Total components and blood-entering components of herba artemisiae scopariae dampness-clearing granules and detection method of total components and blood-entering components

Abstract

The invention discloses a total component and a blood-entering component of a herba artemisiae scopariae dampness-clearing granule and a detection method of the total component and the blood-entering component. The preparation method comprises the following steps: A, decocting the herba artemisiae scopariae dampness-clearing granules, filtering decoction to obtain filtrate, and concentrating the filtrate to obtain concentrated liquid; b, detecting the concentrated solution by adopting liquid chromatography-mass spectrometry; the liquid chromatography conditions are as follows: a reversed-phase C18 chromatographic column is adopted, the column temperature is 40 DEG C, a mobile phase A is formic acid water with the volume fraction of 0.1%, a mobile phase B is acetonitrile, and the flow velocity is 0.3 ml / min. The research investigates all components of the dampness-clearing prescription, i.e., the rhizoma smilacis glabrae dampness-clearing granules, and investigates blood entering components of the dampness-clearing prescription, i.e., the rhizoma smilacis glabrae dampness-clearing granules after long-term intervention in patients with the dampness-syndrome metabolic syndrome in clinical practice. A new modern theoretical system is injected for the research of traditional Chinese medicine dampness syndrome and dampness clearing prescription clinical intervention, and a reference is provided for the research of other syndromes of traditional Chinese medicine.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine, and specifically relates to full ingredients and blood-entering ingredients of Yinling Qushi granules and a detection method thereof. Background Art

[0002] In recent years, scholars have summarized the experience of famous Chinese medicine practitioners in treating dampness through data mining. Meng Peipei and others used association analysis and other methods to find that Mr. Pu Fuzhou used Poria, Licorice, Tongcao, Artemisia capillaris, and Coix seed to treat dampness, emphasizing the importance of removing dampness. Commonly used pairs of drugs include Tongcao-Artemisia capillaris, Tongcao-Coix seed, Pinellia-Poria, etc., and horn drugs include Talc-Tongcao-Apricot kernel, Tongcao-Poria-Coix seed, etc. Mr. Pu added flavors to the classic prescriptions, such as Apricot kernel Talc Decoction with Artemisia capillaris, Alisma orientalis, etc., to clear dampness and heat and harmonize the spleen and stomach. Zhang Xinwen and others summarized Li Shimao's experience in treating dampness. Mr. Li advocated "nourishing yin and removing dampness", and often used Ophiopogon japonicus, Rehmannia glutinosa, Scrophularia ningpoensis, etc. to nourish yin fluid, and Atractylodes macrocephala and Poria cocos to dry and benefit. Zeng Chuiyi summarized Mao Dexi's experience. Mao believed that dampness should be treated according to the three burners: the upper burner clears heat and dries dampness, the middle burner removes dampness with aromatics, and the lower burner promotes diuresis and eliminates dampness. Commonly used drugs include coptis root, scutellaria, patchouli, atractylodes, tuckahoe, umbellate, and oriental herbs. Zhang Junhua and others used complex network analysis technology to find that tuckahoe, atractylodes, oriental herbs, and dried orange peel are the core medicinal flavors for removing dampness, and summarized the classic prescriptions and commonly used treatments for dampness. These studies are of great significance for inheriting the ideas of famous Chinese medicine masters on treating dampness, the rules of medication, and clinical guidance of Chinese medicine medication.

[0003] Yinling Qushi Granules are made from Fuling Zexie Decoction (source: Synopsis of the Golden Chamber) and are made by adding or subtracting ingredients from it. Yinling Qushi Granules are made up of eight herbs: Fuling, Atractylodes macrocephala, Polyporus umbellatus, Cinnamon twig, Dry ginger, Licorice root, Artemisia capillaris, and Adenophora scabra. Its [source], [effects], [main indications], and [prescription explanation] are as follows:

[0004]

Source

[0005] [Effects] Strengthens the spleen, promotes diuresis and eliminates dampness, warms the middle and benefits the stomach, and eliminates dampness and heat.

[0006] [Indications] It is used for dampness blocking the middle part of the body, spleen and stomach deficiency and coldness, which leads to stagnation of the middle burner, diffuse dampness, and concurrent damp-heat symptoms.

[0007] [Explanation of the prescription] The main ingredient in the prescription is Poria cocos, which is sweet, mild and neutral in nature, and can strengthen the spleen, promote diuresis and eliminate dampness. The auxiliary ingredient is cinnamon twig, which is pungent and warm, and can open the bladder to help transform qi and promote diuresis, so as to eliminate dampness.

[0008] The assistant drug Atractylodes macrocephala is sweet, bitter and warm in nature, and can strengthen the middle jiao, helping the main drug to strengthen the ability to remove dampness, so as to eliminate the source of dampness. Polyporus umbellatus is sweet, mild and neutral in nature, and can penetrate dampness and promote diuresis, helping the main drug to remove dampness without hurting the true yin. Dry ginger is pungent and hot, and can strengthen the spleen, warm the middle and eliminate dampness.

[0009] The adjuvant herb Artemisia capillaris is bitter, pungent and slightly cold, which can clear away heat and dampness, and restrain the warm and dry nature of the above herbs. The herb Adenophora scabra is sweet, bitter and slightly cold, and its sweetness can nourish yin and prevent excessive dampness from damaging yin fluid.

[0010] Licorice root is sweet and mild in nature, and can invigorate the spleen, clear away heat and promote diuresis, and harmonize the other medicines. The whole formula can invigorate the spleen, promote diuresis and eliminate dampness, warm the middle and benefit the stomach, and promote dampness and heat. Summary of the invention

[0011] The purpose of the present invention is to provide a complete composition of Yinling Qushi granules and blood-entering components and a detection method thereof.

[0012] The method for detecting all ingredients of Yinling Qushi Granules of the present invention comprises the following steps:

[0013] A. decoct Yinling Qushi granules, filter the decoction to obtain a filtrate, and concentrate the filtrate to obtain a concentrated solution;

[0014] B. detecting the concentrate by liquid chromatography-mass spectrometry;

[0015] The liquid chromatography conditions are: using a reverse phase C18 chromatographic column, a column temperature of 40°C, a mobile phase: A is 0.1% by volume formic acid water, B is acetonitrile, and a flow rate of 0.3 ml / min;

[0016] 0-20min mobile phase A was reduced from 95% to 50%;

[0017] 20-27min mobile phase A was reduced from 50% to 5%;

[0018] 27-32min mobile phase A was maintained at 5%;

[0019] 32-33min, mobile phase A increased from 5% to 95%;

[0020] 33-38min mobile phase A was maintained at 95%;

[0021] The mass spectrometry conditions were as follows: ESI ion source, carrier gas 1 / 2: 50 psi, CUR: 35 psi, temperature: 500 °C, ISVF: 5500 V (+), -4500 V (-), CE and CES: 35 ± 15 eV, m / z: 50-1500 Da, DP: 80 eV;

[0022] The full ingredients of Yinling Qushi Granules include triterpenes, flavonoids, saponins, amino acids, and phenolic acids.

[0023] Preferably, the particle analysis of the full composition is shown in Table 2.

[0024] Preferably, the concentrated solution obtained by concentrating the filtrate is concentrated under reduced pressure to a concentrated solution having a specific gravity of 1.08 to 1.10.

[0025] The second object of the present invention is to provide a method for detecting the blood-entering components of Yinling Qushi Granules, comprising the following steps:

[0026] A. Give Yinling Qushi Granules to patients with dampness syndrome, and collect serum after a predetermined period of time as the experimental group, and use the serum of patients with dampness syndrome who were not given Yinling Qushi Granules as the control group;

[0027] B. The experimental group and the control group were tested by liquid chromatography-mass spectrometry;

[0028] The liquid chromatography conditions are: using a reverse phase C18 chromatographic column, a column temperature of 40°C, a mobile phase: A is 0.1% by volume formic acid water, B is acetonitrile, and a flow rate of 0.3 ml / min;

[0029] 0-20min mobile phase A was reduced from 95% to 50%;

[0030] 20-27min mobile phase A was reduced from 50% to 5%;

[0031] 27-32min mobile phase A was maintained at 5%;

[0032] 32-33min, mobile phase A increased from 5% to 95%;

[0033] 33-38min mobile phase A was maintained at 95%;

[0034] The mass spectrometry conditions were as follows: ESI ion source, carrier gas 1 / 2: 50 psi, CUR: 35 psi, temperature: 550 °C, ISVF 5500 V (+), -4500 V (-), CE and CES: 35 ± 15 eV, m / z: 20-1500 Da, DP: 80 eV;

[0035] The ingredients of Yinling Qushi Granules that enter the blood are: L-Valin, Liquiritigenin, Ononin, Formononetin, Sulfation of Formononetin, Daidzein, calycosin, Sulfation of calycosin, poricioic acid G, Glycyrrhizic acid, Daedaleanic acid B, Poricoic acid B, Poricoic Acid DM, Dehydroeburic acid, α-linolenic acid, linoleic acid, and oleic acid.

[0036] Preferably, the serum is pretreated, and the pretreatment steps are as follows:

[0037] (1) 90 μL of human serum from the patient 4 weeks after administration was taken into a centrifuge tube and 10 μL of IS solution containing 10 μg / mL deuterated naringenin was added;

[0038] (2) Add 1 mL of ethyl acetate solution, vortex for 5 min, and centrifuge at 10,000 rpm at 4 °C for 15 min;

[0039] (3) Take the supernatant, blow dry with nitrogen, and add 100 μL of 80% methanol-water mixed solution;

[0040] (4) Ultrasonication for 5 min, vortexing for 5 min, centrifugation at 13000 rpm at 4°C for 15 min, and testing by a microcomputer.

[0041] This study investigated the full ingredients of the dehumidifying prescription Yinling Qushi Granules, and investigated the blood components of the dehumidifying prescription Yinling Qushi Granules after long-term intervention in patients with metabolic syndrome with dampness syndrome in clinical practice. It is expected to inject a new modern theoretical system into the research of dampness syndrome in traditional Chinese medicine and clinical intervention of dehumidifying prescriptions, and provide reference for the research of other syndromes in traditional Chinese medicine. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 This is the metabolic transformation pathway of the components of Yinling Qushi Granules entering the blood;

[0043] Figure 2 This is the secondary fragmentation diagram of formononetin and its sulfated products, (A) formononetin; (B) formononetin sulfated products. DETAILED DESCRIPTION

[0044] The following examples are provided to further illustrate the present invention, rather than to limit the present invention.

[0045] Embodiment 1:

[0046] 1. Methods

[0047] 1. Analysis of the full ingredients of Yinling Qushi Granules

[0048] 1.1 Preparation of Yinling Qushi Granules and Single Herbal Ingredients

[0049] The preparation method of Yinling Qushi Granules is as follows:

[0050] (1) Poria 77.76g, Atractylodes macrocephala 29.16g, Ginger 12.96g, Polyporus umbellatus 19.44g, Cinnamomum cassia twig 19.44g, Artemisia capillaris 24.3g, Adenophora australis 19.44g, Licorice root 19.44g.

[0051] (2) Mix all the Chinese medicinal materials and boil them. For the first decoction, add 10 times the volume of water, heat to boiling, keep it boiling for 1.5 hours, and filter it (200 mesh sieve).

[0052] (3) For the second decoction, add 10 times the volume of water, heat to boiling, maintain a slight boil for 1.0 h, and filter with suction (200 mesh sieve).

[0053] (4) Combine the two filtrates and concentrate under reduced pressure to a concentrate with a specific gravity of 1.08 to 1.10.

[0054] (5) After spray drying, sieve and mix to obtain the product.

[0055] (6) The extracts of each single herbal medicine were prepared in the same manner as above.

[0056] (7) Accurately measure 1 mL of Yinling Qushi Granule Concentrate and each single herbal concentrate into a 5 mL volumetric flask, add water to the scale line, mix well, and then test on the machine.

[0057] 1.2 Liquid chromatography-mass spectrometry detection conditions

[0058] Liquid phase conditions: a reverse phase C18 chromatographic column (3×150 mm, 2.6 μm) of pheromone (Phenonmenex, USA) was used, the column temperature was 40° C., the mobile phases: A was 0.1% by volume formic acid water, B was acetonitrile, and the elution parameters were shown in Table 1.

[0059] Table 1 Gradient elution program

[0060]

[0061] The 5600plus mass spectrometer system from AB Sciex was used for detection. The optimized parameters are as follows: ESI ion source, carrier gas 1 / 2: 50psi, CUR: 35psi, temperature: 500℃, ISVF: 5500V(+), -4500V(-), CE and CES: 35±15eV, m / z: 50~1500Da, DP: 80eV. The detection mode of IDA was developed by AB Sciex. 1.2 software (AB Sciex, USA) was used for acquisition and detection.

[0062] 1.3 Data Analysis

[0063] The steps for analyzing the full ingredients of Yinling Qushi Granules are as follows

[0064] (1) Self-built database construction: Use PubMed and CNKI to collect the chemical component information of the single medicinal material of Yinling Qushi Granules, and integrate it into a Peakview (version 1.2, AB sciex) compatible data format. The specific steps are as follows: Use the built-in XIC Manager module in Peakview, click "New session" and select the wiff file, select "Asastandard TIC", import the single medicinal material information collected in the early stage into the XIC manager, and save this information in the XIClist suffix file format to obtain the self-built database. Then in the subsequent analysis, you only need to use Peakview to import the data, import this XIClist file, and click show XIC to get the comparison results. Finally, perform the secondary fragment information comparison, delete the components that are both red in Mass and Isotope, and get the final comparison results.

[0065] (2) High-resolution natural product database comparison: using Natural Products HR MS 2 Spectral Library (Version 1.0.3, ABsciex) database for further comparison. The specific steps are as follows: Use Peakview (version 2.2, AB Sciex), which is compatible with the natural product database developed by AB Sciex. Click "File" to import the wiff file, use the "Masterview" module, and the comparison parameters are: Precursor Mass is 0.4Da, Mass Tolerence: 0.4Da, Maximal Number of Hits: 5, Intensity Threshold: 0.05, Minmal Purity: 10%, and the parameters are all default options. Click "Import Compounds from LibraryViewDatabase", and after loading, select the "TCM MS / MS Library" database, and check "Import FragmentMass (Da) for compounds", and start searching after confirmation. Finally, sort the data, delete the components with both Mass and Isotope being red, and retain the compounds with a comparison score above 0.8, and the final comparison results are obtained.

[0066] 2. Analysis of blood components of Yinling Qushi Granules

[0067] 2.1 Preparation of serum samples after long-term intervention with Yinling Qushi Granules

[0068] Thirty patients with metabolic syndrome with dampness were selected clinically and given a daily dose of 137 g of Yinling Qushi Granules for 4 weeks, and their serum was collected.

[0069] (1) Take 90 μL of clinical human serum after 4 weeks of administration into a centrifuge tube and add 10 μL of IS solution (containing 10 μg / mL deuterated naringenin);

[0070] (2) Add 1 mL of ethyl acetate solution, vortex for 5 min, and centrifuge at 4°C for 15 min (10,000 rpm);

[0071] (3) Take the supernatant, blow dry with nitrogen, and add 100 μL of 80% methanol-water mixed solution;

[0072] (4) Ultrasonication for 5 min, vortexing for 5 min, centrifugation at 4°C for 15 min (13000 rpm), and testing on a centrifuge.

[0073] 2.2 Liquid chromatography-mass spectrometry detection conditions

[0074] A pheromone (Phenonmenex, USA) reverse phase C18 chromatographic column (2.1×100 mm, 2.6 μm) was used, and the elution parameters were the same as those in “1.2 Liquid chromatography-mass spectrometry detection under the item of “1. Analysis of all ingredients of Yinling Qushi granules”.

[0075] The 7600 Zeno-TOF mass spectrometer system of AB Sciex was used for detection. The optimized parameters are as follows: ESI was used as the ion source, carrier gas 1 / 2: 50 psi, CUR: 35 psi, temperature: 550 ° C, ISVF was 5500 V (+), -4500 V (-), CE and CES: 35 ± 15 eV, m / z: 20-1500 Da, DP: 80 eV. Data acquisition was performed in IDA mode, and data analysis was performed using SCIEXOS2.0 software (AB Sciex, USA).

[0076] 2.3 Data Analysis

[0077] The analysis steps of the components entering the blood of Yinling Qushi Granules are the same as those in "1.3 Data Analysis under 1. Analysis of All Components of Yinling Qushi Granules". For the identification of metabolites of the components entering the blood, the secondary fragments and the cleavage fragments of the parent nucleus show similar cleavage patterns under mass spectrometry detection, and if a phase I metabolic reaction occurs in the body, such as a methylation (CH2) reaction, the mass of the metabolite is 14Da more than the mass of the parent nucleus; if a phase II metabolic reaction occurs, such as a sulfation (SO3) reaction, the mass of the metabolite is 80Da more than the mass of the parent nucleus. According to the above rules, a self-built library of the in vivo metabolic pathway of each chemical component can be constructed according to the steps of self-built libraries, and the in vivo metabolite results can be obtained by searching and comparing.

[0078] 2. Experimental Results

[0079] In order to comprehensively analyze the chemical composition of Yinling Qushi Granules, the chemical composition of single medicinal materials, including Poria cocos, Polyporus umbellatus, dried ginger and other medicinal materials, were collected by consulting literature.

[0080] UFLC-Q-TOF-MS / MS technology was used to analyze the chemical components of Yinling Qushi Granules and the eight medicinal materials it contained. According to the self-built library information and the secondary fragment information collected in the natural product library, a total of 106 chemical components of Yinling Qushi Granules were identified under positive and negative mode detection. After comparison with single medicinal materials and component attribution, 43 species were found to be derived from Poria cocos, 6 species were derived from dried ginger, and 8 species were derived from Polyporus umbellatus. The number of other medicinal materials detected is shown in Table 2. The chemical components were then structurally classified. The results showed that the components of Yinling Qushi Granules include triterpenes, flavonoids, saponins, amino acids, phenolic acids and other types. The above results show that the components of Junling Qushi Granules are complex and varied, suggesting the characteristics of multi-component Chinese medicine.

[0081] Table 2 Analysis of ingredients in Yinling Qushi Granules

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098] Note: FL: Poria, BZ: Atractylodes macrocephala, ZL: Polyporus umbellatus, GJ: Dry ginger, YC: Artemisia capillaris, GZ: Cinnamomum cassia twig, BSS: Glehnia littoralis, GC: Licorice root

[0099] In previous studies, we conducted a comprehensive analysis of the full composition of Yinling Qushi Granules and constructed a chemical composition database of Yinling Qushi Granules. Based on this database, we identified the blood-entering components of Yinling Qushi Granules in vivo. The results are shown in Table 3. Through comparison with secondary fragments, a total of 17 blood-entering components were detected in the patient's serum, of which 14 were prototype components, which were divided into four categories according to the parent core structure: terpenes, flavonoids, saponins, and amino acids; 3 metabolites (M4-1, M4-2, M5-1).

[0100] Table 3 Blood components and metabolites of Yinling Qushi Granules

[0101]

[0102]

[0103]

[0104] Note: Glc = glucoside; P: prototype compound, M: metabolite; BZ, Atractylodes macrocephala; GC, Licorice root; FL, Poria cocos

[0105] Therefore, we constructed the metabolic pathways of the blood-entering components and their metabolites in the body. Figure 1As shown in the figure, the results show that the terpenoids and saponins in Yinling Qushi Granules mainly exist in the body in the original form, but the abundance is low; while after the flavonoids are absorbed and transformed in the body, some compounds exist in the body in the original form; the other part undergoes metabolic transformation, and the sulfation reaction in phase II metabolism is the main reaction. Among them, P3, P4, and P5 were confirmed to be ononin, formononetin, and calycosin respectively after comparison with the secondary fragments. Unlike terpenoids, these flavonoids undergo extensive phase I and phase II metabolic reactions in the body. Among them, formononetin (P3) is hydrolyzed in the body to remove the glycoside part to form its aglycone component formononetin (P4). Formononetin can generate daidzein (M4-2) through demethylation in the body. On the contrary, daidzein can generate formononetin (P4) under the action of methylation; in addition, formononetin can be dehydroxylated to generate calycosin (P5). Similarly, calycosin generates formononetin (P5) under the action of hydroxylation. For the phase II reaction, the sulfation reaction loses 80Da (SO3) on the mass spectrometry characteristic fragment. Based on this fragmentation pattern, M4-1 and M5-1 were preliminarily identified as the sulfation products of formononetin (P4) and calycosin (P5) in the body, respectively. For example, under negative mode detection, formononetin (Molecular weight: 268.26) generates precursor ions 267.0665 [MH]- under the action of ESI source, then loses CH3 (15Da) to generate 252.0433 [MH-CH3]-, and then continues to cleave to generate 223.0404 [MHO-CO]-, 208.0526 [MHO-CO-CH3]- and 195.0455 characteristic fragments. As a flavonoid compound, formononetin ( Figure 2 A) Under the action of ESI source, a ring-opening reaction (retro Diels-Alder, RDA) occurs to generate characteristic ion fragment 132.0218[1,3B]-. Based on this fragmentation pattern, M4-1 was identified as a sulfated metabolite of formononetin ( Figure 2In the figure, the precursor ion 347.0227[MH]- was generated under the action of ESI source, and then 80Da(SO3) was removed to obtain the fragment ion 267.0666[MH-SO3]-, which was the precursor ion of the original compound of formononetin in negative mode, and then the same characteristic fragments were generated, such as 252.0429[MH-SO3-CH3]-, 223.0397[MH-SO3-O-CO]-, 208.0526[MH-SO3-O-CO-CH3]- and 195.0457. Finally, the formononetin sulfate product (M4-1) also underwent RDA ring-opening reaction and generated the characteristic fragment 132.0233[1,3B]-. In summary, after the administration of Yinling Qushi Granules, the blood-entering components were absorbed into the body and underwent phase I or phase II metabolic reactions, and the metabolites were found to have highly similar secondary fragments and pyrolysis patterns with the corresponding blood-entering components under the detection of high-resolution Zeno-TOF-MS / MS, suggesting that the above-mentioned blood-entering components and metabolites may be the active compounds of Yinling Qushi Granules.

Claims

1. A method for detecting all ingredients of Yinling Qushi Granules, characterized in that: The following steps are involved: A. decoct Yinling Qushi granules, filter the decoction to obtain a filtrate, and concentrate the filtrate to obtain a concentrated solution; B. detecting the concentrate by liquid chromatography-mass spectrometry; The liquid chromatography conditions are: using a reverse phase C18 chromatographic column, a column temperature of 40°C, a mobile phase: A is 0.1% by volume formic acid water, B is acetonitrile, and a flow rate of 0.3 ml / min; 0-20min mobile phase A was reduced from 95% to 50%; 20-27min mobile phase A was reduced from 50% to 5%; 27-32min mobile phase A was maintained at 5%; 32-33min, mobile phase A increased from 5% to 95%; 33-38min mobile phase A was maintained at 95%; The mass spectrometry conditions were as follows: ESI ion source, carrier gas 1 / 2: 50 psi, CUR: 35 psi, temperature: 500 °C, ISVF: 5500 V (+), -4500 V (-), CE and CES: 35 ± 15 eV, m / z: 50-1500 Da, DP: 80 eV; The full ingredients of Yinling Qushi Granules include triterpenes, flavonoids, saponins, amino acids, and phenolic acids.

2. The detection method according to claim 1, characterized in that: The analysis of the full ingredients is shown below.

3. The detection method according to claim 1, characterized in that: The concentrated solution obtained by concentrating the filtrate is concentrated under reduced pressure to a concentrated solution with a specific gravity of 1.08 to 1.

10.

4. A method for detecting blood-entering components of Yinling Qushi Granules, characterized in that: The following steps are involved: A. Give Yinling Qushi Granules to patients with dampness syndrome, and collect serum after a predetermined period of time as the experimental group, and use the serum of patients with dampness syndrome who were not given Yinling Qushi Granules as the control group; B. The experimental group and the control group were tested by liquid chromatography-mass spectrometry; The liquid chromatography conditions are: using a reverse phase C18 chromatographic column, a column temperature of 40°C, a mobile phase: A is 0.1% by volume formic acid water, B is acetonitrile, and a flow rate of 0.3 ml / min; 0-20min mobile phase A was reduced from 95% to 50%; 20-27min mobile phase A was reduced from 50% to 5%; 27-32min mobile phase A was maintained at 5%; 32-33min, mobile phase A increased from 5% to 95%; 33-38min mobile phase A was maintained at 95%; The mass spectrometry conditions were as follows: ESI ion source, carrier gas 1 / 2: 50 psi, CUR: 35 psi, temperature: 550 °C, ISVF 5500 V (+), -4500 V (-), CE and CES: 35 ± 15 eV, m / z: 20-1500 Da, DP: 80 eV; The ingredients of Yinling Qushi Granules that enter the blood are: L-Valin, Liquiritigenin, Ononin, Formononetin, Sulfation of Formononetin, Daidzein, calycosin, Sulfation of calycosin, poricioic acid G, Glycyrrhizic acid, Daedaleanic acid B, Poricoic acid B, Poricoic Acid DM, Dehydroeburic acid, α-linolenic acid, linoleic acid, and oleic acid.

5. The detection method according to claim 4, characterized in that: The serum is pretreated, and the pretreatment steps are as follows: (1) 90 μL of human serum from the patient 4 weeks after administration was taken into a centrifuge tube and 10 μL of IS solution containing 10 μg / mL deuterated naringenin was added; (2) Add 1 mL of ethyl acetate solution, vortex for 5 min, and centrifuge at 10,000 rpm at 4 °C for 15 min; (3) Take the supernatant, blow dry with nitrogen, and add 100 μL of 80% methanol-water mixed solution; (4) Ultrasonication for 5 min, vortexing for 5 min, centrifugation at 13000 rpm at 4°C for 15 min, and testing by a microcomputer.

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