A traditional Chinese medicine composition, its preparation method and application

Abstract

This invention relates to a traditional Chinese medicine composition comprising active ingredients extracted and purified from Artemisia annua, Lonicera japonica, and Gardenia jasminoides. The active ingredients include: neochlorogenic acid (≥2‰), chlorogenic acid (≥5‰), cryptochlorogenic acid (≥2‰), caffeic acid (≥0.07‰), isochlorogenic acid B (≥0.35‰), isochlorogenic acid A (≥0.17‰), isochlorogenic acid C (≥0.19‰), geniposide (≥5.97‰), strychnos nux-oxidase (≥0.07‰), geniposide (≥0.17‰), genipin (≥0.22‰), genipin gentiopicroside (≥0.76‰), and 7-hydroxyswert (≥2.44‰). The composition contains ≥10.3‰ total acid and ≥21.58‰ total glycosides. Based on the experimental content of this invention, traditional Chinese medicine compositions with specific active ingredient content characteristics were screened out, which have stable and excellent therapeutic effects. The above-mentioned active ingredient content characteristics can be used for the quality control of the traditional Chinese medicine composition, thereby ensuring the quality stability and reliable efficacy of the traditional Chinese medicine preparation.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and in particular to a traditional Chinese medicine composition, its preparation method, and its application. Background Technology

[0002] Exogenous wind-heat syndrome occurs when the body's defensive functions are weakened and unable to adapt, allowing pathogenic factors to invade through the mouth, nose, skin, and hair, attacking the lungs and the defensive qi, leading to disharmony between the defensive and defensive qi. It is commonly seen in upper respiratory tract infections, influenza, acute bronchitis, pneumonia, and other diseases. Main symptoms include persistent high fever, slight aversion to wind, nasal congestion, runny nose, headache and body aches, cough, thirst, sore throat, red tongue tip and sides, and a floating and rapid pulse. Statistics show that at least 2 million people worldwide die each year from fever, especially from high fever caused by upper respiratory tract infections and acute bronchitis, which seriously affects people's health, with annual treatment costs reaching billions of yuan.

[0003] There are many causes of high fever, such as viral infections, bacterial infections, mycoplasma infections, and parasitic infections, among which viral or bacterial infections are the most common. Fever is a defensive response of the body. It can accelerate metabolic processes, inhibit the growth and reproduction of pathogens, promote the increase of white blood cells, and enhance the function of the reticuloendothelial system (phagocytic cell activity, antibody production, and liver detoxification capacity, etc.), which is beneficial to disease recovery. The fever pattern is of important reference value for disease diagnosis, judging treatment effectiveness, and prognosis. On the other hand, fever can also deplete physical strength, causing headaches, insomnia, and even convulsions. Hyperpyrexia (above 41°C) or prolonged fever can lead to permanent brain damage and even death. Therefore, it is necessary to use antipyretics appropriately for moderate to severe fever. Western medicine mainly treats fever with antipyretic analgesics (such as nonsteroidal anti-inflammatory drugs like carbaspirin calcium, aspirin lysine, ibuprofen, acetaminophen, naproxen, etc.) and antiviral and antibacterial drugs (such as acyclovir, amantadine, ribavirin, penicillin, erythromycin, etc.). While its fever-reducing properties are rapid, it has significant side effects, primarily manifesting as gastrointestinal reactions such as nausea, vomiting, diarrhea, and abdominal pain, as well as effects on platelets. Occasionally, dizziness, excessive salivation, insomnia, and skin rashes may also occur. Traditional Chinese medicine, on the other hand, is characterized by its high efficacy and minimal side effects in treating this disease. Therefore, developing new, highly effective antiviral, antibacterial, and antipyretic Chinese medicines holds great promise.

[0004] Patent application number 02114995.X discloses a drug for clearing heat and detoxifying, with a formula of Artemisia annua 6-25g, Lonicera japonica 3-15g, and Gardenia jasminoides 3-12g, which has a good therapeutic effect on fever caused by exogenous wind-heat. However, for traditional Chinese medicine compound prescriptions, the raw materials are mostly derived from natural products. Affected by factors such as place of origin, growth period, harvesting season, and planting and processing techniques, the quality of medicinal materials varies greatly. Even if the raw materials all meet the pharmacopoeia requirements, the differences in effective components between different batches of raw materials from different places of origin will still lead to unstable composition and poor quality consistency in different batches of preparations. It is evident that prescriptions based solely on the weight of medicinal materials cannot meet the production requirements of modern traditional Chinese medicine preparations, seriously restricting the stability and controllability of the clinical efficacy of traditional Chinese medicine, and affecting the reproducibility and acceptance of research results. Meanwhile, the effective components in complex systems of traditional Chinese medicine compound prescriptions are not clearly defined. Which main components have a significant impact on efficacy, and what is the reasonable optimal content range for them, all require reasonable control of the range of different effective components. How to ensure the quality stability of traditional Chinese medicine preparations while also ensuring better efficacy is a key issue restricting the development of modern traditional Chinese medicine. Summary of the Invention

[0005] This invention aims to further study a traditional Chinese medicine compound composed of three herbs: Artemisia annua, Lonicera japonica, and Gardenia jasminoides. By combining herbs from different origins and batches, the invention conducts component analysis and efficacy tests on the compound. It focuses on the relationship between the main components with significant content variations in the compound and the efficacy results, identifies specific active ingredients that can affect efficacy and their appropriate content, screens out more potent traditional Chinese medicine compositions, further limits the optimal range of active ingredient combinations, and improves the preparation process to ensure the stable quality and reliable efficacy of the prepared traditional Chinese medicine product.

[0006] Extensive preliminary research on the material basis has isolated 101 compounds from this traditional Chinese medicine composition, including 32 organic acid compounds (neochlorogenic acid, chlorogenic acid, isochlorogenic acid A / B / C, cryptochlorogenic acid, etc.) and 29 iridoid glycosides (geniposide, genipin gentiopicroside, genipin glycoside, geniposide, strychnine, etc.). Literature reports that neochlorogenic acid, chlorogenic acid, isochlorogenic acid A / B / C, and cryptochlorogenic acid all possess antibacterial pharmacological activities; caffeic acid has anti-respiratory syncytial virus activity; strychnine has anti-inflammatory and antiviral pharmacological activities; most iridoid glycosides in gardenia have anti-inflammatory, analgesic, and antiviral effects; in vitro antiviral activity tests have confirmed that genipin compounds have an inhibitory effect on H1N1 influenza virus.

[0007] Based on literature reports, and combining the content control components of three medicinal materials in the pharmacopoeia with the amount of structurally defined components in the finished product of Retoxin Injection, a total of 13 components were selected as indicator component combinations, including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, 7-hydroxyswertia, geniposide, strychnos nux-vomica glycoside, geniposide, genipin glycoside, and genipin gentiopicroside. The pharmacological effects of different combinations of the above indicator components were studied to find the content range and mutual ratio relationship that can more effectively exert the pharmacological effect.

[0008] In view of this, the present invention proposes a traditional Chinese medicine composition, characterized in that, by mass fraction (parts per thousand), the composition comprises, at least 2‰ neochlorogenic acid, at least 5‰ chlorogenic acid, at least 2‰ cryptochlorogenic acid, at least 0.07‰ caffeic acid, at least 0.35‰ isochlorogenic acid B, at least 0.17‰ isochlorogenic acid A, at least 0.19‰ isochlorogenic acid C, at least 5.97‰ geniposide, at least 0.07‰ strychnoside, at least 0.17‰ geniposide, at least 0.22‰ genipin, at least 0.76‰ genipin gentiopicroside, and at least 2.44‰ 7-hydroxyswert; the extract contains at least 10.3‰ total acid and at least 21.58‰ total glycosides.

[0009] Preferably, the active ingredients include: neochlorogenic acid 2.5-5.14‰, chlorogenic acid 5.56-11.11‰, cryptochlorogenic acid 2.36-5.00‰, caffeic acid 0.14-0.26‰, isochlorogenic acid B 0.43-1.00‰, isochlorogenic acid A 0.22-0.46‰, and isochlorogenic acid C. The extract contains 0.38-0.69‰, geniposide 9.53-17.71‰, strychnoside 0.35-3.43‰, geniposide 0.42-0.79‰, genipin glycoside 0.36-0.68‰, genipin gentiopicroside 1.00-1.88‰, and 7-hydroxyswertia 3.31-6.88‰; the total acid content is 13.89-27.78‰, and the total glycoside content is 23.68-50.61‰.

[0010] The ratio of the above active ingredients is as follows:

[0011] Neochlorogenic acid 2.5-5.14%, chlorogenic acid 5.56-11.11%, cryptochlorogenic acid 2.36-5.00%, caffeic acid 0.14-0.26%, isochlorogenic acid B 0.43-1.00%, isochlorogenic acid A 0.22-0.46%, isochlorogenic acid C 0.38-0.69%, geniposide 9.53-17.71%, strychnos nux-vomica 0.35-3.43%, geniposide 0.42-0.79%, genipin glycoside 0.36-0.68%, genipin gentiopicroside 1.00-1.88%, 7-hydroxyswertia 3.31-6.88%; the extract contains 13.89-27.78% total acid and 23.68-50.61% total glycosides. The traditional Chinese medicine composition can be in any optional formulation, and the specific preparation method is as follows:

[0012] Take 750 parts by weight of honeysuckle, add 9-16 times the amount of hot water at 90-100℃, and extract 1-3 times, 1-2 hours each time, to obtain honeysuckle extract. Take 1250 parts by weight of artemisia annua, soak in 3-6 times the amount of water, and steam distill for 5-8 hours to extract artemisia annua volatile oil. Add 3-9 times the amount of water to the residue and extract for 1-3 hours. Concentrate the extract to a relative density of 1.10-1.12 (70-80℃), add ethanol to make the alcohol content 60-80%, let stand for 12 hours, and combine the alcohol precipitation supernatant with the honeysuckle extract. Concentrate under reduced pressure at 60-80℃ to a relative density of 1.05-1.15 to obtain concentrated artemisia annua and honeysuckle extract. Add ethanol to the concentrated extract to make the alcohol content 70%-80%, precipitate alcohol, let stand for 24-48 hours, and concentrate the alcohol precipitation supernatant under reduced pressure. The alcohol-precipitated concentrate was obtained, and the pH was adjusted to 1.5-2.5 with concentrated hydrochloric acid. Extraction was performed using 8-16 times the amount of ethyl acetate. The extract was concentrated under reduced pressure until the ethyl acetate odor was eliminated, and then vacuum dried to obtain the honeysuckle and artemisia extract. 600 parts by weight of gardenia were crushed into coarse powder and extracted 1-3 times with 6-10 times the amount of 60-80% ethanol, each time for 1-3 hours. The extract was concentrated under reduced pressure to a relative density of 1.10-1.20 (60-70℃) to obtain the gardenia extract concentrate. This concentrate was heated to boiling, and the pH was adjusted to 2.5-3.5 with hydrochloric acid. 1% by weight of solid paraffin wax was added, stirred until completely dissolved, cooled to room temperature, and refrigerated for 24 hours. The extract was filtered and extracted with 5-10 times the amount of n-butanol. The extract concentrate was concentrated under reduced pressure until the n-butanol odor was eliminated, and then vacuum dried to obtain the gardenia extract. The honeysuckle and artemisia extract, gardenia extract, and artemisia volatile oil were then mixed using standard procedures to obtain the final product.

[0013] Specifically, the fingerprint spectrum of Artemisia annua and the control fingerprint spectrum are compared using no fewer than 15 fingerprints. Figure 1 The indicated fingerprint peaks are reference correction peaks with a similarity greater than 0.5. Specifically, the peak areas and retention times of the control fingerprint spectrum are as follows:

[0014] Mark Peak Sequence Number Retention time [Min] Peak area 1 3.882885 731.039 2 9.348604 238.769 3 10.29601 271.04 4 12.74284 231.6808 5 15.21369 585.2557 6 16.36838 243.2447 7 19.64414 683.0679 8 25.43884 415.1487 9 26.61364 252.1879 10 28.55216 264.1007 11 32.46935 936.6907 12 33.13216 616.8419 13 34.28907 386.3651 14 36.97964 219.1915 15 38.53798 842.427

[0015] Specifically, the extraction amounts of the four components from honeysuckle are as follows: chlorogenic acid should not be less than 2.0%; neochlorogenic acid should not be less than 0.05%; cryptochlorogenic acid should not be less than 0.05%; and deoxygenated strychnine should not be less than 0.2%; their fingerprint chromatograms compared with the reference chromatogram are as follows: Figure 2 In comparison, the 12 chromatographic peaks identified showed a similarity greater than 0.95. Specifically, the peak areas and retention times of the control fingerprint are as follows:

[0016]

[0017] The fingerprint spectrum of the gardenia and the control fingerprint spectrum are as follows: Figure 3 In comparison, the four chromatographic peaks indicated a similarity greater than 0.9. Specifically, the peak areas and retention times of the control fingerprint are as follows:

[0018]

[0019] Specifically, the traditional Chinese medicine composition may include extracts, decoctions, granules, capsules, soft capsules, pills, oral liquids, tinctures, syrups, suppositories, gels, sprays, and injections.

[0020] Furthermore, when the traditional Chinese medicine composition is an injection, the preparation method further includes: boiling 800-1200 parts by volume of water for injection, adding the honeysuckle and artemisia extract and gardenia extract obtained by the above method, stirring to dissolve the two extracts, maintaining boiling for 3-7 minutes, cooling to room temperature, refrigerating for 20-36 hours, filtering, adjusting the pH of the filtrate to 2-3, heating the filtrate to boiling, adding 1% activated carbon, boiling for 8-14 minutes, refrigerating for 20-36 hours, filtering, taking the filtrate, heating to boiling, adjusting the pH to 5.05, boiling for 9-13 minutes, cooling to room temperature, refrigerating for 40-58 hours, filtering, adding 0.3-0.8 parts by weight of sodium bisulfite, making up to 1000 parts by volume, stirring, taking 50 parts by volume of the solution, heating to 65°C, adding Tween 80 Mix 3-5 volumes of Artemisia annua volatile oil, stir well, filter, and ultrafilter the filtrate using a hollow fiber membrane with a molecular weight of 30,000 to obtain ultrafiltrate; take 30,000 ultrafiltered drug solution and ultrafilter it using a plate membrane with a molecular weight of 10,000 to obtain ultrafiltrate, filter it through a 0.22μm microporous membrane, dispense it, and sterilize it with flowing steam for 35-50 minutes to obtain the final product.

[0021] This invention also proposes the application of the above-mentioned active components of the traditional Chinese medicine composition in the preparation of antipyretic drugs and / or in the preparation of anti-inflammatory drugs. Using a mouse model, this invention has demonstrated that the above-mentioned active ingredients in a specific range of traditional Chinese medicine compositions exhibit excellent antipyretic and anti-inflammatory effects.

[0022] The term "application" refers to administering the above-mentioned extract to a subject with a corresponding disease or a predisposition to such disease, with the aim of imparting a therapeutic effect, such as curing, alleviating, altering, influencing, improving, or preventing the aforementioned disease, its symptoms, or its predisposition. Those skilled in the art can easily determine the specific effective dose based on the type of disease being treated, the route of administration, and the excipients used; this dose may vary due to the concurrent use of other drugs.

[0023] The above-mentioned component content relationship also provides a basis for the quality control of the traditional Chinese medicine composition. That is, the present invention proposes a quality control method for the above-mentioned traditional Chinese medicine composition. The traditional Chinese medicine composition can be optionally an injection prepared from the active ingredient extract obtained by extracting and purifying Artemisia annua, Lonicera japonica, and Gardenia jasminoides. The active ingredients include: neochlorogenic acid ≥ 2‰, chlorogenic acid ≥ 5‰, cryptochlorogenic acid ≥ 2‰, caffeic acid ≥ 0.1‰, isochlorogenic acid B ≥ 0.5‰, isochlorogenic acid A ≥ 0.1‰, isochlorogenic acid C ≥ 0.1‰, geniposide ≥ 10‰, strychnos nux-ointense glycoside ≥ 1‰, strychnos nux-ointense acid ≥ 4‰, geniposide ≥ 0.1‰, genipin gentiopicroside ≥ 0.1‰, genipin gentiopicroside ≥ 0.5‰, 7-hydroxyswert ≥ 2‰, total acid ≥ 10‰, and total glycosides ≥ 15‰.

[0024] Further, the proportions of the active ingredients are as follows: neochlorogenic acid 2.5-5.14, chlorogenic acid 5.56-11.11, cryptochlorogenic acid 2.36-5.00, caffeic acid 0.14-0.26, isochlorogenic acid B 0.43-1.00, isochlorogenic acid A 0.22-0.46, isochlorogenic acid C 0.38-0.69, geniposide 9.53-17.71, strychnos nux-ointense 0.35-3.43, geniposide 0.42-0.79, genipin glycoside 0.36-0.68, genipin gentiopicroside 1.00-1.88, and 7-hydroxyswert 3.31-6.88; the composition contains 13.89-27.78% total acid and 23.68-50.61% total glycosides.

[0025] This invention investigates the effects of active ingredients in traditional Chinese medicine (TCM) compositions on LPS-induced fever and temperature changes in rats, as well as on acetic acid-induced peritoneal capillary permeability in mice. Experimental results show that differences in the effective components of raw materials from different origins and batches can lead to instability and poor quality consistency in different batches of preparations. However, based on the experimental content of this invention, TCM compositions with specific active ingredient content characteristics have been screened and exhibit stable and excellent therapeutic effects. These are preferred TCM compositions, and the aforementioned active ingredient content characteristics can be used for quality control of these compositions, thereby ensuring the stability of TCM preparation quality and reliable efficacy. Attached Figure Description

[0026] Figure 1Fingerprint spectrum of Artemisia annua as a reference standard;

[0027] Figure 2 Fingerprint spectrum of honeysuckle medicinal material;

[0028] Figure 3 Fingerprint spectrum of Gardenia jasminoides medicinal material. Detailed Implementation

[0029] As mentioned above, the present invention aims to provide a traditional Chinese medicine composition, its preparation method, and its application. The following will describe these in detail with reference to experimental examples.

[0030] It is particularly important to note that similar substitutions and modifications made to this invention are obvious to those skilled in the art, and they are all considered to be included in this invention. Those skilled in the art will clearly be able to modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to implement and apply the technology of this invention. Obviously, the described embodiments are only some embodiments of this invention, and not all embodiments.

[0031] Unless otherwise specified, this invention is carried out under conventional conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0032] Example 1: Preparation of Traditional Chinese Medicine Composition

[0033] Multiple batches of traditional Chinese medicine compositions were prepared by combining medicinal materials from different origins and batches. The following methods were used in all batches:

[0034] Take 750 parts by weight of honeysuckle, add 13 times the amount of hot water at 100℃, and extract twice, 2 hours each time, to obtain honeysuckle extract; take 1250 parts by weight of artemisia annua, add 5 times the amount of water to soak, and steam distill for 7 hours to extract artemisia annua volatile oil. Add 6 times the amount of water to the residue and extract for 2 hours. Concentrate the extract and honeysuckle extract under reduced pressure at 60℃ to a relative density of 1.10. Add ethanol to make the alcohol content reach 80%. Let stand for 30 hours, take the supernatant of alcohol precipitation, concentrate under reduced pressure to obtain alcohol precipitation concentrate, add concentrated hydrochloric acid to adjust the pH to 2.2, and extract with 12 times ethyl acetate to obtain extract. Concentrate under reduced pressure until there is no ethyl acetate odor, and vacuum dry to obtain honeysuckle and artemisia annua extract. Take 600 parts by weight of gardenia, crush it into coarse powder, heat it under reflux for 2 hours each time with 7 times the amount of 80% ethanol, concentrate the extract under reduced pressure to a relative density of 1.15 (65℃) to obtain gardenia extract concentrate, heat it to boiling, adjust the pH value to 3.0 with hydrochloric acid, add 1% solid paraffin by weight of gardenia, stir to completely dissolve, cool to room temperature, refrigerate for 24 hours, filter, extract with 5 times the amount of n-butanol, concentrate the extract under reduced pressure until there is no n-butanol odor, and vacuum dry to obtain gardenia extract.

[0035] Boil 900 parts by volume of water for injection, add honeysuckle and artemisia annua extracts and gardenia extracts, stir to dissolve the two extracts, maintain boiling for 6 minutes, cool to room temperature, refrigerate for 28 hours, filter, adjust the pH of the filtrate to 2.5, heat the filtrate to boiling, add 1% activated carbon, boil for 12 minutes, refrigerate for 28 hours, filter, heat the filtrate to boiling, adjust the pH to 5.05, boil for 12 minutes, cool to room temperature, refrigerate for 50 hours, filter, add 0.5 parts by weight of sodium bisulfite, make up to 1000 parts by volume, stir, take 50 parts by volume of the solution, heat to 65°C, add Tween 80 Mix 4 volumes of the mixture, add Artemisia annua volatile oil, mix well, filter, and ultrafilter the filtrate using a hollow fiber membrane with a molecular weight of 30,000 to obtain ultrafiltrate; take 30,000 ultrafiltered drug solution and ultrafilter it using a plate membrane with a molecular weight of 10,000 to obtain ultrafiltrate, filter it through a 0.22μm microporous membrane, dispense it, and sterilize it with flowing steam for 50 minutes to obtain the final product.

[0036] The volume parts / weight parts correspond to ml / g.

[0037] Example 2: Detection method for 13 components

[0038] 1. Instruments and reagents

[0039] Agilent 1290 high-performance liquid chromatograph (Agilent Technologies, Inc.); ME204E electronic analytical balance (Mettler-Toledo Shanghai Co., Ltd.); XS205 electronic analytical balance (Mettler-Toledo Shanghai Co., Ltd.); KH-300DB ultrasonic cleaner (Kunshan Hechuang Ultrasonic Instrument Co., Ltd.); UV-2401PC ultraviolet spectrophotometer (Shimadzu Corporation, Japan); ST16R high-speed refrigerated centrifuge (Thermo Fisher Scientific); HH-4 digital display constant temperature water bath (Guohua Electric Co., Ltd.); electric thermostatic drying oven (Shanghai Jinghong Experimental Equipment Co., Ltd.). Thirteen reference components: genipin, genipin gentiopicroside, genipin gentiopicroside, genipin, strychnoside, 7-hydroxyswertiamarin, neochlorogenic acid, caffeic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C.

[0040] 2. Determination method: The content of 13 components was determined by UPLC method.

[0041] Preparation of reference solutions: Weigh appropriate amounts of each of the following reference standards: genipin, genipin gentioside, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, geniposide, strychnoside, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, and 7-hydroxyswert. Dissolve in 25% methanol and dilute to volume to prepare reference stock solutions. Preparation of mixed reference solutions: Take an appropriate amount of the above reference standard stock solution and prepare a mixed reference solution. Take 1 mL, 2 mL, 4 mL, and 6 mL of the mixed solution and place them in 10 mL volumetric flasks, dilute to the mark, and then take 4 mL of the solution and place it in a 5 mL volumetric flask, dilute to volume, and then dilute to the mark to obtain reference solutions for 13 indicators. See Table 1 for details.

[0042] Table 1. Concentration information of 13 organic acids and iridoid glycosides.

[0043]

[0044] Preparation of test solution: Accurately pipette 1.0 mL of the injection solution prepared in Example 1 into a 25 mL volumetric flask, dilute to near the mark with 25% methanol, sonicate for 20 min, cool, dilute to the mark with 25% methanol, shake well, centrifuge at 12000 r / min for 10 min, and take the supernatant as the test solution.

[0045] Chromatographic conditions: Column: Agilent Eclipise Plus C18 (2.1 mm × 50 mm, 1.8 μm); Mobile phase: Phase A: 0.1% phosphoric acid solution, Phase B: acetonitrile:methanol (3:97); Elution gradient: 0–6 min: 4% B, 6–8 min: 4–5% B, 8–14 min: 5–12% B, 14–22 min: 12–16% B, 22–30 min: 16–24% B, 30–38 min: 24–38% B, 38–40 min: 38–95% B, 40–42 min: 95–4% B; Detection wavelength: 237 nm, 324 nm (detection wavelength for iridoid glycosides: 237 nm, detection wavelength for organic acids: 324 nm); Flow rate: 0.3 ml / min; Column temperature: 30 °C. Accurately pipette 1 μL of the above solution and inject it into the liquid chromatograph for analysis. Take the injection solution from Example 1 and determine the content of 13 components using the above method. The results are shown in Table 2.

[0046] Example 3: Determination of total acid content

[0047] Chlorogenic acid, as the main organic acid component, is the most inexpensive and readily available. Therefore, the experiment used ultraviolet spectrophotometry, with chlorogenic acid as a reference standard, to determine the total acid content in the injection solution prepared in Example 1.

[0048] 1. Instruments and reagents

[0049] UV-2401PC ultraviolet spectrophotometer (Shimadzu Corporation, Japan); ST16R high-speed refrigerated centrifuge (Thermo Fisher Scientific); 0.001 g electronic analytical balance (AL104, Mettler Toledo Shanghai Instruments Co., Ltd.); 0.001 g electronic balance (XS205, Mettler Toledo Shanghai Co., Ltd.); KH2200B ultrasonic cleaner (Kunshan Hechuang Ultrasonic Instruments Co., Ltd.); HH-4 digital display constant temperature water bath (Guohua Electric Appliances Co., Ltd.); electric thermostatic drying oven (Shanghai Jinghong Experimental Equipment Co., Ltd.). Chlorogenic acid reference standard.

[0050] 2. Detection Method

[0051] Within the wavelength range of 200–600 nm, scanning was performed on chlorogenic acid reference standard, finished product, and blank samples lacking three medicinal materials (excipients such as sodium bisulfite and Tween-80). The results showed that chlorogenic acid reference standard and finished product both had maximum absorption at 324 nm, but no significant absorption at 400 nm; the blank sample lacking three medicinal materials had no significant absorption at either 324 nm or 400 nm. Therefore, the determination wavelength was set at 324 nm, and the reference wavelength was set at 400 nm.

[0052] Preparation of reference solution: Accurately weigh 11.04 mg of chlorogenic acid reference standard, place it in a 100 mL volumetric flask, dissolve and dilute with water to the mark, shake well, and the solution is obtained (each 1 mL contains 110.4 μg of chlorogenic acid).

[0053] Preparation of the test solution: Accurately measure 1 mL of this product and place it in a 100 mL volumetric flask. Dilute with water to the mark and shake well. Then accurately measure 2 mL of this product and place it in a 25 mL volumetric flask. Dilute with water to the mark and shake well. The test results are shown in Table 2.

[0054] Example 4: Determination of total glycoside content

[0055] 1. Instruments

[0056] UV-2401PC and UV-2550 UV spectrophotometers (Shimadzu Corporation, Japan); ST16R high-speed refrigerated centrifuge (Thermo Fisher Scientific); 0.001 g electronic analytical balance (AL104, Mettler Toledo Shanghai Instruments Co., Ltd.); 0.001 g electronic balance (XS205, Mettler Toledo Shanghai Co., Ltd.); KH2200B ultrasonic cleaner (Kunshan Hechuang Ultrasonic Instruments Co., Ltd.); HH-4 digital display constant temperature water bath (Guohua Electric Appliances Co., Ltd.); electric thermostatic drying oven (Shanghai Jinghong Experimental Equipment Co., Ltd.). Gardenoside reference standard, vanillin reference standard, glacial acetic acid.

[0057] 2. Detection Method

[0058] Preparation of standard solution: Accurately weigh an appropriate amount of geniposide reference standard dried to constant weight, place it in a 10 mL volumetric flask, dissolve and dilute to the mark with 50% methanol, and shake well to obtain the solution (each 1 mL contains 1891 μg of geniposide).

[0059] Preparation of the test solution: Accurately pipette 1.0 mL of the injection solution prepared in Example 1 into a 25 mL volumetric flask, dilute to near the mark with 50% methanol, sonicate for 30 min, cool, and dilute to the mark with 50% methanol. Shake well, centrifuge at 12,000 r / min for 10 min, and take the supernatant as the test solution.

[0060] Content determination method: Accurately measure 0.05 mL, 0.10 mL, 0.15 mL, 0.20 mL, and 0.30 mL of the reference solution into 10 mL volumetric flasks, evaporate the solvent, add 0.4 mL of 5% vanillin-glacial acetic acid solution to each flask, mix well, quickly add 1.6 mL of perchloric acid, heat in a 60°C water bath for 15 min, then transfer to an ice-water bath and let stand for 3 min. Remove from the water, add 8.0 mL of glacial acetic acid, and shake well. Using the sample without reference solution as a blank, measure the absorbance A at 530 nm. Plot a standard curve with A as the ordinate and content (mg) as the abscissa. Accurately measure 0.40 mL of the test solution into a 10 mL stoppered test tube, evaporate the solvent, and proceed according to the procedure starting from "add 0.4 mL of 5% vanillin-glacial acetic acid solution and mix well". Calculate the total glycoside content in the test solution based on the standard curve. The test results are shown in Table 2.

[0061]

[0062] Example 5. Pharmacological test

[0063] 1. Effects of different batches of traditional Chinese medicine compositions on changes in body temperature in LPS-induced fever in rats

[0064] 1.1 Reagents and Test Samples

[0065] LPS: Extracted from Escherichia coli O55:B5; Ibuprofen sustained-release capsules, specification: each capsule contains 0.3 grams of the main ingredient ibuprofen; a total of 15 batches of the traditional Chinese medicine composition AO of this invention were manufactured by Jiangsu Kangyuan Pharmaceutical Co., Ltd.

[0066] 1.2 Experimental animals:

[0067] 300 male Wistar rats, weighing 150-170g, SPF grade.

[0068] 1.3 Main Instruments:

[0069] Electronic thermometer.

[0070] 1.4 Experimental Methods

[0071] Rat pretreatment:

[0072] Rats were acclimatized to the experimental environment (temperature 20-22℃) for 3 days with free access to food and water. For the first two days of the experiment, rectal temperature was measured twice daily (the thermometer was inserted 2cm into the anus). Rats with a body temperature not exceeding 38.5℃ and a temperature difference of no more than 0.5℃ between two measurements were selected as experimental subjects. On the day of the experiment, body temperature was measured twice consecutively, once every 30 minutes. The average of the two measurements was defined as the basal body temperature of the rats before modeling.

[0073] Preparation of rat fever model and grouping of experimental animals:

[0074] Two hundred and thirty rats selected based on basal body temperature were intraperitoneally injected with LPS saline solution (40 μg / kg, 5 ml / kg by rat weight). Four hours after modeling, their body temperature was measured. Rats with a temperature rise of 0.8-1.6℃ were randomly divided into 17 groups: a model group, an ibuprofen group (54.4 mg / kg), and a traditional Chinese medicine composition AO group (4.68 g crude drug / kg), with ten rats in each group. Immediately after grouping, the rats in the treatment groups received the drug via tail vein injection (4 mL / kg body weight), while the model group and positive control group received the drug via gavage. Body temperature changes were continuously monitored after drug administration. The time was calculated from the start of administration, and observation continued for 4 hours, with body temperature measured at 0.5h, 1h, 2h, 3h, and 4h. The magnitude of temperature rise and fall was calculated.

[0075] 1.5 Experimental Results and Conclusions

[0076] The experimental results are shown in Table 3. It can be seen that there was no significant difference in body temperature among the groups before administration. After administration, the increase in body temperature in each group was lower than that in the model group. Ibuprofen capsules showed rapid onset of action 0.5 hours after administration, significantly reducing the increase in body temperature caused by LPS. Compared with the model group, there were significant differences at 0.5, 1, 2, 3, and 4 hours (p<0.05; p<0.01). Compositions A, B, F, G, H, J, M, and N showed a trend of decreasing body temperature starting 1 hour after administration. Compared with the model group, there were significant differences at 2, 3, and 4 hours (p<0.05; p<0.01). Compositions C, E, I, L, and O showed significant differences only at 4 hours compared with the model group (p<0.05; p<0.01). Composition D showed significant differences only at 3 hours compared with the model group (p<0.05; p<0.01). The cooling effect of composition K was not significantly different from that of the model group.

[0077] Table 3. Effects of different batches of traditional Chinese medicine compositions on changes in body temperature in LPS-induced fever in rats.

[0078]

[0079]

[0080] There was a statistically significant difference compared to the model group. * P < 0.05, ** P < 0.01

[0081] 2. Effects of different batches of traditional Chinese medicine compositions on acetic acid-induced peritoneal capillary permeability in mice 2.1 Reagents and test samples

[0082] Evans Blue, glacial acetic acid, ibuprofen sustained-release capsules, each capsule containing 0.3 grams of ibuprofen as the main ingredient; a total of 15 batches of the traditional Chinese medicine composition AO of this invention were manufactured by Jiangsu Kangyuan Pharmaceutical Co., Ltd.

[0083] 2.2 Laboratory animals:

[0084] 170 ICR mice, half male and half female, weighing 15-17g, SPF grade.

[0085] 2.3 Main Instruments:

[0086] ELISA reader, syringe.

[0087] 2.4 Experimental Methods

[0088] Healthy ICR mice, SPF grade, weighing 16-18g, were randomly divided into 17 groups according to body weight: a model group, an ibuprofen group (78.07mg / kg), and a traditional Chinese medicine composition AO group (6.76g crude drug / kg), with 10 mice in each group, half male and half female. The composition groups were administered via tail vein (0.1ml / 10g body weight), while the model control group and the positive control group were administered via gavage (0.1ml / 10g body weight). Administration was once daily for 7 consecutive days. One hour after the last administration, 0.1ml / 10g of 0.5% Evans blue solution was injected via tail vein, followed immediately by an intraperitoneal injection of 0.1ml / 10g of 0.8% acetic acid. Thirty minutes later, the mice were sacrificed, and 5ml of physiological saline was injected intraperitoneally. The abdomen was gently massaged 50 times, the skin was cut open, and the peritoneal fluid was aspirated using a pipette. After centrifugation, the supernatant was measured at 590nm, and intergroup comparisons were performed using a t-test. The inhibition rate was calculated.

[0089] 2.5 Test Results

[0090] The experimental results are shown in Table 4. Compositions A, B, C, E, F, G, H, I, K, L, M, N and the positive control drug ibuprofen group can significantly inhibit the increase in peritoneal capillary permeability in mice induced by acetic acid, and have significant differences compared with the model group (p<0.01, p<0.05); while compositions D, J, O and the model group have no significant differences.

[0091] Table 4. Effects of different batches of traditional Chinese medicine compositions on acetic acid-induced peritoneal capillary permeability in mice.

[0092]

[0093] There was a statistically significant difference compared to the model group. * P < 0.05, ** P < 0.01

[0094] The results of the experiment on the effect of the compositions on the change of body temperature in LPS-induced fever in rats showed that compositions A, B, F, G, H, J, M, and N began to show a trend of decreasing body temperature 1 hour after administration, and there were significant differences at 2, 3, and 4 hours compared with the model group (p<0.05; p<0.01). However, compositions C, E, I, L, and O showed significant differences only at 4 hours compared with the model group (p<0.05; p<0.01), and composition D showed significant differences only at 3 hours compared with the model group (p<0.05; p<0.01). Furthermore, the results of the experiment on the effect of the compositions on acetic acid-induced peritoneal capillary permeability in mice showed that compositions A, B, C, E, F, G, H, I, K, L, M, and N could all significantly inhibit the increase in peritoneal capillary permeability induced by acetic acid in mice, and there were significant differences compared with the model group (p<0.01, p<0.05). The combined results of the two experiments show that compositions A, B, F, G, H, M, and N have significant antipyretic and anti-inflammatory effects.

[0095] Based on the active ingredient content of batches A, B, F, G, H, M, and N that all showed significant differences (p<0.01) compared to the model group, the optimal range of active ingredient content can be obtained, namely:

[0096] Neochlorogenic acid 2.5-5.14‰, chlorogenic acid 5.56-11.11‰, cryptochlorogenic acid 2.36-5.00‰, caffeic acid 0.14-0.26‰, isochlorogenic acid B 0.43-1.00‰, isochlorogenic acid A 0.22-0.46‰, isochlorogenic acid C 0.38-0.69‰, geniposide 9.53-17.71‰, strychnoside 0.35-3.43‰, geniposide 0.42-0.79‰, genipin glycoside 0.36-0.68‰, genipin gentiopicroside 1.00-1.88‰, 7-hydroxyswertia 3.31-6.88‰, total acid 13.89-27.78‰, total glycosides 23.68-50.61‰.

[0097] The ratio of the above active ingredients is as follows:

[0098] Neochlorogenic acid 2.5-5.14, chlorogenic acid 5.56-11.11, cryptochlorogenic acid 2.36-5.00, caffeic acid 0.14-0.26, isochlorogenic acid B 0.43-1.00, isochlorogenic acid A 0.22-0.46, isochlorogenic acid C 0.38-0.69, geniposide 9.53-17.71, strychnos nux-vomica 0.35-3.43, geniposide 0.42-0.79, genipin glycoside 0.36-0.68, genipin gentiopicroside 1.00-1.88, 7-hydroxyswertia 3.31-6.88; the extract contains 13.89-27.78% total acid and 23.68-50.61% total glycosides. Animal experiments have shown that traditional Chinese medicine compositions with the above-mentioned active ingredient content characteristics have stable and excellent therapeutic effects and are preferred traditional Chinese medicine compositions; the above-mentioned active ingredient content characteristics can also be used for the quality control of the traditional Chinese medicine composition.

[0099] Example 6. Establishing internal control standards and optimizing the preparation process

[0100] Based on the above-mentioned samples with optimal efficacy, we traced back the component content characteristics of the raw materials, established relevant medicinal fingerprint spectra as internal control standards, and improved the preparation process to ensure improved product quality and uniformity.

[0101] 1. Establish internal control standards for Artemisia annua: Add liquid chromatography fingerprinting control items to the pharmacopoeia.

[0102] Liquid phase fingerprinting detection method: The fingerprint spectrum of the test sample and the control fingerprint spectrum 1 are compared with the control fingerprint spectrum 1. The fingerprint peaks as indicated in the control fingerprint spectrum are used as reference correction peaks. The similarity should be greater than 0.5 after calculation by calculation software.

[0103] In this embodiment, the peak areas and retention times of the comparative fingerprint spectrum are as follows:

[0104] Mark Peak Sequence Number Retention time [Min] Peak area 1 3.882885 731.039 2 9.348604 238.769 3 10.29601 271.04 4 12.74284 231.6808 5 15.21369 585.2557 6 16.36838 243.2447 7 19.64414 683.0679 8 25.43884 415.1487 9 26.61364 252.1879 10 28.55216 264.1007 11 32.46935 936.6907 12 33.13216 616.8419 13 34.28907 386.3651 14 36.97964 219.1915 15 38.53798 842.427

[0105] Chromatographic conditions and system suitability tests were conducted using octadecylsilane-bonded silica gel as the stationary phase and a methanol-0.1% phosphoric acid aqueous solution system as the mobile phase; the elution program was as follows:

[0106]

[0107] The flow rate was 0.8 ml / min; the detection wavelength was 225 nm. The theoretical plate number, calculated based on the reference (quercetin) peak, should be no less than 6000.

[0108] Preparation of reference solution: Take an appropriate amount of quercetin standard, accurately weigh it, and add methanol to prepare a solution containing 50 μg per ml.

[0109] Preparation of the test solution: Take Artemisia annua, crush it, take about 1.0g of powder, accurately weigh it, put it in a 100ml flask, add 50ml of water, reflux for 1 hour, filter, and take the filtrate to obtain the test solution.

[0110] Assay: Accurately pipette 10 μl each of the reference solution and the test solution and inject them separately into the liquid chromatograph for determination.

[0111] 2. Establish internal control standards for honeysuckle medicinal materials: Based on the pharmacopoeia standards, add the extraction amounts of chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, and deoxidized strychnine, as well as the liquid phase fingerprint spectrum of the medicinal materials.

[0112] (1) Extraction amounts of four components from honeysuckle: chlorogenic acid should not be less than 2.0%; neochlorogenic acid should not be less than 0.05%; cryptochlorogenic acid should not be less than 0.05%; and oxidized strychnine should not be less than 0.2%. That is, in this embodiment, taking chlorogenic acid as an example, the amount of chlorogenic acid obtained from water extraction of 100g of honeysuckle should not be less than 2g.

[0113] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material, and a methanol-0.1% phosphoric acid aqueous solution system was used as the mobile phase. Gradient elution was performed according to the table below; the flow rate was 0.8 ml / min; the detection wavelength for chlorogenic acid, neochlorogenic acid, and cryptochlorogenic acid was 324 nm, and the detection wavelength for geniposide and strychnoside was 237 nm; the theoretical plate number, calculated based on the chlorogenic acid peak, should not be less than 10,000.

[0114]

[0115] Preparation of reference solution: Take appropriate amounts of chlorogenic acid reference standard and geniposide reference standard, add 50% methanol to prepare a solution containing approximately 0.3 mg of chlorogenic acid and 0.5 mg of geniposide per ml, shake well, and the solution is obtained.

[0116] Preparation of the test solution: Measure 1800 ml of water, place it in a flask, boil, add 100 g of the sample, ensuring the water covers the herb, reflux for 1 hour, filter, add another 1800 ml of water to the residue, boil, reflux for 1 hour, filter while hot, combine the filtrates, cool, accurately measure 1-10 ml of the subsequent filtrate, place it in a 50 ml volumetric flask, add 50% methanol to the mark. Shake well to obtain the test solution.

[0117] Assay: Accurately measure 10 μl each of the reference solution and the test solution, inject them into the liquid chromatograph, and determine the content. Using chlorogenic acid reference standard as a reference, calculate the chlorogenic acid content in the product according to the external standard method; using chlorogenic acid in the product as an internal reference, calculate the content of neochlorogenic acid and cryptochlorogenic acid in the product according to the relative correction factor; using geniposide in the reference solution as an internal reference, calculate the concentration of strychnine oxidase interrupted in the product according to the relative correction factor; determine the peak position of each component by relative retention time.

[0118] The relative correction factors and relative retention times of neochlorogenic acid, cryptochlorogenic acid, and deoxidized strychnine are shown in the table below:

[0119]

[0120] Note: Relative retention time is the ratio of the retention time of the analyte to that of its internal reference.

[0121] Technical parameters (for reference): chromatographic column: phenomenex Luna C18 (4.6×250mm, 5μm) or Kromasil C18 (4.6×250mm, 5μm).

[0122] Calculate using the following formula:

[0123]

[0124] In the formula: f 平均 —The average value of the correction factor;

[0125] A 样 —Peak area of ​​chlorogenic acid in the test solution (average of 2 injections);

[0126] —Dilution volume of the test sample;

[0127] 10 -3 —Unit conversion factor;

[0128] m 样 —Sample size (g) of the test sample

[0129] (2) Liquid phase fingerprint spectrum: The fingerprint spectrum of the test sample should be similar to that of the reference fingerprint spectrum, and the 12 chromatographic peaks should be displayed. The similarity should be greater than 0.95 according to the calculation software.

[0130] In this embodiment, the peak area and retention time of the comparative fingerprint spectrum are as follows: 209020ID

[0131]

[0132] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material (Phenomenex Luna C18 column, 4.6 × 250 mm, 5 μm); methanol-0.1% phosphoric acid aqueous solution was used as the mobile phase; the detection wavelength was 225 nm. The theoretical plate number, calculated based on the reference (chlorogenic acid) peak, should be no less than 6000. The elution program was as follows:

[0133]

[0134] Preparation of reference solution: Take an appropriate amount of chlorogenic acid reference standard, accurately weigh it, and add 50% methanol to prepare a solution containing 50 μg per ml.

[0135] Preparation of the test solution: Take the solution from the chlorogenic acid extraction section, filter it, and take the filtrate to obtain the test solution.

[0136] For the assay, accurately pipette 5 μl each of the reference solution and the test solution and inject them into the liquid chromatograph for determination.

[0137] 3. Establish internal control standards for gardenia medicinal materials: Add liquid chromatography fingerprinting control items to the pharmacopoeia.

[0138] Liquid chromatography fingerprinting detection method: The fingerprint spectrum of the test sample should be similar to that of the reference fingerprint spectrum, and the four marked chromatographic peaks should be displayed. The similarity calculated by the calculation software should be greater than 0.9.

[0139] In this embodiment, the peak areas and retention times of the comparative fingerprint spectrum are as follows:

[0140]

[0141] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase (Phenomenex Luna C18, 4.6 × 250 mm, 5 μm); the mobile phase was methanol-0.1% phosphoric acid aqueous solution, and the elution program was as follows:

[0142] Time (minutes) Methanol (%) 0.1% phosphoric acid aqueous solution (%) 0 20 80 50 60 40 60 60 40

[0143] The flow rate was 0.8 ml / min; the detection wavelength was 225 nm. The theoretical plate number, calculated based on the reference (geniposide) peak, should be no less than 6000.

[0144] Preparation of reference solution: Take an appropriate amount of geniposide reference standard, accurately weigh it, and add methanol to prepare a solution containing 50 μg per ml.

[0145] Preparation of the test solution: Take gardenia medicinal material, crush it, take about 0.1g of powder, accurately weigh it, put it in a 50ml conical flask, accurately add 25ml of methanol, sonicate for 30 minutes, filter, and take the filtrate to obtain the test solution.

[0146] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine them.

[0147] 4. Optimize the preparation process

[0148] In analyzing the impact of the preparation process on the stability of product components, we found that the tannins in Artemisia annua have a significant impact on the product's composition. In the process of Example 1, Artemisia annua extract and honeysuckle extract were combined for alcohol precipitation. However, the tannins in Artemisia annua are not easily precipitated completely during alcohol precipitation and tend to form precipitates with the effective components of honeysuckle, resulting in losses. This is a significant factor contributing to the instability of the finished product's component content. By comparing the processes of one alcohol precipitation of Artemisia annua extract and honeysuckle extract combined, one alcohol precipitation of Artemisia annua extract and honeysuckle extract separately, two alcohol precipitations of Artemisia annua extract separately and one alcohol precipitation of honeysuckle extract separately, and one alcohol precipitation of Artemisia annua extract separately followed by one alcohol precipitation of honeysuckle extract, we found that two alcohol precipitations are required to completely remove the interference of tannins in Artemisia annua. Furthermore, the second alcohol precipitation can be combined with the honeysuckle precipitation and does not cause significant losses to the effective components of honeysuckle. Therefore, in Example 1, after obtaining the Artemisia annua extract, an additional step of alcohol precipitation is added, followed by ethanol precipitation and then combining the Artemisia annua extract with the honeysuckle extract. Specifically, the Artemisia annua extract is concentrated to a relative density of 1.10-1.12 (70-80℃), ethanol is added to make the alcohol content reach 60-80%, and the mixture is allowed to stand for 12 hours. The supernatant of the alcohol precipitation is then combined with the honeysuckle extract.

[0149] 5. Verification Experiment

[0150] Medicinal materials from different origins and batches were selected and tested to meet the above internal control standards. Ten batches of traditional Chinese medicine compositions were prepared by combining them according to the above optimized process, and the content of relevant components was tested.

[0151]

[0152] It is evident that the aforementioned internal control standards for medicinal materials, combined with optimized preparation processes, can ensure that the content of the main active ingredients in the product falls within the optimal range, thereby achieving improved product quality and uniform stability.

[0153] The above is merely an illustrative description of the embodiments of the present invention. It should be noted that those skilled in the art can make several improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition for relieving fever and inflammation, characterized in that, The active ingredients are extracted and purified from Artemisia annua, honeysuckle, and gardenia. On a mass percentage basis (in parts per thousand), the active ingredients include: neochlorogenic acid 2.5-5.14‰, chlorogenic acid 5.56-11.11‰, cryptochlorogenic acid 2.36-5.00‰, caffeic acid 0.14-0.26‰, isochlorogenic acid B 0.43-1.00‰, isochlorogenic acid A 0.22-0.46‰, and isochlorogenic acid C. The composition contains 0.38-0.69‰, geniposide 9.53-17.71‰, strychnoside 0.35-3.43‰, geniposide 0.42-0.79‰, genipin glycoside 0.36-0.68‰, genipin gentiopicroside 1.00-1.88‰, and 7-hydroxyswert 3.31-6.88‰; the composition contains 13.89-27.78‰ total acid and 23.68-50.61‰ total glycosides; among which, The process includes the following steps: Take 750 parts by weight of honeysuckle and extract it 1-3 times with 9-16 times the amount of water at 90-100℃, for 1-2 hours each time, to obtain honeysuckle extract; Take 1250 parts by weight of artemisia annua, soak it in 3-6 times the amount of water, and then steam distill it for 5-8 hours to extract the volatile oil of artemisia annua. Add 3-9 times the amount of water to the residue and extract it for 1-3 hours. Concentrate the extract at 70-80℃ to a relative density of 1.10-1.12, then add ethanol to make the alcohol content reach 60-80%. Let it stand for 12 hours, and combine the supernatant of the alcohol precipitation with the honeysuckle extract. Concentrate it under reduced pressure at 60-80℃ to a relative density of 1.05-1.15 to obtain concentrated artemisia annua and honeysuckle extract. Add ethanol to the concentrated extract to make the alcohol content reach 70%-80%. Let it stand for 24-48 hours after alcohol precipitation, and take the supernatant of the alcohol precipitation. Concentrate it under reduced pressure to obtain concentrated alcohol precipitate, and adjust the concentration with concentrated hydrochloric acid. Adjust the pH to 1.5-2.5, extract with 8-16 times the amount of ethyl acetate, concentrate under reduced pressure until no ethyl acetate odor remains, and vacuum dry to obtain honeysuckle and artemisia extract; take 600 parts by weight of gardenia, crush into coarse powder, add 6-10 times the amount of 60-80% ethanol and extract 1-3 times, 1-3 hours each time, concentrate the extract under reduced pressure at 60-70℃ to a relative density of 1.10-1.20 to obtain gardenia extract concentrate, heat to boiling, adjust the pH to 2.5-3.5 with hydrochloric acid, add 1% by weight of solid paraffin of gardenia, stir to completely dissolve, cool to room temperature, refrigerate for 24 hours, filter, extract with 5-10 times the amount of n-butanol, concentrate the extract concentrate under reduced pressure until no n-butanol odor remains, and vacuum dry to obtain gardenia extract; mix honeysuckle and artemisia extract, gardenia extract and artemisia volatile oil through conventional procedures; The fingerprint spectrum of the Artemisia annua showed a similarity greater than 0.5 with the control fingerprint spectrum. The peak areas and retention times of the control fingerprint spectrum are as follows: The chromatographic conditions for comparing the fingerprint of Artemisia annua with the reference fingerprint include: The elution program used octadecylsilane-bonded silica gel as the packing material and a methanol-0.1% phosphoric acid aqueous solution system as the mobile phase. The flow rate was 0.8 ml / min; the detection wavelength was 225 nm; and the theoretical plate number, calculated based on the peak of the reference substance quercetin, should be no less than 6000. The extraction amounts of the four components from honeysuckle are as follows: chlorogenic acid should not be less than 2.0%; neochlorogenic acid should not be less than 0.05%; cryptochlorogenic acid should not be less than 0.05%; and oxidized strychnine should not be less than 0.2%. The fingerprint spectrum of the honeysuckle is similar to that of the control fingerprint spectrum by more than 0.

95. The peak areas and retention times of the control fingerprint spectrum are as follows: The chromatographic conditions for comparing the fingerprint chromatogram of honeysuckle with the reference fingerprint chromatogram include: Using octadecylsilane-bonded silica gel as the packing material and a methanol-0.1% phosphoric acid aqueous solution system as the mobile phase, gradient elution was performed according to the table below; the flow rate was 0.8 ml / min; the detection wavelength for chlorogenic acid, neochlorogenic acid, and cryptochlorogenic acid was 324 nm, and the detection wavelength for geniposide and strychnine was 237 nm; the theoretical plate number, calculated based on the chlorogenic acid peak, should not be less than 10,000. The fingerprint spectrum of the gardenia showed a similarity greater than 0.9 compared to the control fingerprint spectrum. The peak areas and retention times of the control fingerprint spectrum are as follows: The chromatographic conditions for comparing the fingerprint chromatogram of Gardenia jasminoides with those for the reference fingerprint chromatogram include: The chromatographic column used was octadecylsilane-bonded silica gel, with specifications of phenomenexluna C18, 4.6 × 250 mm, 5 μm; the mobile phase was methanol-0.1% phosphoric acid aqueous solution; the detection wavelength was 225 nm; the theoretical plate number, calculated based on the chlorogenic acid peak, should be no less than 6000; and the elution program was as follows:

2. A traditional Chinese medicine composition for antipyresis and anti-inflammation, characterized in that, It is an active ingredient extracted from Artemisia annua, honeysuckle, and gardenia, and the proportions of the components in the active ingredient are as follows: The composition contains: neochlorogenic acid 2.5-5.14%, chlorogenic acid 5.56-11.11%, cryptochlorogenic acid 2.36-5.00%, caffeic acid 0.14-0.26%, isochlorogenic acid B 0.43-1.00%, isochlorogenic acid A 0.22-0.46%, isochlorogenic acid C 0.38-0.69%, geniposide 9.53-17.71%, strychnos nux-vomica 0.35-3.43%, geniposide 0.42-0.79%, genipin glycoside 0.36-0.68%, genipin gentiopicroside 1.00-1.88%, and 7-hydroxyswertia 3.31-6.88%. The composition also contains 13.89-27.78% total acid and 23.68-50.61% total glycosides. in, The process includes the following steps: Take 750 parts by weight of honeysuckle and extract it 1-3 times with 9-16 times the amount of water at 90-100℃, for 1-2 hours each time, to obtain honeysuckle extract; Take 1250 parts by weight of artemisia annua, soak it in 3-6 times the amount of water, and then steam distill it for 5-8 hours to extract the volatile oil of artemisia annua. Add 3-9 times the amount of water to the residue and extract it for 1-3 hours. Concentrate the extract at 70-80℃ to a relative density of 1.10-1.12, then add ethanol to make the alcohol content reach 60-80%. Let it stand for 12 hours, and combine the supernatant of the alcohol precipitation with the honeysuckle extract. Concentrate it under reduced pressure at 60-80℃ to a relative density of 1.05-1.15 to obtain concentrated artemisia annua and honeysuckle extract. Add ethanol to the concentrated extract to make the alcohol content reach 70%-80%. Let it stand for 24-48 hours after alcohol precipitation, and take the supernatant of the alcohol precipitation. Concentrate it under reduced pressure to obtain concentrated alcohol precipitate, and adjust the concentration with concentrated hydrochloric acid. Adjust the pH to 1.5-2.5, extract with 8-16 times the amount of ethyl acetate, concentrate under reduced pressure until no ethyl acetate odor remains, and vacuum dry to obtain honeysuckle and artemisia extract; take 600 parts by weight of gardenia, crush into coarse powder, add 6-10 times the amount of 60-80% ethanol and extract 1-3 times, 1-3 hours each time, concentrate the extract under reduced pressure at 60-70℃ to a relative density of 1.10-1.20 to obtain gardenia extract concentrate, heat to boiling, adjust the pH to 2.5-3.5 with hydrochloric acid, add 1% by weight of solid paraffin of gardenia, stir to completely dissolve, cool to room temperature, refrigerate for 24 hours, filter, extract with 5-10 times the amount of n-butanol, concentrate the extract concentrate under reduced pressure until no n-butanol odor remains, and vacuum dry to obtain gardenia extract; mix honeysuckle and artemisia extract, gardenia extract and artemisia volatile oil through conventional procedures; The fingerprint spectrum of the Artemisia annua showed a similarity greater than 0.5 with the control fingerprint spectrum. The peak areas and retention times of the control fingerprint spectrum are as follows: The chromatographic conditions for comparing the fingerprint of Artemisia annua with the reference fingerprint include: The elution program used octadecylsilane-bonded silica gel as the packing material and a methanol-0.1% phosphoric acid aqueous solution system as the mobile phase. The flow rate was 0.8 ml / min; the detection wavelength was 225 nm; and the theoretical plate number, calculated based on the peak of the reference substance quercetin, should be no less than 6000. The extraction amounts of the four components from honeysuckle are as follows: chlorogenic acid should not be less than 2.0%; neochlorogenic acid should not be less than 0.05%; cryptochlorogenic acid should not be less than 0.05%; and oxidized strychnine should not be less than 0.2%. The fingerprint spectrum of the honeysuckle is similar to that of the control fingerprint spectrum by more than 0.

95. The peak areas and retention times of the control fingerprint spectrum are as follows: The chromatographic conditions for comparing the fingerprint chromatogram of honeysuckle with the reference fingerprint chromatogram include: Using octadecylsilane-bonded silica gel as the packing material and a methanol-0.1% phosphoric acid aqueous solution system as the mobile phase, gradient elution was performed according to the table below; the flow rate was 0.8 ml / min; the detection wavelength for chlorogenic acid, neochlorogenic acid, and cryptochlorogenic acid was 324 nm, and the detection wavelength for geniposide and strychnine was 237 nm; the theoretical plate number, calculated based on the chlorogenic acid peak, should not be less than 10,000. The fingerprint spectrum of the gardenia showed a similarity greater than 0.9 compared to the control fingerprint spectrum. The peak areas and retention times of the control fingerprint spectrum are as follows: The chromatographic conditions for comparing the fingerprint chromatogram of Gardenia jasminoides with those for the reference fingerprint chromatogram include: The chromatographic column used was octadecylsilane-bonded silica gel, with specifications of phenomenexluna C18, 4.6 × 250 mm, 5 μm; the mobile phase was methanol-0.1% phosphoric acid aqueous solution; the detection wavelength was 225 nm; the theoretical plate number, calculated based on the chlorogenic acid peak, should be no less than 6000; and the elution program was as follows:

3. The traditional Chinese medicine composition according to any one of claims 1-2, characterized in that, This traditional Chinese medicine composition is available in any optional formulation.

4. The traditional Chinese medicine composition according to claim 3, characterized in that, The formulations include decoctions, granules, capsules, pills, oral liquids, tinctures, syrups, suppositories, gels, sprays, and injections.

5. A method for preparing the traditional Chinese medicine composition as described in claim 3, characterized in that, Includes the following steps: Take 750 parts by weight of honeysuckle and extract it 1-3 times with 9-16 times the amount of water at 90-100℃ for 1-2 hours each time to obtain honeysuckle extract. Take 1250 parts by weight of artemisia annua, soak it in 3-6 times the amount of water, and then steam distill it for 5-8 hours to extract the volatile oil of artemisia annua. Add 3-9 times the amount of water to the residue and extract it for 1-3 hours. Concentrate the extract at 70-80℃ to a relative density of 1.10-1.12, then add ethanol to make the alcohol content 60-80%. Let it stand for 12 hours, and combine the supernatant of the alcohol precipitation with the honeysuckle extract. Concentrate it under reduced pressure at 60-80℃ to a relative density of 1.05-1.15 to obtain concentrated artemisia annua and honeysuckle extract. Add ethanol to the concentrated extract to make the alcohol content 70%-80%. Let it stand for 24-48 hours to precipitate the alcohol. Take the supernatant of the alcohol precipitation and concentrate it under reduced pressure to obtain concentrated alcohol precipitate. Adjust the pH value with concentrated hydrochloric acid. To obtain the honeysuckle and artemisia extract, extract with 8-16 times the amount of ethyl acetate, concentrate under reduced pressure until the ethyl acetate odor is gone, and vacuum dry. Take 600 parts by weight of gardenia, crush it into coarse powder, add 6-10 times the amount of 60-80% ethanol and extract 1-3 times, 1-3 hours each time. Concentrate the extract under reduced pressure at 60-70℃ to a relative density of 1.10-1.20 to obtain a gardenia extract concentrate. Heat to boiling, adjust the pH to 2.5-3.5 with hydrochloric acid, add 1% by weight of solid paraffin from gardenia, stir to completely dissolve, cool to room temperature, refrigerate for 24 hours, filter, and extract with 5-10 times the amount of n-butanol. Concentrate the extract under reduced pressure until the n-butanol odor is gone, and vacuum dry to obtain a gardenia extract. Mix the honeysuckle and artemisia extract, gardenia extract, and artemisia volatile oil through conventional procedures.

6. The preparation method according to claim 5, characterized in that, The traditional Chinese medicine composition is an injection. The preparation method further includes: boiling 800-1200 parts by volume of water for injection, adding the honeysuckle and artemisia extract and gardenia extract obtained according to claim 5, stirring to dissolve the two extracts, maintaining boiling for 3-7 minutes, cooling to room temperature, refrigerating for 20-36 hours, filtering, adjusting the pH of the filtrate to 2-3, heating the filtrate to boiling, adding 1% activated carbon, boiling for 8-14 minutes, refrigerating for 20-36 hours, filtering, heating the filtrate to boiling, adjusting the pH to 5.05, boiling for 9-13 minutes, cooling to room temperature, refrigerating for 40-58 hours, filtering, adding 0.3-0.8 parts by weight of sodium bisulfite, making up to 1000 parts by volume, stirring, taking 50 parts by volume of the solution, heating to 65°C, and adding Tween 80. Mix 3-5 volumes of Artemisia annua volatile oil, stir well, filter, and ultrafilter the filtrate using a hollow fiber membrane with a molecular weight of 30,000 to obtain ultrafiltrate; take 30,000 ultrafiltered drug solution and ultrafilter it using a plate membrane with a molecular weight of 10,000 to obtain ultrafiltrate, filter it through a 0.22μm microporous membrane, dispense it, and sterilize it with flowing steam for 35-50 minutes to obtain the final product.

7. The use of the traditional Chinese medicine composition according to claim 1 or 2 in the preparation of antipyretic and anti-inflammatory drugs.

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