Method for detecting premna pubescens leaf flavor substances based on solid-phase microextraction and gas chromatography-mass spectrometry

Abstract

The invention discloses a method for detecting flavor substances in leaves of premna pubescens based on solid-phase microextraction and gas chromatography-mass spectrometry, which can efficiently qualitatively and quantitatively analyze chemical components of volatile flavor substances in the leaves of premna pubescens. The problem that in the prior art, key volatile flavor substance components of the premna pubescens leaves cannot be extracted is solved. The method disclosed by the invention has the advantages of simplicity in operation, low cost, good repeatability, high automation degree and the like.

Description

Field of the Invention

[0001] The invention relates to the technical field of component analysis of oleander leaves, and in particular to a method for detecting flavor substances in oleander leaves based on solid phase microextraction and gas chromatography-mass spectrometry (GC-MS) technology. Background Art

[0002] Premna puberula Pamb. is a perennial shrub of the Verbenaceae family, mainly distributed in Guizhou, Yunnan, Sichuan and other places in my country. At present, the research on pharmacological effects is mainly focused on the same genus Premna microphylla Turcz. The extract has significant anti-inflammatory, tumor prevention, analgesic and other functions, which are closely related to its components. Its leaves are rich in various components. At present, the chemical composition research of the leaves is mainly focused on protein, pectin, flavonoids, etc., and there is little research on its volatile oil components, and there is no research on the volatile oil of the leaves of the Verbenaceae family Premna microphylla.

[0003] The present invention combines gas chromatography with mass spectrometry to efficiently perform qualitative and quantitative analysis on the chemical components of volatile flavor substances in the herb. The volatile components of the herb can complete the entire process of sampling-extraction-analysis and detection, and can achieve the purpose of qualitative and quantitative analysis of multiple volatile flavor substances in the herb in a short time. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide a method for detecting flavor substances in the leaves of the herbaceous wormwood based on solid phase microextraction and gas chromatography-mass spectrometry technology. The method can efficiently perform qualitative and quantitative analysis on the chemical components of volatile flavor substances in the leaves of the herbaceous wormwood, so as to solve the problem that the existing technology cannot extract the key volatile flavor substance components of the leaves of the herbaceous wormwood.

[0005] The present invention is implemented by the following technical solutions:

[0006] A method for detecting flavor substances in leaves of Psoralea corylifolia based on solid phase microextraction and gas chromatography-mass spectrometry technology comprises the following steps:

[0007] a. Pretreatment of the leaf samples of the Chinese yew vine: take the leaves of the Chinese yew vine and clean the surface mud and sand with tap water, rinse with single distilled water for 1-3 times, then soak with 75% ethanol for 20-40 seconds, rinse with sterilized double distilled water for 1-3 times, dry with sterile filter paper and store in liquid nitrogen for 1-24 hours, take out the leaf samples of the Chinese yew vine stored in liquid nitrogen and quickly grind them into a homogenized state, weigh 1-2g of the homogenized sample and place it in the sampling bottle of the solid phase microextraction instrument, add 0.1-0.3μL of the internal standard 2-octanol, and then add 0.5-2.5g of sodium chloride and 1-5mL of ultrapure water to the headspace bottle of the solid phase microextraction to seal the bottle mouth to obtain product A;

[0008] b. Headspace solid phase microextraction: The solid phase microextraction sampling bottle containing product A was equilibrated at 55-65°C for 15-25 minutes, then inserted into a manual injector equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber head, preheated at 55-65°C for 5-15 minutes, and headspace extracted for 30-50 minutes to obtain product B;

[0009] c. Qualitative and quantitative analysis: Remove the extraction head with product B and immediately insert it into the injection port of the gas chromatography-mass spectrometry instrument. After 6 to 10 minutes of thermal desorption, inject the sample for qualitative and quantitative analysis.

[0010] In the above step a, the pretreatment of the leaf sample of the Herba Lycopodii is as follows: take the Herba Lycopodii leaves, clean the mud and sand on the surface with tap water, rinse with single distilled water for 2-3 times, soak with 75% ethanol for 25-35 seconds, and then rinse with sterilized double distilled water for 2-3 times, dry with sterile filter paper and store in liquid nitrogen for 1-24 hours, take out the Herba Lycopodii leaf samples stored in liquid nitrogen and quickly grind them into a homogenate state, weigh 1.2-1.8g of the homogenate sample and place it in a sampling bottle of a solid phase microextraction instrument, add 0.15-0.25μL of the internal standard 2-octanol, and then add 0.8-2.2g of sodium chloride and 1.5-4.5mL of ultrapure water to the solid phase microextraction headspace bottle to seal the bottle mouth to obtain product A.

[0011] Specifically, in the aforementioned step a, the pretreatment of the leaf sample of the Herba Lycopodii is as follows: take the Herba Lycopodii leaves, clean the surface mud and sand with tap water, rinse twice with single distilled water, then soak with 75% ethanol for 30 seconds, rinse twice with sterilized double distilled water, blot dry with sterile filter paper and store in liquid nitrogen for 1-24 hours, take out the Herba Lycopodii leaf samples stored in liquid nitrogen and quickly grind them into a homogenate state, weigh 1.5g of the homogenate sample and place it in the sampling bottle of the solid phase microextraction instrument, add 0.2μL of the internal standard 2-octanol, and then add 1.0-2.0g of sodium chloride and 2-4mL of ultrapure water to the solid phase microextraction headspace bottle to seal the bottle mouth to obtain product A.

[0012] In the above step b, headspace solid phase microextraction: the solid phase microextraction sampling bottle containing product A is equilibrated at 58-62°C for 18-22 minutes, then inserted into a manual injector equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber head, preheated under flat plate heating conditions at 58-62°C for 8-12 minutes, and headspace extraction was performed for 35-45 minutes to obtain product B.

[0013] Specifically, in the above step b, headspace solid phase microextraction: the solid phase microextraction sampling bottle containing product A is equilibrated at 60°C for 20 minutes, then inserted into a manual injector equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber head, preheated under 60°C flat plate heating conditions for 10 minutes, and headspace extraction for 40 minutes to obtain product B.

[0014] In the above step c, the thermal desorption time is 8 minutes.

[0015] In the aforementioned step c, the gas chromatography conditions are as follows: chromatograph: a GC-MS / MS coupling instrument is used; the chromatographic column is an HP-5MS elastic quartz capillary column, 30m*250μm*0.25μm; the initial temperature is 40°C and maintained for 2min, then the temperature is increased to 180°C at 3.5°C / min, and then the temperature is increased to 310°C at 10°C / min, and the running time is 55min; the vaporization chamber temperature is 250°C; the carrier gas is 99.999% high-purity He; the column head pressure is 7.1866psi, the carrier gas flow rate is 1.0mL / min, the split ratio is 5:1, and the solvent delay time is 3min.

[0016] In the above step c, the mass spectrometry conditions are: the ion source is an EI source; the ion source temperature is 230°C; the quadrupole temperature is 150°C; the electron energy is 70eV; the emission current is 34.6μA; the multiplier voltage is 2188V; the interface temperature is 280°C; and the mass range is 29-500amu.

[0017] In the above step c, the qualitative analysis is completed by the automatic deconvolution system AMDIS of the computer mass spectrometry workstation and the reference analysis of the standard mass spectrum library NIST 2020.

[0018] In the above step c, the quantitative analysis adopts the internal standard quantitative analysis method to determine the relative percentage content.

[0019] Beneficial effects of the present invention:

[0020] (1) When headspace-solid phase microextraction and gas chromatography-mass spectrometry are used together, the volatile components in the leaves of P. foetida can complete the entire process of sampling-extraction-analysis and detection, and can achieve qualitative analysis of multiple volatile components in a short period of time. 63 volatile components were detected and identified;

[0021] (2) The present invention uses the internal standard method to quantitatively determine the volatile flavor species in the leaves of the herb, and the deviation is no more than 5%.

[0022] (3) The present invention has the advantages of simple operation, low cost, good repeatability and high degree of automation. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1: is a total ion current diagram for analyzing the volatile components of leaves of Herba Lysimachiae provided in an embodiment of the present invention.

[0024] In order to enable those skilled in the art to better understand and implement the technical solution of the present invention, the present invention is further described below in conjunction with specific embodiments, but the embodiments are not intended to limit the present invention. DETAILED DESCRIPTION

[0025] Embodiment 1:

[0026] The method for detecting the flavor substances of the leaves of the Herba Lysimachiae, comprises the following steps:

[0027] a. Pretreatment of the leaf samples of the Herba Lysimachiae: Take the Herba Lysimachiae leaves and clean the surface mud and sand with tap water, rinse with single distilled water twice, then soak with 75% ethanol for 30 seconds, rinse with sterilized double distilled water twice, dry with sterile filter paper and store in liquid nitrogen for 1-24 hours, take out the Herba Lysimachiae leaf samples stored in liquid nitrogen and quickly grind them into a homogenate state, weigh 1.5g of the homogenate sample and place it in the sampling bottle of the solid phase microextraction instrument, add 0.2μL of the internal standard 2-octanol, and then add 1.0-2.0g of sodium chloride and 2-4mL of ultrapure water to the solid phase microextraction headspace bottle to seal the bottle mouth to obtain product A;

[0028] b. Headspace solid phase microextraction: The solid phase microextraction sampling bottle containing product A was equilibrated at 60℃ for 20 minutes, then inserted into a manual injector equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber head, preheated at 60℃ for 10 minutes, and headspace extraction was performed for 40 minutes to obtain product B.

[0029] c. Qualitative and quantitative analysis: Remove the extraction head with product B and immediately insert it into the injection port of the gas chromatography-mass spectrometry instrument. After 8 minutes of thermal desorption, inject the sample for qualitative and quantitative analysis. The qualitative analysis is completed by the automatic deconvolution system AMDIS of the computer mass spectrometry workstation and the standard mass spectrum library NIST 2020 for reference analysis. The quantitative analysis adopts the internal standard quantitative analysis method.

[0030] Gas chromatography conditions: Chromatographic column: GC-MS / MS coupling instrument (Agilent, USA, 8890 / 7000D) was used. The chromatographic column was HP-5MS (30m*250μm*0.25μm) elastic quartz capillary column, initial temperature 40°C (retained for 2min), heated to 180°C at 3.5°C / min, and then heated to 310°C at 10°C / min, running time: 55min; vaporization chamber temperature 250°C; carrier gas was high-purity He (99.999%); column head pressure 7.1866psi, carrier gas flow rate 1.0mL / min, split ratio 5:1, solvent delay time: 3min.

[0031] Mass spectrometry conditions: ion source is EI source; ion source temperature is 230°C; quadrupole temperature is 150°C; electron energy is 70eV; emission current is 34.6μA; multiplier voltage is 2188V; interface temperature is 280°C; mass range is 29-500amu.

[0032] Embodiment 2:

[0033] The method for detecting the flavor substances of the leaves of the Herba Lysimachiae, comprises the following steps:

[0034] a. Pretreatment of the leaf samples of the Herba Lysimachiae: Take the Herba Lysimachiae leaves and clean the surface mud and sand with tap water, rinse once with single distilled water, then soak with 75% ethanol for 20 seconds, rinse three times with sterilized double distilled water, dry with sterile filter paper and store in liquid nitrogen for 1-24 hours, take out the Herba Lysimachiae leaf samples stored in liquid nitrogen and quickly grind them into a homogenate state, weigh 2g of the homogenate sample and place it in the sampling bottle of the solid phase microextraction instrument, add 0.1μL of the internal standard 2-octanol, and then add 0.5g of sodium chloride and 5mL of ultrapure water to the solid phase microextraction headspace bottle to seal the bottle mouth to obtain product A;

[0035] b. Headspace solid phase microextraction: The solid phase microextraction sampling bottle containing product A was equilibrated at 55°C for 15 minutes, then inserted into a manual injector equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber head, preheated at 65°C for 15 minutes, and headspace extracted for 30 minutes to obtain product B;

[0036] c. Qualitative and quantitative analysis: Remove the extraction head with product B and immediately insert it into the injection port of the gas chromatography-mass spectrometry instrument. After 6 minutes of thermal desorption, inject the sample for qualitative and quantitative analysis. The qualitative analysis is completed by the automatic deconvolution system AMDIS of the computer mass spectrometry workstation and the standard mass spectrum library NIST 2020 for reference analysis. The quantitative analysis adopts the internal standard quantitative analysis method.

[0037] Gas chromatography conditions: Chromatographic column: GC-MS / MS coupling instrument (Agilent, USA, 8890 / 7000D) was used. The chromatographic column was HP-5MS (30m*250μm*0.25μm) elastic quartz capillary column, initial temperature 40°C (retained for 2min), heated to 180°C at 3.5°C / min, and then heated to 310°C at 10°C / min, running time: 55min; vaporization chamber temperature 250°C; carrier gas was high-purity He (99.999%); column head pressure 7.1866psi, carrier gas flow rate 1.0mL / min, split ratio 5:1, solvent delay time: 3min.

[0038] Mass spectrometry conditions: ion source is EI source; ion source temperature is 230°C; quadrupole temperature is 150°C; electron energy is 70eV; emission current is 34.6μA; multiplier voltage is 2188V; interface temperature is 280°C; mass range is 29-500amu.

[0039] Embodiment 3:

[0040] The method for detecting the flavor substances of the leaves of the Herba Lysimachiae, comprises the following steps:

[0041] a. Pretreatment of the leaf samples of the Herba Lysimachiae: Take the Herba Lysimachiae leaves and clean the surface mud and sand with tap water, rinse with single distilled water for 3 times, then soak with 75% ethanol for 40 seconds, rinse with sterilized double distilled water once, dry with sterile filter paper and store in liquid nitrogen for 1-24 hours, take out the Herba Lysimachiae leaves stored in liquid nitrogen and quickly grind them into a homogenate state, weigh 1g of the homogenate sample and place it in the sampling bottle of the solid phase microextraction instrument, add 0.3μL of the internal standard 2-octanol, and then add 2.5g of sodium chloride and 1mL of ultrapure water to the solid phase microextraction headspace bottle to seal the bottle mouth to obtain product A;

[0042] b. Headspace solid phase microextraction: The solid phase microextraction sampling bottle containing product A was equilibrated at 65°C for 25 minutes, then inserted into a manual injector equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber head, preheated at 55-65°C for 5 minutes, and headspace extracted for 50 minutes to obtain product B;

[0043] c. Qualitative and quantitative analysis: Remove the extraction head with product B and immediately insert it into the injection port of the gas chromatography-mass spectrometry instrument. After 10 minutes of thermal desorption, inject the sample for qualitative and quantitative analysis. The qualitative analysis is completed by the automatic deconvolution system AMDIS of the computer mass spectrometry workstation and the standard mass spectrum library NIST 2020 for reference analysis. The quantitative analysis adopts the internal standard quantitative analysis method.

[0044] Gas chromatography conditions: Chromatographic column: GC-MS / MS coupling instrument (Agilent, USA, 8890 / 7000D) was used. The chromatographic column was HP-5MS (30m*250μm*0.25μm) elastic quartz capillary column, initial temperature 40°C (retained for 2min), heated to 180°C at 3.5°C / min, and then heated to 310°C at 10°C / min, running time: 55min; vaporization chamber temperature 250°C; carrier gas was high-purity He (99.999%); column head pressure 7.1866psi, carrier gas flow rate 1.0mL / min, split ratio 5:1, solvent delay time: 3min.

[0045] Mass spectrometry conditions: ion source is EI source; ion source temperature is 230°C; quadrupole temperature is 150°C; electron energy is 70eV; emission current is 34.6μA; multiplier voltage is 2188V; interface temperature is 280°C; mass range is 29-500amu.

[0046] Embodiment 4:

[0047] The method for detecting the flavor substances of the leaves of the Herba Lysimachiae, comprises the following steps:

[0048] a. Pretreatment of the leaf samples of the Herba Lysimachiae: Take the Herba Lysimachiae leaves and clean the surface mud and sand with tap water, rinse with single distilled water twice, then soak with 75% ethanol for 35 seconds, rinse with sterilized double distilled water three times, dry with sterile filter paper and store in liquid nitrogen for 1-24 hours, take out the Herba Lysimachiae leaf samples stored in liquid nitrogen and quickly grind them into a homogenate state, weigh 1.2g of the homogenate sample and place it in the sampling bottle of the solid phase microextraction instrument, add 0.25μL of the internal standard 2-octanol, and then add 1.5g of sodium chloride and 2mL of ultrapure water to the solid phase microextraction headspace bottle to seal the bottle mouth to obtain product A;

[0049] b. Headspace solid phase microextraction: The solid phase microextraction sampling bottle containing product A was equilibrated at 60°C for 18 minutes, then inserted into a manual injector equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber head, preheated at 60°C for 12 minutes, and headspace extracted for 40 minutes to obtain product B;

[0050] c. Qualitative and quantitative analysis: Remove the extraction head with product B and immediately insert it into the injection port of the gas chromatography-mass spectrometry instrument. After 9 minutes of thermal desorption, inject the sample for qualitative and quantitative analysis. The qualitative analysis is completed by the automatic deconvolution system AMDIS of the computer mass spectrometry workstation and the standard mass spectrum library NIST 2020 for reference analysis. The quantitative analysis adopts the internal standard quantitative analysis method.

[0051] Gas chromatography conditions: Chromatographic column: GC-MS / MS coupling instrument (Agilent, USA, 8890 / 7000D) was used. The chromatographic column was HP-5MS (30m*250μm*0.25μm) elastic quartz capillary column, initial temperature 40°C (retained for 2min), heated to 180°C at 3.5°C / min, and then heated to 310°C at 10°C / min, running time: 55min; vaporization chamber temperature 250°C; carrier gas was high-purity He (99.999%); column head pressure 7.1866psi, carrier gas flow rate 1.0mL / min, split ratio 5:1, solvent delay time: 3min.

[0052] Mass spectrometry conditions: ion source is EI source; ion source temperature is 230°C; quadrupole temperature is 150°C; electron energy is 70eV; emission current is 34.6μA; multiplier voltage is 2188V; interface temperature is 280°C; mass range is 29-500amu.

[0053] Embodiment 5:

[0054] The method for detecting the flavor substances of the leaves of the Herba Lysimachiae, comprises the following steps:

[0055] a. Pretreatment of the leaf samples of the Herba Lysimachiae: Take the Herba Lysimachiae leaves and clean the surface mud and sand with tap water, rinse with single distilled water for 3 times, then soak with 75% ethanol for 25 seconds, rinse with sterilized double distilled water for 2 times, dry with sterile filter paper and store in liquid nitrogen for 1-24 hours, take out the Herba Lysimachiae leaves stored in liquid nitrogen and quickly grind them into a homogenate state, weigh 1.8g of the homogenate sample and place it in the sampling bottle of the solid phase microextraction instrument, add 0.15μL of the internal standard 2-octanol, and then add 1.0g of sodium chloride and 3mL of ultrapure water to the solid phase microextraction headspace bottle to seal the bottle mouth to obtain product A;

[0056] b. Headspace solid phase microextraction: The solid phase microextraction sampling bottle containing product A was equilibrated at 62°C for 22 minutes, then inserted into a manual injector equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber head, preheated at 62°C for 8 minutes, and headspace extracted for 45 minutes to obtain product B;

[0057] c. Qualitative and quantitative analysis: Remove the extraction head with product B and immediately insert it into the injection port of the gas chromatography-mass spectrometry instrument. After 7 minutes of thermal desorption, inject the sample for qualitative and quantitative analysis. The qualitative analysis is completed by the automatic deconvolution system AMDIS of the computer mass spectrometry workstation and the standard mass spectrum library NIST 2020 for reference analysis. The quantitative analysis adopts the internal standard quantitative analysis method.

[0058] Gas chromatography conditions: Chromatographic column: GC-MS / MS coupling instrument (Agilent, USA, 8890 / 7000D) was used. The chromatographic column was HP-5MS (30m*250μm*0.25μm) elastic quartz capillary column, initial temperature 40°C (retained for 2min), heated to 180°C at 3.5°C / min, and then heated to 310°C at 10°C / min, running time: 55min; vaporization chamber temperature 250°C; carrier gas was high-purity He (99.999%); column head pressure 7.1866psi, carrier gas flow rate 1.0mL / min, split ratio 5:1, solvent delay time: 3min.

[0059] Mass spectrometry conditions: ion source is EI source; ion source temperature is 230°C; quadrupole temperature is 150°C; electron energy is 70eV; emission current is 34.6μA; multiplier voltage is 2188V; interface temperature is 280°C; mass range is 29-500amu.

[0060] The present invention has done a lot of analysis and verification experiments, and the following are the results of the experimental research of the present invention:

[0061] 1. Preparation of Alcohol Extract from the Leaves of Psoralea corylifolia

[0062] Experimental materials P.puberula were collected from Wuchuan Gelao and Miao Autonomous County, Guizhou Province. The research team has studied and established its seedling breeding technology. In early 2019, the two-year-old seedlings of P.puberula were planted at the scientific research base of Guizhou University (26°25'54"N, 106°40′53″E, with a subtropical humid monsoon climate, an average altitude of 1178m, an average annual temperature of 20.21℃, and an average annual precipitation of 1318.90mm). Leaves were collected on July 30, 2023 for related research.

[0063] According to the detection method of Example 1, the flavor substances of the leaves of the Herba Lysimachiae were qualitatively and quantitatively analyzed. The analysis results are shown in Table 1:

[0064] Table 1 Detection of flavor components of the leaves of Psoralea corylifolia by solid phase microextraction

[0065]

[0066]

[0067]

[0068]

[0069] The invention detects 63 flavor substances in the leaves of the herb of radix scutellariae by solid phase microextraction and gas chromatography-mass spectrometry technology, and the quantitative determination deviation is no more than 5%.

Claims

1. A method for detecting flavor substances in leaves of Psoralea corylifolia based on solid phase microextraction and gas chromatography-mass spectrometry, characterized in that: The following steps are involved: a. Pretreatment of the leaf samples of the Chinese yew vine: take the leaves of the Chinese yew vine and clean the surface mud and sand with tap water, rinse with single distilled water for 1-3 times, then soak with 75% ethanol for 20-40 seconds, rinse with sterilized double distilled water for 1-3 times, dry with sterile filter paper and store in liquid nitrogen for 1-24 hours, take out the leaf samples of the Chinese yew vine stored in liquid nitrogen and quickly grind them into a homogenized state, weigh 1-2g of the homogenized sample and place it in the sampling bottle of the solid phase microextraction instrument, add 0.1-0.3μL of the internal standard 2-octanol, and then add 0.5-2.5g of sodium chloride and 1-5mL of ultrapure water to the headspace bottle of the solid phase microextraction to seal the bottle mouth to obtain product A; b. Headspace solid phase microextraction: The solid phase microextraction sampling bottle containing product A was equilibrated at 55-65°C for 15-25 minutes, then inserted into a manual injector equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber head, preheated at 55-65°C for 5-15 minutes, and headspace extracted for 30-50 minutes to obtain product B; c. Qualitative and quantitative analysis: Remove the extraction head with product B and immediately insert it into the injection port of the gas chromatography-mass spectrometry instrument. After 6 to 10 minutes of thermal desorption, inject the sample for qualitative and quantitative analysis.

2. The method for detecting flavor substances in leaves of Psoralea corylifolia based on solid phase microextraction and gas chromatography-mass spectrometry according to claim 1, characterized in that: In the step a, the pretreatment of the leaf sample of the Herba Lysimachiae is as follows: the Herba Lysimachiae leaves are taken and the mud and sand on the surface are cleaned with tap water, rinsed with single distilled water for 2-3 times, soaked with 75% ethanol for 25-35 seconds, and then rinsed with sterilized double distilled water for 2-3 times, dried with sterile filter paper and stored in liquid nitrogen for 1-24 hours, the Herba Lysimachiae leaf samples stored in liquid nitrogen are taken out and quickly ground into a homogenate state, 1.2-1.8 g of the homogenate sample is weighed and placed in a sampling bottle of a solid phase microextraction instrument, 0.15-0.25 μL of the internal standard 2-octanol is added, and then 0.8-2.2 g of sodium chloride and 1.5-4.5 mL of ultrapure water are added to the solid phase microextraction headspace bottle to seal the bottle mouth to obtain product A.

3. The method for detecting flavor substances of leaves of Psoralea corylifolia based on solid phase microextraction and gas chromatography-mass spectrometry according to claim 2, characterized in that: In the step a, the pretreatment of the leaf sample of the Herba Lycopodii is as follows: the Herba Lycopodii leaves are taken and the mud and sand on the surface are cleaned with tap water, rinsed twice with single distilled water, then immersed in 75% ethanol for 30 seconds, rinsed twice with sterilized double distilled water, blotted dry with sterile filter paper and stored in liquid nitrogen for 1-24 hours, the Herba Lycopodii leaf sample stored in liquid nitrogen is taken out and quickly ground into a homogenate state, 1.5 g of the homogenate sample is weighed and placed in a sampling bottle of a solid phase microextraction instrument, 0.2 μL of the internal standard 2-octanol is added, and then 1.0-2.0 g of sodium chloride and 2-4 mL of ultrapure water are added to the solid phase microextraction headspace bottle to seal the bottle mouth to obtain product A.

4. The method for detecting flavor substances in leaves of Psoralea corylifolia based on solid phase microextraction and gas chromatography-mass spectrometry according to claim 1, characterized in that: In the step b, headspace solid phase microextraction: the solid phase microextraction sampling bottle containing product A is equilibrated at 58-62° C. for 18-22 minutes, then inserted into a manual injector equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber head, preheated under flat plate heating conditions at 58-62° C. for 8-12 minutes, and headspace extraction is performed for 35-45 minutes to obtain product B.

5. The method for detecting flavor substances in leaves of Psoralea corylifolia based on solid phase microextraction and gas chromatography-mass spectrometry according to claim 4, characterized in that: In the step b, headspace solid phase microextraction: the solid phase microextraction sampling bottle containing product A is equilibrated at 60°C for 20 minutes, then inserted into a manual injector equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber head, preheated at 60°C for 10 minutes, and headspace extracted for 40 minutes to obtain product B.

6. The method for detecting flavor substances in leaves of Psoralea corylifolia based on solid phase microextraction and gas chromatography-mass spectrometry according to claim 1, characterized in that: In the step c, the thermal analysis time is 8 minutes.

7. The method for detecting flavor substances in leaves of Psoralea corylifolia based on solid phase microextraction and gas chromatography-mass spectrometry according to claim 1, characterized in that: In the step c, the gas chromatography conditions are as follows: chromatograph: a GC-MS / MS coupling instrument is used; the chromatographic column is an HP-5MS elastic quartz capillary column, 30m*250μm*0.25μm; the initial temperature is 40°C, maintained for 2 minutes, the temperature is increased to 180°C at 3.5°C / min, and then increased to 310°C at 10°C / min, and the running time is 55 minutes; the vaporization chamber temperature is 250°C; the carrier gas is 99.999% high-purity He; the column head pressure is 7.1866psi, the carrier gas flow rate is 1.0mL / min, the split ratio is 5:1, and the solvent delay time is 3min.

8. The method for detecting flavor substances in leaves of Psoralea corylifolia based on solid phase microextraction and gas chromatography-mass spectrometry according to claim 1, characterized in that: In the step c, the mass spectrometry conditions are as follows: the ion source is an EI source; the ion source temperature is 230° C.; the quadrupole temperature is 150° C.; the electron energy is 70 eV; the emission current is 34.6 μA; the multiplier voltage is 2188 V; the interface temperature is 280° C.; and the mass range is 29-500 amu.

9. The method for detecting flavor substances in leaves of Psoralea corylifolia based on solid phase microextraction and gas chromatography-mass spectrometry according to claim 1, characterized in that: In the step c, the qualitative analysis is completed by the automatic deconvolution system AMDIS of the computer mass spectrometry workstation and the standard mass spectrum library NIST 2020 for comparison analysis.

10. The method for detecting the leaves of Psoralea corylifolia based on solid phase microextraction and gas chromatography-mass spectrometry according to claim 1 The method for producing flavor substances is characterized by: In step c, the quantitative analysis adopts the internal standard quantitative analysis method, Determine the relative percentage content.

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