gear speed in the middle is low, the gear speed in the outer side is gradually increased, and each screw rod is connected with the gear through a coupling;

4. The full-automatic fat-soluble vitamin-specific liquid phase mass spectrometry detection system according to claim 3, wherein the pipette comprises a pipette fixture block configured to position a lateral position of the pipette assembly, wherein a Z-axis motor and a Z-axis slide rail set are arranged on the pipette fixture block, and the Z-axis motor and the Z-axis slide rail set are configured to drive the pipette to move up and down;

the pipetting gun further comprises a pipetting motor, the pipetting motor is connected with a pipetting piston through a pipetting piston rod, the pipetting piston is configured to move in a piston cavity to complete pipetting action, the pipetting piston is connected with a pipetting head, a head removing piece is arranged between the pipetting piston and the pipetting head, one side of the head removing piece is fixedly connected with a head removing auxiliary mechanism, and the head removing auxiliary mechanism comprises a metal fixing piece, a spring, an electromagnet and a spring fixing piece; the liquid suction head comprises a rubber sealing ring, a rubber fixing piece and a metal elastic piece.

5. The fully automated fat soluble vitamin specific liquid phase mass spectrometry detection system of claim 1, wherein the pump module comprises: the two mobile phase switching valves are communicated with the degasser, two pipelines of the degasser are respectively connected with two mobile phase pumps, the two mobile phase pumps are connected to the primary mixer, one outlet of the primary mixer is connected to the waste liquid port, and the other outlet of the primary mixer is connected with the mixer.

6. The full-automatic liquid-phase mass spectrum detection system special for fat-soluble vitamins according to claim 1, wherein the sample injection module is communicated with the pump module, the pump module comprises a three-way switching valve, the three-way switching valve is respectively connected with a strong washing liquid inlet, a weak washing liquid inlet and an output end, the three-way switching valve is connected with the three-way switching valve through a switching motor and a piston is used for selecting a communicated valve port, the piston is configured to suck washing liquid through a coupling and the sample injection liquid suction motor and then output the washing liquid from the output end, the output end is connected with a six-way valve module, and the six-way valve module is connected with a liquid suction needle module;

the six-way valve module comprises a six-way valve body, wherein a mobile phase interface, a chromatographic column interface, a liquid suction needle interface and a sample injection ring are arranged on the six-way valve body, and are connected with an output end pipeline;

the liquid suction needle module comprises a liquid suction needle motor seat, the liquid suction needle motor seat is connected with a liquid suction needle connecting sliding block, a liquid suction needle motor is arranged in the liquid suction needle motor seat, the liquid suction needle motor is connected with a screw rod sliding block through a screw rod, and the screw rod sliding block moves in a guide groove and drives the liquid suction needle to move up and down to suck liquid.

7. The full-automatic liquid-phase mass spectrum detection system special for fat-soluble vitamins according to claim 1, wherein the column temperature box module comprises a sample inlet pipe, the sample inlet pipe is led into the column temperature box module, a chromatographic column is connected in the column temperature box module, the chromatographic column is fixedly installed on the temperature control module through a chromatographic column buckle, a ventilation fan is arranged at the rear end of the column temperature box module, a window is arranged at the front end of the column temperature box module, and the periphery of the window is sealed through a sealing ring;

The temperature control module comprises an inner cavity radiator, a rear side radiator, heat preservation cotton, an inner cavity radiator fan, a rear side fan and a semiconductor refrigerating sheet, wherein the semiconductor refrigerating sheet is arranged in a mounting groove of the rear side radiator and the inner cavity radiator, heat conduction silicone grease is coated on two sides of the semiconductor refrigerating sheet, heat preservation cotton is used for preserving heat all around, the rear side radiator and the inner cavity radiator are arranged on a fixing plate, the inner cavity radiator fan, the rear side fan and the semiconductor refrigerating sheet are respectively arranged on the outer sides of the inner cavity radiator and the rear side radiator, and the fan is used for improving the working efficiency of the semiconductor refrigerating sheet.

8. A full-automatic liquid-phase mass spectrometry detection method special for fat-soluble vitamins, which is characterized by adopting the detection system as claimed in any one of claims 1-7 and comprising the following steps:

(1) Sample pretreatment:

(1.1) centrifuging the blood collection tube in the carrier at 3000-8000rpm for 5 minutes;

(1.2) drawing 50-100. Mu.L of serum into a 96-well deep well plate;

(1.4) precisely adding 100-400 mu L of internal standard solution;

(1.5) placing the 96 deep hole plate in a mixer, and swirling for 10 minutes at 600-3000 rpm;

(1.6) centrifuging at 2000-4000rpm for 5-10 minutes;

(3) The sample injection module carries a sample to enter the column temperature box module and finishes the separation of fat-soluble vitamins in the column temperature box module;

(4) The separated fat-soluble vitamins enter a mass spectrum module for analysis, and the analysis result of the quality control is fed back to a computer module.