Gas chromatography-mass spectrometry component determination method for uncaria rhynchophylla leaves

Abstract

The invention discloses a GC-MS (Gas Chromatography-Mass Spectrometry) component determination method for uncaria rhynchophylla leaves, which comprises the following steps of: establishing a content detection method for a premna microphylla alcohol extract by adopting a liquid chromatography-mass spectrometry method, calling original data acquired by mass spectrometry by using software, comparing and analyzing by combining a mass spectrometry information database, and identifying chemical components of the premna microphylla leaf alcohol extract. And finally, 54 chemical components in the alcohol extract are detected, so that a basis is provided for the quality and quality evaluation of the premna microphylla medicinal material.

Description

Technical Field

[0001] This invention relates to a method for determining the components of *Clerodendrum trichotomum*, specifically a method for determining the components of *Uncaria rhynchophylla* leaves using gas chromatography-mass spectrometry (GC-MS). Background Technology

[0002] Uncaria rhynchophylla (Miq.) Miq. ex Havil. is a vine belonging to the Rubiaceae family. Its leaves are papery, elliptical or oblong-elliptical, and brown or reddish-brown when dry, sometimes with a white powdery coating on the underside. Uncaria leaves have the effects of clearing heat and calming the liver, relieving wind and calming convulsions, and lowering blood pressure, and are widely used in Traditional Chinese Medicine. Its effects include clearing heat and calming the liver, relieving wind and calming convulsions, lowering blood pressure, relieving headaches, and improving insomnia.

[0003] Currently, there are no reported methods for detecting the chemical components of Uncaria rhynchophylla leaves; most methods focus on detecting components in the hooked stems and branches. Because the effective components of Uncaria rhynchophylla leaves are not clearly defined, it is difficult to comprehensively evaluate their intrinsic quality. Therefore, establishing a rapid, efficient, and accurate method for determining the main chemical components of Uncaria rhynchophylla leaves, enabling comprehensive control over the quality of these substances, and providing a more sufficient basis for quality evaluation of Uncaria rhynchophylla leaves, is of great significance. Summary of the Invention

[0004] The technical problem to be solved by this invention is to provide a method for determining the chemical components of Uncaria rhynchophylla leaves using gas chromatography-mass spectrometry (GC-MS). This method can perform qualitative analysis of the chemical components in Uncaria rhynchophylla leaves, overcoming the difficulties in detecting chemical components and the uncertainty of effective components in existing technologies.

[0005] This invention is achieved using the following technical solution: a method for determining the components of Uncaria rhynchophylla leaves using gas chromatography-mass spectrometry, comprising the following steps: a. Pre-treatment of Uncaria rhynchophylla leaves: Cut the Uncaria rhynchophylla leaves into 1-3cm pieces to obtain Uncaria rhynchophylla leaf fragments for later use; b. Sample preparation: Take 0.5-1.5g of Uncaria rhynchophylla leaf fragments, place them on the heating platform of the GERSTEL MPS multi-functional autosampler, insert the head equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber, preheat at 40-50℃ for 8-12min, perform headspace extraction for 40-50min, remove the extraction head and immediately insert it into the gas chromatograph injection port, perform thermal desorption for 4-6min to obtain the sample, which is then ready for injection. c. The components of the test sample were separated and detected using a gas chromatography-mass spectrometry (GC-MS) system 8890 / 7000D. d. Data analysis: The peaks in the total ion chromatogram of the mass spectrometer were retrieved and compared with the MassHunter / Library / Nist20 standard mass spectra using the mass spectrometry computer data system to determine the volatile chemical components. The relative mass fraction of each chemical component was determined by the peak area normalization method to identify the chemical components in the Uncaria rhynchophylla leaves.

[0006] In step b above, the sample injection process is as follows: Take 1g of Uncaria rhynchophylla leaf fragments, place them on the heating platform of the GERSTEL MPS multifunctional autosampler, insert the head equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber, preheat at 45℃ for 10min, perform headspace extraction for 45min, remove the extraction head and immediately insert it into the gas chromatograph injection port, perform thermal desorption for 5min, and then the sample can be injected.

[0007] In step c above, the gas chromatography detection conditions were as follows: instrument: 8890 / 7000D GCMSMS coupled system, GERSTELMPS multi-functional autosampler, column: HP-5MS flexible quartz capillary column, specifications: 30 m x 250 μm x 0.25 μm, initial temperature: 40℃ held for 2 min, temperature increased to 180℃ at 3.5℃ / min, then increased to 310℃ at 10℃ / min, run time: 55 min; vaporization chamber temperature: 250℃; carrier gas: 99.999% high purity He; column inlet pressure: 7.0699 psi, carrier gas flow rate: 1.0 mL / min, split ratio: 10:1, solvent delay time: 3 min.

[0008] In step c above, the mass spectrometry detection conditions are as follows: the ion source is an EI source; the ion source temperature is 230℃; the quadrupole temperature is 150℃; the electron energy is 70eV; the emission current is 34.6μA; the multiplier voltage is 2168V; the interface temperature is 280℃; and the mass range is 29~500amu.

[0009] In step d above, a total of 54 chemical components were detected in Uncaria rhynchophylla leaves, including asparagine, D-(−)-aspartic acid, proline, D-(−)-quinic acid, D-gluconic acid, isocitrate, L-valine, shikimic acid, adenine, pyridoxine, citric acid, uridine, benzoic acid, L-tyrosine, L-isoleucine, L-norleucine, guanine, guanosine, L-phenylalanine, panthenol, 6-methylquinoline, neochlorogenic acid, 5′-S-methyl-5′-thioadenosine, and protopanaxadione. Anthocyanin B2, Gardenoside Methyl Ester, Epicatechin, Loganic Acid, Chlorogenic Acid, Cryptochlorogenic Acid, Genipin Acid, Protocatechuic Aldehyde, Catechin, Strychnoside, Dehydrated Monoglycoside, Proanthocyanidin B1, Lovosin, Capryloyl Alk ...

[0010] Beneficial effects of this invention: (1) It overcomes the shortcomings of the lack of chemical composition detection methods for Uncaria rhynchophylla leaves, unclear effective components, and difficulty in evaluating the intrinsic quality of medicinal materials.

[0011] (2) The present invention uses a gas chromatography-mass spectrometry system 8890 / 7000D GCMSMS (Agilent Technologies, USA), a GERSTEL MPS multi-functional autosampler (GERSTEL), and an HP-5MS flexible quartz capillary column. The initial temperature is 40℃ and held for 2 min, then increased to 180℃ at 3.5℃ / min, and then increased to 310℃ at 10℃ / min. The running time is 55 min. The vaporization chamber temperature is 250℃. The carrier gas is 99.999% high-purity He. The column inlet pressure is 7.0699 psi, the carrier gas flow rate is 1.0 mL / min, the split ratio is 10:1, and the solvent delay time is 3 min. The ion source was an EI source; the ion source temperature was 230℃; the quadrupole temperature was 150℃; the electron energy was 70 eV; the emission current was 34.6 μA; the multiplier voltage was 2168 V; the interface temperature was 280℃; and the mass range was 29~500 amu. The specific parameters of the above experiment were obtained after several months of trial and error. If the experimental parameters such as the gas chromatography temperature program, ion source, and mass spectrometry conditions change, the chemical components in Uncaria rhynchophylla leaves will not be detected or will be detected inaccurately.

[0012] (3) The chemical composition detection method of Uncaria rhynchophylla leaves described in this invention can quickly, accurately and sensitively detect 54 chemical components of Uncaria rhynchophylla leaves: asparagine (1), D-(−)-aspartic acid (2), proline (3), D-(−)-quinic acid (4), D-gluconic acid (5), isocitrate (6), L-valine (7), shikimic acid (8), adenine (9), pyridoxine (10), citric acid (11), uridine (12), benzoic acid (13), L-tyrosine (14), L-isoleucine (15), L-norleucine (16), guanine (17), guanosine (18), L-phenylalanine (19), panthenol (20), 6-methylquinoline (21), neochlorogenic acid (22), 5′-S-methyl-5′-thioadenosine (23), proanthocyanidin B2 (24), genistein methyl ester (25) Epicatechin (26), Loganic acid (27), Chlorogenic acid (28), Cryptochlorogenic acid (29), Genipin (30), Protocatechuic aldehyde (31), Catechin (32), Strychnine (33), Dehydrated monoglycoside (34), Proanthocyanidin B1 (35), Lovosin (36), Capryloyl alkaloid (37), Heptylide (38), Yohimbine (39), Brucine I (40), Dehydrouncle The following compounds were used to guide the assessment of the intrinsic quality of Uncaria rhynchophylla leaves: alkaloid (41), conosine (42), isodehydrouncitonin (43), uncaria alkaloid (44), azelaic acid (45), dehydrouncitonin (46), glycyrrhizin (47), atractylodesin A (48), saponin (49), uncaria alkaloid (50), 1-dodecanesulfonic acid (51), triptolide A (52), ursolic acid (53), and erucamide (54). Attached Figure Description

[0013] Figure 1 Total Ion Flow Chromatogram (TIC) of Chemical Components in Uncaria Rhizoma Leaf. Detailed Implementation

[0014] To enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention will be further described below with reference to specific embodiments, but the embodiments are not intended to limit the present invention. Example 1: Gas chromatography-mass spectrometry (GC-MS) method for determining the components of Uncaria rhynchophylla leaves. a. Pre-treatment of Uncaria rhynchophylla leaves: Cut the Uncaria rhynchophylla leaves into 1-3cm pieces to obtain Uncaria rhynchophylla leaf fragments for later use; b. Sample preparation: Take 1g of Uncaria rhynchophylla leaf fragments, place them on the heating platform of the GERSTEL MPS multifunctional autosampler, insert the head equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber, preheat at 45℃ for 10min, perform headspace extraction for 45min, remove the extraction head and immediately insert it into the gas chromatograph injection port, perform thermal desorption for 5min, and then inject the sample. c. The components of the test sample were separated and detected using a gas chromatography-mass spectrometry (GC-MS) system 8890 / 7000D. The gas chromatography detection conditions were as follows: instrument: 8890 / 7000D GCMSMS-MS system, GERSTEL MPS multi-functional autosampler, HP-5MS flexible quartz capillary column (30 m x 250 μm x 0.25 μm), initial temperature 40℃ held for 2 min, temperature increased at 3.5℃ / min to 180℃, then increased at 10℃ / min to 310℃, run time: 55 min; vaporization chamber temperature: 250℃; carrier gas: 99.999% high-purity He; column inlet pressure: 7.0699 psi, carrier gas flow rate: 1.0 mL / min, split ratio: 10:1, solvent delay time: 3 min. The mass spectrometry detection conditions were as follows: EI source; ion source temperature 230℃; quadrupole temperature 150℃; electron energy 70eV; emission current 34.6μA; multiplier voltage 2168V; interface temperature 280℃; mass range 29~500amu. d. Data analysis: The peaks in the total ion chromatogram of the mass spectrometer were retrieved and compared with the MassHunter / Library / Nist20 standard mass spectra using the mass spectrometry computer data system to determine the volatile chemical components. The relative mass fraction of each chemical component was determined by the peak area normalization method to identify the chemical components in the Uncaria rhynchophylla leaves.

[0015] Example 2: Gas chromatography-mass spectrometry (GC-MS) method for component determination of Uncaria rhynchophylla leaves a. Pre-treatment of Uncaria rhynchophylla leaves: Cut the Uncaria rhynchophylla leaves into 1-3cm pieces to obtain Uncaria rhynchophylla leaf fragments for later use; b. Sample preparation: Take 0.5g of Uncaria rhynchophylla leaf fragments, place them on the heating platform of the GERSTEL MPS multi-functional autosampler, insert the head equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber, preheat at 50℃ for 8min, perform headspace extraction for 50min, remove the extraction head and immediately insert it into the gas chromatograph injection port, perform thermal desorption for 6min to obtain the sample, which is then ready for injection. c. The components of the test sample were separated and detected using a gas chromatography-mass spectrometry (GC-MS) system 8890 / 7000D. The gas chromatography detection conditions were as follows: instrument: 8890 / 7000D GCMSMS-MS system, GERSTEL MPS multi-functional autosampler, HP-5MS flexible quartz capillary column (30 m x 250 μm x 0.25 μm), initial temperature 40℃ held for 2 min, temperature increased at 3.5℃ / min to 180℃, then increased at 10℃ / min to 310℃, run time: 55 min; vaporization chamber temperature: 250℃; carrier gas: 99.999% high-purity He; column inlet pressure: 7.0699 psi, carrier gas flow rate: 1.0 mL / min, split ratio: 10:1, solvent delay time: 3 min. The mass spectrometry detection conditions were as follows: EI source; ion source temperature 230℃; quadrupole temperature 150℃; electron energy 70eV; emission current 34.6μA; multiplier voltage 2168V; interface temperature 280℃; mass range 29~500amu. d. Data analysis: The peaks in the total ion chromatogram of the mass spectrometer were retrieved and compared with the MassHunter / Library / Nist20 standard mass spectra using the mass spectrometry computer data system to determine the volatile chemical components. The relative mass fraction of each chemical component was determined by the peak area normalization method to identify the chemical components in the Uncaria rhynchophylla leaves.

[0016] Example 3: Gas chromatography-mass spectrometry (GC-MS) method for component determination of Uncaria rhynchophylla leaves a. Pre-treatment of Uncaria rhynchophylla leaves: Cut the Uncaria rhynchophylla leaves into 1-3cm pieces to obtain Uncaria rhynchophylla leaf fragments for later use; b. Sample preparation: Take 1.5g of Uncaria rhynchophylla leaf fragments, place them on the heating platform of the GERSTEL MPS multi-functional autosampler, insert the head equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber, preheat at 40℃ for 12min, perform headspace extraction for 40min, remove the extraction head and immediately insert it into the gas chromatograph injection port, perform thermal desorption for 4min to obtain the sample, which is then ready for injection. c. The components of the test sample were separated and detected using a gas chromatography-mass spectrometry (GC-MS) system 8890 / 7000D. The gas chromatography detection conditions were as follows: instrument: 8890 / 7000D GCMSMS-MS system, GERSTEL MPS multi-functional autosampler, HP-5MS flexible quartz capillary column (30 m x 250 μm x 0.25 μm), initial temperature 40℃ held for 2 min, temperature increased at 3.5℃ / min to 180℃, then increased at 10℃ / min to 310℃, run time: 55 min; vaporization chamber temperature: 250℃; carrier gas: 99.999% high-purity He; column inlet pressure: 7.0699 psi, carrier gas flow rate: 1.0 mL / min, split ratio: 10:1, solvent delay time: 3 min. The mass spectrometry detection conditions were as follows: EI source; ion source temperature 230℃; quadrupole temperature 150℃; electron energy 70eV; emission current 34.6μA; multiplier voltage 2168V; interface temperature 280℃; mass range 29~500amu. d. Data analysis: The peaks in the total ion chromatogram of the mass spectrometer were retrieved and compared with the MassHunter / Library / Nist20 standard mass spectra using the mass spectrometry computer data system to determine the volatile chemical components. The relative mass fraction of each chemical component was determined by the peak area normalization method to identify the chemical components in the Uncaria rhynchophylla leaves.

[0017] Example 4: a. Pre-treatment of Uncaria rhynchophylla leaves: Cut the Uncaria rhynchophylla leaves into 1-3cm pieces to obtain Uncaria rhynchophylla leaf fragments for later use; b. Sample preparation: Take 0.8g of Uncaria rhynchophylla leaf fragments, place them on the heating platform of the GERSTEL MPS multi-functional autosampler, insert the head equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber, preheat at 45℃ for 11min, perform headspace extraction for 45min, remove the extraction head and immediately insert it into the gas chromatograph injection port, perform thermal desorption for 4.5min to obtain the sample, which is then ready for injection. c. The components of the test sample were separated and detected using a gas chromatography-mass spectrometry (GC-MS) system 8890 / 7000D. The gas chromatography detection conditions were as follows: instrument: 8890 / 7000D GCMSMS-MS system, GERSTEL MPS multi-functional autosampler, HP-5MS flexible quartz capillary column (30 m x 250 μm x 0.25 μm), initial temperature 40℃ held for 2 min, temperature increased at 3.5℃ / min to 180℃, then increased at 10℃ / min to 310℃, run time: 55 min; vaporization chamber temperature: 250℃; carrier gas: 99.999% high-purity He; column inlet pressure: 7.0699 psi, carrier gas flow rate: 1.0 mL / min, split ratio: 10:1, solvent delay time: 3 min. The mass spectrometry detection conditions were as follows: EI source; ion source temperature 230℃; quadrupole temperature 150℃; electron energy 70eV; emission current 34.6μA; multiplier voltage 2168V; interface temperature 280℃; mass range 29~500amu. d. Data analysis: The peaks in the total ion chromatogram of the mass spectrometer were retrieved and compared with the MassHunter / Library / Nist20 standard mass spectra using the mass spectrometry computer data system to determine the volatile chemical components. The relative mass fraction of each chemical component was determined by the peak area normalization method to identify the chemical components in the Uncaria rhynchophylla leaves.

[0018] This invention has undergone extensive analytical verification experiments, and the following are the results of the experimental research of this invention: 1. Preparation of Uncaria leaves The experimental material, Uncaria rhynchophylla (Miq.) Miq. ex Havil, was collected from Jianhe County, Qiandongnan Prefecture, Guizhou Province (N 26°44′00″E 108°27′04″). The seedlings were cultivated and propagated at the Guizhou University Research Base (26°25'54"N.106°40'53"E, located in a subtropical humid monsoon climate zone with an average altitude of 1178m, an average annual temperature of 20.21℃, and an average annual precipitation of 1318.90mm). Leaves from one-year-old plants were collected on November 23, 2025, for related research.

[0019] Pre-treatment of Uncaria rhynchophylla leaves: Cut the Uncaria rhynchophylla leaves into 1-3cm pieces to obtain Uncaria rhynchophylla leaf fragments for later use.

[0020] 2. Sample preparation for testing: Take 1g of Uncaria rhynchophylla leaf fragments, place them on the heating platform of the GERSTEL MPS multifunctional autosampler, insert the head equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber, preheat at 45℃ for 10min, perform headspace extraction for 45min, remove the extraction head and immediately insert it into the gas chromatograph injection port, perform thermal desorption for 5min, and then inject the sample.

[0021] 3. Instruments and Equipment The components of the test sample were separated and detected using an 8890 / 7000D GC-MS-MS system.

[0022] 4. Gas chromatography detection conditions are as follows: The instrument used was an 8890 / 7000D GCMSMS system with a GERSTEL MPS multi-functional autosampler. The column was an HP-5MS flexible quartz capillary column with dimensions of 30 m x 250 μm x 0.25 μm. The initial temperature was 40℃ and held for 2 min, then increased to 180℃ at 3.5℃ / min, and then increased to 310℃ at 10℃ / min. The run time was 55 min. The vaporization chamber temperature was 250℃. The carrier gas was 99.999% high-purity He. The column inlet pressure was 7.0699 psi, the carrier gas flow rate was 1.0 mL / min, the split ratio was 10:1, and the solvent delay time was 3 min.

[0023] 5. Mass spectrometry detection conditions The ion source is an EI source; the ion source temperature is 230℃; the quadrupole temperature is 150℃; the electron energy is 70eV; the emission current is 34.6μA; the multiplier voltage is 2168V; the interface temperature is 280℃; and the mass range is 29~500amu.

[0024] 6. Qualitative chemical composition The peaks in the total ion chromatogram of the mass spectrometer were retrieved and compared with the MassHunter / Library / Nist20 standard mass spectra using a mass spectrometry computer data system to determine the volatile chemical components. The relative mass fraction of each chemical component was determined by peak area normalization to identify the chemical components in the Uncaria rhynchophylla leaves. Figure 1 See Table 1.

[0025] Table 1. Detailed information on the chemical composition of Uncaria rhynchophylla leaves Serial Number Retention time name Chinese name Chemical formula quality error molecular weight Maximum peak area 1 0.832 Asparagine Asparagine <![CDATA[C4H8N2O3]]> -0.38 132.05344 269164621.1 2 0.845 D-(-)-Aspartic acid D-(−)-Aspartic acid <![CDATA[C4H7NO4]]> -0.92 133.03739 32886065.13 3 0.913 Proline proline <![CDATA[C5H9NO2]]> 1.3 115.06348 490440205.8 4 0.949 D-(-)-Quinic acid D-(−)-quinic acid <![CDATA[C7H 12 O6]]> -4.04 192.06261 5983754815 5 1.049 D-Saccharic acid D-gluconic acid (mucolytic acid) <![CDATA[C6H 10 O8]]> -2.84 210.03697 969512932.6 6 1.083 Isocitric acid Isocitrate <![CDATA[C6H8O7]]> -3.87 192.02626 3283558449 7 1.14 L-Valine L-valine <![CDATA[C5H 11 NO2]]> 1.26 117.07913 554732775.6 8 1.144 Shikimic acid Shikimic acid <![CDATA[C7H 10 O5]]> -5.17 174.05192 459643494.7 9 1.161 Adenine adenine <![CDATA[C5H5N5]]> -0.82 135.05438 1125265913 10 1.477 Pyridoxine <![CDATA[Pyridoxine (vitamin B6)]]> <![CDATA[C8H 11 NO3]]> -1.76 169.07359 120121994.1 11 1.522 Citric acid Citric acid <![CDATA[C6 H 8 O7]]> -3.21 192.02639 13186575944 12 1.854 Uridine Ureidine <![CDATA[C9H 12 N2O6]]> -0.95 244.0693 92094374.86 13 1.882 Benzoic acid benzoic acid <![CDATA[C7H6O2]]> 1.03 122.03691 127576468.6 14 1.882 L-Tyrosine L-tyrosine <![CDATA[C9H 11 NO3]]> -4.62 181.07306 47685921.66 15 2.034 L-Isoleucine L-Isoleucine <![CDATA[C6H 13 NO2]]> -0.52 131.09456 479568109.2 16 2.199 L-Norleucine L-norleucine <![CDATA[C6H 13 NO2]]> -0.52 131.09456 658057067.3 17 2.96 Guanine Guanine <![CDATA[C5H5N5O]]> -1.01 151.04926 87665417.98 18 2.977 Guanosine Guanosine <![CDATA[C 10 H 13 N5O5]]> -0.73 283.09146 169447655 19 4.3 L-Phenylalanine L-phenylalanine <![CDATA[C9H 11 NO2]]> -1.18 165.07878 1565310405 20 7.984 Panthenol <![CDATA[Panthenol (provitamin B5)]]> <![CDATA[C9H 19 NO4]]> -1.31 205.13114 792540018.1 21 9.289 6-Methylquinoline 6-Methylquinoline <![CDATA[C 10 H9N]]> -0.93 143.07337 1488447963 22 9.534 Neochlorogenic acid Neochlorogenic acid <![CDATA[C 16 H 18 O9]]> -0.46 354.09492 1151825706 23 10.505 5'-S-Methyl-5'-thioadenosine 5′-S-methyl-5′-thioadenosine <![CDATA[C 11 H 15 N5O3S]]> -0.63 297.08937 139907124.8 24 10.639 Procyanidin B2 Proanthocyanidins B2 <![CDATA[C 30 H 26 O 12 ]]> 0.77 578.14287 65724696.83 25 10.971 Shanzhiside methyl ester Gardenoside Methyl Ester <![CDATA[C 17 H 26 O 11 ]]> 113272.78 452.15297 23717786.36 26 11.379 Epicatechin Catechin <![CDATA[C 15 H 14 O6]]> -0.13 290.079 1448759202 27 11.628 Loganic acid Strychnos nuciferic acid <![CDATA[C 16 H 24 O 10 ]]> 0.05 376.13697 460001688.2 28 11.807 Chlorogenic acid chlorogenic acid <![CDATA[C 16 H 18 O9]]> -0.55 354.09489 8873661506 29 12.106 Cryptochlorogenic acid cryptochlorogenic acid <![CDATA[C 16 H 18 O9]]> -0.4 354.09494 2802763875 30 12.278 Geniposidic acid Genipin acid <![CDATA[C 16 H 22 O 10 ]]> -0.18 374.12123 122569545.3 31 12.934 Protocatechualdehyde Protocatechuic aldehyde <![CDATA[C7H6O3]]> -1.66 138.03147 53803930.69 32 12.935 Cianidanol Catechins ((+)-catechins) <![CDATA[C 15 H 14 O6]]> -0.16 290.07899 3502519059 33 13.175 Loganin Strychnoside <![CDATA[C 17 H 26 O 10 ]]> 117916.29 436.15794 47049112.21 34 14.126 Sarracenin Dehydrated monoglycoside <![CDATA[C 11 H 14 O5]]> -1.69 226.08374 196348667.7 35 14.513 Procyanidin B1 Proanthocyanidins B1 <![CDATA[C 30 H 26 O 12 ]]> 0.35 578.14263 116726127.9 36 14.696 Rauwolscine Lauwsin (α-yohimbine) <![CDATA[C 21 H 26 N2O3]]> -0.92 354.19402 2825923662 37 15.03 Mitraphylline Cap post wood alkali <![CDATA[C 21 H 24 N2O4]]> -0.76 368.17333 128592083.5 38 15.193 Heptanophenone Heptylacetone <![CDATA[C 13 H 18 O]]> 94723.59 208.14611 90065613.88 39 15.245 Yohimbine Yuhimbin <![CDATA[C 21 H 26 N2O3]]> -0.92 354.19402 88682885.83 40 15.261 Kaempferol 3-glucorhamnoside Bai Rui Cao Su I <![CDATA[C 27 H 30 O 15 ]]> 0.4 594.1587 53307461.23 41 15.321 Corynoxeine Dehydrounerin <![CDATA[C 22 H 26 N2O4]]> -83797.55 350.16274 1.21152E+11 42 15.649 Corynoxine Knosin <![CDATA[C 22 H 28 N2O4]]> -83357.82 352.17842 60063043779 43 15.659 Isocorynoxeine Isodehydrounergine <![CDATA[C 22 H 26 N2O4]]> -0.81 382.18895 57575227053 44 15.922 Rhynchophylline Uncariae <![CDATA[C 22 H 28 N2O4]]> -0.7 384.20464 21438431323 45 16.091 Azelaic acid azelaic acid <![CDATA[C9H 16 O4]]> -3.91 188.10412 169265254.3 46 17.368 Hirsuteine Dehydrocrassaline <![CDATA[C 22 H 26 N2O3]]> -0.89 366.19402 2493865988 47 17.641 Glabrolide glycyrrhizin <![CDATA[C 30 H 44 O4]]> -0.71 468.32363 387073171.1 48 18.204 Atractyloside Atractylodes A <![CDATA[C 21 H 36 O 10 ]]> 102638.02 494.23637 158147346.6 49 19.06 Quillaic acid soap acid <![CDATA[C 30 H 46 O5]]> -0.48 486.33429 269804233.1 50 19.128 Hirsutine rhubarb alkaloid <![CDATA[C 22 H 28 N2O3]]> -0.79 368.2097 2911230910 51 27.597 1-Dodecanesulfonic acid 1-Dodecanesulfonic acid <![CDATA[C 12 H 26 O3S]]> -1.11 250.15999 186694286.3 52 30.61 Wilforlide A Tripterygium wilfordii lactone A <![CDATA[C 30 H 46 O3]]> -1.3 454.3441 86743530.55 53 36.505 Ursolic acid Ursolic acid <![CDATA[C 30 H 48 O3]]> -0.16 456.36027 163305595.2 54 43.01 Erucamide Erucamide <![CDATA[C 22 H 43 NO]]> -1.37 337.334 221468560.3

Claims

1. A method for determining the components of Uncaria rhynchophylla leaves using gas chromatography-mass spectrometry (GC-MS), characterized in that: Includes the following steps: a. Pre-treatment of Uncaria rhynchophylla leaves: Cut the Uncaria rhynchophylla leaves into 1-3cm pieces to obtain Uncaria rhynchophylla leaf fragments for later use; b. Sample preparation: Take 0.5-1.5g of Uncaria rhynchophylla leaf fragments, place them on the heating platform of the GERSTEL MPS multi-functional autosampler, insert the head equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber, preheat at 40-50℃ for 8-12min, perform headspace extraction for 40-50min, remove the extraction head and immediately insert it into the gas chromatograph injection port, perform thermal desorption for 4-6min to obtain the sample, which is then ready for injection. c. The components of the test sample were separated and detected using a gas chromatography-mass spectrometry (GC-MS) system 8890 / 7000D. d. Data analysis: The peaks in the total ion chromatogram of the mass spectrometer were retrieved and compared with the MassHunter / Library / Nist20 standard mass spectra using the mass spectrometry computer data system to determine the volatile chemical components. The relative mass fraction of each chemical component was determined by the peak area normalization method to identify the chemical components in the Uncaria rhynchophylla leaves.

2. The method for determining the components of Uncaria rhynchophylla leaves using gas chromatography-mass spectrometry according to claim 1, characterized in that: In step b, the sample injection process is as follows: Take 1g of Uncaria rhynchophylla leaf fragments, place them on the heating platform of the GERSTEL MPS multifunctional autosampler, insert the head equipped with a 2cm-50 / 30μm DVB / CAR / PDMS StableFlex fiber, preheat at 45℃ for 10min, perform headspace extraction for 45min, remove the extraction head and immediately insert it into the gas chromatograph injection port, perform thermal desorption for 5min, and then the sample can be injected.

3. The method for determining the components of Uncaria rhynchophylla leaves using gas chromatography-mass spectrometry according to claim 1, characterized in that: In step c, the gas chromatography detection conditions are as follows: instrument: 8890 / 7000D GCMSMS coupled system, GERSTEL MPS multi-functional autosampler, column: HP-5MS flexible quartz capillary column, specifications: 30 m x 250 μm x 0.25 μm, initial temperature: 40℃ held for 2 min, temperature increased to 180℃ at 3.5℃ / min, then increased to 310℃ at 10℃ / min, run time: 55 min; vaporization chamber temperature: 250℃; carrier gas: 99.999% high-purity He; column inlet pressure: 7.0699 psi, carrier gas flow rate: 1.0 mL / min, split ratio: 10:1, solvent delay time: 3 min.

4. The method for determining the components of Uncaria rhynchophylla leaves using gas chromatography-mass spectrometry according to claim 1, characterized in that: In step c, the mass spectrometry detection conditions are as follows: the ion source is an EI source; the ion source temperature is 230℃; the quadrupole temperature is 150℃; the electron energy is 70eV; the emission current is 34.6μA; the multiplier voltage is 2168V; the interface temperature is 280℃; and the mass range is 29~500amu.

5. The method for determining the components of Uncaria rhynchophylla leaves using gas chromatography-mass spectrometry according to claim 1, characterized in that, In step d, 54 chemical components were detected in the leaves of Uncaria rhynchophylla, including asparagine, D-(−)-aspartic acid, proline, D-(−)-quinic acid, D-gluconic acid, isocitrate, L-valine, shikimic acid, adenine, pyridoxine, citric acid, uridine, benzoic acid, L-tyrosine, L-isoleucine, L-norleucine, guanine, guanosine, L-phenylalanine, panthenol, 6-methylquinoline, neochlorogenic acid, 5′-S-methyl-5′-thioadenosine, and protopanaxadione. Anthocyanin B2, Gardenoside Methyl Ester, Epicatechin, Loganic Acid, Chlorogenic Acid, Cryptochlorogenic Acid, Genipin Acid, Protocatechuic Aldehyde, Catechin, Strychnoside, Dehydrated Monoglycoside, Proanthocyanidin B1, Lovosin, Capryloyl Alk ...

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