Disposable Sampling Device for Biological Tissue, Mass Spectrometry Analysis System, and Analysis Method

Abstract

The present invention discloses a disposable sampling device for biological tissues, a mass spectrometry analysis system and an analysis method. The device comprises a capillary extraction electrospray unit and a sampling unit. The capillary extraction electrospray unit includes a first connector and a first capillary, and the first capillary is disposed in a first through hole of the first connector; the sampling unit includes a second connector, a second capillary, a cutting part and a gland. The second connector is used for connecting with the first connector, so that the second capillary penetrates into the first through hole to communicate with the first capillary. The other end of the second connector is a sample carrier plate. The second capillary is disposed in a second through hole of the second connector. The cutting part is on the end face, and the gland is buckled outside the outer end of the sample carrier plate. Through the detachable connection of the first connector and the second connector, the sampling unit can be quickly coupled with the capillary extraction electrospray unit, and the sampling unit can be discarded after mass spectrometry analysis, thereby eliminating the cleaning work of the sampling unit, and having the advantages of high efficiency, rapidity, accuracy and simple operation.

Description

Technical Field

[0001] The present invention relates to the technical field of mass spectrometry analysis, and in particular to a disposable sample introduction device for biological tissues, a mass spectrometry analysis system and an analysis method, which can be used for analyzing and detecting clenbuterol in biological tissues, especially meat tissues. Background Art

[0002] The detection of target substances in biological tissue samples using electrospray ionization combined with mass spectrometry is a commonly used technology in inspection and quarantine.

[0003] At present, the detection methods for target substances such as clenbuterol in biological tissue samples are mainly divided into two categories: instrument detection methods and immunological analysis methods. The most commonly used instrument detection methods are gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS), which mainly use the principle of chromatographic separation to separate target substances such as clenbuterol from complex matrices and detect their content through a mass spectrometer detector. However, this technology requires sample crushing, sampling, extraction, separation and other processes before the sample to be tested is put on the machine for testing, which makes the sample pretreatment process more cumbersome, and after the sample is tested, the sample introduction device and pipeline need to be cleaned to eliminate the memory effect. Therefore, a sample introduction device for biological tissue mass spectrometry analysis that does not require cleaning and is easy to operate is needed, which is of great significance for convenient mass spectrometry analysis of biological tissues. Summary of the invention

[0004] Based on the above-mentioned defects, one object of the present invention is to provide a disposable sampling device for mass spectrometry analysis of biological tissues which does not require cleaning and is easy to operate. Another object of the present invention is to provide a mass spectrometry analysis system including the above-mentioned disposable sampling device. Another object of the present invention is to provide a mass spectrometry analysis method for target substances in biological tissues, such as clenbuterol in meat tissues, using the above-mentioned mass spectrometry analysis system.

[0005] The above object of the present invention is achieved through the following technical solutions:

[0006] A disposable sample injection device for biological tissue mass spectrometry analysis comprises a capillary extraction electrospray unit and a sample injection unit, wherein:

[0007] The capillary extraction electrospray unit comprises a first joint and a first capillary, wherein the first joint is a columnar body having a first through hole along the axial direction, and a high-voltage power line interface is provided on the side wall of the first joint, wherein the high-voltage power line interface is used to penetrate the high-voltage power line to apply ion source high voltage electricity to the extraction liquid in the first capillary; the first capillary is arranged in the first through hole;

[0008] The sample injection unit includes a second connector, a second capillary, a cutting part, and a gland. The second connector is a columnar body having a second through hole along the axis, and one end of the second connector is used for detachably connecting to the first connector, so that the second capillary penetrates into the first through hole to communicate with the first capillary, and at the other end of the second connector is a sample carrier plate along the radial direction; the second capillary is arranged in the second through hole, and its end penetrates through the sample carrier plate; the cutting part is fixed circumferentially on the end face of the outer end of the sample carrier plate; the gland is buckled on the outer end of the sample carrier plate to block the cutting part, and there is a third through hole coaxial with the second through hole on the gland for exposing the biological tissue sample clamped between the sample carrier plate and the gland.

[0009] Optionally, the first through hole is in a form where the diameters at both ends are larger than the middle diameter, and the first capillary and the second capillary communicate in the middle of the first through hole.

[0010] Optionally, the first capillary and the second capillary are any one of a quartz capillary and a metal capillary, and the inner diameters of the first capillary and the second capillary are 0.1 - 0.32 mm, the length range of the first capillary is 0.5 - 1 m, and the length range of the second capillary is 1 - 5 cm.

[0011] Optionally, a threaded hole is provided at one end of the first connector away from the second connector, the first capillary passes through and is fixed in a nut, and the nut is screwed into the threaded hole so that the first capillary is fixed in the first through hole.

[0012] Optionally, the first connector is made of a conductive metal, and an insulating sleeve is sleeved on the side wall of the first connector.

[0013] The first capillary and the second capillary are metal capillaries.

[0014] Optionally, the first capillary and the second capillary communicate in a form where there is a distance between their ends.

[0015] Optionally, when the gland is buckled on the second connector, the second capillary passes through the sample carrier plate but does not pass through the third through hole of the gland.

[0016] The present invention also provides a mass spectrometry analysis system for biological tissues, including a mass spectrometer and the disposable sample injection device for mass spectrometry analysis of biological tissues described above.

[0017] The present invention also provides a method for mass spectrometry analysis of target substances in biological tissues, which uses the mass spectrometry analysis system described above to perform the following steps:

[0018] A biological tissue sample is obtained in a biological tissue by a cutting part. A gland is buckled to clamp and fix the biological tissue sample between the sample carrier plate and the gland, and the outlet end of the second capillary of the sample injection unit is made to enter the biological tissue sample, so that an external conical surface is formed on the biological tissue sample.

[0019] The outlet of the second capillary of the sample injection unit is aligned with the sample inlet of the mass spectrometer.

[0020] An extraction liquid is introduced into the first capillary, and at the same time, a high-voltage voltage of the ion source is loaded through the high-voltage power supply line interface.

[0021] The charged extraction liquid enters the biological tissue sample and ionizes it, and an electrospray is formed between the biological tissue sample and the sample inlet of the mass spectrometer, so as to perform mass spectrometry detection.

[0022] Optionally, the biological tissue is meat tissue, and the target substance is clenbuterol; the extraction liquid is a mixed solution formed by methanol and pure water.

[0023] For the disposable sample injection device for mass spectrometry analysis of biological tissues of the present invention, through the detachable connection of the first connector and the second connector, the sample injection unit can be quickly coupled with the capillary extraction electrospray unit, and the cutting part of the sample injection unit is used to realize the immediate use of the biological tissue sample. After mass spectrometry analysis, the sample injection unit can be discarded, and a new sample injection unit is replaced for sampling analysis during the next mass spectrometry analysis, thereby eliminating the cleaning work of the sample injection unit, and having the advantages of high efficiency, rapidity, accuracy, and simple operation. Description of the Drawings

[0024] By describing its embodiments in combination with the following drawings, the above features and technical advantages of the present invention will become clearer and easier to understand.

[0025] Figure 1 is a schematic exploded view of the structure of the sample injection device according to an embodiment of the present invention;

[0026] Figure 2 is an axial sectional view of the first connector according to an embodiment of the present invention;

[0027] Figure 3 is a schematic structural diagram of the mass spectrometry analysis system according to an embodiment of the present invention;

[0028] Figure 4 is a mass spectrometry analysis spectrum of pork tissue in Application Example 1;

[0029] Figure 5 is a mass spectrometry analysis spectrum of pork tissue in Application Example 2. Detailed Embodiments

[0030] Embodiments of the present invention will be described below with reference to the accompanying drawings. Those of ordinary skill in the art can recognize that the described embodiments can be modified in various different ways or combinations thereof without departing from the spirit and scope of the present invention. Therefore, the drawings and the description are illustrative in nature and are not used to limit the scope of protection of the claims. In addition, in this specification, the drawings are not drawn to scale, and the same reference numerals represent the same parts.

[0031] Figure 1 is a schematic exploded view showing the structure of a disposable sampling device for mass spectrometry analysis of biological tissues according to an embodiment of the present invention. The sampling device includes a capillary extraction electrospray unit 10 and a sampling unit 20. Among them, the capillary extraction electrospray unit 10 includes a first connector 12, a first capillary 11, and a high-voltage power supply line interface 13, and the sampling unit 20 includes a second connector 27 and a second capillary 21.

[0032] Specifically, as Figure 2 shown, the first connector 12 is a columnar body having a first through hole 121 along the axis. The first capillary 11 is disposed in the first through hole 121, and a high-voltage power supply line interface is provided in the radial direction of the first connector 12. Optionally, the high-voltage power supply line interface is a threaded interface to facilitate the installation and disassembly of the high-voltage power line. The high-voltage power supply line interface is used to penetrate the high-voltage power line 122 to apply the ion source high voltage to the extraction liquid in the first capillary 11. The high-voltage power line can be directly connected to the high-voltage power line on the mass spectrometer to provide the ion source high voltage. The extraction liquid can be injected into the inlet of the first capillary through an extraction liquid injector such as a syringe.

[0033] The second connector 27 of the sampling unit 20 is also a columnar body having a second through hole 271 along the axis. The second capillary 21 is fixed in the second through hole 271. In this embodiment, the first through hole 121 and the second through hole 271 are both provided along the axis of the columnar body. One end of the second connector 27 is used for detachably connecting to the first connector 12. The detachable connection can be in the form of a snap connection, a threaded connection, an adhesive connection, etc. Such that the second capillary 21 penetrates into the first through hole 121 to communicate with the first capillary 11. It should be noted that the communication between the first capillary 11 and the second capillary 21 means that the extraction liquid can be transmitted between the first capillary 11 and the second capillary 21. The first capillary 11 and the second capillary 21 can be in end-to-end contact, or end-to-end insertion, or there can be a certain distance between the ends. All can achieve the purpose of transmitting the extraction liquid. And preferably, there is a certain distance between the ends.

[0034] At the other end of the second joint 27 is a radially extending sample carrier plate 23 for carrying and fixing a biological tissue sample. The end of the second capillary 21 passes through the sample carrier plate 23 so that the outlet end of the second capillary of the sample introduction unit can enter the biological tissue sample, thereby forming an external conical surface on the biological tissue sample.

[0035] By introducing an extraction solution into the first capillary 11 and applying a high voltage to the first joint via the high-voltage power line 122, the extraction solution enters the second capillary 21 through the first capillary 11. The high voltage is applied to the extraction solution, causing the charged extraction solution at the other end of the second capillary 21 in the second joint to extract and ionize the biological tissue when it enters the biological tissue, thereby forming an electrospray between the external conical surface of the biological tissue sample and the inlet of the mass spectrometer. The inlet 30 of the mass spectrometer is used to receive the electrospray formed by the extraction solution mixed with the target substance, thus realizing rapid mass spectrometry detection.

[0036] Furthermore, the sample introduction unit 20 further includes a cutting part 22 and a gland 24. The cutting part 22 can be annular and is fixed circumferentially on the end face of the outer end of the sample carrier plate. It can be integrally formed with the second joint and the sample carrier plate. The cutting part 22 can have a cutting edge, and the cutting edge faces axially outward. The distance between the cutting edge and the sample carrier plate is 3 mm. The cutting part can easily cut out the biological tissue sample. The gland 24 includes a flat plate 241 and a receiving groove 242 in the circumferential direction of the flat plate. The gland 24 can be snapped onto the outer end of the sample carrier plate 23 so that the cutting part is received in the receiving groove 242 to shield the cutting part 22. And there is a third through-hole 243 on the gland 24 that is coaxial with the second through-hole 271 for exposing the biological tissue sample clamped between the sample carrier plate 23 and the gland 24. And, preferably, when the gland is snapped onto the second joint, the second capillary passes through the sample carrier plate but does not pass through the third through-hole 243 of the gland. The second joint, the sample carrier plate, the cutting part, and the gland can be integrally formed and are preferably made of plastic.

[0037] Optionally, the first through-hole 121 has a form where the diameters at both ends are larger than the middle diameter, and the first capillary 11 and the second capillary 21 communicate in the middle of the first through-hole 121.

[0038] Optionally, the first capillary 11 and the second capillary 21 can be either quartz capillaries or metal capillaries. And an insulating layer can be sleeved at the exposed part of the metal capillary. For example, the first joint 12 is made of conductive metal, and an insulating sleeve is sleeved on the side wall of the first joint. The first capillary 11 and the second capillary 21 can be metal capillaries, or both the first capillary 11 and the second capillary 21 can be quartz capillaries, or one can be a quartz capillary and the other can be a metal capillary. If both are quartz capillaries, the ends of the two capillaries should be spaced apart by a certain distance so that the high-voltage electricity of the ion source can be loaded onto the extraction solution. If they are metal capillaries, it doesn't matter whether the ends are spaced or not, and the high-voltage electricity of the ion source can be loaded onto the extraction solution.

[0039] Optionally, a threaded hole is provided at one end of the first joint away from the second joint. The first capillary passes through and is fixed in a nut, and the nut is screwed into the threaded hole so that the first capillary penetrates into the first through hole.

[0040] Optionally, the outer diameters of the first capillary and the second capillary are 0.2 - 0.5 mm; the inner diameters are 0.1 - 0.32 mm. And the length range of the first capillary is 0.5 - 1 m, for example, 0.6 - 0.9 m, and the length range of the second capillary is 1 - 5 cm.

[0041] According to another aspect of the present invention, the present invention provides a mass spectrometry analysis system for biological tissues, including the above-mentioned disposable sampling device for biological tissue mass spectrometry and a mass spectrometer.

[0042] Furthermore, the mass spectrometry analysis system further includes a three-dimensional adjustment frame (not shown). There is no limitation on the three-dimensional adjustment frame, and it can be commercially available. For example, it can be purchased from Beijing Lianying Precision Machinery Technology Co., Ltd. Through the three-dimensional adjustment frame, the position of the sampling device can be adjusted in three directions, namely, front and back, left and right, and up and down, so that during mass spectrometry analysis, the outlet of the second capillary enters the surface of the biological tissue, and this outlet is accurately aligned with the inlet of the mass spectrometer, effectively improving the detection efficiency of the mass spectrometry and the accuracy of the analysis method.

[0043] According to yet another aspect of the present invention, the present invention further provides a method for mass spectrometry detection of target substances in biological tissues, which uses the above-mentioned mass spectrometry analysis system to perform the following steps:

[0044] Obtain a biological tissue sample in the biological tissue through the cutting part, clamp and fix the biological tissue sample between the sample carrier plate 23 and the pressing cover 24 by buckling the pressing cover, and make the outlet end of the second capillary of the sampling unit enter the biological tissue sample, so that the biological tissue sample forms an external conical surface;

[0045] Align the outlet of the second capillary of the sample introduction unit with the inlet of the mass spectrometer;

[0046] Introduce the extraction solution into the first capillary, and at the same time apply the high-voltage of the ion source through the high-voltage power supply interface;

[0047] Under the action of high voltage, form an electrospray between the outer conical surface of the biological tissue sample and the inlet of the mass spectrometer with the extraction solution containing the target substance in the biological tissue sample, and perform mass spectrometry detection.

[0048] Optionally, the extraction solution is a mixed solution formed by methanol and pure water. Preferably, the volume ratio of methanol to pure water is 50:50. The injection volume of the extraction solution can be 50 - 1005 μL.

[0049] Application Example 1

[0050] Detect the content of salbutamol (C13H21NO3) in pork tissue

[0051] Use the mass spectrometry analysis system of the present invention to detect the content of salbutamol in pork tissue.

[0052] Materials, Instruments, Conditions

[0053] Pork tissue: Take out the pork from the 4 °C refrigerator, and use a disposable sampling device to sample the pork tissue sample;

[0054] Extraction solution: Prepare a mixed solution with a volume ratio of 50:50 of methanol and pure water;

[0055] Thermo LTQ XL linear ion trap mass spectrometer;

[0056] Ion source voltage: 4.5 kV;

[0057] Ion transfer tube temperature: 200 °C;

[0058] Capillary voltage: 10 V;

[0059] Lens voltage: 100 V;

[0060] Sheath gas flow rate: 10 Arb;

[0061] Positive ion detection mode;

[0062] Mass scanning range: m / z 50 - 2000;

[0063] Flow rate: 5 μL / min.

[0064] Adopt Figure 3 The mass spectrometry analysis system shown performs mass spectrometry analysis on the sliced pork tissue, including the following steps:

[0065] Take the pork tissue out of the 4°C environment, obtain a biological tissue sample from the biological tissue through the cutting part, clamp and fix the biological tissue sample between the sample carrier plate and the gland cover, and make the outlet end of the second capillary of the sampling unit enter the biological tissue sample, so that the biological tissue sample forms an external conical surface; align the outlet of the second capillary of the sampling unit with the inlet of the mass spectrometer; introduce the extraction solution into the first capillary, and at the same time load the high-voltage of the ion source through the high-voltage power supply interface; the charged extraction solution enters the biological tissue and ionizes it, forming an electrospray between the biological tissue and the inlet of the mass spectrometer, so as to perform mass spectrometry detection and obtain the mass spectrum of this site, as Figure 4 shown. It can be found from this mass spectrum that the present method can detect substances such as signals in pork tissue, and these substances can be used as potential biomarker molecules for judging whether clenbuterol is contained in pork tissue to a certain extent.

[0066] Application Example 2: Detection of the content of clenbuterol hydrochloride (C 12 H 18 C 12 N 2 O) in pork tissue

[0067] Materials, Instruments, Conditions

[0068] Pork tissue: Take the pork out of the 4 °C refrigerator and sample the pork tissue sample using a disposable sampling device;

[0069] Extraction solution: Prepare a mixed solution with a volume ratio of 50:50 of methanol and pure water;

[0070] Thermo LTQ XL linear ion trap mass spectrometer;

[0071] Ion source voltage: 4.5 kV;

[0072] Ion transfer tube temperature: 200 °C;

[0073] Capillary voltage: 10 V;

[0074] Lens voltage: 100 V;

[0075] Sheath gas flow rate: 10 Arb;

[0076] Positive ion detection mode;

[0077] Mass scanning range: m / z 50 - 2000;

[0078] Flow rate: 5 μL / min.

[0079] Adopt Figure 3 the mass spectrometry analysis system shown to perform mass spectrometry analysis on the sliced pork tissue, including the following steps:

[0080] Take the pork tissue out of the 4°C environment, obtain a biological tissue sample from the biological tissue through the cutting part, clamp and fix the biological tissue sample between the sample carrier plate and the gland cover, and make the outlet end of the second capillary of the sample injection unit enter the biological tissue sample, so that the biological tissue sample forms an external conical surface; align the outlet of the second capillary for sample injection with the inlet of the mass spectrometer; introduce the extraction solution into the first capillary, and at the same time load the high-voltage of the ion source through the high-voltage power supply line interface; the charged extraction solution enters the biological tissue and ionizes it, forming an electrospray between the biological tissue and the inlet of the mass spectrometer, so as to perform mass spectrometry detection to obtain the mass spectrum of this site, as Figure 5 shown. It can be found from this mass spectrum that the present method can detect substances such as signals in the pork tissue, and such substances can be used to a certain extent as potential biomarker molecules for judging whether clenbuterol is contained in the pork tissue.

[0081] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention can have various changes and modifications. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A disposable sampling device for mass spectrometry analysis of biological tissues, characterized in that, it includes a capillary extraction electrospray unit and a sampling unit, wherein, the capillary extraction electrospray unit includes a first connector and a first capillary. The first connector is a columnar body having a first through hole along the axis, and a high-voltage power supply interface is provided on the side wall of the first connector. The high-voltage power supply interface is used to penetrate a high-voltage power supply wire to apply ion source high-voltage electricity to the extraction liquid in the first capillary; the first capillary is arranged in the first through hole; the sampling unit includes a second connector, a second capillary, a cutting part and a gland. The second connector is a columnar body having a second through hole along the axis, and one end of the second connector is used for detachable connection with the first connector, so that the second capillary penetrates into the first through hole to communicate with the first capillary, and at the other end of the second connector is a sample carrier plate along the radial direction; the second capillary is arranged in the second through hole, and its end passes through the sample carrier plate; the cutting part is fixed on the outer end surface of the outer end of the sample carrier plate along the circumferential direction; the gland is buckled on the outer end of the sample carrier plate to block the cutting part, and a third through hole coaxial with the second through hole is provided on the gland for exposing the biological tissue sample clamped between the sample carrier plate and the gland.

2. The disposable sampling device for mass spectrometry analysis of biological tissues according to claim 1, characterized in that, the first through hole is in a form where the diameters at both ends are larger than the middle diameter, and the first capillary and the second capillary communicate in the middle of the first through hole.

3. The disposable sampling device for mass spectrometry analysis of biological tissues according to claim 1, characterized in that, the first capillary and the second capillary are any one of quartz capillaries and metal capillaries, and the inner diameters of the first capillary and the second capillary are 0.1 - 0.32 mm, the length range of the first capillary is 0.5 - 1 m, and the length range of the second capillary is 1 - 5 cm.

4. The disposable sampling device for mass spectrometry analysis of biological tissues according to claim 1, characterized in that, a threaded hole is provided at one end of the first connector away from the second connector, the first capillary passes through and is fixed in a nut, and the nut is screwed into the threaded hole so that the first capillary is fixed in the first through hole.

5. The disposable sampling device for mass spectrometry analysis of biological tissues according to claim 1, characterized in that, the first connector is made of conductive metal, and an insulating sleeve is sleeved on the side wall of the first connector, the first capillary and the second capillary are metal capillaries.

6. The disposable sampling device for mass spectrometry analysis of biological tissues according to claim 1, characterized in that, the first capillary and the second capillary communicate in a form where there is a distance between their ends.

7. The disposable sampling device for mass spectrometry analysis of biological tissues according to claim 2, characterized in that, when the gland is buckled on the second connector, the second capillary passes through the sample carrier plate but does not pass through the third through hole of the gland.

8. A mass spectrometry analysis system for biological tissues, characterized in that, it includes a mass spectrometer and a disposable sampling device for mass spectrometry analysis of biological tissues according to any one of claims 1 to 7.

9. A method for mass spectrometry analysis of target substances in biological tissues, characterized in that, using the mass spectrometry analysis system according to claim 8 to perform the following steps: obtaining a biological tissue sample in the biological tissue through the cutting part, clamping and fixing the biological tissue sample between the sample carrier plate and the gland by buckling the gland, and making the outlet end of the second capillary of the sampling unit enter the biological tissue sample, so that the biological tissue sample forms an external conical surface; aligning the outlet of the second capillary of the sampling unit with the inlet of the mass spectrometer; introducing an extraction solution into the first capillary, and simultaneously loading the high voltage of the ion source through the high voltage power supply line interface; the charged extraction solution enters the biological tissue sample and ionizes it, forming an electrospray between the biological tissue sample and the inlet of the mass spectrometer, thereby performing mass spectrometry detection.

10. The method for mass spectrometry analysis of target substances in biological tissues according to claim 9, characterized in that, the biological tissue is meat tissue, the target substance is clenbuterol; the extraction solution is a mixed solution formed by methanol and pure water.

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