Method for characterizing volatile components in compound pseudo-ginseng and salvia miltiorrhiza capsules based on gas chromatography-mass spectrometry

Abstract

The invention discloses a method for characterizing volatile components in a compound pseudo-ginseng and salvia miltiorrhiza capsule based on gas chromatography-mass spectrometry, and relates to the technical field of compound pseudo-ginseng and salvia miltiorrhiza capsule detection, and the method comprises the following steps: (1) preparing a compound pseudo-ginseng and salvia miltiorrhiza capsule test solution; and (2) detecting the volatile components in the traditional Chinese medicine compound test solution by adopting a triple quadrupole gas chromatograph-mass spectrometer. According to the method, volatile oil of a trace component of rhizoma cyperi in a compound preparation is fully retained and extracted by pre-treating the compound pseudo-ginseng and salvia miltiorrhiza capsule, and after detection by the method, a quasi-molecular ion peak can be obtained, and a fragment ion peak containing rich structure information can be obtained, so that the structure information of the compound can be quickly and accurately obtained.

Description

Technical Field

[0001] This application relates to the field of detection technology for Compound Panax notoginseng and Salvia miltiorrhiza capsules, and in particular to a method for characterizing volatile components in Compound Panax notoginseng and Salvia miltiorrhiza capsules based on gas chromatography-mass spectrometry. Background Technology

[0002] The approval number for Compound Panax notoginseng and Salvia miltiorrhiza Capsules is Dianyaozhizi (Z) 20082438A; Specification: 0.34g / capsule, 60 capsules / bottle; Indications: Promotes blood circulation and removes blood stasis, soothes the liver and regulates qi, calms the mind and soothes the nerves; Dosage and administration: Oral administration, 2-3 capsules three times a day. It is an in-house preparation prepared by the Pharmaceutical Preparation Room of the Affiliated Hospital of Yunnan University. It is composed of Panax notoginseng, Salvia miltiorrhiza, Astragalus membranaceus and Cyperus rotundus. It is used for chest pain caused by qi stagnation and blood stasis, with symptoms such as chest tightness and stabbing pain in the precordial region; and for patients with coronary heart disease, hypertension, hyperlipidemia and other conditions with the above symptoms. Panax notoginseng has the effects of dispersing blood stasis, stopping bleeding, reducing swelling and relieving pain, while Salvia miltiorrhiza has a strong effect of promoting blood circulation and regulating qi. When the two are combined, they complement each other, which can greatly enhance the effects of promoting blood circulation, removing blood stasis, clearing the channels and relieving pain. It is a commonly used drug pair in traditional Chinese medicine for the clinical treatment of cardiovascular diseases. Salvia miltiorrhiza and Astragalus membranaceus are often used as a drug pair for invigorating qi and promoting blood circulation. Salvia miltiorrhiza mainly promotes blood circulation, removes blood stasis and regulates menstruation, while Astragalus membranaceus has the effect of tonifying qi, raising yang and lifting prolapse. When the two drugs are combined, they work together to invigorate qi and nourish blood, promote blood circulation and regulate menstruation.

[0003] Compound Danshen preparations are one of the major traditional Chinese medicine products in my country. They are commonly used by patients with coronary heart disease and angina pectoris, and are included in the National Essential Medicines List and the National Basic Medical Insurance, Work Injury Insurance, and Maternity Insurance Drug List. They have a large clinical usage and definite efficacy. Compound Danshen tablets were successfully developed by Shanghai No. 2 Pharmaceutical Factory in 1977, approved for production in 1981, and first included in the Pharmacopoeia of the People's Republic of China in 1985.

[0004] Currently, research on the chemical components of traditional Chinese medicine compound preparations containing Panax notoginseng and Salvia miltiorrhiza mostly employs liquid chromatography-mass spectrometry (LC-MS / MS), focusing primarily on hydrophilic salvianolic acid and saponins. Reports on methods and applications of characterizing volatile components using gas chromatography-mass spectrometry (GC-MS / MS) are limited to the determination of the content of three fat-soluble components (stigmasterol, stigmasterol, and β-sitosterol) in Panax notoginseng preparations (publication number CN201810817184.9) and two fat-soluble components (ferrous sulfate and eucalyptol) (publication number CN201810626977.2), as well as the determination of the content of volatile components derived from borneol in compound Salvia miltiorrhiza preparations. Unlike compound Salvia miltiorrhiza preparations with similar efficacy, Compound Panax notoginseng and Salvia miltiorrhiza Capsules use Cyperus rotundus instead of borneol, utilizing traditional Chinese medicinal materials, demonstrating definite clinical efficacy and safety. The Compendium of Materia Medica states that "Cyperus rotundus has a neutral and non-cold nature, is fragrant and can penetrate deeply, and its taste is mostly pungent and can disperse, slightly bitter and can descend, and slightly sweet and can harmonize." It has the effects of regulating qi and relieving depression, regulating menstruation and relieving pain. It is an excellent product for soothing the liver and regulating qi, and is known as "the general manager of qi diseases and the commander-in-chief of gynecology".

[0005] The volatile oil is the main active component of Cyperus rotundus, and more than 140 volatile components, including monoterpenes, sesquiterpenes, and their oxides, have been identified. Among them, α-cyperol, α-cyperone, cyperene, and cyperane are characteristic components. Cyperus rotundus volatile oil is often combined with other traditional Chinese medicines to treat diseases of the nervous, cardiovascular, and digestive systems. Compound Sanqi Danshen Capsules use Cyperus rotundus instead of borneol in its formulation. Because it uses traditional Chinese medicine, the volatile components of Cyperus rotundus in Compound Sanqi Danshen Capsules are present in low concentrations, and trace components are easily masked, making conventional methods difficult for quality control analysis.

[0006] The triple quadrupole gas chromatograph-mass spectrometer (GC-MS) adds mass spectrometry information of daughter ions to GC-MS, enhancing structural resolution and qualitative analysis capabilities. It is widely used in the structural analysis of gas-phase ions and the qualitative and quantitative analysis of complex matrices. Through multiple reaction monitoring (MRM) mode, it achieves highly sensitive detection of trace compounds in complex sample matrices. By analyzing and comparing the chromatographic and mass spectrometric characteristics of known compounds, it can perform qualitative and quantitative analysis of components with known and unknown structures in traditional Chinese medicine compound preparations. It can simultaneously obtain rich information such as retention time, molecular weight, and characteristic structural fragments of compounds, making it a powerful research tool for the separation and analysis of complex and trace samples.

[0007] Current detection methods do not utilize gas chromatography-mass spectrometry (GC-MS) for the comprehensive characterization of volatile components in compound Danshen preparations. When existing methods are applied to characterize the volatile components of compound Sanqi Danshen capsules, the complex composition and abundance of trace components in this compound preparation make further analysis difficult. After pretreatment using existing methods, the levels of many volatile components in the compound Sanqi Danshen capsules are even lower, making detection using current methods challenging and hindering further analysis.

[0008] The information disclosed in the background section is intended only to enhance the understanding of the overall background of the invention and should not be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art. Summary of the Invention

[0009] This application addresses the aforementioned technical problem by providing a method for characterizing volatile components in Compound Panax notoginseng and Salvia miltiorrhiza capsules using gas chromatography-mass spectrometry (GC-MS). This method can effectively extract trace amounts of volatile oil components from Cyperus rotundus and accurately obtain the volatile oil components in Compound Panax notoginseng and Salvia miltiorrhiza capsules through GC-MS detection.

[0010] This application provides a method for characterizing volatile components in compound Panax notoginseng and Salvia miltiorrhiza capsules using gas chromatography-mass spectrometry (GC-MS), comprising the following steps: (1) Preparation of the test solution of compound Panax notoginseng and Salvia miltiorrhiza capsules; The preparation method of the test solution includes: adding a first solvent and zeolite to the compound Sanqi Danshen capsule traditional Chinese medicine compound preparation, the mass-to-volume ratio of the traditional Chinese medicine compound preparation and the first solvent is 1:8~40, after thorough mixing, connecting the volatile oil extractor and the reflux condenser, placing it in an electric heating mantle and slowly heating to the boiling point, and maintaining a gentle boil for 1~5 hours until the amount of oil in the extractor no longer increases, stopping the heating, allowing it to stand and separate to obtain an orange-red solid oil layer, adding a second solvent to mix and dissolve, and obtaining the traditional Chinese medicine compound test solution; (2) The volatile components in the test solution of the traditional Chinese medicine compound were detected by a triple quadrupole gas chromatography-mass spectrometry (GC-MS) instrument. The retention time, molecular weight and characteristic structural fragment information of the volatile components contained in the test solution were obtained. The volatile components were determined by comparing them with the chromatographic and mass spectrometric characteristics of known compounds. The chromatographic conditions for detection were as follows: a capillary column was used, with the following column temperature program: initial temperature 100℃, held for 10 min; temperature increased to 150℃ at a rate of 10℃ / min, held for 20 min; then increased to 200℃ at a rate of 3℃ / min, held for 10 min; then increased to 300℃ at a rate of 10℃ / min, held for 6 min; the total analysis time was 77 min; the injection port temperature was set to 200~300℃, the column flow rate was 0.5~2 mL / min, the total flow rate was 15~35 mL / min, and the tail gas flow rate was 1~10 mL / min. The mass spectrometry conditions during detection were as follows: ion source temperature was set to 250℃, interface temperature was set to 250℃, solvent removal time was 3 min, photomultiplier tube voltage was 0.1~0.8 kV, data export started 3 min later, mass scan range was set to 50~1200 amu, and scan interval was 0.20 s.

[0011] Preferably, the chromatographic column used is a Shimadzu SH-I-5Si MS capillary column.

[0012] Preferably, the chromatographic column has the following specifications: column length 30m, inner diameter 0.25mm, and particle size 0.25μm.

[0013] Preferably, the sample injection method during detection is split injection, and the split ratio of the split injection is 8~40.

[0014] Preferably, the split ratio for split injection is 20.

[0015] Preferably, the first solvent is water; the second solvent is any one of ethyl acetate, chloroform, or n-hexane.

[0016] Preferably, the mass-to-volume ratio of the traditional Chinese medicine compound preparation to the first solvent is 1:20.

[0017] Preferably, the mixing volume ratio of the second solvent to the orange-red solid oil layer is 1~20:1.

[0018] The beneficial effects that this application can produce include: 1) The method for characterizing volatile components in Compound Panax notoginseng and Salvia miltiorrhiza capsules based on gas chromatography-mass spectrometry provided in this application fully retains and extracts the volatile oil of Cyperus rotundus, a trace component in the compound preparation, through pretreatment of the capsules. After detection by this method, quasi-molecular ion peaks can be obtained, as well as fragment ion peaks containing rich structural information, thereby rapidly and accurately obtaining compound structural information, providing rich qualitative identification information, automatically analyzing samples, and exhibiting high detection efficiency, high sensitivity, and good reproducibility. All of these aspects are significantly superior to the detection results of existing conventional national standard identification techniques.

[0019] 2) The method for characterizing volatile components in Compound Panax notoginseng and Danshen Capsules based on gas chromatography-mass spectrometry (GC-MS) provided in this application uses GC-MS fingerprinting to characterize the volatile component composition of the compound preparation of Compound Panax notoginseng and Danshen Capsules. The triple quadrupole GC-MS technology of this invention provides an effective approach for quality evaluation and quality control of Compound Panax notoginseng and Danshen Capsules, and has significant scientific implications for the development of this compound preparation. Attached Figure Description

[0020] Figure 1 This is a gas chromatography-mass spectrometry (GC-MS) chromatogram of the test solution obtained by the volatile oil extraction method in Example 1 of this application; Figure 2 This is a gas chromatography-mass spectrometry (GC-MS) chromatogram of the test solution obtained by the volatile oil extraction method in Example 2 of this application; Figure 3 This is a gas chromatography-mass spectrometry (GC-MS) chromatogram of the test solution obtained by the volatile oil extraction method in Example 3 of this application; Figure 4 This is the total ion current spectrum of the compound preparation of Panax notoginseng and Salvia miltiorrhiza containing Cyperus rotundus obtained under the GC-MS / MS analysis conditions in Example 4 of this application; Figure 5 This is the total ion current spectrum of the compound preparation of Panax notoginseng and Salvia miltiorrhiza containing Cyperus rotundus obtained under the GC-MS / MS analysis conditions in Example 5 of this application; Figure 6 This is the total ion current spectrum of the compound preparation of Panax notoginseng and Salvia miltiorrhiza containing Cyperus rotundus obtained under the GC-MS / MS analysis conditions in Example 6 of this application; Figure 7 This is the total ion current spectrum of the compound preparation of Panax notoginseng and Salvia miltiorrhiza containing Cyperus rotundus obtained under the GC-MS / MS analysis conditions in Example 7 of this application; Figure 8This is the total ion current spectrum of the compound preparation of Panax notoginseng and Salvia miltiorrhiza containing Cyperus rotundus obtained under the GC-MS / MS analysis conditions in Example 8 of this application; Figure 9 This is the total ion current spectrum of volatile components in Compound Panax notoginseng and Salvia miltiorrhiza Capsules, batch number 2406046, obtained in Example 9 of this application.

[0021] Figure 10 The total ion current spectrum of volatile components in Compound Panax notoginseng and Salvia miltiorrhiza Capsules, batch number 2408048, obtained in Example 9 of this application; Figure 11 The total ion current spectrum of volatile components in Compound Panax notoginseng and Salvia miltiorrhiza Capsules, batch number 2505023, obtained in Example 9 of this application; Detailed Implementation

[0022] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The components of the embodiments of the present invention described and shown in the accompanying drawings can generally be arranged and designed in various different configurations.

[0023] Therefore, the following detailed description of the embodiments of the invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the invention without inventive effort are within the scope of protection of the invention.

[0024] Technical means not detailed in this application and not used to solve the technical problems of this application are all set according to common general knowledge in the field, and multiple common general knowledge setting methods can be implemented.

[0025] The method for characterizing volatile components in Compound Panax notoginseng and Salvia miltiorrhiza Capsules based on gas chromatography-mass spectrometry (GC-MS) provided in this application includes the following steps: (1) Preparation of compound Panax notoginseng and Salvia miltiorrhiza capsule test solution; The preparation method of the test solution includes: weighing an appropriate amount of compound Panax notoginseng and Salvia miltiorrhiza capsule traditional Chinese medicine compound preparation and placing it in a container, adding the first solvent (water) and zeolite, the mass / volume (mg / mL) ratio between the traditional Chinese medicine compound preparation and the first solvent is 8~40, after thorough mixing, connecting the volatile oil extractor and the reflux condenser, placing it in the electric heating mantle and slowly heating to the boiling point, and maintaining a gentle boil for 1~5 hours until the amount of oil in the extractor no longer increases, stopping the heating, allowing it to stand and separate to obtain an orange-red solid oil layer, adding the second solvent and mixing and dissolving to obtain the traditional Chinese medicine compound test solution; (2) The volatile components in the test solution of the traditional Chinese medicine compound were detected by a triple quadrupole gas chromatography-mass spectrometry (GC-MS) instrument. The retention time, molecular weight and characteristic structural fragment information of the volatile components contained in the test solution were obtained. The volatile components were determined by comparing them with the chromatographic and mass spectrometric characteristics of known compounds. The chromatographic conditions for detection were as follows: a capillary column was used, with the following column temperature program: initial temperature set at 100℃, held for 10 min; temperature increased to 150℃ at a rate of 10℃ / min, held for 20 min; then increased to 200℃ at a rate of 3℃ / min, held for 10 min; then increased to 300℃ at a rate of 10℃ / min, held for 6 min; the total analysis time was 77 min; the injection port temperature was set at 200~300℃, the column flow rate was 0.5~2 mL / min, the total flow rate was 15~35 mL / min, and the tail gas flow rate was 1~10 mL / min. The mass spectrometry conditions for detection were as follows: ion source temperature was set to 250℃, interface temperature was set to 250℃, solvent removal time was 3 min, photomultiplier tube voltage was 0.1~0.8 kV, data export started 3 min later, mass scan range was set to 50~1200 amu, and scan interval was 0.20 s.

[0026] Preferably, the chromatographic column is a Shimadzu SH-I-5Si MS capillary column; Preferably, the chromatographic column has the following specifications: column length 30m, inner diameter 0.25mm, and particle size 0.25μm; Preferably, the sampling method is split injection; More preferably, the split ratio of the split injection is 8 to 40, for example, 20.

[0027] Preferably, the first solvent is water; the second solvent is any one of ethyl acetate, chloroform, or n-hexane. Preferably, the mass-to-volume ratio of the traditional Chinese medicine compound preparation to the first solvent is 1:20.

[0028] Preferably, the mixing volume ratio of the second solvent to the orange-red solid oil layer is 1~20:1.

[0029] Preferably, the dosage form of the traditional Chinese medicine compound preparation is capsules; Preferably, after drying, the sample is stored in a brown sample bottle, sealed, and kept in a refrigerator at 4°C.

[0030] This invention provides a method for analyzing and detecting volatile components in compound Panax notoginseng and Salvia miltiorrhiza capsules using gas chromatography-mass spectrometry (GC-MS). This method provides a technique for characterizing volatile components in compound Panax notoginseng and Salvia miltiorrhiza capsules using GC-MS, which can quickly and accurately obtain qualitative characterization results of volatile components in compound Panax notoginseng and Salvia miltiorrhiza capsules.

[0031] The test solution was prepared according to Method A of "2204 Determination of Volatile Oils" in Part IV of the Pharmacopoeia of the People's Republic of China.

[0032] The triple quadrupole gas chromatography-mass spectrometry (GC-MS) technology of this invention provides an effective approach for the quality evaluation and control of compound Panax notoginseng and Salvia miltiorrhiza capsules, and has important guiding significance for the development of compound Panax notoginseng and Salvia miltiorrhiza capsules.

[0033] The features and performance of the present invention will be further described in detail below with reference to embodiments. Unless otherwise specified, all materials and instruments used in the following embodiments are commercially available; and all detection methods used are existing methods unless otherwise specified.

[0034] In the following embodiments: 1. Materials 1.1 Instruments and Reagents GCMS-TQ8040 NX (SHIMADZU); Electron ionization source (EI); NIST20 mass spectrometry library; Volatile oil extractor; Precisa 125A electronic analytical balance (Precisa GmbH, Switzerland); KQ-250DB CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); Ethyl acetate, n-hexane, and anhydrous sodium sulfate were all analytical grade (Sinopharm Chemical Reagent Co., Ltd.); Acetonitrile was chromatographic grade (Merck).

[0035] 1.2 Samples Six batches of compound Sanqi Danshen capsules were randomly selected as traditional Chinese medicine compound samples (batch numbers: 2406046 / 2408035 / 2408048 / 2503012 / 2505023 / 2507036).

[0036] Example 1: Preparation of the test sample Take 2.0g of the compound Sanqi Danshen capsule powder and place it in a 50mL Erlenmeyer flask. Add 20mL of ethyl acetate, shake well, and extract by ultrasonication 3 times for 30min each time. Filter, combine the filtrates, let stand at room temperature, take the supernatant, filter it through a 0.25μm filter membrane, place it in a brown sample bottle, seal and store it in a 4℃ refrigerator for testing.

[0037] Example 2: Preparation of the test sample The difference from Example 1 is that n-hexane was used for extraction, and the sample was sealed and stored in a refrigerator at 4°C for testing.

[0038] Example 3: Preparation of the test sample Take 2.0g of the compound Sanqi Danshen capsule powder and place it in a 250mL round-bottom flask. Add 40mL of distilled water and boiling stones, mix thoroughly, and then connect the volatile oil extractor to the reflux condenser. Place the flask in a heating mantle and slowly heat it to the boiling point, maintaining a gentle boil for 5 hours until the amount of oil in the extractor no longer increases. Stop heating and allow it to stand to separate, obtaining an orange-red solid oil layer. Add a second solvent (ethyl acetate) and mix to dissolve the oil. The volume ratio of the second solvent to the orange-red solid oil layer is 1:1. The resulting compound Chinese medicine test solution is dried with anhydrous sodium sulfate and then sealed and stored in a refrigerator at 4℃ for testing.

[0039] The GC-MS detection conditions for the test samples obtained by the methods provided in Examples 1-3 are all the same; GC conditions: Shimadzu SH-I-5Si MS capillary column (30m length, 0.25mm inner diameter, 0.25μm particle size), column temperature program: initial temperature set to 100℃, hold for 10 min; increase to 150℃ at a rate of 10℃ / min, hold for 20 min; then increase to 200℃ at a rate of 3℃ / min, hold for 10 min; then increase to 300℃ at a rate of 10℃ / min, hold for 6 min; total analysis time was 77 min; injection port temperature set to 250℃, column flow rate was 1.0 mL / min, total flow rate was 24 mL / min; make-up gas flow rate was 3 mL / min; split ratio was 20; high-pressure injection, pressure 250.0 kPa, time 1.0 min.

[0040] MS conditions: Ion source temperature set to 250℃, interface temperature set to 250℃, solvent delay time set to 3 min, photomultiplier tube voltage set to 0.3 kV, data export started 3 min later, mass scan range set to 50~1000 amu, scan interval set to 0.20 s.

[0041] Examples 1-3 provide chromatographic detection results of the test samples obtained by the method as follows: Figures 1-3 As shown, through comparison Figures 1-3 It was found that the volatile components extracted using the method in Example 3 yielded richer chromatographic information and better separation of chemical components, as shown in the results. Figure 3 As shown.

[0042] Example 4: Optimization of Mass Spectrometry Analysis Conditions The test sample was prepared according to the method in Example 3 and then detected by GC-MS. GC conditions: Shimadzu SH-I-5Si MS capillary column (30m length, 0.25mm inner diameter, 0.25μm particle size), column temperature program: initial temperature set to 80℃, hold for 10 min; increase to 300℃ at a rate of 5℃ / min, hold for 10 min; total analysis time was 64 min; injection port temperature set to 250℃, column flow rate was 1.02 mL / min, total flow rate was 24.4 mL / min; make-up gas flow rate was 3 mL / min; split ratio was 20; high-pressure injection, pressure 250.0 kPa, time 1.0 min.

[0043] MS conditions: Ion source temperature set to 250℃, interface temperature set to 250℃, solvent delay time set to 3 min, photomultiplier tube voltage set to 0.2 kV, data export started 3 min later, mass scan range set to 50~1000 amu, scan interval set to 0.20 s.

[0044] Example 5: Optimization of Mass Spectrometry Analysis Conditions The test sample was prepared according to the method in Example 3 and then detected by GC-MS. GC conditions: Shimadzu SH-I-5Si MS capillary column (30m length, 0.25mm inner diameter, 0.25μm particle size), column temperature program: initial temperature set to 100℃, hold for 10min; increase to 300℃ at a rate of 3℃ / min, hold for 5min; total analysis time is 81min; injection port temperature set to 250℃, column flow rate is 1.00mL / min, total flow rate is 24.0mL / min; tail gas flow rate is 3mL / min; split ratio is 20; high pressure injection, pressure 250.0kPa, time 1.0min.

[0045] MS conditions: Ion source temperature set to 250℃, interface temperature set to 250℃, solvent delay time set to 3 min, photomultiplier tube voltage set to 0.2 kV, data export started 3 min later, mass scan range set to 50~1000 amu, scan interval set to 0.20 s.

[0046] Example 6: Optimization of Mass Spectrometry Analysis Conditions The test sample was prepared according to the method in Example 3 and then detected by GC-MS. GC conditions: Shimadzu SH-I-5Si MS capillary column (30m length, 0.25mm inner diameter, 0.25μm particle size), column temperature program: initial temperature set to 100℃, hold for 10 min; increase to 150℃ at a rate of 10℃ / min, hold for 20 min; then increase to 200℃ at a rate of 10℃ / min, hold for 20 min; then increase to 300℃ at a rate of 10℃ / min, hold for 5 min; total analysis time was 75 min; injection port temperature set to 250℃, column flow rate was 1.0 mL / min, total flow rate was 24 mL / min; make-up gas flow rate was 3 mL / min; split ratio was 20; high-pressure injection, pressure 250.0 kPa, time 1.0 min.

[0047] MS conditions: Ion source temperature set to 250℃, interface temperature set to 250℃, solvent delay time set to 3 min, photomultiplier tube voltage set to 0.2 kV, data export started 3 min later, mass scan range set to 50~1000 amu, scan interval set to 0.20 s.

[0048] Example 7: Optimization of Mass Spectrometry Analysis Conditions The test sample was prepared according to the method in Example 3 and then detected by GC-MS. GC conditions: Shimadzu SH-I-5Si MS capillary column (30m length, 0.25mm inner diameter, 0.25μm particle size), column temperature program: initial temperature set to 100℃, hold for 10 min; increase to 150℃ at a rate of 10℃ / min, hold for 20 min; then increase to 200℃ at a rate of 3℃ / min, hold for 10 min; then increase to 300℃ at a rate of 10℃ / min, hold for 6 min; total analysis time was 77 min; injection port temperature set to 250℃, column flow rate was 1.0 mL / min, total flow rate was 24 mL / min; make-up gas flow rate was 3 mL / min; split ratio was 20; high-pressure injection, pressure 250.0 kPa, time 1.0 min.

[0049] MS conditions: Ion source temperature set to 250℃, interface temperature set to 250℃, solvent delay time set to 3 min, photomultiplier tube voltage set to 0.2 kV, data export started 3 min later, mass scan range set to 50~1000 amu, scan interval set to 0.20 s.

[0050] Example 8: Optimization of different mass spectrometry analysis conditions The test sample was prepared according to the method in Example 3 and then detected by GC-MS. GC conditions: Shimadzu SH-I-5Si MS capillary column (30m length, 0.25mm inner diameter, 0.25μm particle size), column temperature program: initial temperature set to 100℃, hold for 10 min; increase to 150℃ at a rate of 10℃ / min, hold for 20 min; then increase to 200℃ at a rate of 3℃ / min, hold for 10 min; then increase to 300℃ at a rate of 10℃ / min, hold for 6 min; total analysis time was 77 min; injection port temperature set to 250℃, column flow rate was 1.0 mL / min, total flow rate was 24 mL / min; make-up gas flow rate was 3 mL / min; split ratio was 20; high-pressure injection, pressure 250.0 kPa, time 1.0 min.

[0051] MS conditions: Ion source temperature set to 250℃, interface temperature set to 250℃, solvent delay time set to 3 min, photomultiplier tube voltage set to 0.3 kV, data export started 3 min later, mass scan range set to 50~1000 amu, scan interval set to 0.20 s.

[0052] The detection results of Examples 4-8 are as follows: Figures 4-8 As shown, a comprehensive comparison is made from multiple aspects, including molecular ion peak response, number and intensity of ion peaks, overall characteristic peak layout, baseline stability, and noise interference. Figures 4-8 The results showed that the chromatographic conditions used in Example 8 were the optimal analytical conditions, and the analytical time was set at 77 min.

[0053] Example 9: Analysis and Characterization of Volatile Components in Compound Panax Notoginseng and Salvia Miltiorrhiza Capsules After preparing test samples from six batches of Compound Panax notoginseng and Salvia miltiorrhiza capsules according to the method in Example 3, GC-MS detection was performed under the conditions in Example 8 to establish a qualitative characterization method. Representative total ion chromatograms of Compound Panax notoginseng and Salvia miltiorrhiza capsules with batch numbers 2406046, 2408048, and 2505023 are shown below. Figures 9-11 As shown.

[0054] Fifty-six representative chemical components were selected from the common peaks of six batches of compound preparations. After comparison with the NIST20 mass spectrometry library and a self-built database, these components included the main active ingredients from Cyperus rotundus (such as caryophyllene, β-serinene, caryophyllin, cyperone, and α-cyperone), the main active ingredients from Salvia miltiorrhiza (such as isopyraldehyde and ferricyl alcohol), and the main active ingredients from Panax notoginseng (such as ginsenoside alcohol and ginsenoside epoxide). Mass spectrometry data and compound names are shown in Tables 1-4.

[0055] Table 1 Qualitative Characterization Results

[0056] Table 2 Qualitative Characterization Results

[0057] Table 3 Qualitative Characterization Results

[0058] Table 4 Qualitative Characterization Results

[0059] From Tables 1-4 and Figures 9-11 It is evident that the method provided in this application can accurately obtain the characteristic substances of the volatile components of trace amounts of Cyperus rotundus contained in the Compound Panax notoginseng and Salvia miltiorrhiza Capsules in the sample to be tested, and realize the accurate detection of Cyperus rotundus components mixed in Compound Panax notoginseng and Salvia miltiorrhiza Capsules as well as various volatile components contained in Compound Panax notoginseng and Salvia miltiorrhiza Capsules, with accurate detection results.

[0060] Example 10 The difference from the preparation of the test sample in Example 3 is as follows: the mass-to-volume ratio of the first solvent is 1:20; the volume ratio of the second solvent to the orange-red solid oil layer is 20:1. The chromatographic conditions differed from those in Example 8 as follows: the injection port temperature was set to 200°C, the column flow rate was 0.5 mL / min, the total flow rate was 15 mL / min, the tail gas flow rate was 1 mL / min, and the split ratio for split injection was 8. The difference in quality conditions compared to Example 8 is that the photomultiplier tube voltage is 0.1 kV, data export begins 3 minutes later, and the quality scan range is set to 50 amu.

[0061] Example 11 The difference from the preparation of the test sample in Example 3 is that the mixing volume ratio of the second solvent to the orange-red solid oil layer is 1:1; The chromatographic conditions differ from those in Example 8 as follows: the injection port temperature is set to 300°C, the column flow rate is 2 mL / min, the total flow rate is 35 mL / min, the tail gas flow rate is 10 mL / min, and the split ratio for split injection is 40. The difference in quality conditions compared to Example 8 is that the photomultiplier tube voltage is 0.8 kV, data export starts 3 minutes later, and the quality scan range is set to 1200 amu.

[0062] Example 12 The chromatographic conditions differed from those in Example 3 as follows: the mass-to-volume ratio of the traditional Chinese medicine compound preparation to water was 1:40; the volume ratio of the second solvent (chloroform) to the orange-red solid oil layer was 16:1; and the split ratio for split injection was 20.

[0063] Example 13 The chromatographic conditions differed from those in Example 3 as follows: the mass-to-volume ratio of the traditional Chinese medicine compound preparation to water was 1:8; and the volume ratio of the second solvent (n-hexane) to the orange-red solid oil layer was 1:1.

[0064] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for characterizing volatile components in Compound Panax notoginseng and Salvia miltiorrhiza Capsules using gas chromatography-mass spectrometry (GC-MS), characterized in that, Includes the following steps: (1) Preparation of the test solution of compound Panax notoginseng and Salvia miltiorrhiza capsules; The preparation method of the test solution includes: adding a first solvent and zeolite to the compound Sanqi Danshen capsule traditional Chinese medicine compound preparation, the mass-to-volume ratio of the traditional Chinese medicine compound preparation and the first solvent is 1:8~40, after thorough mixing, connecting the volatile oil extractor and the reflux condenser, placing it in an electric heating mantle and slowly heating to the boiling point, and maintaining a gentle boil for 1~5 hours until the amount of oil in the extractor no longer increases, stopping the heating, allowing it to stand and separate to obtain an orange-red solid oil layer, adding a second solvent to mix and dissolve, and obtaining the traditional Chinese medicine compound test solution; (2) The volatile components in the test solution of the traditional Chinese medicine compound were detected by a triple quadrupole gas chromatography-mass spectrometry (GC-MS) instrument. The retention time, molecular weight and characteristic structural fragment information of the volatile components contained in the test solution were obtained. The volatile components were determined by comparing them with the chromatographic and mass spectrometric characteristics of known compounds. The chromatographic conditions for detection were as follows: a capillary column was used, with the following column temperature program: initial temperature 100℃, held for 10 min; temperature increased to 150℃ at a rate of 10℃ / min, held for 20 min; then increased to 200℃ at a rate of 3℃ / min, held for 10 min; then increased to 300℃ at a rate of 10℃ / min, held for 6 min; the total analysis time was 77 min; the injection port temperature was set to 200~300℃, the column flow rate was 0.5~2 mL / min, the total flow rate was 15~35 mL / min, and the tail gas flow rate was 1~10 mL / min. The mass spectrometry conditions during detection were as follows: ion source temperature was set to 250℃, interface temperature was set to 250℃, solvent removal time was 3 min, photomultiplier tube voltage was 0.1~0.8 kV, data export started 3 min later, mass scan range was set to 50~1200 amu, and scan interval was 0.20 s.

2. The method according to claim 1, characterized in that, The chromatographic column used was a Shimadzu SH-I-5Si MS capillary column.

3. The method according to claim 1, characterized in that, The specifications of the chromatographic column are: column length 30m, inner diameter 0.25mm, particle size 0.25μm.

4. The method according to claim 1, characterized in that, The sample is injected using a split injection method with a split ratio of 8 to 40.

5. The method according to claim 4, characterized in that, The split ratio for split injection is 20.

6. The method according to claim 1, characterized in that, The first solvent is water; the second solvent is any one of ethyl acetate, chloroform, or n-hexane.

7. The method according to claim 1, characterized in that, The mass-to-volume ratio of the traditional Chinese medicine compound preparation to the first solvent is 1:

20.

8. The method according to claim 1, characterized in that, The mixing volume ratio of the second solvent to the orange-red solid oil layer is 1~20:1.

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