Handheld solid sample in-situ extraction device

Abstract

The utility model discloses a handheld solid sample in-situ extraction device which is an off-line in-situ extraction device and comprises a handheld extraction system A, a liquid supply system for providing a solvent for the handheld extraction system A and a negative pressure system for driving an extracting solution to flow in the handheld extraction system A, or the device is an online in-situ extraction device and comprises a handheld extraction system B and a liquid supply system for providing a solvent for the handheld extraction system B. According to the handheld solid sample in-situ extraction device provided by the utility model, chemical information of a target sample can be rapidly obtained in an off-line extraction or on-line analysis mode by matching with various mass spectrometry technologies, so that the flexibility and the application mode of the in-situ extraction detection device are enhanced, and pollution caused by approaching to mass spectrometry is reduced.

Description

Technical Field

[0001] This utility model belongs to the field of machinery and relates to a sample chemical composition extraction device, specifically a handheld solid sample in-situ extraction device that can be applied to mass spectrometry detection. Background Technology

[0002] Mass spectrometry (MS) boasts excellent sensitivity, specificity, analytical speed, and molecular coverage, and is widely used in food and drug safety, environmental monitoring, forensic identification, and clinical analysis. While mass spectrometry offers powerful analytical capabilities, traditional mass spectrometry analysis typically requires coupling with instruments such as gas chromatography (GC) or liquid chromatography (LC), resulting in lengthy analysis times and demanding sample pretreatment requirements, thus failing to meet the needs of in-situ analysis. In-situ mass spectrometry avoids complex sample processing, enabling rapid and efficient sample collection and detection.

[0003] Ambient ionization techniques can analyze samples with complex chemical compositions under natural conditions. They require very few sample pretreatment steps, offering significant advantages such as simplicity, speed, and versatility. Ambient ionization techniques also combine direct sampling and ionization of tiny regions on solid samples, thus enabling controlled in-situ sampling and ionization. Examples include desorption electrospray ionization (DESI), liquid microjunction-surface sampling (LMJ-SSP), and liquid extraction surface analysis (LESA). However, the cleanliness requirements of the sampling end and the mass spectrometer end in atmospheric pressure ionization technology are drastically different: due to the complexity of the samples being tested, the sampling end faces a relatively harsh working environment, while the mass spectrometer end is a vacuum environment with very high cleanliness requirements. The above-mentioned atmospheric pressure ionization technology usually places the sampling end and the mass spectrometer end in close proximity, which can easily cause mass spectrometry contamination, errors, and instrument wear and tear when analyzing complex samples. On the other hand, the close coupling between the sampling end and the mass spectrometer end reduces the portability of the sampling end, and the resulting steric hindrance further limits the size and distance of the samples. This lack of flexibility limits its application in clinical sampling, field sampling, and other fields.

[0004] This invention is proposed to achieve in-situ, portable extraction of solid samples and reduce sample contamination of the mass spectrometer. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of the prior art and provide a handheld solid sample in-situ extraction device and its application in mass spectrometry detection.

[0006] The above-mentioned objective of this utility model is achieved through the following technical solution:

[0007] A handheld solid sample in-situ extraction device is disclosed. This device is an offline in-situ extraction device, wherein the offline extraction refers to the extraction liquid being tested and analyzed after extraction is completed. The offline in-situ extraction device includes a handheld extraction system A, a liquid supply system for providing solvent to the handheld extraction system A, and a negative pressure system for driving the extraction liquid to flow in the handheld extraction system A.

[0008] The handheld extraction system A includes: a handheld casing A1, a sampling pipe A2 penetrating the handheld casing A1, two tee fittings A3 and A4 arranged along the sampling pipe A2 and sleeved outside the sampling pipe A2 and fixed inside the handheld casing A1, and a collection pipe A5 for collecting liquid in the sampling pipe A2; one end of the sampling pipe A2 is a sampling end, and the other end is a collection end, both of which extend outside the handheld casing A1; the tee fittings A3 and A4 are respectively located near the sampling end and near the collection end of the sampling pipe A2; the collection pipe A5 is located at the collection end of the sampling pipe A2;

[0009] The main pipe of the three-way fitting A3 is fitted onto the sampling pipe A2 near the sampling end. The branch pipe is equipped with a suitable plug A33. The plug A33 is tightly fitted with a pipe A8 for transporting the solvent in the liquid supply system A9 to the three-way fitting A3. The main pipe is equipped with a suitable plug A32 near the collection end. The plug A32 is tightly fitted with a fixing sleeve for fixing the sampling pipe A2. The fixing sleeve is tightly fitted onto the outside of the sampling pipe A2. The main pipe is equipped with a suitable plug A31 near the sampling end. The plug A31 is tightly fitted with a nested tube A6 that is fitted onto the outside of the sampling pipe A2. The inner wall of the nested tube A6 and the outer wall of the sampling pipe A2 are separated to allow the solvent transported to the three-way fitting A3 to flow out to the sampling end of the sampling pipe A2.

[0010] The main pipe of the tee fitting A4 is fitted onto the sampling pipe A2 near the collection end, and the branch pipe is equipped with a suitable plug A43. A pipe A10 for drawing gas from the tee fitting A4 to the negative pressure system A11 is tightly fitted onto the plug A43. A suitable plug A41 is fitted onto the main pipe near the sampling end, and a fixing sleeve for fixing the sampling pipe A2 is tightly fitted onto the outside of the sampling pipe A2. A suitable plug A42 is fitted onto the main pipe near the collection end, and a fixing sleeve for fixing the sampling pipe A2 is tightly fitted onto the branch pipe A43. The sampling pipe A2 is surrounded by a nested tube A7, which extends out of the handheld housing A1 and is tightly inserted into the collection tube A5. The inner wall of the nested tube A7 and the outer wall of the sampling pipe A2 are separated to allow the negative pressure system to draw the gas in the collection tube A5 through the three-way fitting A4 and create a negative pressure environment in the collection tube A5. The end of the sampling pipe A2 and the inner wall of the collection tube A5 are connected in a wall-fitting manner to facilitate the flow of the extract in the sampling pipe A2 into the collection tube A5 under the drive of the gas pressure difference.

[0011] Preferably, the liquid supply system A9 includes a syringe and an injection pump that controls the injection of solvent by the syringe.

[0012] Preferably, sampling pipe A2 is a quartz capillary tube.

[0013] Preferably, the nested tubes A6 and A7 are ETFE tubes.

[0014] Preferably, the fixing sleeve is a PEEK pipe.

[0015] A handheld solid sample in-situ extraction device is disclosed. This device is an online in-situ extraction device, wherein the online extraction refers to the direct delivery of the extract to a mass spectrometry system for analysis and detection while extracting. The online in-situ extraction device includes a handheld extraction system B and a liquid supply system that provides solvent to the handheld extraction system B.

[0016] The handheld extraction system B includes: a handheld shell B1, a sampling pipe B2 penetrating inside the handheld shell B1, a three-way fitting B3 and a two-way fitting B4 arranged along the sampling pipe B2, sleeved outside the sampling pipe B2, and fixed inside the handheld shell B1; one end of the sampling pipe B2 is a sampling end, and the other end is a mass spectrometry connection end, both of which extend outside the handheld shell B1; the three-way fitting B3 is located near the sampling end of the sampling pipe B2, and the two-way fitting B4 is located near the mass spectrometry connection end of the sampling pipe B2;

[0017] The main pipe of the three-way fitting B3 is fitted onto the sampling pipe B2 near the sampling end, and the branch pipe is equipped with a suitable plug B33. The plug B33 is tightly fitted with a pipe B6 for transporting the solvent in the liquid supply system B7 to the three-way fitting B3. The main pipe is equipped with a suitable plug B32 near the mass spectrometer connection end. The plug B32 is tightly fitted with a fixing sleeve for fixing the sampling pipe B2. The fixing sleeve is tightly fitted onto the outside of the sampling pipe B2. The main pipe is equipped with a suitable plug B31 near the sampling end. The plug B31 is tightly fitted with a nested tube B5 that is fitted onto the outside of the sampling pipe B2. The inner wall of the nested tube B5 and the outer wall of the sampling pipe B2 leave a cavity for the solvent transported to the three-way fitting B3 to flow out to the sampling end of the sampling pipe B2.

[0018] The two-way fitting B4 is sleeved on the sampling pipe B2 near the mass spectrometer connection end. The sampling end and the mass spectrometer connection end of the two-way fitting B4 are respectively provided with matching plugs B41 and B42. The plugs B41 and B42 are respectively tightly provided with fixing sleeves for fixing the sampling pipe B2. The fixing sleeves are tightly sleeved on the outside of the sampling pipe B2 inside the mass spectrometer system B8, driven by the pressure difference formed by the negative pressure in the mass spectrometer system.

[0019] Preferably, the liquid supply system includes a syringe and an injection pump that controls the injection of solvent by the syringe.

[0020] Preferably, sampling pipe B2 is a quartz capillary tube.

[0021] Preferably, the nested tube B5 is an ETFE tube.

[0022] Preferably, the fixing sleeve is a PEEK pipe.

[0023] Beneficial effects:

[0024] 1. This utility model provides a detection device for in-situ analysis of the surface of solid samples, which is divided into an offline in-situ extraction device and an online in-situ extraction device. The device can obtain the chemical information of the target sample quickly by means of offline extraction or online analysis, combined with a variety of mass spectrometry techniques.

[0025] 2. This utility model utilizes two handheld devices to achieve two different detection modes, enhancing the flexibility and application of the in-situ extraction detection device, and reducing contamination caused by proximity to the mass spectrometer. Attached Figure Description

[0026] Figure 1 , Figure 2 This is a diagram of an offline in-situ extraction device and its structure.

[0027] Figure 3 , Figure 4 This is a diagram of the online in-situ extraction device and its structure.

[0028] Figure 5 The representative spectra A and B of the blank matrix 2,5-dihydroxybenzoic acid are used for the detection of buspirone standard spotted on a glass slide by an offline in-situ extraction device combined with MALDI-TOF MS.

[0029] Figure 6 , Figure 7 An offline in-situ extraction device combined with MALDI-TOF MS was used to detect representative mass spectra in rat brain slices, in which... Figure 7 Image A shows lipid signals in the mass-to-charge ratio range of 750-850. Figure 7 B shows signals of small molecule metabolites with a mass-to-charge ratio in the range of 100-180;

[0030] Figure 8 This document presents representative physical spectra of an offline in-situ extraction device combined with MALDI-TOF MS for detecting various chemical components in multiple traditional Chinese medicine decoction pieces, including extracts of honeysuckle, chrysanthemum, rose, and coptis. A represents the representative physical spectrum of honeysuckle extract; B represents the representative physical spectrum of chrysanthemum extract; C represents the representative physical spectrum of rose extract; and D represents the representative physical spectrum of coptis extract.

[0031] Figure 9 Representative spectral data for detecting safflower and stained chrysanthemum using an offline in-situ extraction device combined with MALDI-TOF MS are shown. A represents the spectral data for detecting hydroxysafflower yellow pigment A standard reference; B represents the spectral data for detecting the safflower standard extract; C represents the spectral data for detecting the extract of analyte 1; and D represents the spectral data for detecting the extract of analyte 2.

[0032] Figure 10 , Figure 11 This describes the use of an online in-situ extraction device combined with HESI-Q Exactive MS to detect representative mass spectra of rat brain slices. Figure 11 Image A shows lipid signals in the mass-to-charge ratio range of 720-850. Figure 11 B shows signals of small molecule metabolites with a mass-to-charge ratio in the range of 120-200;

[0033] Figure 12 The following are representative mass spectra of safflower and stained chrysanthemum obtained by combining an online in-situ extraction device with HESI-Q Exactive MS: A represents the ion signal of analyte 1 in the mass-to-charge ratio range of 405-430; B represents the ion signal of analyte 1 in the mass-to-charge ratio range of 606-618; C represents the ion signal of analyte 2 in the mass-to-charge ratio range of 405-430; and D represents the ion signal of analyte 2 in the mass-to-charge ratio range of 606-618.

[0034] Figure 13 This document presents representative property spectra of hydroxysaffron yellow A standard sample points on glass slides and hydroxysaffron yellow A from safflower decoction pieces, obtained using an online in-situ extraction device combined with HESI-Q Exactive MS. Spectra A represent the property of hydroxysaffron yellow A standard sample points detected in full scan mode; Spectra B represent the property of hydroxysaffron yellow A standard sample points detected in ion fragmentation mode; Spectra C represent the property of safflower decoction pieces detected in full scan mode; Spectra D represent the property of hydroxysaffron yellow A from safflower decoction pieces detected in ion fragmentation mode; and E represents the hypothesized ion fragmentation mechanism of hydroxysaffron yellow A.

[0035] Wherein: A is the offline handheld extraction system, A1 is the handheld shell, A2 is the sampling pipe, A3 is the tee fitting, A4 is the tee fitting, A5 is the liquid collection pipe, A6 is the nested pipe, A7 is the nested pipe, A8 is the liquid supply pipe, A9 is the liquid supply system, A10 is the negative pressure pipe, A11 is the negative pressure system, and A31, A32, A33, A41, A42, and A43 are plugs; B is the online handheld extraction system, B1 is the handheld shell, B2 is the sampling pipe, B3 is the tee fitting, B4 is the two-way fitting, B5 is the nested pipe, B6 is the liquid supply pipe, B7 is the liquid supply system, B8 is the mass spectrometry system, and B31, B32, B33, B41, and B42 are plugs. Detailed Implementation

[0036] The substantive content of this utility model is described in detail below with reference to the embodiments, but this is not intended to limit the scope of protection of this utility model.

[0037] Example 1: Offline in-situ extraction device

[0038] The method for constructing and using the handheld solid sample in-situ extraction device for offline in-situ extraction provided by this utility model is as follows: assemble the handheld extraction system A, install a liquid supply system to provide solvent for the handheld extraction system A, install a negative pressure system to drive the extraction liquid to flow in the handheld extraction system A, apply the handheld solid sample in-situ extraction device to realize offline in-situ extraction, and perform detection and analysis on the extraction liquid after extraction is completed.

[0039] The specific operation and usage methods are as follows:

[0040] (1) Assemble the handheld extraction system A: such as Figure 1 and Figure 2As shown, the device includes a handheld casing A1, a sampling pipe A2 penetrating the handheld casing A1, two tee fittings A3 and A4 arranged along the sampling pipe A2 and fitted over the sampling pipe A2 and fixed inside the handheld casing A1, and a collection pipe A5 for collecting liquid from the sampling pipe A2. One end of the sampling pipe A2 is the sampling end, and the other end is the collection end, both of which extend outside the handheld casing A1; the tee fittings A3 and A4 are respectively located near the sampling end and near the collection end of the sampling pipe A2; the collection pipe A5 is located at the collection end of the sampling pipe A2. The main pipe of the tee fitting A3 is fitted onto the sampling pipe A2 near the sampling end. The branch pipe is equipped with a suitable plug A33. The plug A33 is tightly fitted with a pipe A8 for conveying the solvent in the liquid supply system A9 to the tee fitting A3. The main pipe is equipped with a suitable plug A32 near the collection end. The plug A32 is tightly fitted with a fixing sleeve for fixing the sampling pipe A2. The fixing sleeve is tightly fitted onto the outside of the sampling pipe A2. The main pipe is equipped with a suitable plug A31 near the sampling end. The plug A31 is tightly fitted with a nested tube A6 that is fitted onto the outside of the sampling pipe A2. There is a cavity between the inner wall of the nested tube A6 and the outer wall of the sampling pipe A2 for the solvent conveyed to the tee fitting A3 to flow out to the sampling end of the sampling pipe A2. The main pipe of the tee fitting A4 is fitted onto the sampling pipe A2 near the collection end. A branch pipe has a suitable plug A43, and the plug A43 is tightly fitted with a pipe A10 for drawing gas from the tee fitting A4 to the negative pressure system A11. The main pipe near the sampling end has a suitable plug A41, and the plug A41 is tightly fitted with a fixing sleeve for securing the sampling pipe A2. This fixing sleeve is tightly fitted onto the outside of the sampling pipe A2. The main pipe near the collection end has a suitable plug A42, and the plug A42 is tightly fitted onto the sampling pipe A11. The sampling pipe A2 is surrounded by a nested tube A7, which extends out of the handheld housing A1 and is tightly inserted into the collection tube A5. The inner wall of the nested tube A7 and the outer wall of the sampling pipe A2 are separated to allow the negative pressure system to draw the gas in the collection tube A5 through the three-way fitting A4 and create a negative pressure environment in the collection tube A5. The end of the sampling pipe A2 and the inner wall of the collection tube A5 are connected in a wall-fitting manner to facilitate the flow of the extract in the sampling pipe A2 into the collection tube A5 under the drive of the gas pressure difference.

[0041] The handheld outer shell A1 is manufactured using computer software modeling combined with 3D printing technology. It consists of two parts: a front part and a back part. The back part is approximately planar with a wall thickness of 2.5mm and features grooves and screw holes for fixing tee fittings A3 and A4. The front part is approximately a hollow semi-cylinder with a wall thickness of 2.5mm. Near the sampling end, it has an approximately conical structure with a hole at the tip for the sampling tube A2 and the nested tube A6 to pass through. The front and back parts are fixed together with screws through the screw holes. An extended section on the left side of the handheld shell accommodates a branch tube for tee fittings A3 and A4. All screw holes are 3mm, and M3 screws and nuts are used for fixing. The sampling tube A2 is a quartz capillary tube with an outer diameter of 362μm, an inner diameter of 150μm, and a length of approximately 160mm.

[0042] During assembly, first, insert tee fittings A3 and A4 into the grooves on the opposite sides and secure them with screws. Pass the sampling pipe A2 through the main pipes of tee fittings A3 and A4. Install a suitable plug A32 at the near-collection end of the main pipe of tee fitting A3. A fixing sleeve for securing the sampling pipe A2 is tightly fitted onto plug A32. This fixing sleeve is made of PEEK tubing with an outer diameter of 1 / 16 inch, an inner diameter of 405 μm, and a length of approximately 25 mm. Since the inner diameter of the fixing sleeve (405 μm) is slightly larger than the outer diameter of the sampling pipe A2 (362 μm), a slight elastic deformation is created in the fixing sleeve by tightening the plug A32, thereby sealing and securing the sampling pipe A2. A suitable plug A31 is located at the near-sampling end of the main pipe of tee fitting A3. A nested tube A6 is tightly fitted onto plug A31 and fitted onto the outside of the sampling pipe A2. Nested tube A6 is an ETFE tube with an outer diameter of 1 / 16 inch, an inner diameter of 1000 μm, and a length of approximately 40 mm. Since the inner diameter of nested tube A6 (1000 μm) is much larger than the outer diameter of sampling pipe A2 (362 μm), a cavity is left between the inner wall of nested tube A6 and the outer wall of sampling pipe A2 to allow solvent transported to the tee fitting A3 to flow out to the sampling end of sampling pipe A2. Taking advantage of the characteristic that the plug A32 only seals and secures sampling pipe A2 when tightened, the relative position of sampling pipe A2 and nested tube A6 can be adjusted when loosened. The sampling tube A2 and the nesting tube A6 can have two positional relationships: When the sampling tube A2 does not extend beyond the nesting tube A6 and is approximately 1-2 mm away from the sampling end of the nesting tube A6, the nesting tube A6 can protect the sampling tube A2 and form a solvent backflow from the nesting tube A6 to the sampling tube A2 within the nesting tube A6. However, if the negative pressure and flow rate are unstable or mismatched, there may be droplet interruption and solvent overflow, resulting in extraction failure or sample contamination. When the sampling tube A2 extends beyond the nesting tube A6 and is approximately 1-2 mm away from the sampling end of the nesting tube A6, only sufficient negative pressure from the negative pressure supply system is needed to allow the solvent to flow out of the nesting tube A6 and contact the analyte. Driven by the pressure difference, the solvent then enters the sampling tube A2, reducing the matching requirements of the liquid flow rate and negative pressure and simplifying the process of adjusting the flow rate and negative pressure. The sampling tube A2 and the nesting tube A6 are passed through the hole at the conical tip of the positive part of the handheld casing A1 near the sampling end, and adjusted so that the nesting tube A6 extends approximately 2-3 mm beyond the hole at the conical tip. Take a PEEK pipe with an outer diameter of 1 / 16 inch, an inner diameter of 500 μm, and a length of about 30 cm as pipe A8. Connect one end of pipe A8 to the liquid supply system A9, and the other end is tightly fixed to the branch pipe of tee fitting A3 through a matching plug A33.

[0043] A suitable plug A41 is installed near the sampling end of the main pipe of the tee fitting A4. A fixing sleeve for securing the sampling pipe A2 is tightly fitted onto the plug A41. The fixing sleeve is made of PEEK tubing with an outer diameter of 1 / 16 inch, an inner diameter of 405 μm, and a length of approximately 25 mm. Since the inner diameter of the fixing sleeve (405 μm) is slightly larger than the outer diameter of the sampling pipe A2 (362 μm), a slight elastic deformation is caused by tightening the plug A41, thereby sealing and securing the sampling pipe A2. A suitable plug A42 is installed near the collection end of the main pipe of the tee fitting A4. A nested tube A7 is tightly fitted onto the plug A42, extending out of the handheld casing A1 and tightly inserted into the collection tube A5. The nested tube A7 is a 1 / 16-inch outer diameter, a 1000μm inner diameter, and approximately 35mm long PEEK tube. Since the 1000μm inner diameter of the nested tube A7 is much larger than the 362μm outer diameter of the sampling pipe A2, a cavity is left between the inner wall of the nested tube A7 and the outer wall of the sampling pipe A2. This cavity allows the negative pressure system to draw gas from the collection pipe A5 through the tee fitting A4, creating a negative pressure environment within the collection pipe A5. The end of the sampling pipe A2 is connected to the inner wall of the collection pipe A5 in a wall-mounted manner to facilitate the flow of the extract from the sampling pipe A2 into the collection pipe A5 under the pressure difference. Utilizing the characteristic that the plug A41 only seals and secures the sampling pipe A2 when tightened, the length of the sampling pipe A2 extending beyond the nested tube A7 is adjusted by approximately 20-25mm by loosening the plug A41. Take an ETFE pipe with an outer diameter of 1 / 16 inch, an inner diameter of 1000 μm, and a length of about 30 cm as pipe A10. One end of pipe A10 is connected to the negative pressure supply system A11, and the other end is tightly fixed to the branch pipe of tee fitting A4 through a matching plug A43.

[0044] Before sampling, a pre-drilled EP tube with a hole approximately 1.6 mm in diameter is used as the collection tube A5. The nesting tube A7 is tightly inserted into the collection tube A5. Since the hole diameter of approximately 1.6 mm is close to 1 / 16 inch of the outer diameter of the nesting tube A7, the collection tube A5 and the nesting tube A7 can be tightly connected and sealed. The position of the collection tube A5 is adjusted so that the end of the sampling tube A2 forms a wall-to-wall connection with the inner wall of the collection tube A5. This allows the extract in the sampling tube A2 to flow into the collection tube A5 along the inner wall under the pressure difference. Furthermore, the nesting tube A7 extends approximately 15-20 mm into the collection tube A5, thus preventing the extract from entering the negative pressure system. Similarly, the collection tube A5 can be replaced with a vial equipped with a cap and a gasket. When the nesting tube A7 is tightly inserted into the collection tube A5, the gasket of the vial can still maintain a seal after puncture, thus eliminating the need for drilling.

[0045] (2) Install a liquid supply system for supplying solvent to the handheld extraction system A: The liquid supply system A9 includes a syringe and an injection pump that controls the injection of solvent by the syringe, as well as a fixing assembly that connects the syringe to the pipeline A8.

[0046] (3) Install a negative pressure system to drive the flow of extractant in handheld extraction system A: negative pressure system A11 includes a small vacuum pump and a regulating component for controlling the output voltage of the vacuum pump, as well as a fixing component for connecting the small vacuum pump to pipe A10.

[0047] (4) Offline in-situ extraction is achieved using a handheld solid sample in-situ extraction device: First, use a syringe to draw a suitable amount of solvent and fix the syringe on the syringe pump; then push the syringe until the nested tube A6 is filled with solvent and free of air bubbles; then prepare the analyte, and bring the sampling end of the handheld extraction system A close to the analyte until the sampling tube A2 is about to contact the analyte; then turn on the negative pressure system and run the syringe pump to push the syringe to the tip of the probe to form a liquid condensate, and start sampling when the liquid condensate forms; when enough extract has been collected in the collection tube A5, lift the handheld extraction system A away to end the sampling. At this time, the liquid condensate is still forming, and clean solvent flows in and washes the sampling tube A2 to prevent extract residue; after washing is completed, turn off the syringe pump, and then turn off the negative pressure system, and the offline in-situ extraction is completed. In addition, the handheld extraction system A can also achieve negative pressure concentration of extracts of small amounts of low-boiling-point solvents. The specific operation is as follows: after washing is completed, turn off the syringe pump and invert the handheld extraction system A. You can see the extract flowing to the bottom of the tube and gradually evaporating and concentrating under the action of negative pressure.

[0048] (5) Analysis of the extract: After the offline in-situ extraction is completed, the extract can be analyzed. Various analytical instruments can be used to analyze the extract, including liquid chromatography-mass spectrometry (LC-MS) and matrix-assisted laser desorption / ionization mass spectrometry (MALDI-MS). Among these, MALDI-MS has the advantages of high throughput, high sensitivity, wide applicability, and predominance of single-charge molecular ion peaks, making it suitable for the analysis of multi-component samples. Furthermore, MALDI-MS has less stringent sample preparation requirements and is simple to operate. Therefore, this invention uses MALDI-MS to detect the extract.

[0049] Taking matrix-assisted laser desorption / ionization mass spectrometry (MALDI-MS) analysis as an example, it requires spotting the sample onto an indium tin oxide (ITO) coated glass slide and adding a matrix. Matrix components include, for example, aminoacridine (9-AA) as the negative ion detection matrix and 2,5-dihydroxybenzoic acid (DHB) as the positive ion detection matrix. The spotting experiment involves two sample loading methods combined with matrix addition: matrix reconstitution spotting and matrix spraying. In the matrix reconstitution spotting method, 10 μL of prepared matrix solution is used to directly reconstitute the evaporated sample extract. In the matrix spraying method, 10 μL of pure solvent is used to reconstitute the sample extract, and then the matrix is ​​uniformly deposited onto the ITO glass slide using a matrix spraying instrument. Using a commercial MALDI mass spectrometer, the UltrafleXtreme MALDI TOF / TOF MS, the ITO glass slide is placed on the target plate, the target plate is inserted, and the software is opened according to the general operating procedures for a MALDI mass spectrometer to determine the chemical components in the extract. After the aforementioned extract processing, a good signal can be obtained.

[0050] Example 2: Online In-situ Extraction Device

[0051] The method for constructing and using the handheld solid sample in-situ extraction device for online in-situ extraction provided by this utility model is as follows: assemble the handheld extraction system B, install the liquid supply system that provides solvent for the handheld extraction system B, and apply the handheld solid sample in-situ extraction device to realize online in-situ extraction and detection analysis.

[0052] The specific operation and usage methods are as follows:

[0053] (1) Assemble the handheld extraction system B: such as Figure 3 and 4 As shown, it includes a handheld casing B1, a sampling pipe B2 passing through the handheld casing B1, a three-way fitting B3 and a two-way fitting B4 arranged along the sampling pipe B2, sleeved outside the sampling pipe B2 and fixed inside the handheld casing B1; one end of the sampling pipe B2 is the sampling end, and the other end is the mass spectrometry connection end, both of which extend outside the handheld casing B1; the three-way fitting B3 is located near the sampling end of the sampling pipe B2, and the two-way fitting B4 is located near the mass spectrometry connection end of the sampling pipe B2.

[0054] The main pipe of the three-way fitting B3 is fitted onto the sampling pipe B2 near the sampling end. The branch pipe is equipped with a suitable plug B33. The plug B33 is tightly fitted with a pipe B6 for conveying the solvent in the liquid supply system B7 to the three-way fitting B3. The main pipe is equipped with a suitable plug B32 near the mass spectrometer connection end. The plug B32 is tightly fitted with a fixing sleeve for fixing the sampling pipe B2. The fixing sleeve is tightly fitted onto the outside of the sampling pipe B2. The main pipe is equipped with a suitable plug B31 near the sampling end. The plug B31 is tightly fitted with a nested tube B5 outside the sampling pipe B2. The inner wall of the nested tube B5 and the outer wall of the sampling pipe B2 have a cavity for the solvent conveyed to the three-way fitting B3 to flow out to the sampling end of the sampling pipe B2.

[0055] Two-way fitting B4 is fitted onto the sampling pipe B2 near the mass spectrometer connection end. The sampling end and the mass spectrometer connection end of the two-way fitting B4 are respectively equipped with matching plugs B41 and B42. The plugs B41 and B42 are respectively tightly fitted with fixing sleeves for fixing the sampling pipe B2. The fixing sleeves are tightly fitted onto the outside of the sampling pipe B2 inside the mass spectrometer system B8, driven by the pressure difference formed by the negative pressure in the mass spectrometer system.

[0056] The handheld outer shell B1 is manufactured using computer software modeling combined with 3D printing technology. It consists of two parts: a front part and a back part. The back part is approximately planar with a wall thickness of 2.5mm and features grooves for fixing the tee fitting B3 and the two-way fitting B4, as well as screw holes for fixing these fittings. The front part is approximately a hollow semi-cylinder with a wall thickness of 2.5mm, slightly narrowing near the mass spectrometer connection end. Near the sampling end, it has an approximately conical structure with a hole at the tip for the sampling tube B2 and the nested tube B5. Both the front and back parts are secured with screws through the through screw holes. An extended section on the left side of the handheld outer shell accommodates a branch tube for the tee fitting B3. All screw holes are 3mm in diameter, and M3 screws and nuts are used for fixing.

[0057] During assembly, first, insert the tee fitting B3 and the two-way fitting B4 into the grooves on the opposite side, and then secure them with screws. A quartz capillary tube with an outer diameter of 362 μm, an inner diameter of 150 μm, and a length of approximately 75 cm is used as the sampling conduit B2. The sampling end of the sampling conduit B2 is passed through the main pipes of the tee fitting B3 and the two-way fitting B4. A suitable plug B32 is provided near the mass spectrometer connection end of the main pipe of the tee fitting B3. A fixing sleeve for securing the sampling conduit B2 is tightly fitted onto the plug B32. A PEEK tube with an outer diameter of 1 / 16 inch, an inner diameter of 405 μm, and a length of approximately 25 mm was selected as the fixing sleeve. Since the inner diameter of the fixing sleeve (405 μm) is slightly larger than the outer diameter of the sampling pipe B2 (362 μm), a slight elastic deformation of the fixing sleeve was caused by a hand-tightening plug B32, thereby sealing and fixing the sampling pipe B2. A matching plug B31 was provided near the sampling end of the main pipe of the tee fitting B3, and a nesting tube B5 was tightly fitted onto the plug B31, which was fitted over the sampling pipe B2. An ETFE tube with an outer diameter of 1 / 16 inch, an inner diameter of 1000 μm, and a length of approximately 40 mm was selected as the nesting tube B5. Since the inner diameter of the nesting tube B5 (1000 μm) is much larger than the outer diameter of the sampling pipe B2 (362 μm), a cavity was left between the inner wall of the nesting tube B5 and the outer wall of the sampling pipe B2 for the solvent transported to the tee fitting B3 to flow out to the sampling end of the sampling pipe B2. Utilizing the characteristic that the plug B32 only seals and secures the sampling tube B2 when tightened, the relative position of the sampling tube B2 and the nesting tube B5 can be adjusted when loosened. There are two possible positional relationships between the sampling tube B2 and the nesting tube B5: When the sampling tube B2 does not extend beyond the nesting tube B5 and is approximately 1-2 mm from the sampling end of the nesting tube B5, the nesting tube B5 protects the sampling tube B2 and creates a backflow of solvent from the nesting tube B5 to the sampling tube B2 within the nesting tube B5. However, if the negative pressure and flow rate are unstable or mismatched, droplet interruption and solvent overflow may occur, leading to extraction failure or sample contamination. When the sampling tube B2 extends approximately 1-2 mm beyond the nesting tube B5, sufficient negative pressure from the negative pressure supply system allows the solvent to flow out of the nesting tube B5, contact the analyte, and then enter the sampling tube B2 under the drive of the pressure difference. This reduces the matching requirements of the liquid flow rate and negative pressure, simplifying the process of adjusting the flow rate and negative pressure. Pass the sampling conduit B2 and the nested tube B5 through the hole at the tip of the cone near the sampling end of the positive section. Adjust the hole so that the nested tube B5 extends about 2-3 mm beyond the tip of the cone. Use a PEEK tube with an outer diameter of 1 / 16 inch, an inner diameter of 500 μm, and a length of about 30 cm as conduit B6. Connect one end of conduit B6 to the liquid supply system B7, and secure the other end to the branch pipe of the tee fitting B3 using a suitable plug B33.

[0058] Sampling conduit B2 is passed through the main pipe of the two-way fitting B4. Adaptive plugs B41 and B42 are installed at the sampling end and mass spectrometer connection end of the main pipe, respectively. Plugs B41 and B42 are each tightly fitted with a fixing sleeve for securing sampling conduit B2. This fixing sleeve is tightly fitted onto the external sampling conduit B2 within the mass spectrometry system, driven by the pressure difference created by the negative pressure within the system. A PEEK tube with an outer diameter of 1 / 16 inch and an inner diameter of 405 μm is selected as the fixing sleeve. The fixing sleeve, approximately 25 mm long, is tightly fixed to plug B41 at the sampling end. Since the inner diameter of the fixing sleeve (405 μm) is slightly larger than the outer diameter of sampling conduit B2 (362 μm), a slight elastic deformation is achieved by hand-tightening plug B41, thereby sealing and securing sampling conduit B2. A retaining sleeve, tightly fixed to the plug B42 near the mass spectrometer connection end, extends out of the handheld housing B1, but its length does not exceed the distance from the sampling tube B2 extending from the plug B42 to the mass spectrometer connection end. The two-way fitting B4 is used to create a handheld structure that is easy to grip and to fix and protect the sampling tube B2.

[0059] During sampling, driven by the pressure difference created by the negative pressure within the mass spectrometry system, the extract in sampling pipe B2 is transported to mass spectrometry system B8. Taking a commercially available thermo-electrospray ionization (H-ESI) source and a QExactive combined quadrupole Orbitrap mass spectrometer as an example, mass spectrometry system B8 includes the H-ESI source and the QExactive combined quadrupole Orbitrap mass spectrometer, as well as a fixing assembly connecting the H-ESI source and sampling pipe B2.

[0060] (2) Install a liquid supply system for supplying solvent to the handheld extraction system B: The liquid supply system B7 includes a syringe and an injection pump that controls the injection of solvent by the syringe, as well as a fixing assembly that connects the syringe to the pipeline B6.

[0061] (3) Online in-situ extraction and analysis of solid samples using a handheld solid sample in-situ extraction device: First, use a syringe to draw a suitable amount of solvent and fix the syringe on the syringe pump; then push the syringe forward until the nested tube B5 is filled with solvent and free of air bubbles; taking a commercially available quadrupole Orbitrap mass spectrometer equipped with a heated electrospray ionization (H-ESI) ion source and a QExactive combination as an example, operate the software interface of mass spectrometer B8 and switch the mass spectrometer status from "standby" to "on". At this time, the HESI ion source starts to run and generates negative pressure inside the ion source due to the Venturi effect, running the syringe pump to push the syringe to the probe tip to form a liquid condensation. Observe whether the mass spectrometry signal is stable, and start acquiring the signal when the total ion current is stable. Prepare the analyte. Bring the sampling end of the handheld extraction system B close to the analyte until the sampling tube B2 is about to contact it. At this point, the liquid condensate contacts the analyte and sampling begins. Once sufficient signal has been collected, remove the handheld extraction system B to end the sampling. The liquid condensate is still forming. Clean solvent flows in and washes the sampling tube B2 to prevent residue of the extract. After washing is complete, turn off the syringe pump. After the mass spectrometry signal gradually disappears, switch the mass spectrometer to "standby" mode. The online in-situ extraction is now complete.

[0062] Example 3: Application of Offline In-situ Extraction Device

[0063] (1) An offline in-situ extraction device combined with MALDI-TOF MS was used to detect buspirone standards spotted on glass slides.

[0064] To evaluate the detection capability and accuracy of the offline in-situ extraction device for small molecule drug components, this embodiment employs an offline in-situ extraction device combined with MALDI-TOF MS to detect buspirone standards spotted on a glass slide. Buspirone belongs to the azaspirocyclodecanedione class of compounds and possesses anti-anxiety effects. Due to its good ionization efficiency, it is often used to evaluate the detection performance of mass spectrometry analysis.

[0065] First, prepare the buspirone sample: Take buspirone standard (Mw 385.2472) and prepare a solution of 1 pg / ml. Use a pipette to spot the buspirone standard solution onto a glass slide. The sample volume is 1 μL. After evaporation, it is ready for use.

[0066] Subsequently, sampling was performed using an offline in-situ extraction device: According to the in-situ extraction device provided in Example 1, and in accordance with Example 1-(4), the injection parameters were as follows: methanol:water = 8:2 (v / v) was used as the extraction solvent, the injection pump flow rate was 30 μL / min, the extraction volume was 100 μL, the washing solvent was 50 μL, and the spotting and extraction were repeated 6 times; a pre-drilled 1.5 mL EP tube was selected as the collection tube A5, and the handheld liquid microjunction surface extraction probe device was aimed at the evaporated buspirone standard sample spot on the glass slide for offline in-situ extraction.

[0067] The extract was then analyzed according to the procedures in Examples 1-(5): Matrix-assisted laser desorption / ionization mass spectrometry (MALDI-MS) was used, with 2,5-dihydroxybenzoic acid (DHB) as the detection matrix in positive ion mode. The matrix was deposited using a matrix spray method. Specifically, the extract was concentrated by evaporation under nitrogen blowing, reconstituted with 10 μL of pure solvent, and spotted onto an indium tin oxide (ITO) coated slide with a sample volume of 1 μL. The matrix was then uniformly deposited onto the ITO slide using a matrix spray method. The operation was performed according to the MALDI-MS instrument instructions, with detection in positive ion mode. Representative MALDI-MS mass spectra are shown below. Figure 5 Representative spectra of buspirone standard samples were obtained. Figure 5 On plate A, buspirone [M+H] can be seen at m / z 386.2. + Addition peaks and isotopic peak clusters. And representative spectra in the background matrix. Figure 5 There is no signal at m / z 386.2 in B.

[0068] (2) An offline in-situ extraction device combined with MALDI-TOF MS was used to detect endogenous small molecule metabolites and lipids in rat brain slices.

[0069] To examine the ability of the offline in situ extraction device to detect endogenous small molecule metabolites and lipids in biological tissues, this embodiment uses the offline in situ extraction device combined with MALDI-TOF MS to detect sagittal sections of rat brain.

[0070] First, prepare sagittal sections of the rat brain: Take a rat brain stored at -80℃, cut it along the sagittal plane, and slice it on a cryostat to a thickness of 10μm. Transfer the slices to a glass slide using the fusion mounting method. The sagittal sections of the rat brain are now ready.

[0071] Sampling was then performed using an offline in-situ extraction device: The in-situ extraction device provided in Example 1 was used, and the operation was performed according to Example 1-(4). The injection parameters were as follows: 0.1% trifluoroacetic acid in methanol:water = 7.5:2.5 (v / v) was used as the extraction solvent, the injection pump flow rate was 30 μL / min, the extraction volume was 100 μL, the washing solvent was 50 μL, and the extraction was repeated 5 times; a pre-drilled 1.5 mL EP tube was selected as the collection tube A5, and the handheld liquid microjunction surface extraction probe device was aimed at the sagittal section of rat brain on the glass slide for offline in-situ extraction.

[0072] The extract was then analyzed according to the procedures in Examples 1-(5): Matrix-assisted laser desorption / ionization mass spectrometry (MALDI-MS) was used, with 2,5-dihydroxybenzoic acid (DHB) as the detection matrix in positive ion mode. The matrix was deposited using a matrix spray method. Specifically, the extract was concentrated by evaporation under nitrogen blowing, reconstituted with 10 μL of pure solvent, and spotted onto an indium tin oxide (ITO) coated slide with a sample volume of 1 μL. The matrix was then uniformly deposited onto the ITO slide using a matrix spray method. The operation was performed according to the MALDI-MS instrument instructions, with detection in positive ion mode. Representative MALDI-MS mass spectra are shown below. Figure 6 and Figure 7 ,from Figure 7 In sample A, lipid signals distributed in the mass-to-charge ratio range of 750-850 can be observed, including glycerophospholipids (GP): phosphatidylcholine (PC), phosphatidylethanolamine (PE), etc., and sphingolipids (SP): sphingomyelin (SM), etc.; from Figure 7 In B, signals of small molecule metabolites distributed in the mass-to-charge ratio range of 100-180 can be observed, including γ-aminobutyric acid (GABA), glutamate (Glu), acetylcholine (Ach), etc.

[0073] (3) An offline in-situ extraction device combined with MALDI-TOF MS was used to detect various chemical components in a variety of Chinese herbal medicine slices, including honeysuckle, chrysanthemum, rose, and coptis.

[0074] To examine the ability of the offline in-situ extraction device to detect various chemical components in traditional Chinese medicine decoction pieces, this embodiment uses an offline in-situ extraction device combined with MALDI-TOF MS to detect various chemical components in traditional Chinese medicine decoction pieces such as honeysuckle, chrysanthemum, rose, and coptis.

[0075] First, prepare the Chinese herbal medicine slices: Take honeysuckle, chrysanthemum, rose and coptis slices purchased from a Chinese medicine pharmacy and weigh about 0.5mg of the slices on a balance. Place the 0.5mg slices in a 35mm x 12mm petri dish and spread them evenly. The Chinese herbal medicine slices honeysuckle, chrysanthemum, rose and coptis are now ready.

[0076] Subsequently, sampling was performed using an offline in-situ extraction device: According to the in-situ extraction device provided in Example 1, and in accordance with Example 1-(4), the injection parameters were as follows: methanol:water = 8:2 (v / v) was used as the extraction solvent, the injection pump flow rate was 30 μL / min, the extraction volume was 100 μL, the washing solvent was 50 μL, and each type of medicinal slice was extracted 5 times. Among them, the alkaloid content of Coptis chinensis slices was high and had a strong ionization efficiency. The extraction volume was increased to 50 μL, and the washing volume was increased to 300 μL to completely wash the residue in the sampling pipe A2. A pre-drilled 1.5 mL EP tube was selected as the collection tube A5, and the handheld liquid microjunction surface extraction probe device was aimed at the medicinal slices to be tested for offline in-situ extraction.

[0077] The extract was then analyzed according to the procedures in Examples 1-(5): Matrix-assisted laser desorption / ionization mass spectrometry (MALDI-MS) was used, with 2,5-dihydroxybenzoic acid (DHB) as the detection matrix in positive ion mode. The matrix was deposited using a matrix spray method. Specifically, the extract was concentrated by evaporation under nitrogen blowing, reconstituted with 10 μL of pure solvent, and spotted onto an indium tin oxide (ITO) coated slide with a sample volume of 1 μL. The matrix was then uniformly deposited onto the ITO slide using a matrix spray method. The operation was performed according to the MALDI-MS instrument instructions, with detection in positive ion mode. Representative MALDI-MS mass spectra are shown below. Figure 8 The following characteristic chemical components of various types of Chinese herbal medicines can be seen from the spectrum:

[0078] (a) Representative spectrum of extracts from honeysuckle slices Figure 8 A sample A contains a variety of characteristic chemical components, including various flavonoids and flavonoid glycosides, such as rutin, in an addition form [M+H]. + The mass-to-charge ratio is m / z 611.1; kaempferol-rutin glycoside, in the addition form [M+H]. + The mass-to-charge ratio is m / z 595.1; luteolin-glucoside, in the addition form [M+H]. + The mass-to-charge ratio is m / z 499.1, etc.

[0079] (b) Representative spectrum of chrysanthemum extract. Figure 8 A variety of characteristic chemical components can be detected in B, including various flavonoids and flavonoid glycosides, such as robinin, in an addition form of [M+H]. + The mass-to-charge ratio is m / z 285.1; quercetin, in additive form [M+H] + The mass-to-charge ratio is m / z 465.2; luteolin-glucoside, in the addition form [M+H]. + The mass-to-charge ratio is m / z 499.1, etc.

[0080] (c) Representative spectrum of rose extract Figure 8 A variety of characteristic chemical components can be detected in C, including various organic acids and phenolic compounds, such as gallic acid, in the addition form [M+H]. + The mass-to-charge ratio is m / z 171.1; the proanthocyanidins are in the additive form of [M+H]. + The mass-to-charge ratio is m / z 579.1, etc.

[0081] (d) Representative spectrum of extracts from Coptis chinensis slices Figure 8 A variety of characteristic chemical components can be detected in D, including various alkaloid compounds, such as berberine, in an addition form [M+H]. + The mass-to-charge ratio is m / z 336.1; berberine, in the addition form [M+H] + The mass-to-charge ratio is m / z 322.1; berberine, in the addition form [M+H] + The mass-to-charge ratio is m / z 321.1; corydaline, in the addition form [M+H]. + The mass-to-charge ratio is m / z 342.2, etc.

[0082] (4) An offline in-situ extraction device combined with MALDI-TOF MS was used to detect and distinguish safflower from dyed chrysanthemum based on chemical composition.

[0083] To examine the application capability of offline in-situ extraction devices in identifying the authenticity of traditional Chinese medicine decoction pieces, this embodiment uses an offline in-situ extraction device combined with MALDI-TOF MS to detect and distinguish safflower from dyed chrysanthemum based on chemical composition. Dyed chrysanthemum has been reported as an adulterant of safflower.

[0084] First, prepare the medicinal herbs: Take safflower slices and chrysanthemum slices dyed with Sunset Yellow solution purchased from a Chinese medicine pharmacy, grind them in advance in an electric grinder so that the two types of herbs cannot be directly distinguished by appearance, and weigh about 0.5 mg of the herbs on a balance. Place the 0.5 mg of herbs in a 35 mm x 12 mm petri dish and spread them evenly. The preparation of the safflower and dyed chrysanthemum herbs is now complete. Then, randomly shuffle the petri dishes and label the two types of medicinal herbs to be tested as Test 1 and Test 2.

[0085] Subsequently, sampling was performed using an offline in-situ extraction device: According to the in-situ extraction device provided in Example 1, and in accordance with Example 1-(4), the injection parameters were as follows: methanol:water = 8:2 (v / v) was used as the extraction solvent, the injection pump flow rate was 30 μL / min, the extraction volume was 100 μL, the washing solvent was 50 μL, and each type of medicinal slice was extracted 5 times; a pre-drilled 1.5 ml EP tube was selected as the collection tube A5, and the handheld liquid microjunction surface extraction probe device was aimed at the medicinal slice to be tested for offline in-situ extraction.

[0086] Simultaneously, 0.5 mg of safflower slices were weighed on a balance and extracted with 500 μL of methanol:water = 8:2 (v / v) as the extraction solvent. The extract was ultrasonically extracted for 30 minutes to obtain the standard herbal extract. Subsequently, a standard solution containing 0.5 μg / mL of hydroxysafflower yellow A (HSYA) was prepared using methanol:water = 8:2 (v / v) as the solvent.

[0087] The extract was then analyzed according to the procedures in Examples 1-(5): Matrix-assisted laser desorption / ionization mass spectrometry (MALDI-MS) was used, with aminoacridine (9-AA) as the detection matrix in negative ion mode. The matrix was deposited using a matrix spray method. Specifically, the extract was concentrated by evaporation under nitrogen blowing, and then reconstituted with 10 μL of pure solvent. The solution was then spotted onto an indium tin oxide (ITO) coated slide with a loading volume of 1 μL. Simultaneously, the standard medicinal extract and the standard hydroxysaffron yellow A (HSYA) were spotted onto the same ITO coated slide for comparison, with a loading volume of 1 μL. The matrix was then uniformly deposited onto the ITO slide using a matrix spray method. The operation was performed according to the MALDI-MS instrument instructions, with detection in negative ion mode. Representative MALDI-MS mass spectra are shown below. Figure 9 The following characteristic chemical components of various types of Chinese herbal medicines can be seen from the spectrum:

[0088] (a) Representative spectrum of extracts from standard medicinal materials Figure 9 Representative spectra of standard reference standards for A and hydroxysaffron yellow A (HSYA) Figure 9 The characteristic component hydroxysaffron yellow A, in the addition form [MH], can be detected in B. - The mass-to-charge ratio is m / z 611.1.

[0089] (b) From the representative spectrum of analyte 1 Figure 9 The characteristic component of safflower, hydroxysafflower yellow A, can be detected in C, in an additive form of [MH]. - A signal with a mass-to-charge ratio of m / z 611.1 was observed, and sunset yellow pigment was undetectable in analyte 1. According to existing literature, the characteristic peak of sunset yellow pigment contains an additive form of [M-2Na+H].- The signal with a mass-to-charge ratio of m / z 407.3 and the summation form are [M-Na]. - The signal has a mass-to-charge ratio of m / z 429.3. Based on the above chemical information, analyte 1 may be safflower slices.

[0090] (c) Representative spectrum of analyte 2 detected Figure 9 Hydroxysafflower yellow A, a characteristic component of safflower, could not be detected in analyte D, while the characteristic peak of sunset yellow pigment, containing the additive form [M-2Na+H], could be detected in analyte 2. - The signal with a mass-to-charge ratio of m / z 407.3 and the summation form are [M-Na]. - The signal has a mass-to-charge ratio of m / z 429.3. Based on the above chemical information, analyte 2 may be dyed chrysanthemum slices, i.e., adulterated safflower.

[0091] Example 4: Application of Online In-situ Extraction Device

[0092] (1) An online in-situ extraction device combined with HESI-Q Exactive MS was used to detect endogenous small molecule metabolites and lipids in rat brain slices.

[0093] To examine the ability of the online in-situ extraction device to detect endogenous small molecule metabolites and lipids in biological tissues, this embodiment uses the online in-situ extraction device combined with HESI-Q Exactive MS to detect sagittal sections of rat brain.

[0094] First, prepare sagittal sections of the rat brain: Take a rat brain stored at -80°C, cut it along the sagittal plane, and slice it on a cryostat to a thickness of 10 μm. Transfer the slices to a glass slide using the fusion mounting method. The sagittal sections of the rat brain are now ready.

[0095] Subsequently, online in-situ extraction device was used for sampling and analysis: According to the in-situ extraction device provided in Example 2 and the operation in Example 2-(3), the injection parameters were as follows: 0.1% trifluoroacetic acid in methanol:water = 7.5:2.5 (v / v) was used as the extraction solvent, and the injection pump flow rate was 10 μL / min; the heating electrospray ionization (H-ESI) ion source was set with the following parameters: sheath gas pressure 15 psi, ion source voltage 4000V, ion transmission tube temperature 320℃, ion source nozzle heating temperature 300℃, and detection mode positive ion mode; the mass spectrometer status was switched to "on" and the injection pump was run to push the syringe to the probe tip to form a liquid knot. After the mass spectrometry signal stabilized, the handheld liquid microknot surface extraction probe device was aligned with the tissue slice and the mass spectrometry was set to start sampling; the sampling parameters were as follows: 30 μL of extract was collected and detected, 50 μL of washing solvent was used, and the extraction was repeated 5 times. The detection data of each time was saved and analyzed. The representative mass spectrometry spectrum is shown in Figure 10 and Figure 11 ,from Figure 11 In sample A, lipid signals distributed in the mass-to-charge ratio range of 700-850 can be observed, including lipids such as glycerophospholipids (GP), phosphatidylcholine (PC), and phosphatidylethanolamine (PE); similarly, from... Figure 11 In sample B, signals of small molecule metabolites with a mass-to-charge ratio in the range of 100-180 can be observed, including tyrosine, in the addition form [M+H]. + The mass-to-charge ratio is m / z 180.0658; phenylalanine, in the addition form [M+H]. + The mass-to-charge ratio is m / z 164.0706; the dobar concentration is [M+H] in additive form. + The mass-to-charge ratio is m / z 196.0632, etc.

[0096] (2) An online in-situ extraction device combined with HESI-Q Exactive MS was used to detect and distinguish safflower from dyed chrysanthemum based on chemical composition.

[0097] To examine the application capability of online in-situ extraction devices in identifying the authenticity of traditional Chinese medicine decoction pieces, this embodiment uses an online in-situ extraction device combined with HESI-Q Exactive MS to detect and differentiate safflower from dyed chrysanthemum based on chemical composition. Dyed chrysanthemum has been reported as an adulterant of safflower.

[0098] First, prepare the medicinal herbs: Take safflower slices and chrysanthemum slices dyed with Sunset Yellow solution purchased from a Chinese medicine pharmacy, grind them in advance in an electric grinder so that the two types of herbs cannot be directly distinguished by appearance, and weigh about 0.5 mg of the herbs on a balance. Place the 0.5 mg of herbs in a 35 mm x 12 mm petri dish and spread them evenly. The preparation of the safflower and dyed chrysanthemum herbs is now complete. Then, randomly shuffle the petri dishes and label the two types of medicinal herbs to be tested as Test 1 and Test 2.

[0099] Subsequently, sampling and analysis were performed using an online in-situ extraction device: According to the in-situ extraction device provided in Example 2, and following the operation in Example 2-(3), the injection parameters were as follows: 0.1% trifluoroacetic acid in methanol:water = 7.5:2.5 (v / v) was used as the extraction solvent, and the injection pump flow rate was 10 μL / min; the heating electrospray ionization (H-ESI) ion source was set with the following parameters: sheath gas pressure 15 psi, ion source voltage 3500 V, ion transmission tube temperature 320 °C, ion source nozzle heating temperature 300 °C, and detection mode negative ion mode; the mass spectrometer was switched to "on" and the injection pump was run to push the syringe to the probe tip to form a liquid junction. After the mass spectrometry signal stabilized, the handheld liquid microjunction surface extraction probe device was aligned with the sample to be tested, and the mass spectrometer was set to start sampling; the sampling parameters were as follows: 30 μL of extract was collected and detected, 50 μL of washing solvent was used, and the extraction was repeated 5 times. The detection data of each time was saved and analyzed. The representative mass spectrometry spectrum is shown in Figure 12 The following are characteristic chemical components of various types of Chinese herbal medicine slices:

[0100] (a) From the representative spectrum of analyte 1 Figure 12 Hydroxysafflower yellow A, a characteristic component of safflower, can be detected in A and 12B, in an additive form [MH]. - A signal with a mass-to-charge ratio of m / z 611.1586 was observed, but sunset yellow pigment was undetectable in analyte 1. According to existing literature, the characteristic peak of sunset yellow pigment contains an additive form of [M-2Na+H]. - The signal with a mass-to-charge ratio of m / z 407.3 and the summation form are [M-Na]. - The signal has a mass-to-charge ratio of m / z 429.3. Based on the above chemical information, analyte 1 may be safflower slices.

[0101] (b) From the representative spectrum of analyte 2 Figure 12 Hydroxysafflower yellow A, a characteristic component of safflower, could not be detected in analytes C and 12D, while the characteristic peak of sunset yellow pigment, containing the additive form [M-2Na+H], could be detected in analyte 2. - The signal with a mass-to-charge ratio of m / z 407.3 and the summation form are [M-Na].- The signal has a mass-to-charge ratio of m / z 429.3. Based on the above chemical information, analyte 2 may be dyed chrysanthemum slices, i.e., adulterated safflower.

[0102] (3) The online in-situ extraction device combined with HESI-Q Exactive MS was used to detect the MS and MS / MS results of hydroxysaffron yellow A standard sample spots on glass slides and hydroxysaffron yellow A in safflower decoction pieces.

[0103] To examine the ability of the online in-situ extraction device to detect characteristic chemical components of traditional Chinese medicine decoction pieces and obtain secondary mass spectrometry information, this embodiment uses the online in-situ extraction device combined with HESI-Q Exactive MS to detect the MS and MS / MS results of hydroxysaffron yellow A standard on a glass slide and hydroxysaffron yellow A in safflower decoction piece.

[0104] First, prepare the safflower decoction pieces: Weigh approximately 0.5 mg of safflower decoction pieces purchased from a Chinese medicine pharmacy using a balance. Place the 0.5 mg pieces evenly in a 35 mm x 12 mm petri dish. After preparing the safflower decoction piece sample, prepare a standard solution containing 0.5 μg / mL of hydroxysafflower yellow A (HSYA) standard using methanol:water = 8:2 (v / v) as the solvent. Spot the prepared hydroxysafflower yellow A (HSYA) standard solution onto a glass slide for comparison. The sample volume is 1 μL, serving as the standard sample spot for hydroxysafflower yellow A (HSYA).

[0105] Subsequently, an online in-situ extraction device was used for sampling and analysis: According to the in-situ extraction device provided in Example 2, and in accordance with the operation in Example 2-(3), the injection parameters were as follows: 0.1% trifluoroacetic acid in methanol:water = 7.5:2.5 (v / v) was used as the extraction solvent, and the injection pump flow rate was 10 μL / min; the heating electrospray ionization (H-ESI) ion source was set with the following parameters: sheath gas pressure 15 psi, ion source voltage 3500 V, ion transmission tube temperature 320 °C, ion source nozzle heating temperature 300 °C, and detection mode negative ion mode; the mass spectrometer status was switched from "on" and the injection pump was run to push the syringe to the probe tip to form a liquid junction. After the mass spectrometry signal stabilized, the handheld liquid microjunction surface extraction probe device was aligned with the sample to be tested, and the mass spectrometer was set to start sampling; when the target compound signal was observed (in this example, the signal of hydroxysaffron yellow A (HSYA) at m / z 611.1586), the mass spectrometry acquisition mode was adjusted from "Full" to "Full". "Scan" was performed in "MS / MS" mode, using a preset m / z 611.1 precursor ion detection window to collect secondary ion fragments of the target. Sampling parameters were as follows: a total of 50 μL of extract was collected and analyzed, followed by 50 μL of washing solvent. Extraction was repeated 5 times, and the detection data from each extraction was saved and analyzed. Representative spectral data are shown below. Figure 13 The following are the characteristic chemical components and secondary ion fragment information of the samples to be tested:

[0106] (a) Characteristic spectra from standard sample points of HSYA Figure 13 A high-response HSYA signal can be detected in A, with the summation form being [MH]. - The mass-to-charge ratio is m / z 611.1586, and the MS / MS scanning mode spectrum is shown. Figure 13 Characteristic secondary ion fragments of HSYA, including m / z 325.07, m / z 473.11 and m / z 491.12, can be detected in B.

[0107] (b) Characteristic spectrum from safflower slices Figure 13 C can detect the characteristic component HSYA signal, which is in additive form [MH]. - The mass-to-charge ratio is m / z 611.1586, and the MS / MS scanning mode spectrum is shown. Figure 13 Characteristic secondary ion fragments of HSYA, including m / z 325.07, m / z 473.11 and m / z 491.12, can be detected in D.

[0108] Based on existing literature and structural information of HSYA, the ion fragmentation pathway of HSYA can be determined as follows: Figure 13 E.

[0109] The purpose of the above embodiments is to specifically introduce the substantive content of this utility model. However, those skilled in the art should know that the protection scope of this utility model should not be limited to the specific embodiments.

Claims

1. A handheld solid sample in-situ extraction device, wherein the device is an offline in-situ extraction device, wherein the offline extraction refers to the subsequent detection and analysis of the extract after extraction, characterized in that: The off-line in-situ extraction device comprises a handheld extraction system A, a liquid supply system for providing solvent for the handheld extraction system A, and a negative pressure system for driving the flow of the extraction liquid in the handheld extraction system A; The handheld extraction system A comprises a handheld shell A1, a sampling pipeline A2 penetrating through the handheld shell A1, a tee fitting A3 and a tee fitting A4 arranged along the sampling pipeline A2 and sleeved outside the sampling pipeline A2 and fixed in the handheld shell A1, and a liquid collecting pipe A5 for collecting liquid in the sampling pipeline A2; one end of the sampling pipeline A2 is a sampling end, and the other end is a collecting end, both of which extend out of the handheld shell A1; the tee fitting A3 and the tee fitting A4 are respectively arranged near the sampling end and near the collecting end of the sampling pipeline A2; the liquid collecting pipe A5 is arranged at the collecting end of the sampling pipeline A2; The main pipe of the tee fitting A3 is sleeved on the sampling pipeline A2 near the sampling end, the branch pipe is provided with a matched plug A33, the plug A33 is tightly provided with a pipeline A8 for conveying the solvent in the liquid supply system A9 into the tee fitting A3; the near-collecting end of the main pipe is provided with a matched plug A32, the plug A32 is tightly provided with a fixing sleeve for fixing the sampling pipeline A2, and the fixing sleeve is tightly sleeved outside the sampling pipeline A2; the near-sampling end of the main pipe is provided with a matched plug A31, the plug A31 is tightly provided with a nested pipe A6 sleeved outside the sampling pipeline A2, and a cavity is left between the inner wall of the nested pipe A6 and the outer wall of the sampling pipeline A2 for the solvent conveyed into the tee fitting A3 to flow out to the sampling end of the sampling pipeline A2; The main pipe of the tee fitting A4 is sleeved on the sampling pipeline A2 near the collecting end, the branch pipe is provided with a matched plug A43, the plug A43 is tightly provided with a pipeline A10 for extracting the gas in the tee fitting A4 to the negative pressure system A11; the near-sampling end of the main pipe is provided with a matched plug A41, the plug A41 is tightly provided with a fixing sleeve for fixing the sampling pipeline A2, and the fixing sleeve is tightly sleeved outside the sampling pipeline A2; the near-collecting end of the main pipe is provided with a matched plug A42, the plug A42 is tightly provided with a nested pipe A7 sleeved outside the sampling pipeline A2, the nested pipe A7 extends out of the handheld shell A1 and is tightly inserted into the liquid collecting pipe A5, a cavity is left between the inner wall of the nested pipe A7 and the outer wall of the sampling pipeline A2 for the negative pressure system to extract the gas in the liquid collecting pipe A5 through the tee fitting A4, so as to form a negative pressure environment in the liquid collecting pipe A5, and the tail end of the sampling pipeline A2 and the inner wall of the liquid collecting pipe A5 are in a wall-attached connection mode for facilitating the extraction liquid in the sampling pipeline A2 to flow into the liquid collecting pipe A5 along the inner wall of the liquid collecting pipe A5 under the driving of the air pressure difference.

2. The in-situ extraction device of claim 1, wherein: The liquid supply system comprises a syringe and an injection pump for controlling the injection of the syringe.

3. The in-situ extraction device of claim 1, wherein: The sampling pipeline A2 is a quartz capillary.

4. The in-situ extraction device of claim 1, wherein: The nested pipes A6 and A7 are ETFE pipes.

5. The in-situ extraction device of claim 1, wherein: The fixing sleeve is a PEEK pipe.

6. A hand-held solid sample in-situ extraction device, which is an on-line in-situ extraction device, the on-line indicating that the extraction liquid is directly delivered to a mass spectrometry system for analysis and detection, characterized in that: The on-line in-situ extraction device comprises a handheld extraction system B and a liquid supply system for providing solvent for the handheld extraction system B; The handheld extraction system B comprises a handheld shell B1, a sampling pipe B2 penetrating the handheld shell B1, a tee pipe B3 and a two-way pipe B4 arranged along the sampling pipe B2 and sleeved outside the sampling pipe B2 and fixed in the handheld shell B1; one end of the sampling pipe B2 is a sampling end, and the other end is a mass spectrometry connecting end, both of which extend outside the handheld shell B1; the tee pipe B3 is arranged near the sampling end of the sampling pipe B2, and the two-way pipe B4 is arranged near the mass spectrometry connecting end of the sampling pipe B2; The main pipe of the tee pipe B3 is sleeved on the sampling pipe B2 near the sampling end, the branch pipe is provided with a matched plug B33, the plug B33 is closely provided with a pipe B6 for conveying the solvent in the liquid supply system B7 into the tee pipe B3; the main pipe near the mass spectrometry connecting end is provided with a matched plug B32, the plug B32 is closely provided with a fixing sleeve for fixing the sampling pipe B2, and the fixing sleeve is closely sleeved outside the sampling pipe B2; the main pipe near the sampling end is provided with a matched plug B31, the plug B31 is closely provided with a nested pipe B5 sleeved outside the sampling pipe B2, and a cavity is left between the inner wall of the nested pipe B5 and the outer wall of the sampling pipe B2 for conveying the solvent flowing out of the tee pipe B3 to the sampling end of the sampling pipe B2; The two-way pipe B4 is sleeved on the sampling pipe B2 near the mass spectrometry connecting end, the two-way pipe B4 is provided with matched plugs B41 and B42 near the sampling end and the mass spectrometry connecting end respectively, the plugs B41 and B42 are closely provided with fixing sleeves for fixing the sampling pipe B2, and the fixing sleeves are closely sleeved outside the sampling pipe B2 to convey the extraction liquid to the sampling pipe B2 outside the mass spectrometry system B8 under the driving of the air pressure difference formed by the negative pressure in the mass spectrometry system.

7. The in-situ extraction device of claim 6, wherein: The liquid supply system comprises a syringe and an injection pump for controlling the injection of the syringe.

8. The in-situ extraction device of claim 6, wherein: The sampling pipe B2 is a quartz capillary.

9. The in-situ extraction device of claim 6, wherein: The nested pipe B5 is an ETFE pipe.

10. The in-situ extraction device of claim 6, wherein: The fixing sleeve is a PEEK pipe.

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