CN218121875U - Double-spray rapid evaporation-based liquid chromatography-mass spectrometry device capable of analyzing salt-containing sample - Google Patents
Double-spray rapid evaporation-based liquid chromatography-mass spectrometry device capable of analyzing salt-containing sample Download PDFInfo
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Abstract
The utility model discloses a but assay contains liquid chromatography-mass spectrometry device of salt sample based on two spray flash evaporation, include: a base module; the spraying fixing and control module and the electric heating piece fixing and control module are arranged on the base module; the sample spray generating module and the organic solvent spray generating module are arranged on the spray fixing and controlling module; the electric heating piece module is arranged on the electric heating piece fixing and controlling module; the jet orifice of the sample spray generation module and the jet orifice of the organic solvent spray generation module both face the emission end of the electric heating piece module. The device provided by the invention does not need to apply high voltage or other forms of high-energy to sample spray or organic solvent spray, only needs two sprays to interact with the electric heating sheet, realizes high-efficiency ionization and desalination in the process of rapid evaporation of the two sprays, and solves the problems of transmission system blockage, ion inhibition and the like in the conventional mass spectrometry detection of salt-containing solutions.
Description
Technical Field
The utility model relates to a liquid chromatogram-mass spectrometry allies oneself with technical field, concretely relates to but direct analysis contains liquid chromatography-mass spectrometry device of salt sample based on two spray flash evaporation.
Background
The liquid chromatography-mass spectrometry technology is widely applied to important fields such as medicine and life science by combining the excellent separation capability of liquid chromatography and the high-sensitivity analysis capability of mass spectrometry. The technology has the main advantages that samples with different polarities in the mixed solution can be separated through liquid chromatography, then mass analysis is carried out through high-sensitivity mass spectrometry, and qualitative and quantitative analysis of complex samples is realized through the combined action of the chromatography and the mass spectrometry.
In the medical field, biological samples are generally required to be preserved in high-salt systems such as buffer salts to maintain physiological activity, and in addition, active ingredients in many medicaments are basic organic substances, and the compounds generally have the phenomena of peak tailing or poor selective separation in reverse phase chromatographic separation, so that a non-volatile buffer or an ion pair reagent is required to be added to a mobile phase. When the mass spectrometry is used, on one hand, salt can suppress signals of an object to be detected, so that the sensitivity of mass spectrometry detection is greatly reduced, and on the other hand, salt in a high-salt solution can be deposited in a pipeline, so that a sample injection capillary, a taper hole and the like are blocked, and serious damage is brought to an instrument. Therefore, high salt samples or samples with high salt buffer salts as the mobile phase generally cannot be directly analyzed using mass spectrometry, and both problems need to be solved by design.
Due to the fact that the open type ion source is easy to operate, the sample solution can not directly act on a mass spectrum inlet through design, and the problem that a high-salt solution is prone to blocking is solved. In order to solve the problem of signal suppression, researchers use methods such as desalting by using different electrophoretic capacities of salt and a sample, suppressing a salt signal by adding other substances, and performing sample desalting by using a two-dimensional liquid phase method. However, the current methods exist more or less: the continuous detection can not be realized, the structure is complex, the efficiency is low and the like.
SUMMERY OF THE UTILITY MODEL
The utility model provides a but direct analysis contains liquid mass spectrometer of salt sample based on two spray flash evaporation (heat-assisted short hollow spraying mass spectrometry, HADSI for short for the following), the device need not to apply the high energy of high voltage or other forms to sample spraying or organic solvent spraying, only need two strands of spraying and electric heat piece interact, realize high-efficient ionization and desalination at two strands of spraying flash evaporation's in-process, solved the transmission system that conventional mass spectrometry detected and contained salt solution and face and blockked up and the ionic suppression scheduling problem. The device can realize liquid chromatogram and use together, fuses the high salt tolerance of liquid chromatogram high efficiency separation characteristic and HADSI, high efficiency desalination characteristic.
The utility model provides a but direct analysis contains liquid chromatography-mass spectrometry device of salt sample based on two spray flash evaporation, include:
a base module;
the spray fixing and control module and the electric heating piece fixing and control module are arranged on the base module;
the sample spray generating module and the organic solvent spray generating module are arranged on the spray fixing and controlling module;
the electric heating piece module is arranged on the electric heating piece fixing and controlling module;
the spray opening of the sample spray generation module and the spray opening of the organic solvent spray generation module face the emission end of the electric heating piece module.
The base module comprises a base, a fixed electric heating piece structure and a fixed spraying structure, wherein the fixed electric heating piece structure and the fixed spraying structure are fixed on the base, the electric heating piece fixing and controlling module is installed on the fixed electric heating piece structure, and the spraying fixing and controlling module is installed on the fixed spraying structure.
The sample spray generation module comprises a spray generation module comprising: the liquid phase atomizer comprises a sample liquid phase atomizer body, wherein an inlet of the sample liquid phase atomizer body is connected with a gas path pipeline and a liquid path pipeline, an outlet of the sample liquid phase atomizer body is connected with a liquid path sprayer and a gas path sprayer, and the gas path sprayer is wrapped on the liquid path sprayer to form mixed spraying.
The organic solvent spray generation module comprises: the liquid phase atomizer comprises an organic solvent liquid phase atomizer, wherein an inlet of the organic solvent liquid phase atomizer is connected with a gas path pipeline and a liquid path pipeline, an outlet of the organic solvent liquid phase atomizer is connected with a liquid path nozzle and a gas path nozzle, and the gas path nozzle is wrapped on the liquid path nozzle to form mixed injection.
The spray fixing and control module is a double-hole accessory which can be fixed, and liquid phase atomizers in the sample spray generation module and the organic solvent spray generation module respectively penetrate through two small holes to be tightly sleeved with the spray fixing and control module. Namely, the sample liquid phase atomizer is tightly sleeved in one hole, and the organic solvent liquid phase atomizer is tightly sleeved in the other hole.
The utility model discloses in, sample spraying generation module atomizes sample solution and organic solvent and sprays to the electric heat piece simultaneously to gasified rapidly. The substance to be measured in the sample solution and the salt solution are crystallized on the high-temperature electric heating sheet, the substance to be measured is selectively distributed on the outer surface of the salt crystal, and the substance to be measured enters the gas phase under the combined action of the electric heating sheet and the organic solvent through the combined action of thermal desorption and solvent elution. The organic solvent is quickly evaporated and converted into charged solvent ions under the action of the high-temperature electric heating sheet, and the charged solvent ions and the molecules of the object to be detected in the gas phase are subjected to charge transfer/ion exchange, so that the object to be detected is ionized. And the ionized substance to be detected is absorbed into the mass spectrum under the action of negative pressure at the mass spectrum inlet to realize detection.
The electric heating plate module is one of important modules for realizing sample ionization, and comprises: the electric heating device comprises a heat generating and adjusting device and an electric heating piece connected with the heat generating and adjusting device; the types of electrothermal sheets include, but are not limited to: electric iron, ceramic electric heating sheet, etc.; the shape of the electric heating piece includes but is not limited to: long knife type, rectangular type, triangular type, circular type, etc.; the electric heating piece is arranged between the sample spray generation module and the mass spectrum inlet; the heat generating and adjusting device is connected with a national standard 220V voltage, and the temperature of the electric heating piece can be continuously adjusted at 0-1000 ℃ through adjustment and can be quickly and stably; the electric heating piece fixing and control module is a single-hole accessory which can be fixed, and the electric heating piece in the electric heating piece module penetrates through the small hole to be tightly sleeved with the electric heating piece fixing and control module.
The angle and the distance between the electric heating piece module and the liquid phase generation module and the mass spectrum inlet can be adjusted by adjusting the angle and the distance between the fixed electric heating piece structure on the base module and the angle and the distance between the fixed spraying structure and the mass spectrum inlet.
Further, the fixed electric heating piece structure and the mass spectrum inlet are kept on the same axis; the distance between the fixed electric heating piece structure and the mass spectrum inlet is d1, and d1 is more than or equal to 0mm and less than or equal to 300mm; the included angle between the fixed spraying structure and the mass spectrum inlet is A1, and A1 is more than or equal to 0 degrees and less than or equal to 90 degrees; the distance between the fixed spraying structure and the mass spectrum inlet is d2, and d2 is more than or equal to 0mm and less than or equal to 300mm;
furthermore, the spray fixing and control module and the fixed double-hole accessory have the first hole with the diameter D7, and the diameter D7 is more than or equal to 6mm and less than or equal to 6.5mm; the diameter of the second hole of the double-hole accessory which can be fixed is D8, and D8 is more than or equal to 6mm and less than or equal to 6.5mm; the sample spray generation module and the organic solvent spray generation module) respectively pass through the first round hole and the second round hole and are tightly sleeved; the double holes are distributed on the same straight line, including but not limited to: vertical lines, horizontal lines, etc.; the included angle between the two holes is A2, and A2 is more than or equal to 0 degrees and less than or equal to 90 degrees; the distance between the centers of the two holes is d3, and d3 is more than or equal to 0mm and less than or equal to 20mm; the spray fixing and control module is fixedly connected with the fixed spray structure and can slide; the distance between the spray fixing and control module and the mass spectrum inlet is d4, and d4 is more than or equal to 0mm and less than or equal to 50mm;
furthermore, the diameter of the single hole of the electric heating piece fixing and controlling module is D9, and D9 is more than or equal to 6mm and less than or equal to 6.5mm; the distance between the electric heating piece fixing and control module and the mass spectrum inlet is d5, and d5 is more than or equal to 0mm and less than or equal to 20mm;
furthermore, the liquid pipeline interface is of a metal pipe structure, the inner diameter is D1, and D1 is more than or equal to 0.5mm and less than or equal to 1 mm; the outer diameter is D2, and D2 is more than or equal to 1mm and less than or equal to 1.5mm; the gas path pipeline interface is of a metal pipe structure, the inner diameter is D3, and D3 is more than or equal to 5.5mm and less than or equal to 6mm; the outer diameter is D4, and D4 is more than or equal to 7mm and less than or equal to 7.5mm; the liquid path spray head and the gas path spray head are millimeter-scale apertures to prevent salt solution blockage, the aperture of the liquid path spray head is D5, D4 is more than or equal to 0.5mm and less than or equal to 3mm, the aperture of the gas path spray head is D6, D6 is more than or equal to 1mm and less than or equal to 5mm; the liquid path nozzle and the gas path nozzle are made of quartz glass materials so as to prevent salt solution corrosion.
Further, the gas flow rate connected with the sample spray generating module is continuously adjustable within the range of 0L/min to 10L/min, and the degree and speed of sample spray are adjusted by adjusting the gas flow rate loaded on the sample spray generating module.
Furthermore, the flow rate of the gas connected with the sample spray generation module and the organic solvent spray generation module is continuously adjustable within the range of 0L/min to 10L/min.
The utility model also provides a method for using of the LC-MS device that contains the salt sample of direct analysis based on two spray flash evaporations, including following step:
1) Double spray generation: the salt-containing sample solution enters a sample spray generation module (1) and is atomized to form a first spray; the organic solvent is pumped to the organic solvent spray generation module (2) through a peristaltic pump and the like, and a second spray is formed through atomization;
2) Double spray focusing: sample spray and organic solvent spray are focused on the electric heating sheet through the spray fixing and control module (3), namely two sprays simultaneously form interaction with the electric heating sheet;
the two spray focusing directions include, but are not limited to: one above and one below, one left and one right, and two sprays are focused at a certain angle;
3) Desalting and ionizing: two sprays are sprayed on the electric heating sheet, salt in the sample solution spray is crystallized on the surface of the electric heating sheet at high temperature, and the object to be measured is deposited on the surface of the salt crystal. Under the combined action of thermal desorption of the electric heating plate and spray elution of the organic solvent, the object to be detected enters a gas phase. The organic solvent is converted into charged solvent ions in the process of rapid evaporation of the contact electric heating sheet, and the charged solvent ions and molecules of the object to be detected in the gas phase carry out charge transfer, so that the object to be detected is ionized to realize detection.
Compared with the prior art, the utility model has the advantages of as follows:
1) The utility model has low energy consumption, is different from the conventional desalination method, and does not need to apply high voltage to two sprays or direct the two sprays to certain forms of high-energy such as electric arc, plasma and the like. The organic solvent spray and the sample spray face the electric heating piece together, and then the high-efficiency ionization and high-performance desalting effect can be realized.
2) The utility model provides a salt-tolerant interface arrangement compares with conventional method, and this device desalination effect is stronger, still has outstanding detection effect to the sample in the high concentration salt solution. Meanwhile, the salt tolerance is high; when the conventional desalination method is used in the face of liquid-mass combination, a mass spectrum transmission system can still be blocked by high-concentration salt solution due to long-time work of a liquid phase relative to a mass spectrum, and the device solves the problem through the design that an atomizer with a millimeter-scale aperture and double-spray orientation deflect at a mass spectrum inlet and the like.
3) The utility model discloses the device is simple, and the flow of sample spraying, organic solvent spraying and atomizing air current isoparametric all can adjust at great within range, consequently possesses very strong suitability, can be applicable to the detection under the different conditions.
Drawings
FIGS. 1 and 2 are a dual spray rapid evaporation-based LC/MS apparatus for direct analysis of salt-containing samples according to the present invention;
FIG. 3 is a perspective view of a sample spray generation module, an organic solvent spray generation module;
FIG. 4 is a mass spectrogram of the device detecting 50 μmol/L reserpine dissolved in different kinds of buffered saline solutions: FIG. 4 (A) is 20mmol/L borate buffer; (B) 20mmol/L MES buffer; (C) 20mmol/L Tris-acetate buffer; (D) 20mmol/L HEPES buffer; figure 5 is the utility model discloses the device detects the mass spectrogram that 50 mu mol/L reserpine dissolves in different concentration PBS buffer salt solution: FIG. 5 (A) is 20mmol/LPBS buffer; (B) is 50mmol/L PBS buffer; (C) is 100mmol/L PBS buffer; (D) is 200mmol/L PBS buffer;
FIG. 6 is a diagram of the mass spectrum of 50 μmol/L rhodamine B dissolved in different buffer salt solutions detected by the device of the utility model: FIG. 6 (A) is 20mmol/L borate buffer; (B) 20mmol/L MES buffer; (C) 20mmol/L Tris-acetate buffer; (D) 20mmol/L HEPES buffer;
FIG. 7 is the mass spectrogram that the device of the utility model detects 50 μmol/L rhodamine B dissolved in PBS buffer salt solution with different concentrations: FIG. 7 (A) is 20mmol/LPBS buffer; (B) is 50mmol/L PBS buffer; (C) is 100mmol/L PBS buffer; (D) is 200mmol/L PBS buffer;
fig. 8 is a mass spectrum diagram of the device of the present invention for detecting 50 μmol/L olanzapine dissolved in different buffer salt solutions: FIG. 8 (A) is 20mmol/L borate buffer; (B) 20mmol/L MES buffer; (C) 20mmol/L Tris-acetate buffer; (D) 20mmol/L HEPES buffer solution;
fig. 9 is a mass spectrum diagram of the device of the present invention for detecting 50 μmol/L olanzapine dissolved in PBS buffer solutions of different concentrations: FIG. 9 (A) is 20mmol/LPBS buffer; (B) is 50mmol/L PBS buffer; (C) 100mmol/L PBS buffer; (D) is 200mmol/L PBS buffer;
FIG. 10 shows the mass spectrum of Roxithromycin 50 μmol/L dissolved in different buffer salt solutions: FIG. 10 (A) is 20mmol/L borate buffer; (B) 20mmol/L MES buffer; (C) 20mmol/L Tris-acetate buffer; (D) 20mmol/L HEPES buffer;
figure 11 is the mass spectrogram that the device of the utility model detects 50 mu mol/L roxithromycin dissolved in PBS buffer salt solution with different concentrations: FIG. 11 (A) is 20mmol/LPBS buffer; (B) is 50mmol/L PBS buffer; (C) is 100mmol/L PBS buffer; (D) is 200mmol/L PBS buffer;
FIG. 12 is a mass spectrum of the device of the present invention for detecting the liquid chromatography-mass spectrometry of 50. Mu. Mol/L erythromycin, roxithromycin, azithromycin, clarithromycin, midecamycin mixed solution;
FIG. 13 is a total ion flow graph of the mixture of example 5;
the numerical designations in the drawings are respectively:
1: a sample spray generation module; 2: an organic solvent spray generation module; 3: a spray fixing and control module; 4: an electric heating piece module; 5: the electric heating piece fixing and control module; 6: base module
11: a sample liquid phase atomizer; 12: a gas path pipeline; 13: a liquid path conduit;
41: an electric heating piece generating and adjusting device; 42: an electric heating sheet;
61: a base; 62: fixing the electric heating sheet structure; 63: fixed spraying structure
111: a liquid path spray head; 112: a gas circuit nozzle; 113: a liquid line pipe interface; 114: and a gas pipeline interface.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, the present invention is described in detail with reference to the accompanying drawings and the following detailed description. It should be noted that the following examples are intended to facilitate understanding of the present invention, and do not limit the present invention in any way.
First, experimental operation
As shown in fig. 1 and fig. 2, a dual-spray rapid evaporation-based liquid chromatography-mass spectrometry device for directly analyzing a salt-containing sample mainly comprises: the device comprises a sample spray generation module 1, an organic solvent spray generation module 2, a spray fixing and control module 3, an electric heating piece module 4, an electric heating piece fixing and control module 5 and a base module 6; the sample spray generation module 1 comprises a sample liquid phase atomizer 11, an air path pipeline 12 and a liquid path pipeline 13; one end of the liquid pipeline 13 is communicated with the liquid atomizer 11 through a liquid pipeline interface 113, and the other end of the liquid pipeline 13 is communicated with the liquid chromatogram; one end of the gas pipeline 12 is connected with the liquid phase atomizer 11 through a gas pipeline interface 114, and the other end of the gas pipeline 12 is connected with the atomizing gas. The gas path nozzle 112 is wrapped on the liquid path nozzle 111 to form mixed injection.
The organic solvent spray generation module 2 comprises a sample liquid phase atomizer 21, a gas path pipeline 22 and a liquid path pipeline 23; one end of the liquid pipeline 23 is communicated with the liquid atomizer 21 through a liquid pipeline interface 213, and the other end of the liquid pipeline 23 is communicated with the liquid chromatogram; one end of the gas pipeline 22 is connected with the liquid phase atomizer 21 through a gas pipeline interface 214, and the other end of the gas pipeline 22 is connected with the atomizing gas. The gas path spray head 212 is wrapped on the liquid path spray head 211 to form mixed spray;
the spray fixing and control module 3 is a double-hole fitting which can be fixed; the liquid phase atomizer 11 is tightly sleeved on a double-hole fitting which can be fixed through a round hole;
the electric heating piece module 4 comprises an electric heating piece generating and adjusting device 41 and an electric heating piece 42 connected with the electric heating piece generating and adjusting device 41; the types of electrothermal sheets include, but are not limited to: electric iron, ceramic electric heating sheet, etc.; the shape of the electric heating sheet 42 includes but is not limited to: long knife type, rectangular type, triangular type, circular type, etc.;
the electric heating piece fixing and control module 5 is a single-hole accessory which can be fixed; the electric heating sheet 42 is tightly sleeved on the single-hole fitting which can be fixed through the round hole;
the base module 6 comprises a base 61, and a fixed electric heating sheet structure 62 and a fixed spraying structure 63 which are fixed on the base 61. The electric heating piece fixing and control module 5 is installed on the fixed electric heating piece structure 62, and the spraying fixing and control module 3 is installed on the fixed spraying structure 63.
The use method of the liquid chromatography-mass spectrometry device capable of directly analyzing the salt-containing sample based on double-spray rapid evaporation comprises the following steps:
1) Sample atomization: adjusting the spraying directions of the sample spraying generation module 1 and the organic solvent spraying generation module 2, wherein the spray heads point to the electric heating sheet 42; after being separated by liquid chromatography, a sample solution enters a liquid path pipeline 13 of a sample spray generation module and flows out of a liquid path spray head 111, atomizing gas is sprayed out of a gas path spray head 112 through a gas path pipeline 12, and under the action of the atomizing gas, the sample in the liquid path spray head is atomized into small droplets; similarly, the other organic solvent is pumped to the liquid channel pipeline 23 of the organic solvent spray generation module 2 through the peristaltic pump and flows out of the liquid channel spray head 211, the atomized gas is sprayed out of the gas channel spray head 212 through the gas channel pipeline, and the organic solvent in the liquid channel spray head is atomized into small droplets under the action of the atomized gas;
2) Double spray focusing: the sample spray and the organic solvent spray are focused on the electric heating sheet 42 through the spray fixing and control module 3, namely two sprays simultaneously form interaction with the electric heating sheet 42; the two spray focusing directions include, but are not limited to: one above and one below, one left and one right, and two sprays are focused at a certain angle;
3) Desalting and ionizing: the two sprays are sprayed on a 42 electric heating plate, salt in the sample solution spray is crystallized on the surface of the high-temperature electric heating plate 42, and an object to be measured is deposited on the surface of the salt crystal. Under the combined action of thermal desorption of the electric heating sheet 42 and spray elution of the organic solvent, the object to be detected enters a gas phase. The organic solvent is converted into charged solvent ions in the process of rapid evaporation of the contact electric heating sheet, and the charged solvent ions and molecules of the object to be detected in the gas phase carry out charge transfer, so that the object to be detected is ionized to realize detection.
Example analysis
Example 1:
in order to test the effect of the present invention, experimental studies were carried out on reserpine in saline solution using the device provided in example 1 and an ultimate UPLC/LTQ Orbitrap mass spectrometer (seidel feishire usa, the same below).
1. Experimental Material
1.1 analyte to be detected
Reserpine standard (analytically pure, shanghai Deng-resistant Biotech Co., ltd., the same applies below), lot number: and B20722. The acetone is firstly made into 10mmol/L mother liquor which is placed in a refrigerator for proper storage for experiments. 1.2 reserpine solution in different buffer solutions
80. Mu.L of 0.5mol/L borate buffer (Shanghai-derived Phyllobiosciences Co., ltd., the same shall apply hereinafter) was aspirated, and 1920. Mu.L of water was added to obtain 2mL20 mmol/L borate buffer. Adding 10 mu L of 10mmol/L reserpine mother liquor, and ultrasonically mixing for 5min to obtain 50 mu mol/L reserpine solution (dissolved in 20mmol/L borate buffer solution).
80. Mu.L of 0.5mol/L MES buffer (Shanghai-derived Ye Biotech Co., ltd., the same shall apply hereinafter) was aspirated, and 1920. Mu.L water was added thereto to obtain 2mL20 mmol/L MES buffer. Adding 10 μ L of 10mmol/L reserpine mother liquor, and ultrasonically mixing for 5min to obtain 50 μmol/L reserpine solution (dissolved in 20mmol/L MES buffer solution).
80. Mu.L of 0.5mol/L Tris-acetate buffer (Shanghai-derived leaf Biotech Co., ltd., the same shall apply hereinafter) was aspirated, and 1920. Mu.L of water was added to obtain 2mL20 mmol/LTris-acetate buffer. Adding 10 mu L of 10mmol/L reserpine mother liquor, and ultrasonically mixing for 5min to finally prepare 50 mu mol/L reserpine solution (dissolved in 20mmol/L Tris-acetic acid buffer solution).
80. Mu.L of 0.5mol/L HEPES buffer (Shanghai-derived leaf Biotech Co., ltd., the same applies hereinafter) was aspirated, and 1920. Mu.L of water was added to obtain 2mL20 mmol/LHEPES buffer. Adding 10 μ L of 10mmol/L reserpine mother liquor, and ultrasonically mixing for 5min to obtain 50 μmol/L reserpine solution (dissolved in 20mmol/L HEPES buffer solution).
1.3 reserpine solutions in PBS buffers of different concentrations
0.5mol/L PBS buffer (Shanghai-derived leaf Biotech Co., ltd., the same applies hereinafter) was used as the PBS stock solution.
mu.L of PBS stock was aspirated, 1920. Mu.L of water was added, and 2mL20 mmol/LPBS buffer was obtained. Adding 10 μ L of 10mmol/L reserpine mother liquor, and ultrasonically mixing for 5min to obtain 50 μmol/L reserpine solution (dissolved in 20mmol/L PBS buffer solution).
mu.L of PBS was aspirated and 1200. Mu.L of water was added to obtain 2mL of 50mmol/LPBS buffer. Adding 10 μ L of 10mmol/L reserpine mother solution, and mixing for 5min by ultrasonic wave to obtain 50 μmol/L reserpine solution (dissolved in 200mmol/L PBS buffer solution).
1.4 organic solvents
Purchasing methanol as an organic solvent (HPLC grade, merck, USA, the same below)
2 method of experiment
Sample introduction conditions are as follows:
organic solvent: methanol at a flow rate of 0.2mL/min
Sample solution: the above reserpine salt solution has a flow rate of 50 μ L/min
Atomizing: organic solvent spray module flow rate: 3L/min; sample spray generation module flow rate: 3L/min
Mass spectrum conditions:
a positive ion detection mode; the scanning mode is positive ion FULL SCAN (FULL SCAN)
HADSI ion source parameters: temperature of the electric heating sheet: 400 ℃;
3. results of the experiment
FIG. 4 is a mass spectrum of reserpine dissolved in 20mmol/L of a different buffer solution according to the present embodiment, obtained by using the apparatus of the present invention. FIG. 4 (A) is a 50. Mu. Mol/L solution of reserpine dissolved in 20mmol/L borate buffer solution, in which the addition peak of reserpine to proton [ M + H ] is clearly visible] + The mass-to-charge ratio (m/z) was 609.2821. FIG. 4 (B) is a 50. Mu. Mol/L reserpine solution dissolved in 20mmol/L MES buffer solution, in which the hydrogenation peak of reserpine [ M + H ] is clearly visible] + The mass-to-charge ratio (m/z) was 609.2843. FIG. 4 (C) is a 50. Mu. Mol/L reserpine solution dissolved in 20mmol/L Tris-acetate buffer solution, in which the hydrogen reduction peak [ M-H ] of reserpine is clearly visible] + The mass-to-charge ratio (m/z) was 607.2647. FIG. 4 (D) is a 50. Mu. Mol/L reserpine solution dissolved in 20mmol/L HEPES buffer solution, in which the hydrogen reduction peak [ M-H ] of reserpine is clearly visible] + The mass-to-charge ratio (m/z) was 607.2650.
In order to further embody the desalting capability of the device of the utility model, 50 mu mol/L reserpine solution dissolved in 20, 50, 100 and 200mmol/L PBS buffer solution is detected by using the device provided by the utility model. As can be seen from Table 1, in the HADSI device, as the salt concentration increased, the addition peak of reserpine and proton [ M + H ]] + The strength and desalting efficiency did not change significantly. As can be seen from the attached figure 5, when the device provided by the utility model is used for detecting reserpine dissolved in PBS buffer solution with different concentrations, a mass spectrum spectrogram always keeps clean and clear and is not influenced by salt cluster ions.
TABLE 1 average signal intensity and desalting efficiency of reserpine in PBS buffer of various concentrations by HADSI detection
Example 2:
in order to test the effect of the present invention, the device provided in example 1 and the ultimate UPLC/LTQ Orbitrap mass spectrometer were used to perform an experimental study on rhodamine B in saline solution.
1. Experimental Material
1.1 analyte to be detected
Rhodamine B standard (analytically pure, hangzhou bang yi ji ltd, same below), lot number: r006869. Firstly, methanol is used to prepare 10mmol/L mother liquor which is placed in a refrigerator for proper storage for experiments.
1.2 rhodamine B solutions in PBS buffers of different concentrations
PBS mother liquor is 0.5mol/L PBS buffer solution. 50 μmol/L rhodamine B solution (dissolved in 20, 50, 100, 200mmol/L PBS buffer) was prepared as in example 1
1.3 rhodamine B solutions in different buffer solutions
50 μmol/L rhodamine B solution (dissolved in 20mmol/L borate, MES, tris-acetic acid, HEPES buffer) was prepared as in example 1
1.4 organic solvents
Purchasing methanol as an organic solvent
2 method of experiment
Sample introduction conditions are as follows:
organic solvent: methanol at a flow rate of 0.2mL/min
Sample solution: the flow rate of the rhodamine B salt solution is 50 mu L/min
Atomizing: organic solvent spray module flow rate: 3L/min; sample spray generation module flow rate: 3L/min
Mass spectrum conditions:
a positive ion detection mode; the scanning mode is positive ion FULL SCAN (FULL SCAN)
HADSI ion source parameters: temperature of the electric heating sheet: 400 ℃;
3. results of the experiment
FIG. 8 shows the mass spectrum of rhodamine B dissolved in 20mmol/L buffer solution of different kinds obtained by using the device of the present inventionFigure (a). FIG. 6 (A) is a 50. Mu. Mol/L rhodamine B solution dissolved in 20mmol/L borate buffer solution, in which the addition peak [ M + H ] of rhodamine B and proton is clearly seen] + The mass-to-charge ratio (m/z) was 443.2343. FIG. 6 (B) is a 50. Mu. Mol/L rhodamine B solution dissolved in 20mmol/L MES buffer solution, in which the addition peak [ M + H ] of rhodamine B and proton is clearly seen] + The mass/charge ratio (m/z) was 443.2343. FIG. 6 (C) is a 50. Mu. Mol/L rhodamine B solution dissolved in 20mmol/L Tris-acetic acid buffer solution, in which the addition peak [ M + H ] of rhodamine B and proton is clearly seen] + The mass/charge ratio (m/z) was 443.2343. FIG. 6 (D) is a 50. Mu. Mol/L rhodamine B solution dissolved in 20mmol/L HEPES buffer solution, in which the addition peak [ M + H ] of rhodamine B and proton is clearly seen] + The mass-to-charge ratio (m/z) was 443.2315.
In order to further embody the desalting capability of the device of the utility model, 50 mu mol/L rhodamine B solution dissolved in 20, 50, 100 and 200mmol/L PBS buffer solution is detected by using the device provided by the utility model. As shown in Table 2, in the HADSI device, as the salt concentration increases, the addition peak [ M + H ] of rhodamine B and proton] + The strength and desalting efficiency did not change significantly. As can be seen from the attached figure 7, the device provided by the utility model is used for detecting rhodamine B dissolved in PBS buffer solution with different concentrations, and the mass spectrum spectrogram is kept clean and clear all the time and is not influenced by salt cluster ions.
TABLE 2 average Signal intensity and desalting efficiency of rhodamine B in PBS buffers of different concentrations by HADSI detection
Example 3:
to test the effectiveness of the present invention, experimental studies were conducted on olanzapine in saline solution using the apparatus provided in example 1 and an ultimate UPLC/LTQ Orbitrap mass spectrometer.
1. Experimental materials
1.1 analyte to be detected
Olanzapine standards (analytical purity, hangzhou Chentong Biochemical technology Co., ltd., the same below), lot number: s61086-5g. Firstly, methanol is used to prepare 10mmol/L mother liquor which is placed in a refrigerator for proper storage for experiments.
1.2 solution of olanzapine in PBS buffer of various concentrations
PBS mother liquor is 0.5mol/L PBS buffer solution. A50. Mu. Mol/L solution of olanzapine (dissolved in 20, 50, 100, 200mmol/L PBS buffer) was prepared as in example 1
1.3 solution of olanzapine in different kinds of buffers
A50. Mu. Mol/L solution of olanzapine (dissolved in 20mmol/L borate, MES, tris-acetic acid, HEPES buffer) was prepared as in example 1
1.4 organic solvents
Purchasing methanol as an organic solvent
2 Experimental methods
Sample introduction conditions are as follows:
organic solvent: methanol flow rate of 0.2mL/min
Sample solution: the olanzapine salt solution has a flow rate of 50 μ L/min
Atomizing: organic solvent spray module flow rate: 3L/min; sample spray generation module flow rate: 3L/min
Mass spectrum conditions:
a positive ion detection mode; the scanning mode is positive ion FULL SCAN (FULL SCAN)
HADSI ion source parameters: temperature of the electric heating sheet: 400 ℃;
3. results of the experiment
FIG. 8 shows the mass spectrum of olanzapine dissolved in 20mmol/L of a different buffer solution as described in this example, obtained using the apparatus of the present invention. FIG. 8 (A) is a 50. Mu. Mol/L solution of olanzapine dissolved in a 20mmol/L borate buffer solution, in which the addition peak of olanzapine to protons [ M + H ] is clearly visible] + The mass-to-charge ratio (m/z) was 313.1461. FIG. 8 (B) is a 50. Mu. Mol/L solution of olanzapine dissolved in 20mmol/L MES buffer solution, in which the addition peak [ M + H ] of olanzapine to proton is clearly seen] + The mass-to-charge ratio (m/z) was 313.1468. FIG. 8 (C) is a 50. Mu. Mol/L solution of olanzapine dissolved in 20mmol/L Tris-acetate bufferThe addition peak [ M + H ] of olanzapine and proton is clearly seen in the figure] + The mass/charge ratio (m/z) was 313.1465. FIG. 8 (D) is a 50. Mu. Mol/L solution of olanzapine dissolved in 20mmol/L HEPES buffer solution, in which the addition peak [ M + H ] of olanzapine to proton is clearly seen] + The mass-to-charge ratio (m/z) was 313.1468.
To further demonstrate the desalting capacity of the device of the present invention, a 50 μmol/L solution of olanzapine dissolved in 20, 50, 100, 200mmol/L PBS buffer solution was tested using the device provided by the present invention. As can be seen from Table 3, in the HADSI device, as the salt concentration increased, the addition peak of olanzapine and proton [ M + H ]] + The strength and desalting efficiency did not change significantly. As can be seen from the attached figure 9, when the device provided by the utility model is used for detecting olanzapine dissolved in PBS buffer solution with different concentrations, the mass spectrum always keeps clean and clear and is not influenced by salt cluster ions.
TABLE 3 average Signal Strength and desalting efficiency of olanzapine in PBS buffer of various concentrations as measured by HADSI
Example 4:
in order to test the effect of the present invention, experimental studies were carried out on roxithromycin in salt solution using the device provided in example 1 and an ultimate UPLC/LTQ Orbitrap mass spectrometer.
1. Experimental Material
1.1 analyte to be detected
Roxithromycin standards (analytically pure, my shanghai mairei chemical technologies, ltd., same below), lot number: 80214-83-1. The acetone is firstly made into 10mmol/L mother liquor which is placed in a refrigerator for proper storage for experiments.
1.2 Roxithromycin solutions in PBS buffer solutions of different concentrations
0.5mol/L PBS buffer solution is used as PBS mother solution. 50 μmol/L rhodamine B solution (dissolved in 20, 50, 100, 200mmol/L PBS buffer) was prepared as in example 1
1.3 Roxithromycin solutions in different buffer solutions
50 μmol/L rhodamine B solution (dissolved in 20mmol/L borate, MES, tris-acetic acid, HEPES buffer) was prepared as in example 1
1.4 organic solvents
Purchasing methanol as an organic solvent
2 Experimental methods
Sample introduction conditions are as follows:
organic solvent: methanol at a flow rate of 0.2mL/min
Sample solution: the flow rate of the roxithromycin salt solution is 50 mu L/min
Atomizing: organic solvent spray module flow rate: 3L/min; sample spray generation module flow rate: 3L/min
Mass spectrum conditions:
a positive ion detection mode; the scanning mode is positive ion FULL SCAN (FULL SCAN)
HADSI ion source parameters: temperature of the electric heating sheet: 400 ℃;
3. results of the experiment
FIG. 10 shows the mass spectrum of roxithromycin dissolved in 20mmol/L buffer solution of different types, obtained by using the device of the present invention. FIG. 10 (A) is a 50. Mu. Mol/L roxithromycin solution dissolved in 20mmol/L borate buffer solution, in which the addition peak [ M + H ] of roxithromycin and proton is clearly seen] + The mass-to-charge ratio (m/z) was 837.5392. FIG. 10 (B) is a 50. Mu. Mol/L solution of roxithromycin dissolved in 20mmol/L MES buffer solution, in which the addition peak [ M + H ] of roxithromycin to proton is clearly seen] + The mass-to-charge ratio (m/z) was 837.5321. FIG. 10 (C) is a 50. Mu. Mol/L solution of roxithromycin dissolved in 20mmol/L Tris-acetate buffer solution, in which the addition peak [ M + H ] of roxithromycin and proton is clearly seen] + The mass/charge ratio (m/z) was 837.5395. FIG. 10 (D) is a 50. Mu. Mol/L solution of roxithromycin dissolved in 20mmol/L HEPES buffer solution, in which the addition peak [ M + H ] of roxithromycin to protons is clearly visible] + The mass/charge ratio (m/z) was 837.5396.
In order to further embody the desalting capability of the device of the utility model, the utility model providesThe apparatus detects a 50. Mu. Mol/L solution of roxithromycin dissolved in 20, 50, 100, 200mmol/L PBS buffer. As can be seen from Table 4, in the HADSI device, as the salt concentration increases, the addition peak [ M + H ] of roxithromycin and proton] + The strength and desalting efficiency did not change significantly. As can be seen from the attached figure 11, when the device provided by the utility model is used for detecting the roxithromycin dissolved in PBS buffer solutions with different concentrations, the mass spectrum chart is always kept clean and clear and is not influenced by salt cluster ions.
TABLE 4 average signal intensity and desalting efficiency of HADSI detection of Roxithromycin in PBS buffers of different concentrations
Example 5:
in order to test the utility model discloses detect the actual effect of unified kind medicine, adopt the device that embodiment 1 provided and ultate UPLC/LTQ Orbitrap mass spectrometer to carry out experimental study to erythromycin, roxithromycin, azithromycin, clarithromycin, midecamycin mixture in the salt solution.
1. Experimental Material
1.1 analyte to be detected
Erythromycin, roxithromycin, azithromycin, clarithromycin and midecamycin are respectively prepared into 10mmol/L mother liquor by methanol, and the mother liquor is placed in a refrigerator for proper storage for experiments.
1.2 erythromycin, roxithromycin, azithromycin, clarithromycin and midecamycin standard substance mixed solution
Respectively sucking 10 mu L of the five drug mother solutions into a 2mL centrifuge tube, and adding 1960 mu L of methanol to prepare a mixed solution of the four standard substances with the concentration of 50 mu mol/L.
1.3 organic solvents
Purchasing methanol as an organic solvent
2. Experimental methods
Sample introduction conditions are as follows:
organic solvent: methanol at a flow rate of 2mL/min
Atomizing: organic solvent spray module flow rate: 3L/min; sample spray generation module flow rate: 3L/min
Liquid phase conditions:
and (3) chromatographic column: eclipse XDB-C 18 (5mm,4.6μm 250mm)
Column temperature: 21 deg.C
Mobile phase: a is 1.8g/L NaH2PO4 (pH adjusted to 6.2 with NaOH), B is acetonitrile; a: B =70
Elution time: 30min
Flow rate: 1mL/min
Sample injection amount: 20 μ L
Mass spectrum conditions:
a positive ion detection mode; the scanning mode is positive ion FULL SCAN (FULL SCAN)
HADSI ion source parameters: temperature of the electric heating sheet: 500 ℃;
3. results of the experiment
Fig. 13 is a total ion flow diagram of the above experiment, and it can be seen that through the liquid phase, the five species are well separated.
Table 5 shows the highest signal intensities and corresponding retention times for the five substances detected. The attached diagram is a corresponding mass spectrogram when the five substances are detected.
TABLE 5 retention times and detection intensities for the four substances in example 5
As can be seen from the combination of Table 5 and FIG. 12, the method provided by the present invention can still realize efficient desalination when the salt solution is used as mobile phase, and on one hand, five macrolide antibiotics all exhibit adduction peak [ M + H ] with proton] + The relative signal intensity is high enough, and the interference of salt cluster ion peaks is avoided, so that the direct observation can be realized; on the other hand, when the salt-containing solution is continuously detected, the vacuum degree in the mass spectrum is not obviously reduced, which shows that the method can avoid the salt substances from blocking parts such as mass spectrum capillaries.
3. Conclusion
The utility model provides a but direct analysis contains liquid chromatography-mass spectrometry device of salt sample based on two spray flash evaporation. The device realizes high-efficient ionization and desalination effect through the mode that two strands of neutral sprays that do not apply high voltage direct action on the electric heat piece, and then realizes the direct analysis who contains salt solution or use the liquid phase eluant of salt solution as mobile phase. The device solves the problems that a transmission system is easy to block and an object to be detected is subjected to signal suppression of a salt cluster ion peak and the like when the conventional electrospray mass spectrometry is used for analyzing a salt solution. The device can directly analyze the substances to be detected in the buffer solutions with different concentrations and different types, can also realize the combination of liquid chromatography and mass spectrometry, and directly analyze the eluent with the salt solution as the mobile phase. Compared with other desalting methods, the utility model has the advantages of simple device, low energy consumption, good desalting effect, etc. In this meantime the utility model discloses a millimeter level gas circuit shower nozzle atomizer, and two strands of sprays act on the electric heat piece and not the mass spectrum entry, and these settings make this device have extremely strong salt tolerance, can hardly be blockked up by high concentration salt solution. In conclusion, the device provides a good solution for analyzing high-salt samples and provides a new reference for analysis in the fields of medicine, life science and the like in the future.
The above embodiments should not be understood as limiting the scope of application of the present invention, which is defined by the appended claims, and any modification based on the claims is the protection scope of the present invention.
Claims (8)
1. A dual spray flash evaporation based LC MS apparatus for analyzing salt-containing samples, comprising:
a base module;
the spraying fixing and control module and the electric heating piece fixing and control module are arranged on the base module;
the sample spray generating module and the organic solvent spray generating module are arranged on the spray fixing and controlling module;
the electric heating piece module is arranged on the electric heating piece fixing and controlling module;
the spray opening of the sample spray generation module and the spray opening of the organic solvent spray generation module face the emission end of the electric heating piece module.
2. The dual spray flash evaporation-based LC-MS device for analyzing salt-containing samples as claimed in claim 1, wherein the base module comprises a base and a fixed electric heating plate structure and a fixed spraying structure fixed on the base.
3. The dual spray rapid evaporation-based combined liquid and mass analysis device for salt-containing samples according to claim 2, wherein the electric heating plate fixing and control module is installed on the fixed electric heating plate structure, and the spray fixing and control module is installed on the fixed spray structure.
4. The dual spray flash evaporation based LC MS for analyzing salt-containing samples as claimed in claim 1, wherein said sample spray generation module comprises a spray generation module comprising: the liquid phase atomizer comprises a sample liquid phase atomizer, wherein an inlet of the sample liquid phase atomizer is connected with a gas path pipeline and a liquid path pipeline, an outlet of the sample liquid phase atomizer is connected with a liquid path sprayer and a gas path sprayer, and the gas path sprayer is wrapped on the liquid path sprayer.
5. The apparatus for LC-MS and MS for analyzing salt-containing sample based on dual-spray flash evaporation of claim 4, wherein the organic solvent spray generation module comprises: the device comprises an organic solvent liquid phase atomizer, wherein an inlet of the organic solvent liquid phase atomizer is connected with a gas path pipeline and a liquid path pipeline, an outlet of the organic solvent liquid phase atomizer is connected with a liquid path spray head and a gas path spray head, and the gas path spray head is wrapped on the liquid path spray head.
6. The dual spray rapid evaporation-based LC-MS device for analyzing salt-containing samples as claimed in claim 5, wherein the spray fixing and control module is a dual-hole fitting, the sample liquid phase atomizer is tightly sleeved in one hole, and the organic solvent liquid phase atomizer is tightly sleeved in another hole.
7. The dual spray flash evaporation-based integrated liquid chromatography-mass spectrometry device for analyzing salt-containing samples according to claim 1, wherein the electric heating plate module comprises: the electric heating piece is connected with the heat generating and adjusting device;
the electric heating piece is arranged between the sample spray generation module and the mass spectrum inlet.
8. The dual spray rapid evaporation-based combined liquid and mass spectrometer capable of analyzing salt-containing samples as claimed in claim 1, wherein the electric heating plate fixing and control module is a single-hole fitting, and the electric heating plate in the electric heating plate module is installed through the installation hole of the single-hole fitting.
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