Aerosol component analysis device and method in traditional Chinese medicinal material processing process

Abstract

The invention discloses an aerosol component analysis device and method in a traditional Chinese medicinal material processing process, and relates to the technical field of traditional Chinese medicinal material component analys.The aerosol component of a traditional Chinese medicinal material sample in the processing process is monitored through an ND-APCI-MS method, and metabolic difference substances in aerosol are screened in combination with a statistical means; components capable of reflecting the processing process are obtained and can be used for judging the processing end point, and rapid, in-situ and nondestructive analysis of the traditional Chinese medicinal material processing process is realized. The method is expected to be used for accurately predicting the end point of the traditional Chinese medicinal material processing process, is a very promising aerosol analysis method, and can be used for quality monitoring of the traditional Chinese medicinal material processing process.

Description

Technical Field

[0001] The present invention relates to the technical field of Chinese medicinal material component analysis, and in particular to an aerosol component analysis device and method in a Chinese medicinal material processing process.

[0002] Technical term: ND-APCI-MS refers to neutral desorption atmospheric pressure chemical ionization mass spectrometry. Background Art

[0003] Processing is a unique processing method for Chinese medicine. The efficacy of different medicinal materials after different processing methods is very different. For example, Rehmannia root, as a traditional Chinese medicinal material, has a long history of medicinal use and important clinical application value. The nature, taste, meridians and efficacy of its different processed products are obviously different. This is because Rehmannia root has a complex chemical composition.

[0004] At present, the commonly used methods for analyzing the components of traditional Chinese medicine and monitoring the processing process are mainly based on a series of analytical techniques developed based on the principles of spectroscopy, chromatography, and mass spectrometry. For example, high performance liquid chromatography (HPLC) (Yang J, et al. Molecules, 2024, 29, 705-717), high performance liquid chromatography mass spectrometry (HPLC-MS) (Zeng G, et al. Rapid Commu. Mass Spectrom. 2006, 20, 499-506), gas chromatography mass spectrometry (GC-MS) (Feng X, et al. Chromatographia, 2014, 77, 619-628), infrared spectroscopy (IR) (Yu L, et al. Spectrochim. Acta Part A: Mol. Bio. Spectrosc. 2005, 62, 22-29) and other analytical techniques have the advantages of high sensitivity, high separation efficiency, and good selectivity. However, these methods require sample pretreatment before analysis, which is time-consuming, easily leading to changes or even loss of substances, and cannot be monitored in real time, which greatly limits the practicality and application of these analytical techniques in the analysis of Chinese herbal medicine processing. Currently, there are few studies on the analysis of aerosol components in Chinese herbal medicines, and aerosol analysis can achieve rapid, in-situ, and non-destructive analysis of components, so it has broad application prospects in the field of Chinese herbal medicine processing detection. Summary of the invention

[0005] The purpose of the present invention is to solve at least one of the technical problems existing in the prior art and to provide an aerosol component analysis device and method for the preparation process of traditional Chinese medicine.

[0006] The technical solution of the present invention is as follows:

[0007] An aerosol component analysis device for a Chinese herbal medicine processing process, comprising a constant temperature heating platform, a processing container, an ionization device and a mass spectrometer;

[0008] The processing container is located on a constant temperature heating platform, and an air inlet pipe and an air outlet pipe are provided on the processing container;

[0009] The ionization device is used to ionize the substances at the gas outlet into ions so that the ions react with the analyte molecules, and after the reaction, the ions enter the mass spectrometer for detection and analysis.

[0010] As a preferred embodiment of the present invention, a filter screen is provided in the processing container.

[0011] As a preferred embodiment of the present invention, a sealing cover is provided on the preparation container, and an air inlet pipe and an air outlet pipe are located on the sealing cover.

[0012] As a preferred embodiment of the present invention, the ionization device is a discharge needle capable of applying high voltage, which is located between the gas outlet of the gas outlet pipe and the mass spectrometer port of the mass spectrometer.

[0013] The present invention also discloses a method for analyzing aerosol components in a process of preparing Chinese medicinal materials, which is applied to any of the above-mentioned devices, wherein Chinese medicinal materials are placed in a preparation container, a solvent is added, a sealing cover is put on, a constant temperature heating platform is started, nitrogen is introduced into an air inlet pipe at intervals, and an ionization device is started to ionize nitrogen and solvent molecules at an outlet so that the ions react with analyte molecules to obtain ionized aerosols, which are then sent into a mass spectrometer for detection and analysis.

[0014] As a preferred embodiment of the present invention, the detection and analysis specifically comprises the following steps:

[0015] S1: Construct the primary fingerprints of different preparation times to obtain the changes of aerosol metabolites; based on the changes of aerosol components, preliminarily screen out the substances with the most obvious changes;

[0016] S2: Construct an aerosol VIP score graph, and screen out differential components based on VIP>1 and P<0.05. Combine the signal changes and the substances obtained in S1 to determine the markers of the end point of Chinese medicinal material processing, thereby achieving quality monitoring of the processing process.

[0017] As a preferred embodiment of the present invention, the flow rate of the nitrogen is 0.1-0.2Mpa.

[0018] As a preferred solution of the present invention, the discharge needle is connected to a voltage of 3-4 kV.

[0019] The beneficial effects of the present invention are:

[0020] (1) The method of the present invention does not require sample pretreatment and can rapidly, in situ, and non-destructively analyze aerosol components during the preparation of traditional Chinese medicine.

[0021] (2) The processing container designed by the present invention can be applied to most traditional Chinese medicine samples that need to be steamed or calcined. Therefore, the present invention has strong universality.

[0022] (3) The method of the present invention has no restrictions on the detection environment and can be carried out at room temperature and pressure. The method has high sensitivity, fast analysis speed, and no interference with the preparation process.

[0023] (4) The present invention can determine the endpoint marker of the Chinese herbal medicine processing process, and is used for quality monitoring of the Chinese herbal medicine processing process, and is particularly suitable for quality monitoring of the Chinese herbal medicine Rehmannia glutinosa processing process. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a diagram of the aerosol component analysis device during the preparation of Rehmannia glutinosa;

[0025] 1-constant temperature heating platform, 2-filter screen, 3-processing container, 4-Chinese medicinal materials, 5-inlet pipe, 6-outlet pipe, 7-discharge needle, 8-mass spectrometer.

[0026] Figure 2 The ND-APCI-MS test results of aerosol samples at different time points during the processing of Rehmannia glutinosa; (a)-(m) are the primary fingerprint spectra of samples taken every 2 hours during the 24-hour processing;

[0027] Figure 3 The aerosol component analysis diagrams of the Rehmannia glutinosa processing process: (a) OPLS-DA score scatter plot of the aerosol sample after 24 hours of processing; (b) VIP score plot of the OPLS-DA dispersion scatter plot; (c) MS / MS spectrum of m / z 249 of the blank sample in the aerosol; (d) MS / MS spectrum of m / z 249 of the sample in the aerosol after 6 hours;

[0028] Figure 4 This is a diagram of the aerosol component analysis device during the preparation of raw rehmannia charcoal;

[0029] Figure 5 The aerosol component analysis diagrams during the processing of raw rehmannia charcoal: (a) total ion flow diagram of aerosol components during the processing of raw rehmannia charcoal; (b) m / z 249 ion flow diagram of aerosol components during the processing of raw rehmannia charcoal; (c) OPLS-DA score scatter diagram of aerosol samples processed for 20 minutes; (d) VIP score diagram of OPLS-DA dispersion scatter diagram;

[0030] In the figure, NL represents the ionic strength. DETAILED DESCRIPTION

[0031] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention. If no specific techniques or conditions are specified in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used that do not specify the manufacturer are all conventional products that can be obtained commercially.

[0032] This section will describe in detail the specific embodiments of the present invention. The preferred embodiments of the present invention are shown in the accompanying drawings. The purpose of the accompanying drawings is to supplement the description of the text part of the specification with graphics, so that people can intuitively and vividly understand each technical feature and the overall technical solution of the present invention, but it cannot be understood as a limitation on the scope of protection of the present invention.

[0033] In the description of the present invention, it should be understood that descriptions involving orientations, such as up, down, front, back, left, right, etc., and orientations or positional relationships indicated are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present invention.

[0034] In the description of the present invention, "several" means one or more, "more" means more than two, "greater than", "less than", "exceed" etc. are understood as not including the number itself, and "above", "below", "within" etc. are understood as including the number itself. If there is a description of "first" or "second", it is only used for the purpose of distinguishing the technical features, and cannot be understood as indicating or implying the relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0035] In the description of the present invention, unless otherwise clearly defined, terms such as setting, installing, connecting, etc. should be understood in a broad sense, and technicians in the relevant technical field can reasonably determine the specific meanings of the above terms in the present invention based on the specific content of the technical solution.

[0036] An aerosol component analysis device for the processing of traditional Chinese medicine, referring to Figure 1 , including a constant temperature heating platform 1, a preparation container 3, an ionization device and a mass spectrometer 8;

[0037] The processing container 3 is located on the constant temperature heating platform 1, and is provided with an air inlet pipe 5 and an air outlet pipe 6. Specifically, the air inlet pipe is 10 cm long and has an inner diameter of 1 mm, and the air outlet pipe is 7 cm long and has an inner diameter of 1 mm;

[0038] The ionization device is used to ionize the substances at the gas outlet into ions so that the ions react with the analyte molecules, and after the reaction, the ions enter the mass spectrometer 8 for detection and analysis.

[0039] Specifically, a filter screen 2 is provided in the processing container 3 and is made of stainless steel.

[0040] More specifically, a sealing cover is provided on the preparation container 3, and the air inlet pipe 5 and the air outlet pipe 6 are located on the sealing cover.

[0041] In some other embodiments, the ionization device is a discharge needle 7 capable of applying high voltage, which is located between the outlet of the outlet pipe 6 and the mass spectrometer port of the mass spectrometer 8. More specifically, the discharge needle 7 is 0.5 cm away from the outlet of the outlet pipe 6 and is located in the middle of the channel, and 1.5 cm away from the mass spectrometer port.

[0042] There are also some embodiments, a method for analyzing aerosol components in the process of preparing Chinese medicinal materials, which is applied to the device as described above, wherein Chinese medicinal materials 4 are placed in a preparation container 3, a solvent is added, a sealing cover is put on, the constant temperature heating platform 1 is started, nitrogen is introduced into the air inlet pipe 5 at intervals, and an ionization device is started to ionize the nitrogen and solvent molecules at the outlet so that the ions react with the analyte molecules to obtain ionized aerosol, which then enters the mass spectrometer 8 for detection and analysis.

[0043] Preferably, the flow rate of the nitrogen is 0.1-0.2Mpa.

[0044] Preferably, the discharge needle is connected to a voltage of 3-4 kV.

[0045] The specific steps include:

[0046] S1, put water into the processing container and put the Chinese medicinal materials on the stainless steel filter, or directly put the Chinese medicinal materials into the processing container, and continuously heat and process them using the constant temperature heating platform 1;

[0047] S2. During the processing of Chinese herbal medicine, its internal volatile and non-volatile components jointly generate aerosol components;

[0048] S3, N 2 The aerosol components to be tested are introduced through the air inlet pipe 5 and discharged through the air outlet pipe 6;

[0049] S4. Connect the discharge needle 7 placed at the outlet pipe 6 to a 3-4 kV high voltage to ionize some neutral molecules in the air and generate H 3 O + , N 2 + , O 2 + and O + Plasma, solvent molecules will also be ionized; these ions react with aerosol component molecules, transfer energy and exchange charges, and ionize the molecules in the aerosol components;

[0050] S5. The ionized aerosol components enter the mass spectrometer 8 for detection.

[0051] The technical solution of the present invention is further illustrated by specific experiments below.

[0052] Embodiment 1:

[0053] Experimental setup:

[0054] Aerosol component analysis device during the preparation of Rehmannia glutinosa Figure 1 As shown, the instrument consists of a constant temperature heating platform 1, a processing container 3 using a processing metal can, a Rehmannia root sample, water, a stainless steel filter, a discharge needle 7 and a mass spectrometer 8.

[0055] During the experiment, 70 ml of water was placed in the metal can, and the weighed Rehmannia sample (4 g) was placed in a stainless steel filter and the filter was fixed in the metal can. The container was sealed with a lid, and the container was heated at 100 °C using a constant temperature heating platform 1. The processing process lasted for 24 hours. As the reaction progressed during the heating, the volatile and non-volatile components produced during the processing of Rehmannia formed aerosol components. N was added from the air inlet every 2 hours. 2 The air is passed into the air inlet pipe 5, and the discharge needle 7 at the air outlet is connected to a high voltage of +3kV to ionize the neutral molecules near the air outlet. At the same time, the solvent molecules will also be ionized. These ions will react with the analyte molecules to ionize the analyte molecules and enter the mass spectrometer detection.

[0056] Figure 2 These are the ND-APCI-MS test results of aerosol samples at different time points during the processing of Rehmannia glutinosa. Figure 2 (a)-(m) are the primary fingerprints of samples taken every 2 hours during the 24 hours of processing. It can be seen from the figure that the peak intensities of m / z 205, m / z 249, m / z 279 and m / z 295 are significantly lower at the end of processing than during the processing, among which m / z 249 changes the most significantly. Figure 2 In (a), it can be seen that no m / z 249 signal peak was observed at the beginning of the preparation. Figure 2 In (d), we can see that the m / z 249 signal peak appears and reaches the highest value. Then, as time goes by, its signal intensity gradually decreases to the peak at 24h. Figure 2 In the middle (m), the signal peak almost completely disappeared. This shows that during the preparation process, the changes in aerosol components have certain regularity and trends, and the m / z 249 signal peak may be the key component.

[0057] In order to further obtain the possible markers of aerosol components in the processing process, multivariate statistical analysis of the processing process was performed, such as Figure 3 shown. Figure 3(a) is the scatter plot of OPLS-DA of aerosol samples during the preparation of Rehmannia glutinosa. The cumulative explanation rate R 2 X and R 2 Y are 0.984 and 0.764 respectively, and the cumulative predictive ability Q 2 The value is 0.774, indicating that the model has a good fitting effect. As can be seen from the figure, there is a certain separation trend between samples in different time periods, which indicates that the aerosol components change significantly during the preparation process. Figure 3 (b) is the variable importance projection (VIP) diagram of each component, which is used to measure the influence and discrimination ability of each component on the classification and discrimination of samples. 16 differential components can be screened out with VIP>1 and P<0.05 as the standard, among which m / z 93, m / z 249, m / z 148, m / z 391 and m / z 109 have VIP>2, indicating that these substances have a stronger discriminative effect in distinguishing the aerosol components of Rehmannia glutinosa. Among them, m / z 249 has the most obvious change in the fingerprint spectrum, so it was subjected to tandem mass spectrometry analysis. Figure 3 (c) and Figure 3 (d) in the figure are the tandem mass spectrometry results of m / z 249 of the aerosol components at 0h and 6h during the preparation process. Figure 3 The fragment ion of m / z 249 when no sample was added in (c) is shown in Figure 2. Figure 3 In (d), m / z 249 has some new major fragment ions such as m / z 231, m / z 213, m / z 151 and m / z 135, indicating that m / z 249 is a new substance produced during the processing process and is not always present in the air. The experimental results show that the method of processing Rehmannia glutinosa for 24 hours is feasible, and the signal intensity of m / z 249 changes significantly during the processing process and almost disappears at the end of the processing. When the VIP value is high, it shows that it has a strong ability to distinguish samples of aerosol components during the processing process. Therefore, this example uses it as a potential processing end point marker for the processing process of Rehmannia glutinosa.

[0058] The results show that the traditional Chinese medicine aerosol component analysis method of the present invention can be used for quality monitoring of the processing process of Rehmannia glutinosa, with fast analysis speed and no influence on the processing process.

[0059] Embodiment 2:

[0060] Experimental setup:

[0061] The aerosol component analysis device during the preparation of raw rehmannia charcoal is as follows Figure 4 As shown, the instrument consists of a constant temperature heating platform 1, a processing container 3 using a processing metal can, a Rehmannia root sample, a discharge needle 7 and a mass spectrometer 8.

[0062] During the experiment, the container was heated at 200°C for 5 min using a constant temperature heating platform 1, and then the weighed raw Rehmannia root sample (4 g) was placed in the metal can, and the container was sealed with a lid. The processing process lasted for 20 min. As the reaction progressed, the volatile and non-volatile components produced during the processing of raw Rehmannia root charcoal formed aerosol components, and N was continuously introduced from the air inlet. 2 , connect the discharge needle 7 at the outlet to a +3kV high voltage to ionize the neutral molecules near the outlet, and the solvent molecules will also be ionized. These ions will react with the analyte molecules to ionize the analyte molecules and enter the mass spectrometry detection.

[0063] Figure 5 This is an analysis chart of aerosol components during the processing of raw Rehmannia charcoal. Figure 5 (a) and Figure 5 (b) are the total ion flow diagram of the aerosol components in the processing process of raw rehmannia charcoal and the ion flow diagram of m / z 249, respectively. It can be seen from the figure that the total ion flow diagram shows a trend of first rising and then falling during the processing process from 8min to 28min and decreases to the lowest value at 28min, indicating that the content of aerosol components decreases and tends to be stable at this time, and it is judged that the reaction endpoint has been reached. The overall trend of the ion flow diagram of m / z 249 is basically consistent with the total ion flow diagram, and it is also a marker of the processing endpoint in Example 1, so it is judged that there is a similar reaction or a type of reaction in the two processing processes, and the reaction produces a signal peak of m / z 249, which can judge the degree of reaction and thus monitor the processing endpoint of raw rehmannia charcoal.

[0064] Figure 5 (c) is the scatter plot of OPLS-DA of aerosol samples during the processing of raw earth charcoal. The cumulative explanation rate R 2 X and R 2 Y are 0.991 and 0.979 respectively, and the cumulative predictive ability Q 2 The value is 0.926, indicating that the model has a good fitting effect. As can be seen from the figure, there is a slight separation trend between samples in different time periods, which indicates that the aerosol components change significantly during the preparation process. Figure 5 (d) is the variable importance projection (VIP) diagram of each component, which is used to measure the influence and discrimination ability of each component on sample classification. 14 differential components can be screened out with VIP>2 and P<0.05 as the standard. Compared with the differential components in the processing of Rehmannia glutinosa, m / z 249 appeared again, indicating that it also has a discriminative effect in the aerosol component differentiation during the processing of Rehmannia glutinosa charcoal. This result once again verifies the feasibility of the m / z 249 signal peak as a marker for distinguishing the end point of Rehmannia glutinosa and Rehmannia glutinosa charcoal processing.

[0065] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. An aerosol component analysis device for the processing of traditional Chinese medicine, characterized in that: It comprises a constant temperature heating platform (1), a preparation container (3), an ionization device and a mass spectrometer (8); The processing container (3) is located on the constant temperature heating platform (1), and an air inlet pipe (5) and an air outlet pipe (6) are provided on the processing container (3); The ionization device is used to ionize the substances at the gas outlet into ions so that the ions react with the analyte molecules and then enter the mass spectrometer (8) for detection and analysis.

2. The aerosol component analysis device for the processing of traditional Chinese medicine according to claim 1, characterized in that: A filter screen (2) is arranged inside the processing container (3).

3. The aerosol component analysis device for the processing of traditional Chinese medicine according to claim 1, characterized in that: The preparation container (3) is provided with a sealing cover, and an air inlet pipe (5) and an air outlet pipe (6) are located on the sealing cover.

4. The aerosol component analysis device for the processing of traditional Chinese medicine according to claim 1, characterized in that: The ionization device is a discharge needle (7) capable of applying high voltage, which is located between the gas outlet of the gas outlet pipe (6) and the mass spectrometer port of the mass spectrometer (8).

5. A method for analyzing aerosol components in the process of preparing Chinese medicinal materials, applied to the device as described in any one of claims 1 to 4, characterized in that: Chinese medicinal materials (4) are placed in a processing container (3), a solvent is added, a sealing cover is put on, a constant temperature heating platform (1) is started, nitrogen is introduced into the air inlet pipe (5) at intervals, and an ionization device is started to ionize the nitrogen and solvent molecules at the outlet so that the ions react with the analyte molecules to obtain ionized aerosol, which then enters a mass spectrometer (8) for detection and analysis. The metabolic differences in the aerosol are screened by combining statistical methods to obtain components that can reflect the processing process and can be used to determine the processing end point.

6. The method for analyzing aerosol components in the processing of traditional Chinese medicine according to claim 5, characterized in that: The detection analysis specifically comprises the following steps: S1: Construct the primary fingerprints of different preparation times to obtain the changes of aerosol metabolites; based on the changes of aerosol components, preliminarily screen out the substances with the most obvious changes; S2: Construct an aerosol VIP score graph, and screen out differential components based on VIP>1 and P<0.

05. Combine the signal changes and the substances obtained in S1 to determine the markers of the end point of Chinese medicinal material processing.

7. The method for analyzing aerosol components in the processing of Chinese medicinal materials according to claim 5, characterized in that: The flow rate of the nitrogen is 0.1-0.2Mpa.

8. The method for analyzing aerosol components in the processing of traditional Chinese medicine according to claim 5, characterized in that: The discharge needle is connected to a voltage of 3-4 kV.

Images