Drug rehabilitation traditional Chinese medicine composition as well as preparation method, application and quality detection method thereof

Abstract

The invention provides a drug rehabilitation traditional Chinese medicine composition. The drug rehabilitation traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 9.6 to 14.4 parts of American ginseng, 17.6 to 26.4 parts of astragalus membranaceus, 10.4 to 15.6 parts of bighead atractylodes rhizome, 10.4 to 15.6 parts of poria cocos, 8 to 12.5 parts of liquorice, 9.6 to 14.4 parts of angelica sinensis, 8 to 12.5 parts of rhizoma picrorhizae, 12 to 18 parts of honeysuckle, 8.8 to 13.2 parts of husks of millet, 8 to 12 parts of catechu, 10.4 to 15.6 parts of durac, 10.4 to 15.6 parts of radix rehmanniae, 10.4 to 15.6 parts of gentiana macrophylla, 10.4 to 15.6 parts of fructus alpiniae oxyphyllae and 17.6 to 26.4 parts of corydalis tuber. The invention further provides a preparation method, application and a quality detection method of the traditional Chinese medicine composition. Experiments prove that the traditional Chinese medicine composition is small in toxic and side effects, stable and controllable in curative effect and low in resorption rate, the acting point is that a large number of tonifying medicines are used for rescuing people firstly, and the symptom is treated by using the symptom-treating medicines, so that the body is recovered as soon as possible, and the treatment compliance of a user who eliminates the abstinence symptom is improved.

Description

Technical Field

[0001] This invention relates to a traditional Chinese medicine composition for drug addiction treatment, its preparation method, uses, and quality testing methods, belonging to the field of traditional Chinese medicine. Background Technology

[0002] Current mainstream drug addiction treatment programs mostly employ substitution therapy with chemically synthesized drugs such as methadone and buprenorphine, supplemented by symptomatic supportive treatment. However, this strategy has significant limitations: the substitution drugs themselves are addictive, and long-term use may lead to secondary dependence; at the same time, these drugs are often accompanied by adverse reactions such as respiratory depression and constipation, and their intervention effect on psychological cravings and protracted withdrawal symptoms is limited.

[0003] Traditional Chinese medicine (TCM) has a long history of application in the field of drug rehabilitation and has shown unique advantages with the deepening of modern research. Existing studies have shown that scopolamine, a component of TCM, can accelerate morphine excretion by increasing morphine metabolism and promoting subcutaneous absorption of morphine (Zhou Wenhua, Zhu Bo, Chen Qijun, et al. Effects of scopolamine on serum and urinary morphine concentrations in rats [J]. Chinese Medical Journal, 1995, (01): 5-7+60.). Single herbs such as Datura stramonium, Datura stramonium, and Atropine have been proven to effectively combat morphine withdrawal reactions. Other TCM herbs such as Corydalis yanhusuo, Chelidonium majus, Sinomenium acutum, Saposhnikovia divaricata, Citrus reticulata, Paeonia lactiflora, and Cinnamomum cassia possess analgesic effects and are also effective in improving adverse reactions after drug rehabilitation. Furthermore, the alcoholic extract of Jasmine root has central sedative, hypnotic, and analgesic effects, and can alleviate or eliminate major withdrawal symptoms such as anxiety, irritability, pain, and insomnia that occur during drug rehabilitation. In summary, the Chinese herbal medicines currently used for drug addiction treatment mainly cover the following categories[6]: ① sedative and tranquilizing; ② tonifying and strengthening; ③ detoxifying and expelling pathogens; ④ symptomatic treatment.

[0004] However, drug addiction leads to complex pathological changes affecting multiple systems and organs in the body, triggering both acute and prolonged withdrawal symptoms. Compared to single-herb preparations, traditional Chinese medicine compound preparations, through the combination of principal, assistant, and adjuvant herbs, can exert synergistic effects on multiple components and targets, showing greater potential in terms of comprehensive efficacy and medication safety.

[0005] Application No. 201810743205.7, Invention Title: A Traditional Chinese Medicine Composition for Drug Detoxification and its Preparation Method, discloses a traditional Chinese medicine composition for drug detoxification and its preparation method. The composition includes 1-100 parts of American ginseng, 1-100 parts of stir-fried Atractylodes macrocephala, 1-100 parts of Angelica sinensis, 1-100 parts of Phellodendron chinense, 1-100 parts of Coptis chinensis, 1-200 parts of processed Astragalus membranaceus, 1-100 parts of processed Glycyrrhiza uralensis, 1-100 parts of Citrus reticulata peel, 1-100 parts of Bupleurum chinense, 1-100 parts of Gastrodia elata, 1-50 parts of Aucklandia lappa, 0.5-50 parts of Cimicifuga foetida, 0.1-10 parts of Datura stramonium, 1-100 parts of vinegar-processed Corydalis yanhusuo, 1-100 parts of stir-fried Ziziphus jujuba var. spinosa, 0.1-50 parts of Aquilaria sinensis, and 0.1-100 parts of processed Aconitum carmichaelii. This prescription is mainly used for spleen and kidney deficiency, qi deficiency and yang sinking in opium addiction. Summary of the Invention

[0006] This invention provides a traditional Chinese medicine composition for drug addiction treatment, its preparation method, uses, and quality testing method.

[0007] This invention provides a traditional Chinese medicine composition for drug addiction treatment, which is prepared from the following raw materials in the indicated weight ratios:

[0008] American ginseng 9.6-14.4 parts, Astragalus membranaceus 17.6-26.4 parts, Atractylodes macrocephala 10.4-15.6 parts, Poria cocos 10.4-15.6 parts, Glycyrrhiza uralensis 8-12.5 parts, Angelica sinensis 9.6-14.4 parts, Picrorhiza scrophulariiflora 8-12 parts, Lonicera japonica 12-18 parts, Papaver somniferum husk 8.8-13.2 parts, Catechu 8-12 parts, Dula 10.4-15.6 parts, Rehmannia glutinosa 10.4-15.6 parts, Gentiana macrophylla 10.4-15.6 parts, Alpinia oxyphylla 10.4-15.6 parts, Corydalis yanhusuo 17.6-26.4 parts.

[0009] The raw material "Dula" is the tuberous root of Aconitum episcopale Leveille, a plant of the Ranunculaceae family (see literature, Yan Zhuyun, et al., Investigation and identification of varieties of Yi medicine "Dula", Journal of Chengdu University of Traditional Chinese Medicine, June 2007, Vol. 30, No. 2).

[0010] Preferably, it is prepared from the following raw materials in the indicated weight ratios:

[0011] American ginseng 12 parts, Astragalus membranaceus 22 parts, Atractylodes macrocephala 13 parts, Poria cocos 13 parts, Glycyrrhiza uralensis 10 parts, Angelica sinensis 12 parts, Picrorhiza scrophulariiflora 10 parts, Lonicera japonica 15 parts, Papaver somniferum husk 11 parts, Catechu 10 parts, Dula 13 parts, Rehmannia glutinosa 13 parts, Gentiana macrophylla 13 parts, Alpinia oxyphylla 13 parts, Corydalis yanhusuo 22 parts.

[0012] The traditional Chinese medicine composition of the present invention is prepared into a commonly used oral preparation by adding pharmaceutically acceptable excipients or auxiliary ingredients, using the raw drug powder, water or organic solvent extract as the active ingredient.

[0013] The oral preparations mentioned above are tablets, capsules, pills, granules, and oral liquids.

[0014] The pills are honey pills containing at least 0.019 mg / g of tarazine; at least 0.082 mg / g of cryptochlorogenic acid, at least 0.565 mg / g of catechin, at least 0.291 mg / g of berberine II, at least 0.085 mg / g of isochlorogenic acid C, at least 0.110 mg / g of berberine I, and the total content of cryptochlorogenic acid, catechin, berberine II, isochlorogenic acid C, and berberine I is at least 1.134 mg / g.

[0015] The present invention also provides a method for preparing the aforementioned traditional Chinese medicine composition for drug addiction treatment, which includes the following steps:

[0016] a. Weigh the raw materials according to their respective weight ratios;

[0017] b. Take Astragalus membranaceus, Angelica sinensis, Lonicera japonica, Picrorhiza scrophulariiflora, Rehmannia glutinosa, Gentiana macrophylla, and Corydalis yanhusuo, add water and decoct to concentrate;

[0018] c. Grind the remaining raw materials into powder, add the concentrate from step b, and then add pharmaceutically acceptable excipients or auxiliary ingredients to prepare a commonly used oral pharmaceutical preparation.

[0019] The present invention also provides the use of the aforementioned traditional Chinese medicine combination in the preparation of a drug for drug addiction treatment.

[0020] The aforementioned drug is used to treat the coexistence of deficiency of vital energy and excess of pathogenic toxins during addiction withdrawal, or to treat the extreme weakness of qi, blood, yin and yang caused by withdrawal symptoms from taking opioid drugs.

[0021] This invention also provides a method for detecting the aforementioned traditional Chinese medicine composition for drug addiction treatment. This method uses high-performance liquid chromatography (HPLC) to detect tarazine in the composition, with the following chromatographic conditions:

[0022] Octadecylsilane-bonded silica gel was used as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile was used as mobile phase A, and 0.1% triethylamine solution was used as mobile phase B, with gradient elution performed according to the specifications in the table below; the flow rate was 1.0 ml per minute; the column temperature was 35 °C; the detection wavelength was 235 nm; the theoretical plate number, calculated based on the Tarrasque peak, should not be less than 5000.

[0023] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0～25 40→60 60→40

[0024] Specifically, it includes the following steps:

[0025] a. Preparation of reference solution

[0026] Prepare a reference solution by adding methanol to tarassamine reference standard.

[0027] b. Preparation of the test solution

[0028] Take an appropriate amount of the test sample, add ammonia solution and ether, seal tightly and soak for 24 hours, then sonicate, cool, shake well, filter, and take the filtrate to obtain the sample.

[0029] c. Detect according to the chromatographic conditions described.

[0030] This invention also provides a method for detecting the aforementioned traditional Chinese medicine composition for drug addiction treatment, using high performance liquid chromatography (HPLC) to detect the contents of cryptochlorogenic acid, catechins, berberine II, isochlorogenic acid C, and berberine I. The chromatographic conditions are as follows:

[0031] Using octadecylsilane-bonded silica gel as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); using acetonitrile as mobile phase A and 0.1% formic acid solution (containing 1.62 g ammonium formate per 1000 ml) as mobile phase B, gradient elution was performed according to the specifications in the table below; the flow rate was 1.0 ml per minute; the column temperature was 30 °C; the detection wavelength was 280 nm; the theoretical plate number calculated based on the cryptochlorogenic acid peak should not be less than 5000;

[0032] Specifically, it includes the following steps:

[0033] a. Preparation of reference solution

[0034] Prepare a reference solution by adding methanol to cryptochlorogenic acid reference standard, catechin reference standard, berberine II, isochlorogenic acid C, and berberine I reference standards.

[0035] b. Preparation of the test solution

[0036] Take the sample to be tested, add 70% methanol, and sonicate for 30 minutes to obtain the sample.

[0037] c. Detect according to the chromatographic conditions described.

[0038] This invention's formula primarily treats extreme weakness of Qi, blood, Yin, and Yang caused by withdrawal symptoms from narcotics. The formula uses ingredients that greatly replenish Qi and blood, strengthen the spleen and nourish the liver, protecting the body's foundation and effectively alleviating withdrawal symptoms caused by Qi deficiency. It also effectively clears residual "toxic evil" (expelling pathogenic factors) and the "internal heat" caused by metabolic disorders. Therefore, American ginseng is used to replenish Qi and nourish Yin, clear heat and generate fluids; astragalus replenishes Qi and raises Yang, consolidates the exterior and stops sweating, working together to enhance physical constitution and improve immunity. This aligns with the principle in the *Neijing* (Inner Canon of Medicine) that when the body's vital energy is abundant, pathogenic factors cannot invade, and one righteous force can suppress a hundred pathogenic factors. This fully embodies the TCM philosophy of saving the body first, then treating the disease, hence they are the principal herbs. Simultaneously, American ginseng, along with Poria cocos, Atractylodes macrocephala, and licorice, constitute the famous Sijunzi Decoction (Four Gentlemen Decoction) for tonifying the lungs and spleen, enhancing spleen and stomach function. The spleen and stomach are the foundation of acquired constitution and the source of Qi and blood; when the spleen and stomach are strong, the body's vital energy naturally recovers. Astragalus combined with Angelica sinensis is a famous combination used in Angelica sinensis blood-nourishing soup, achieving a dual effect of replenishing both qi and blood to repair the body and enhance its self-healing ability. Therefore, Poria cocos, Atractylodes macrocephala, Glycyrrhiza uralensis, and Angelica sinensis are used as assistant herbs. Coptis chinensis clears heat and dries dampness, reducing deficiency heat, and is particularly effective at clearing deep-seated, lingering damp-heat toxins. Lonicera japonica clears heat and detoxifies, dispersing wind-heat, and can clear relatively superficial heat toxins. These two herbs, one clearing internal heat and the other external heat, work together to clear internal and external heat toxins. Dula is a special Yi medicine widely used by locals for detoxification, alcohol poisoning, opium poisoning, and poisoning by Aconitum carmichaelii and Aconitum kusnezoffii, and is very effective for the "residual toxins" in withdrawal symptoms. Gentiana macrophylla dispels wind and dampness, relaxes muscles and relieves pain, invigorates blood and removes blood stasis, and also has immune-regulating properties. Rehmannia glutinosa clears heat and cools blood, nourishes yin and generates fluids, and is particularly suitable for dry mouth and tongue, internal heat and irritability caused by toxic damage to yin, and for the qi and yin deficiency commonly seen during withdrawal. Alpinia oxyphylla tonifies the spleen and reduces saliva production, specifically designed to treat drooling during withdrawal symptoms. Catechu clears heat and resolves phlegm, astringes dampness and heals sores, and promotes saliva production and quenches thirst. Catechu plays a unique role here, both assisting in clearing heat and detoxifying, and also acting as an astringent, effectively treating diarrhea, ulcers, and thirst that may occur during withdrawal. Poppy shell astringes the lungs, intestines, and relieves pain, while Corydalis has a broad-spectrum analgesic effect. These are two crucial and special herbs in this formula. They contain trace amounts of morphine-like alkaloids, possessing mild analgesic, sedative, and astringent effects. In this formula, they directly target the most painful withdrawal symptoms, such as severe pain, anxiety, diarrhea, and cough, relieving pain and facilitating a smooth transition—a typical example of the "gradual tapering" approach to drug rehabilitation. Providing the body with effective time for repair is key to effectively addressing the symptoms of withdrawal, allowing patients to gradually recover their bodily functions within a tolerable pain range, relying on restorative medications, rather than relapsing due to unbearable pain. Therefore, the herbs *Dula*, *Rehmannia glutinosa*, *Gentiana macrophylla*, *Catechu*, *Papaver somniferum*, and *Alpinia oxyphylla* collectively address different symptoms of withdrawal and serve as adjuvant herbs. *Glycyrrhiza uralensis* clears heat and detoxifies, nourishes the spleen and stomach, and harmonizes the effects of the other herbs, acting as the guiding herb. This formula is ingeniously conceived, particularly targeting the common characteristics of withdrawal in addictive patients: a coexistence of deficiency of vital energy and excess of pathogenic toxins. By supporting the body's resistance and eliminating pathogenic factors, treating both the root cause and symptoms, it effectively alleviates the physical and psychological pain of withdrawal, promotes the recovery of bodily functions, and thus eliminates withdrawal symptoms. The ultimate goal is to completely break free from drug dependence.

[0039] The compound honey pills of traditional Chinese medicine involved in this invention are scientifically formulated from 15 medicinal materials, including Astragalus membranaceus, American ginseng, Angelica sinensis, honeysuckle, catechu, Picrorhiza scrophulariiflora, and the Yi medicine "Dula". This formula follows the principles of supporting the body's resistance and eliminating pathogenic factors, clearing heat and detoxifying, and eliminating withdrawal symptoms. It not only adheres to the essence of traditional Chinese medicine's theory of drug rehabilitation but also incorporates the characteristics of ethnic medicine. American ginseng, Astragalus membranaceus, and Angelica sinensis work synergistically to invigorate qi and nourish blood, aiming to repair the qi and blood deficiency caused by addiction. This aligns with the principle of "tonifying qi and blood to strengthen the foundation" in the medical text *Jiu Mi Liang Fang* (Effective Prescriptions for Saving the Lost). Catechu, Picrorhiza scrophulariiflora, and honeysuckle work together to clear heat and detoxify, targeting inflammation and metabolic disorders caused by internal accumulation of toxins.

[0040] This invention is based on the principle of "treating both the symptoms and the root cause" in traditional Chinese medicine. It is not only about "detoxification" but also about strengthening the body's vital energy (expelling pathogens). It is mainly used to treat the root cause of the extreme weakness of Qi, blood, Yin and Yang caused by long-term use of narcotics during withdrawal symptoms. According to the different symptoms of withdrawal, it selects targeted Chinese medicines to treat the symptoms, thereby eliminating withdrawal symptoms, restoring the body, and achieving the goal of rejuvenating life.

[0041] The beneficial effects of this invention are:

[0042] (1) Experiments have shown that the herbal composition of this invention has few toxic side effects, stable and controllable efficacy, and a low relapse rate. The focus is on using a large amount of tonifying medicine to save the patient first, and using symptomatic medicine to treat the symptoms, thus working together to restore the body as soon as possible. This improves the treatment compliance of users who have eliminated withdrawal symptoms. At the same time, by supporting the body's resistance and eliminating pathogens, and treating both the root cause and the symptoms, it can effectively alleviate the physiological and psychological pain during the withdrawal period and promote the recovery of bodily functions, thereby achieving the goal of eliminating withdrawal symptoms. The ultimate goal is to completely get rid of drug dependence.

[0043] (2) Establish a method for determining the content of the reference sample of the traditional Chinese medicine composition honey pill (Life Rejuvenation Pill) of the present invention, so as to provide an overall quality control means for the compound, control its intrinsic quality from a macroscopic perspective, ensure the efficacy of the drug in drug addiction treatment, and provide a basis for rational market development.

[0044] (3) A method for determining the content of the compound honey pill of this traditional Chinese medicine was established, using tarasamine, catechin, cryptochlorogenic acid, berberine II, berberine I, and isochlorogenic acid C as indicators. Compared with existing traditional Chinese medicine quality control methods or other literature, this invention is the first to incorporate tarasamine, the hallmark component of the Yi medicine "Dula," and several components related to the efficacy of drug addiction treatment into the quality control system, which is of great significance to the quality control indicators of this drug addiction treatment compound.

[0045] (4) The method of the present invention is simple, convenient, stable, precise, reproducible and easy to master. Attached Figure Description

[0046] Figure 1Appearance diagram of the drug of this invention;

[0047] Figure 2 Mobile phase selection diagram;

[0048] Figure 3 Wavelength selection diagram;

[0049] Figure 4 Flow velocity study results;

[0050] Figure 5 Column temperature investigation results;

[0051] Figure 6 Injection volume investigation;

[0052] Figure 7 Tarasamin standard curve;

[0053] Figure 8 Mobile phase selection diagram;

[0054] Figure 9 Wavelength selection diagram;

[0055] Figure 10 Flow velocity study results;

[0056] Figure 11 Column temperature investigation results;

[0057] Figure 12 Injection volume investigation;

[0058] Figure 13 Cryptochlorogenic acid standard curve;

[0059] Figure 14 Catechin standard curve;

[0060] Figure 15 Standard curve of berberine II;

[0061] Figure 16 Isochlorogenic acid C standard curve;

[0062] Figure 17 Standard curve of berberine I;

[0063] Figure 18 Effects of Rensheng Zaizao Dan on mouse behavior (left figure: model group; right figure: high-dose Rensheng Zaizao Dan group);

[0064] Figure 19 Bar chart of jump counts (Note: Compared with the model control group, **P < 0.01, *P < 0.05);

[0065] Figure 20 Weight change graph;

[0066] Figure 21 Bar chart of total withdrawal symptom scores (Note: Compared with the model control group, **P < 0.01, *P < 0.05). Detailed Implementation

[0067] Example 1: Preparation method of the traditional Chinese medicine composition of the present invention

[0068] Prescription composition:

[0069] Take 22 parts of Astragalus membranaceus, 12 parts of American ginseng, 12 parts of Angelica sinensis, 15 parts of honeysuckle, 10 parts of catechu, 10 parts of Picrorhiza scrophulariiflora, 13 parts of Yi medicine "Dula", 13 parts of Alpinia oxyphylla, 11 parts of millet husk, 13 parts of Gentiana macrophylla, 13 parts of Rehmannia glutinosa, 13 parts of Poria cocos, 13 parts of Atractylodes macrocephala, 10 parts of Glycyrrhiza uralensis, and 22 parts of Corydalis yanhusuo.

[0070] Functions: Strengthens the body's resistance and eliminates pathogens, clears heat and detoxifies, and relieves withdrawal symptoms.

[0071] Indications: Withdrawal symptoms caused by opioid drugs, resulting from drug dependence and extreme deficiency of Qi, blood, Yin and Yang, leading to symptoms such as shortness of breath, fatigue, sweating, salivation, palpitations, abdominal pain, diarrhea, anxiety, insomnia, muscle aches, runny nose and tears, and lethargy.

[0072] 1. Weighing: Accurately weigh each processed medicinal material according to the above prescription ratio, and set aside for later use.

[0073] 2. Preparation: The preparation of this honey pill adopts the classic process of "water decoction extraction → concentration into paste → mixing with powder → pill making".

[0074] 2.1 Decoction Extraction: Place the prescribed amounts of Astragalus membranaceus, Angelica sinensis, Lonicera japonica, Corydalis yanhusuo, Picrorhiza scrophulariiflora, Rehmannia glutinosa, Gentiana macrophylla, and "Dula" slices into a multi-functional extraction vessel. Add 12 times the amount of water and soak for 40 minutes. Heat over high heat to boiling, then reduce to a simmer and decoct for 2 hours. Filter the first decoction through a 200-mesh sieve. Add 10 times the amount of water to the dregs, bring to a boil over high heat, then reduce to a simmer and continue decoction for 1.5 hours. Filter the second decoction. Combine the two decoctions, filter, and obtain the filtrate.

[0075] 2.2 Concentration: The above filtrate is concentrated to a thick paste with a relative density of 1.25 to 1.30 (60℃) under conditions of ≤70℃ and vacuum degree of -0.05 to -0.08 MPa, and then set aside for use.

[0076] 2.3 Grinding: Grind the prescribed amounts of American ginseng, catechu, and millet husks separately, and pass them through a 100-mesh sieve to obtain fine powder. Mix the American ginseng powder, catechu powder, and millet husk powder evenly to obtain a mixed powder.

[0077] 2.4 Honey Refining and Forming: Take an appropriate amount of purified honey and heat it to a medium consistency (refining temperature 116-118℃, producing glossy, pale yellow bubbles; it is sticky when rubbed between the fingers, but there are no white threads when two fingers are separated). Place the thick paste and mixed powder together in a kneading machine, and while it is still hot, slowly add the specified amount of refined honey, stirring and kneading thoroughly to form soft, firm, and well-plasticized balls (forming).

[0078] 2.5 Pill Making and Drying: Place the pill blocks in a honey pill machine, roll them into strips, and form them into pills, each weighing 9g. Place the formed honey pills evenly on a clean tray and place them in a low-temperature drying oven. Dry them with forced air at a temperature ≤50℃ until the surface of the pills is hard and the internal moisture content meets the standard (usually ≤15%).

[0079] 2.6 Packaging and Storage: After the dried honey pills pass inspection, they should be packaged in waxed paper or aluminum-plastic blister packs and sealed. Store in a cool, dry place.

[0080] Preparation of the drug of this invention (hereinafter referred to as "Life Rejuvenation Pill") (batch numbers: FFMW-01, FFMW-02, FFMW-03), samples are shown below. Figure 1 .

[0081] Example 2: Detection method for traditional Chinese medicine composition of the present invention

[0082] To elucidate the pharmacodynamic material basis and mechanism of action of this compound, the chemical components were systematically analyzed using ultra-high performance liquid chromatography-time-of-flight mass spectrometry (UPLC-TOF-MS) in the early stages of this invention, and network pharmacology studies were conducted based on this analysis. Using UPLC-TOF-MS, a total of 18 major chemical components were identified, including 9 flavonoids (such as naringenin-7-O-glucoside, catechins, verrucoside, quercetin, etc.), 3 phenolic acids (ferulic acid, cryptochlorogenic acid, isochlorogenic acid C), 2 terpenoids (berberine II, berberine I), 1 alkaloid (tarasamine), and other components.

[0083] Further network pharmacology studies revealed that the 18 components correspond to 470 potential targets, which intersect with 1740 drug addiction-related disease targets, yielding 272 common targets. By constructing a "traditional Chinese medicine-active ingredient-target-pathway" network, it was found that tarassamine, catechins, cryptochlorogenic acid, berberine II, berberine I, and isochlorogenic acid C may be key pharmacologically active substances in the detoxification effect of this compound. Protein interaction network analysis identified SRC, PIK3R1, STAT3, HSP90AA1, and AKT1 as core target genes. GO and KEGG enrichment analyses indicated that the mechanism of action of this compound may be closely related to the regulation of PI3K-Akt, Rap1, HIF-1, and MAPK signaling pathways, as well as the influence on EGFR tyrosine kinase inhibitor resistance.

[0084] Based on the above research results, in order to promote the development and application of Rensheng Rejuvenation Pill and ensure its quality control, this invention has for the first time screened out six key indicator components: tarasamine, catechin, cryptochlorogenic acid, berberine II, berberine I, and isochlorogenic acid C, and established a set of quality control and testing methods specifically for this compound honey pill of traditional Chinese medicine for drug addiction.

[0085] Based on the screening of the mechanism of action of this traditional Chinese medicine composition in the clinical application of drug addiction treatment, six key active ingredients, namely tarassamine, catechin, cryptochlorogenic acid, berberine II, berberine I, and isochlorogenic acid C, were specifically screened. A multi-component content determination method was established using high performance liquid chromatography, which is intended to be used for the quality control of this traditional Chinese medicine product for drug addiction treatment, so as to ensure the reliability of its efficacy and the stability of product quality.

[0086] 1. Experimental instruments and materials

[0087] High Performance Liquid Chromatography (HPLC): Thermo Fisher Scientific HPLC system;

[0088] Electronic balances: ME204E / 02, XPE26 (Mettler-Toledo Instruments Ltd.);

[0089] Ultrapure water system: Cellular type 1810A (Shanghai Moler Scientific Instruments Co., Ltd.);

[0090] Ultrasonic cleaner: KQ600DB model (600W, 40KHz; Kunshan Ultrasonic Instrument Co., Ltd.);

[0091] Column: ZORBAX Eclipse XDB-C18 4.6×250mm 5µm;

[0092] Acetonitrile, methanol, and triethylamine were of chromatographic grade, water was ultrapure water, and all other reagents were of analytical grade.

[0093] Tarasamine (Chengdu Glip Biotechnology Co., Ltd., batch number: 24070203, content calculated as 99.68%)

[0094] Cryptochlorogenic acid (Chengdu Pusi Biotechnology Co., Ltd., batch number: PS020981, content calculated as 98.61%)

[0095] Catechins (China National Institutes for Food and Drug Control, batch number: 110877-202306, content calculated as 96.6%).

[0096] Berberine II (China National Institutes for Food and Drug Control, batch number: 111596-202206, content calculated as 98.3%)

[0097] Berberine I (China National Institutes for Food and Drug Control, batch number: 111727-201702, content calculated as 95.6%)

[0098] Isochlorogenic acid C (Chengdu Maisike Pharmaceutical Technology Co., Ltd., batch number: L2016001, content calculated as 99.78%).

[0099] Samples of Life Rejuvenation Pills (honey pills): FFMW-01, FFMW-02, FFMW-03.

[0100] 2. Experimental conditions for the determination of tarasamine content

[0101] 2.1 Chromatographic conditions and system suitability test

[0102] Tarrassamine was prepared using octadecylsilane-bonded silica gel as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile was used as mobile phase A, and 0.1% triethylamine solution was used as mobile phase B, with gradient elution performed according to the specifications in the table below; the flow rate was 1.0 mL / min; the column temperature was 35 °C; and the detection wavelength was 235 nm. The theoretical plate number, calculated based on the tarrassamine peak, should be no less than 5000.

[0103]

[0104] 2.2 Preparation of reference solution

[0105] Take an appropriate amount of tarazine reference standard, accurately weigh it, and add methanol to prepare a solution containing 1 mg of tarazine per 1 ml.

[0106] 2.3 Preparation of the test solution

[0107] Take an appropriate amount of this product, cut it into small pieces, mix it well, take 3g, accurately weigh it, place it in a stoppered conical flask, accurately add 3ml of ammonia solution and 50ml of ether, seal tightly and soak for 24 hours, after soaking, sonicate for 30 minutes, cool, shake well, filter, and take the filtrate to obtain the test solution.

[0108] 2.4 Determination Method

[0109] Accurately pipette 10 μl each of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0110] 3. Establishment of a method for determining the content of tarasamine

[0111] 3.1 Chromatographic conditions and system suitability test

[0112] 3.1.1 Selection of mobile phase

[0113] Based on the above-planned experimental conditions, acetonitrile-0.1% triethylamine, acetonitrile-0.1% formic acid, and methanol-0.1% formic acid were selected for investigation. (See attached diagram.) Figure 2 .

[0114] The results showed that the target peak shape and separation effect were good when acetonitrile-0.1% triethylamine was used as the mobile phase. Therefore, acetonitrile-0.1% triethylamine solution was selected as the mobile phase for further investigation.

[0115] 3.1.2 Wavelength Selection

[0116] Based on the above-planned experimental conditions, the test solution was scanned across the entire wavelength range using a diode array detector, and chromatograms of the test solution at wavelengths of 218 nm, 235 nm, and 260 nm were extracted. (See...) Figure 3 .

[0117] By comparing the spectra, it was found that the peaks of the tarazanamine index showed significant absorption at a detection wavelength of 235 nm. Considering all factors, the baseline of the chromatogram was more stable, so the detection wavelength was determined to be 235 nm.

[0118] 3.1.3 Flow velocity assessment

[0119] Under the proposed experimental conditions, the flow rates of 0.8 ml / min, 1.0 ml / min, and 1.2 ml / min were investigated. (See...) Figure 4 .

[0120] The results showed that the chromatogram peak shape and separation effect were better when the flow rate was 1.0 ml / min, and the proposed flow rate was 1.0 ml / min.

[0121] 3.1.4 Column Temperature Investigation

[0122] Based on the above-specified experimental conditions, the results were investigated at column temperatures of 25℃, 30℃, and 35℃. (See...) Figure 5 .

[0123] The results showed that the chromatogram peak shape and separation were better when the column temperature was 30℃. The column temperature was tentatively set at 30℃.

[0124] 3.1.5 Sample Injection Volume Examination

[0125] Based on the above-established experimental conditions, the effects were investigated at injection volumes of 5 μl, 10 μl, and 15 μl. Figure 6 As shown.

[0126] The results showed that the chromatogram peak shape was good and the resolution was moderate when the injection volume was 10 μl. Therefore, the injection volume was determined to be 10 μl.

[0127] The tentative method for determining the content of Talasamine in Life Regeneration Pill is as follows:

[0128] Chromatographic conditions and system usability test: Octadecylsilane-bonded silica gel was used as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile was used as mobile phase A, and 0.1% triethylamine solution was used as mobile phase B, with gradient elution performed according to the specifications in the table below; the flow rate was 1.0 mL / min; the column temperature was 35 °C; and the detection wavelength was 235 nm. The theoretical plate number, calculated based on the Tarrassamine peak, should be no less than 5000.

[0129]

[0130] The assay involves precisely pipetting 10 μl each of the reference solution and the test solution into a liquid chromatograph and measuring the results.

[0131] 3.2 Methodological Examination

[0132] 3.2.1 Precision Examination

[0133] Five consecutive injections of tarazamin reference solution were performed, and the peak areas were recorded. The RSD values ​​were calculated. The results are shown in Table 1.

[0134] Table 1 Precision test results

[0135]

[0136] As can be seen from the above, the peak area RSD values ​​of each reference standard in the precision test were all less than 2.0%, indicating that the instrument precision was good.

[0137] 3.2.2 Linear Relationship

[0138] Accurately pipette the stock solution of tarazapine reference standard to prepare reference solutions of different concentrations. Inject 10 μl of each solution into the liquid chromatograph and obtain the peak area. Plot a response curve with reference standard concentration (X, µg / ml) on the x-axis and peak area (Y) on the y-axis. The results are shown in Table 2 and [Table data would be inserted here]. Figure 7 .

[0139] Table 2. Results of Tarasamin Standard Curve Analysis

[0140]

[0141] The results showed that the linear regression equation for tarassamine was y = 1.1161x + 2.3279, with a correlation coefficient R² = 0.9994, indicating a good linear relationship between tarassamine concentration and peak area within the concentration range of 23.0380416 µg / ml to 115.190208 µg / ml.

[0142] 3.2.3 Repeatability

[0143] Take an appropriate amount of this product (batch number: FFMW-01), cut it into small pieces, mix it well, take 3g, accurately weigh 6 portions, and have the same operator prepare the test solution according to the determined method. Accurately pipette 10μl of each test solution and inject it into the liquid chromatograph. Calculate the content of tarazine reference standard in the 6 samples. The results are shown in Table 3.

[0144] Table 3 Results of repeatability experiments

[0145]

[0146] The results show that the RSD value of the index content is less than 3%, indicating that the method has good repeatability.

[0147] 3.2.4 Intermediate Precision

[0148] Based on the above-planned experimental conditions, different personnel conducted measurements at different times. An appropriate amount of the compound honey pill sample (batch number: FFMW-01) was accurately weighed, chopped, mixed thoroughly, and 3g was taken to prepare the test sample for measurement. The calculated content of tarazamin reference standard is shown in Table 4.

[0149] Table 4 Results of intermediate precision testing

[0150]

[0151] The results show that the RSD value of the index content is less than 3%, indicating that the intermediate precision of this method is good.

[0152] 3.2.5 Recovery rate

[0153] Take a known concentration of the test sample (batch number: FFMW, tarazamin content: 0.019 mg / g), cut it into small pieces, mix well, and take 1.5 g of each sample. Divide the mixture into six portions, weigh them accurately, and add a certain amount of tarazamin reference standard. Prepare the test solution according to the prescribed method and determine the recovery rate. The results are shown in Table 5. The calculation formula is as follows:

[0154]

[0155] Table 5 Results of the spiking recovery experiment

[0156] The results showed that the recoveries of the indicator components were all between 90% and 108%. The method had good accuracy.

[0157] 3.2.6 Stability Test

[0158] According to the experimental conditions proposed above, a test solution was prepared, and the peak area of ​​each index was measured at 0h, 2h, 4h, 8h, 12h and 24h respectively. The results are shown in Table 6.

[0159] Table 6 Stability test results

[0160]

[0161] The results showed that under the experimental conditions, the RSD% of the index component was below 3%, and the test solution had good stability within 24 hours.

[0162] 3.3 Validation of Sample Content Determination

[0163] Three batches of compound honey pills were tested according to the proposed method, and the results are shown in Table 7.

[0164] Table 7. Results of content determination in three batches of compound honey pills

[0165]

[0166] As can be seen from the above, the method for determining the content of tarasamine in Rensheng Zaizao Dan honey pills can effectively detect compound honey pill samples, and the method is stable and feasible.

[0167] Example 3: Experiment on the determination of multiple components in the traditional Chinese medicine composition of the present invention

[0168] 1. Chromatographic conditions and system suitability test

[0169] Using octadecylsilane-bonded silica gel as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); using acetonitrile as mobile phase A and 0.1% formic acid solution (containing 1.62 g ammonium formate per 1000 ml) as mobile phase B, gradient elution was performed according to the specifications in the table below; the flow rate was 1.0 ml per minute; the column temperature was 30 °C; and the detection wavelength was 280 nm. The theoretical plate number, calculated based on the cryptochlorogenic acid peak, should be no less than 5000.

[0170]

[0171] 2. Preparation of reference solution

[0172] Take appropriate amounts of cryptochlorogenic acid reference standard, catechin reference standard, berberine II, isochlorogenic acid C, and berberine I reference standard, accurately weigh them, and add methanol to prepare a mixed solution containing 0.35 mg of cryptochlorogenic acid reference standard, 0.2 mg of catechin reference standard, 0.3 mg of berberine II, 0.2 mg of isochlorogenic acid C, and 80 μg of berberine I per ml.

[0173] 3. Preparation of the test solution

[0174] Take an appropriate amount of this product, cut it into small pieces, mix it well, take 3g, accurately weigh it, place it in a stoppered conical flask, accurately add 50ml of 70% methanol, seal it tightly, weigh it, sonicate it for 30 minutes, cool it, weigh it again, make up the lost weight with 70% methanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0175] 4. Determination method

[0176] Accurately pipette 5 μl each of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0177] 5. Establishment of a method for determining the content of multiple components

[0178] 5.1 Chromatographic conditions and system suitability test

[0179] 5.1.1 Selection of mobile phase

[0180] Based on the above-planned experimental conditions, acetonitrile-0.1% formic acid (containing 1.62g ammonium formate per 1000ml), acetonitrile-water, and methanol-0.1% formic acid (containing 1.62g ammonium formate per 1000ml) were selected for investigation. (See attached text.) Figure 8 .

[0181] The results showed that when acetonitrile-0.1% formic acid solution (containing 1.62g ammonium formate per 1000ml) was used as the mobile phase, the peak shape and separation effect of the target peak were good. Therefore, acetonitrile-0.1% formic acid solution (containing 1.62g ammonium formate per 1000ml) was selected as the mobile phase for further investigation.

[0182] 5.1.2 Wavelength Selection

[0183] Based on the above-planned experimental conditions, the test solution was scanned across the entire wavelength range using a diode array detector, and chromatograms of the test solution were extracted at wavelengths of 210 nm, 230 nm, 254 nm, 280 nm, and 300 nm. See [link / reference]. Figure 9 .

[0184] By comparing the spectra, it was found that the target peaks all showed significant absorption at a detection wavelength of 280 nm. Considering all factors, the chromatogram baseline was more stable, so the detection wavelength was determined to be 280 nm.

[0185] 5.1.3 Flow velocity assessment

[0186] Under the proposed experimental conditions, the flow rates of 0.8 ml / min, 1.0 ml / min, and 1.2 ml / min were investigated. (See...) Figure 10 .

[0187] The results showed that the chromatogram peak shape and separation effect were better when the flow rate was 1.0 ml / min, and the proposed flow rate was 1.0 ml / min.

[0188] 5.1.4 Column Temperature Investigation

[0189] Based on the above-specified experimental conditions, the results were investigated at column temperatures of 25℃, 30℃, and 35℃. (See...) Figure 11 .

[0190] The results showed that the chromatogram peak shape and separation were better when the column temperature was 30℃. The column temperature was tentatively set at 30℃.

[0191] 5.1.5 Injection Volume Examination

[0192] Based on the above-established experimental conditions, the effects were investigated at injection volumes of 5 μl, 10 μl, and 15 μl. Figure 12 As shown.

[0193] The results showed that the chromatogram peak shape was good and the resolution was moderate when the injection volume was 5 μl. Therefore, the injection volume was determined to be 5 μl.

[0194] The method for determining the content of Rensheng Zaizao Danmi Pills is tentatively set as follows:

[0195] Chromatographic conditions and system usability test: Octadecylsilane-bonded silica gel was used as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile was used as mobile phase A, and 0.1% formic acid solution (containing 1.62 g ammonium formate per 1000 ml) was used as mobile phase B, with gradient elution as specified in the table below; the flow rate was 1.0 ml / min; the column temperature was 30℃; and the detection wavelength was 280 nm. The theoretical plate number, calculated based on the cryptochlorogenic acid peak, should not be less than 5000.

[0196]

[0197] The assay involves precisely pipetting 5 μl of both the reference solution and the test solution into a liquid chromatograph and measuring the results.

[0198] 5.2 Methodological Examination

[0199] 5.2.1 Precision Examination

[0200] Five consecutive injections were performed using a mixed solution of cryptochlorogenic acid reference standard, catechin reference standard, berberine II, isochlorogenic acid C, and berberine I reference standard. The peak areas were recorded, and the RSD values ​​were calculated. The results are shown in Table 8.

[0201] Table 8 Precision test results

[0202]

[0203] As can be seen from the above, the peak area RSD values ​​of each reference standard in the precision test were all less than 1.0%, indicating that the instrument precision was good.

[0204] 5.2.2 Linear Relationship

[0205] Precisely pipette the mixed stock solution of cryptochlorogenic acid, catechin, berberine II, isochlorogenic acid C, and berberine I reference standards to prepare reference solutions of different concentrations. Inject 5 μl of each solution into the liquid chromatograph and obtain the peak area. Plot a response curve with reference standard concentration (X, µg / ml) on the x-axis and peak area (Y) on the y-axis. The results are shown in Tables 9-13. Figure 13 , 14 15, 16, 17.

[0206] Table 9. Results of Cryptochlorogenic Acid Standard Curve Analysis

[0207] Reference standard concentration (X, µg / ml) 35.1891300 52.783695 70.37826 87.972825 175.945650 Peak area (Y) 252.02614 373.47954 503.07875 632.04425 1289.083

[0208] Table 10 Results of catechin standard curve analysis

[0209] Reference standard concentration (X, µg / ml) 123.4258200 185.138730 246.85164 308.564550 617.1291 Peak area (Y) 425.80129 636.48652 847.80561 1064.31703 2124.56095

[0210] Table 11. Results of the standard curve analysis of berberine II

[0211] Reference standard concentration (X, µg / ml) 67.0700900 100.605135 134.14018 167.675225 335.35045 Peak area (Y) 251.02661 376.21305 500.06696 629.17252 1253.99738

[0212] Table 12 Results of isochlorogenic acid C standard curve analysis

[0213] Reference standard concentration (X, µg / ml) 26.6412600 39.961890 53.28252 66.603150 133.2063 Peak area (Y) 202.67876 296.38011 406.80245 511.24358 1081.75636

[0214] Table 13 Results of the standard curve analysis of berberine I

[0215] Reference standard concentration (X, µg / ml) 39.1768800 58.765320 78.35376 97.942200 195.8844 Peak area (Y) 201.30653 750.20646 997.46528 1256.91443 2503.99293

[0216] The results showed that the linear regression equation for cryptochlorogenic acid was y = 7.3963x - 14.7060, with a correlation coefficient R² = 0.9999, indicating a good linear relationship between cryptochlorogenic acid concentration and peak area within the concentration range of 35.18913 µg / ml to 175.94565 µg / ml.

[0217] The linear regression equation for catechins was y = 3.4431x - 0.1335, with a correlation coefficient R² = 1.0000, indicating a good linear relationship between catechin concentration and peak area within the concentration range of 123.42582 µg / ml to 617.12910 µg / ml.

[0218] The linear regression equation for berberine II was y = 3.7401 + 0.0529, with a correlation coefficient R² = 1.0000, indicating a good linear relationship between the concentration of berberine II and the peak area within the concentration range of 67.07009 µg / ml to 335.35045 µg / ml.

[0219] The linear regression equation for isochlorogenic acid C was y = 8.3216x - 32.3047, with a correlation coefficient R² = 0.9993, indicating a good linear relationship between isochlorogenic acid C concentration and peak area within the concentration range of 26.64126 µg / ml to 133.20630 µg / ml.

[0220] The linear regression equation for berberine I was y = 12.7891x - 0.5149, with a correlation coefficient R² = 1.0000, indicating a good linear relationship between the concentration of berberine I and the peak area within the concentration range of 39.17688 µg / ml to 195.88440 µg / ml.

[0221] 3.3.3.4 Repeatability

[0222] Take an appropriate amount of this product (batch number: FFMW-01), cut it into small pieces, take 3g, accurately weigh 6 portions, and have the same operator prepare the test solution according to the determined method. Accurately pipette 5μl of each test solution and inject it into the liquid chromatograph. Calculate the contents of cryptochlorogenic acid reference standard, catechin reference standard, berberine II reference standard, isochlorogenic acid C reference standard and berberine I reference standard in the 6 samples. The results are shown in Table 14.

[0223] Table 14 Results of Repeatability Experiments

[0224] Indicator Components 1 2 3 4 5 6 Average content (mg / g) RSD% Cryptochlorogenic acid content (mg / g) 0.081 0.078 0.081 0.082 0.081 0.082 0.081 1.8 Catechin content (mg / g) 0.537 0.540 0.536 0.551 0.538 0.550 0.542 1.2 Berberine II content (mg / g) 0.281 0.281 0.280 0.280 0.280 0.280 0.280 0.2 Isochlorogenic acid C content (mg / g) 0.081 0.085 0.081 0.084 0.081 0.084 0.083 2.3 Berberine I content (mg / g) 0.107 0.107 0.106 0.106 0.106 0.106 0.106 0.5

[0225] The results showed that the RSD values ​​of each indicator were all less than 3%, indicating that the method had good repeatability.

[0226] 5.2.3 Intermediate Precision

[0227] Based on the above-established experimental conditions, different personnel conducted measurements at different times. An appropriate amount of the compound honey pill sample (batch number: FFMW-01) was accurately weighed, cut into small pieces, and 3g was used to prepare the test sample for determination. The contents of cryptochlorogenic acid reference standard, catechin reference standard, berberine II reference standard, isochlorogenic acid C reference standard, and berberine I reference standard were calculated, and the results are shown in Table 15.

[0228] Table 15 Results of Intermediate Precision Testing

[0229] Indicator Components AT1-1 AT1-2 AT2-1 AT2-2 BT1-1 BT1-2 Average content (mg / g) RSD% Cryptochlorogenic acid content (mg / g) 0.082 0.082 0.082 0.083 0.082 0.083 0.082 0.6 Catechin content (mg / g) 0.528 0.540 0.542 0.550 0.551 0.553 0.544 1.7 Berberine II content (mg / g) 0.282 0.281 0.282 0.281 0.281 0.282 0.282 0.2 Isochlorogenic acid C content (mg / g) 0.082 0.084 0.082 0.084 0.084 0.084 0.083 1.2 Berberine I content (mg / g) 0.107 0.107 0.107 0.107 0.107 0.107 0.107 0.0

[0230] The results showed that the RSD values ​​of the contents were all less than 2%, indicating that the intermediate precision of this method was good.

[0231] 5.2.4 Recovery rate

[0232] Take a known sample (batch number: FFMW-01, cryptochlorogenic acid content 0.081 mg / g, catechin content 0.542 mg / g, berberine II content 0.280 mg / g, isochlorogenic acid C 0.083 mg / g, berberine I content 0.106 mg / g), cut it into small pieces, take about 1.5g, divide into 6 portions, accurately weigh them, and accurately add a certain amount of cryptochlorogenic acid reference standard, catechin reference standard, berberine II reference standard, isochlorogenic acid C reference standard, and berberine I reference standard to prepare the test solution according to the proposed method and determine the recovery rate. The results are shown in Table 16. The calculation formula is as follows:

[0233]

[0234] Table 16 Results of the spiking recovery experiment

[0235]

[0236] The results showed that the recovery rates of all indicators were between 90% and 120%. The method demonstrated good accuracy.

[0237] 5.2.5 Stability Test

[0238] According to the experimental conditions proposed above, a test solution was prepared, and the peak area of ​​each index was measured at 0h, 2h, 4h, 8h, 12h and 24h respectively. The results are shown in Table 17.

[0239] Table 17 Stability test results

[0240] Analysis time (h) 0 2 4 8 12 24 RSD (%) cryptochlorogenic acid 354.98184 355.42997 355.72741 356.74073 356.09862 356.6643 0.2 Catechins 1230.9528 1230.67363 1232.2914 1232.96693 1230.13882 1232.135 0.1 Berberine II 586.00874 586.5999 589.44755 589.15416 585.64094 588.3283 0.3 Isochlorogenic acid C 470.66341 470.50837 471.96926 473.51522 472.37858 473.4779 0.3 Berberine I 828.89645 830.3705 833.15591 831.18957 829.93092 834.316 0.2

[0241] The results showed that under the experimental conditions, the RSD% of each index was below 2%, and the test solution had good stability within 24 hours.

[0242] 5.3 Validation of Sample Content Determination

[0243] The compound honey pills were validated using the proposed method, and the results are shown in Table 18.

[0244] Table 18. Results of content determination in three batches of compound honey pills.

[0245] Serial Number batch number Cryptochlorogenic acid content (mg / g) Catechin content (mg / g) Berberine II content (mg / g) Isochlorogenic acid C content (mg / g) Berberine I content (mg / g) Total content (mg / g) 1 FFMW-01 0.082 0.565 0.291 0.085 0.111 1.134 2 FFMW-02 0.082 0.571 0.291 0.087 0.110 1.141 3 FFMW-03 0.082 0.572 0.292 0.087 0.111 1.144

[0246] As can be seen from the above, the method for determining the content of Rensheng Zaizao Dan can effectively detect compound honey pill samples, and the method is stable and feasible.

[0247] To further demonstrate the effectiveness of this formula, the following pharmacodynamic tests were conducted to prove the beneficial effects of the present invention.

[0248] Experimental Example 1: Pharmacological evaluation of the traditional Chinese medicine composition of the present invention in inhibiting oxidative stress, anti-inflammation, neuroprotection, and immunomodulation.

[0249] Drug abuse damages the body through multiple mechanisms, primarily involving central nervous system damage, oxidative stress, inflammatory response activation, and metabolic disorders. To verify the overall efficacy of this compound formula, given the objective limitations of directly conducting drug addiction model experiments, this invention uses an alternative model that can simulate core withdrawal symptoms and key damage mechanisms for evaluation. Specifically, its analgesic effect (to simulate the relief of withdrawal pain) is evaluated using the hot plate test, and its role in reducing oxidative stress damage is comprehensively evaluated using the WST-8 assay to detect superoxide dismutase (SOD), an antioxidant enzyme marker in brain tissue, and the thiobarbituric acid (TBA) assay to determine malondialdehyde (MDA), a lipid oxidation product marker in brain tissue. This provides experimental evidence for the detoxification efficacy of this traditional Chinese medicine compound formula.

[0250] 1. Experiment Content

[0251] 1.1 Drug materials and drugs

[0252] SPF grade male C57BL / 6 mice;

[0253] RM-200 Intelligent Hot Plate Tester;

[0254] The Life Rejuvenation Elixir, prepared by the method in Example 1, batch number: FFMW-01;

[0255] D-galactose (used to construct a chronic oxidative damage model);

[0256] Vitamin E (an antioxidant positive drug);

[0257] Aspirin (a positive analgesic drug);

[0258] Superoxide dismutase (SOD) assay kit (WST-8 method); Malondialdehyde (MDA) assay kit (TBA method);

[0259] 0.5% sodium carboxymethyl cellulose (CMC-Na) solution;

[0260] 1.2 Dosage Design

[0261] The inventors used the total content (100%) of tarassamine, cryptochlorogenic acid, catechin, berberine II, berberine I, and isochlorogenic acid C in the reference sample of the Life Regeneration Pill as the experimental high dose. Samples with 85% and 70% of these contents were designated as medium and low doses, respectively. Before use, each dose sample was uniformly suspended in 0.5% CMC-Na solution and administered to animals via gavage.

[0262] 1.3 Pre-treatment of compound honey pills made from traditional Chinese medicine before administration

[0263] Take the compound honey pills of the present invention and grind them thoroughly into a fine powder. Accurately weigh an appropriate amount of the honey pill powder and place it in a stoppered conical flask. Add 0.5% sodium carboxymethyl cellulose (CMC-Na) solution and vortex thoroughly to suspend it evenly, thus preparing a suspension of the required concentration. Prepare the suspension immediately before use to ensure uniform drug distribution during administration.

[0264] 2. Grouping and Modeling

[0265] 2.1 Hot plate test to induce pain in mice

[0266] C57BL / 6J mice were acclimatized to their new environment for one week using a smart hot plate apparatus, with the temperature controlled at (55±0.5) ℃. After preheating for 10 min, mice were placed on the hot plate, and those that licked their hind paws for <5 s or >30 s or jumped were discarded. Fifty mice were randomly divided into a model group (administered an equal volume of 0.5% CMC-Na solution via gavage), an aspirin (positive control group), and high, medium, and low dose groups of a traditional Chinese medicine compound honey pill. Administered once daily for three consecutive days. One hour after the last administration, the mice were placed on a 55℃ hot plate apparatus, and the latency of licking the hind paws or jumping was observed and recorded.

[0267] 2.2 Chronic Oxidative Damage Model

[0268] Sixty C57BL / 6J mice were acclimatized for one week and then randomly divided into six groups of ten mice each: a control group, a chronic oxidative damage model group, a vitamin E (positive control) group, and high, medium, and low dose groups of a traditional Chinese medicine compound honey pill. Except for the control group, mice in the model and treatment groups were intraperitoneally injected with D-galactose daily for 6-8 weeks to establish an aging mouse model. Simultaneously, the treatment groups began gavage therapy once daily for 6-8 weeks. The control group received intraperitoneal injections of physiological saline and gavage with an equal volume of 0.5% CMC-Na solution daily.

[0269] 2.3 Sample Collection and Processing for Chronic Oxidative Damage Model

[0270] Mice were fasted and deprived of water for 12 hours, then anesthetized. The hippocampus and part of the cerebral cortex were rapidly separated on an ice tray and rinsed with pre-cooled physiological saline. The tissue was blotted dry with filter paper and accurately weighed. Pre-cooled phosphate buffer was added at a weight-to-volume ratio (e.g., 1:9), and the mixture was mechanically homogenized under ice bath conditions. The homogenate was then centrifuged at 12000 rpm for 10-15 minutes at 4°C. The supernatant was carefully aliquoted and aliquoted into EP tubes and stored at -80°C for later analysis.

[0271] 3. Experimental Results

[0272] 3.1 Effect on pain induced by the hot plate test in mice

[0273] To evaluate the analgesic effect of the traditional Chinese medicine compound of this invention, a mouse pain model induced by the hot plate test was used. As shown in Table 19, before administration, there was no significant difference in the baseline pain threshold latency among the groups, indicating that the groups were balanced. After administration, no significant change was observed in the latency of the control group mice.

[0274] Compared with the model control group, the pain threshold latency was significantly prolonged in mice in both the aspirin positive control group and the high- and medium-dose groups of the traditional Chinese medicine compound of this invention, and the differences were statistically significant (P<0.01 or P<0.05). The analgesic effect of the high-dose group was comparable to that of the aspirin positive control group. While the low-dose group showed a certain trend towards prolonged latency, there was no statistically significant difference. These results demonstrate that the traditional Chinese medicine compound of this invention can significantly inhibit pain responses induced by heat stimulation, and the effect is dose-dependent.

[0275] Table 19 Effects of Rensheng Rejuvenation Pill on Pain Induced by Hot Plate Test in Mice (x̄±s, n=10)

[0276]

[0277] Note: Compared with the model group, #P<0.05, ##P<0.01.

[0278] 3.2 Effects on MDA content and SOD activity in brain tissue of mice with oxidative damage model

[0279] By establishing a D-galactose-induced chronic oxidative damage model, we further evaluated the antioxidant capacity of the traditional Chinese medicine compound of this invention. The evaluation was conducted by measuring the MDA content and SOD activity in mouse brain tissue, and the results are shown in Table 20.

[0280] Compared with the blank control group, the MDA content in the brain tissue of mice in the model control group was significantly increased (P<0.01), while the SOD activity was significantly decreased (P<0.01), indicating that the oxidative damage model was successfully established. Compared with the model control group, the vitamin E positive control group and the high- and medium-dose groups of the traditional Chinese medicine compound of this invention could significantly reverse the above abnormal indicators, specifically: the MDA content was significantly decreased (P<0.01 or P<0.05), while the SOD activity was significantly increased (P<0.01 or P<0.05). Among them, the high-dose group had the most significant antioxidant effect, approaching the level of the vitamin E positive control group. The low-dose group had no significant improvement effect on MDA and SOD indicators.

[0281] The above results demonstrate that the high and medium doses of the traditional Chinese medicine compound of this invention can effectively reduce the level of lipid peroxidation in the body and enhance the activity of antioxidant enzymes, thus having a clear protective effect against oxidative stress damage, and this effect is dose-dependent.

[0282] Table 20 Effects of Rensheng Zaizao Dan on MDA and SOD in mouse brain tissue (x̄±s, n=10)

[0283]

[0284] Note: Compared with the blank control group, *P<0.05, **P<0.01; compared with the model group, #P<0.05, ##P<0.01.

[0285] The traditional Chinese medicine compound of this invention exhibited significant activity in two pharmacological evaluation models. In the hot plate analgesia test, the compound significantly prolonged the pain latency in model mice in a dose-dependent manner, confirming its definite analgesic efficacy. In the chronic oxidative damage model, the compound also showed dose-dependent antioxidant activity, effectively increasing the activity of antioxidant enzymes in the brain tissue of model mice and significantly inhibiting the formation of lipid peroxidation products. These results collectively indicate that the traditional Chinese medicine compound of this invention possesses both analgesic and antioxidant pharmacological effects, providing core pharmacodynamic evidence for its application in alleviating drug withdrawal symptoms and protecting against neurological damage.

[0286] Experimental Example 2: Pharmacodynamic Study of the Effects of the Invention's Life Regeneration Pill on Withdrawal Symptoms in a Mouse Dependence Model

[0287] I. Experimental Objective

[0288] Main objective: To evaluate whether Life Regeneration Pill can effectively alleviate or eliminate acute withdrawal symptoms induced by naloxone in morphine-dependent mouse models.

[0289] II. Experimental Materials and Methods

[0290] 1. Experimental animals: 60 KM mice (half male and half female, 18 - 22 g), were raised in a laboratory environment with a temperature of 20 - 26 °C, relative humidity of 40% - 70%, and a light - dark cycle. License number for the use of experimental animals: SCXK (Beijing) 2019 - 0010.

[0291] 2. Experimental drugs: Extract of Rensheng Zaizao Dan (2.78 g / ml), morphine, naloxone, normal saline.

[0292] 3. Experimental design and grouping:

[0293] A randomized, blinded, controlled design was adopted. The mice were randomly divided into 6 groups, with 10 mice in each group (half male and half female):

[0294] Negative control group (Saline + Vehicle): Subcutaneous injection of normal saline, and normal saline was given before withdrawal induction.

[0295] Model control group (Morphine + Vehicle): Establish a morphine - dependent model, and normal saline was given before withdrawal induction.

[0296] Positive control group (Morphine + Clonidine): Establish a morphine - dependent model, and clonidine (1 mg / kg) was given before withdrawal induction.

[0297] Low - dose group of Rensheng Zaizao Dan (Morphine + 27.8 g / kg): Establish a morphine - dependent model, and 27.8 g / kg of Rensheng Zaizao Dan was given before withdrawal induction.

[0298] Medium - dose group of Rensheng Zaizao Dan (Morphine + 55.6 g / kg): Establish a morphine - dependent model, and 55.6 g / kg of Rensheng Zaizao Dan was given before withdrawal induction.

[0299] High - dose group of Rensheng Zaizao Dan (Morphine + 111.2 g / kg): Establish a morphine - dependent model, and 111.2 g / kg of Rensheng Zaizao Dan was given before withdrawal induction.

[0300] III. Experimental content:

[0301] 1. The first stage: Establishment of the drug - dependence model (cycle: 10 days)

[0302] In all morphine-dependent model mice, an escalating dose regimen was used, with subcutaneous injections of morphine hydrochloride twice daily (12-hour intervals). The specific dosage regimen was as follows: Days 1-2: 10 mg / kg; Days 3-4: 20 mg / kg; Days 5-6: 40 mg / kg; Days 7-10: 60 mg / kg. The 20th morphine injection was completed on day 10, successfully establishing a stable morphine-dependent mouse model. The negative control group received a subcutaneous injection of an equal volume (equal to the volume of the morphine group solution) of physiological saline at the same time points.

[0303] 2. Second phase: Accelerated withdrawal and efficacy evaluation (conducted 4 hours after the 20th morphine injection on day 10)

[0304] Pre-drug administration (intervention with the test drug):

[0305] Thirty minutes prior to intraperitoneal injection of naloxone to induce withdrawal, the following substances were administered via gavage according to experimental groups:

[0306] Low-dose group of Rensheng Zaizao Dan: 27.8 g / kg Rensheng Zaizao Dan extract solution.

[0307] Medium-dose group of Rensheng Zaizao Dan: 55.6 g / kg Rensheng Zaizao Dan extract solution.

[0308] High-dose group of Rensheng Zaizao Dan: 111.2 g / kg Rensheng Zaizao Dan extract solution.

[0309] Positive control group: 1 mg / kg clonidine solution.

[0310] Model control group and negative control group: equal volume of physiological saline.

[0311] Inducing withdrawal (inducing withdrawal symptoms):

[0312] Thirty minutes after pre-administration, all mice were intraperitoneally injected with naloxone (2 mg / kg).

[0313] This procedure aims to completely block opioid receptors, thereby inducing severe acute withdrawal symptoms in opioid-dependent mouse models. The negative control group was also injected with naloxone to rule out nonspecific effects of naloxone itself on normal animals.

[0314] IV. Experimental Results

[0315] 1. Behavioral observation

[0316] After administration of Rensheng Zaizao Dan (especially in the medium and high dose groups), mice exhibited a brief period of quiet and reduced activity even before naloxone stimulation. Following stimulation withdrawal, compared to the model control group, the abnormal postures and irritable behaviors of the mice in the Rensheng Zaizao Dan group were significantly reduced. These observed behavioral inhibitory effects are consistent with those observed in previous acute toxicity experiments and are presumably one of the mechanisms by which Rensheng Zaizao Dan exerts its anti-withdrawal effect (see...). Figure 18 ).

[0317] 2. Core withdrawal behaviors (number of skips)

[0318] Jumping is one of the most typical behaviors during morphine withdrawal. Life Regeneration Pill significantly reduced the number of jumps in mice (P < 0.01) and showed a dose-dependent effect, with the highest effect observed in the high-dose group (see...). Figure 19 ).

[0319] 3. Weight changes

[0320] Withdrawal leads to significant weight loss. Compared with the model control group, the percentage of weight loss in mice in the medium and high dose groups of Rensheng Zaizao Dan was significantly reduced (P < 0.05), indicating that Rensheng Zaizao Dan alleviates the physiological stress and consumption caused by withdrawal (see...). Figure 20 ).

[0321] Table 21 Mouse body weight data

[0322] serial number Experimental Groups Day 1 Day 5 Day 10 (before quitting) Day 10 (After quitting) Percentage of weight loss after withdrawal 1 negative control group 20.1 ± 0.5 24.5 ± 0.6 28.8 ± 0.7 28.4 ± 0.7 1.4 ± 0.3% 2 Model control group 20.2 ± 0.4 23.9 ± 0.5 27.5 ± 0.6 25.1 ± 0.6 8.7 ± 0.6% 3 Positive control group 20.0 ± 0.6 24.1 ± 0.7 27.9 ± 0.8 26.8 ± 0.8 3.9 ± 0.4% ** 4 Low-dose group of Life Regeneration Pill 19.9 ± 0.5 23.8 ± 0.6 27.3 ± 0.7 25.3 ± 0.7 7.3 ± 0.5% 5 Medium-dose group of Life Regeneration Pill 20.3 ± 0.5 24.2 ± 0.5 27.7 ± 0.6 26.2 ± 0.6 5.4 ± 0.4% * 6 High-dose group of Life Regeneration Pill 20.1 ± 0.4 24.3 ± 0.5 27.8 ± 0.6 26.6 ± 0.6 4.3 ± 0.4% **

[0323] Note: Compared with the model control group, **P < 0.01, *P < 0.05.

[0324] 4. Overall score of withdrawal symptoms

[0325] Compared with the model control group, all dosage groups of Rensheng Zaizao Dan reduced the total score of withdrawal symptoms, and the effect was dose-dependent. The high-dose group significantly reduced the score (P < 0.01), with an effect comparable to that of the positive control drug clonidine (see...). Figure 21 ).

[0326] Table 22 Total score data of withdrawal symptoms in mice

[0327] serial number Experimental Groups Total score of withdrawal severity Number of jumps Percentage of weight loss 1 negative control group 2.5 0.2 1.5% 2 Model control group 48.6 15.8 8.7% 3 Positive control group 15.3 ** 3.5 ** 3.9% ** 4 Low-dose group of Life Regeneration Pill 38.4 12.1 7.3% 5 Medium-dose group of Life Regeneration Pill 25.7 ** 7.3 ** 5.4% * 6 High-dose group of Life Regeneration Pill 17.2 ** 4.2** 4.1%**

[0328] 5. Organ-protective effects of Rensheng Rejuvenation Pill on withdrawal model mice

[0329] The study found that morphine dependence and withdrawal processes themselves cause significant physiological stress to the body. The liver, spleen, and kidney indices in the model control group were significantly higher than those in normal animals (P < 0.01), indicating stress-induced changes at the organ level. Treatment with Rensheng Zaizao Dan, especially the high-dose group, showed significant organ-protective effects, dose-dependently reversing the increases in liver, spleen, and kidney indices caused by morphine dependence (P < 0.05 or P < 0.01 compared to the model control group), restoring them to near-normal levels. There were no significant changes in heart and lung indices among the groups.

[0330] Table 23 Animal Organ Index Table (mg / g)

[0331] Experimental Groups heart liver spleen lung kidney negative control group 4.16 54.32 2.98 6.12 13.58 Model control group 4.25 62.47 ## 3.65 ## 6.38 15.89 ## Positive control group 4.18 57.15 * 3.12 * 6.25 14.23 * Low-dose group of Life Regeneration Pill 4.23 60.26 3.48 6.35 15.45 Medium-dose group of Life Regeneration Pill 4.20 58.34 * 3.25 * 6.28 14.78 * High-dose group of Life Regeneration Pill 4.18 55.21** 3.01** 6.23 13.62**

[0332] Compared with the negative control group, ## P < 0.01; compared with the model control group, *P < 0.05; compared with the model control group, **P < 0.01

[0333] V. Conclusion

[0334] 1. Rensheng Zaizao Dan exhibits significant anti-withdrawal efficacy: In a morphine-dependent mouse model, Rensheng Zaizao Dan significantly alleviated acute withdrawal symptoms induced by naloxone in a dose-dependent manner. Specifically, it significantly reduced the total withdrawal symptom score, decreased characteristic jumping behavior, and effectively inhibited withdrawal-induced weight loss. The high-dose Rensheng Zaizao Dan (111.2 g / kg) showed an anti-withdrawal effect comparable to that of the positive control drug clonidine.

[0335] 2. The Life Regeneration Pill exhibits organ protective effects: This invention discovered that morphine dependence and withdrawal processes can lead to a significant increase in the indices of the liver, spleen, and kidneys, indicating stress-induced damage at the organ level. Treatment with the Life Regeneration Pill, especially the high-dose group, can dose-dependently reverse the abnormal changes in the aforementioned organs.

[0336] 3. Life Rejuvenation Pill demonstrates good safety: Throughout the experiment, no animal deaths or abnormal toxic reactions occurred in any of the dosage groups of Life Rejuvenation Pill. At the dosage that exerts significant efficacy, it has no adverse effects on major organs (heart and lungs), confirming its high therapeutic safety.

[0337] Experimental Example 3: Acute toxicity test of the present invention's Life Rejuvenation Pill (oral administration)

[0338] This study investigated acute toxicity in animals after single or multiple administrations of Rensheng Zaizao Dan (a traditional Chinese medicine) within 24 hours, observing the acute poisoning reactions caused by overdose. This research aimed to design dosages for repeated-dose toxicity studies, analyze the main target organs of toxicity, and analyze potential toxicity in humans under overdose. The study was conducted by a GLP-qualified third party. Based on the experimental results, the LD50 of Rensheng Zaizao Dan could not be determined. 50 The maximum single dose was 111.2 g / Kg; the maximum experimental dose was 222.4 g / Kg, and no toxic reactions were observed in either case, providing reference information for clinical dose selection and the design of observation indicators.

Claims

1. A traditional Chinese medicine composition for drug addiction treatment, characterized in that: It is prepared from the following raw materials in the indicated weight ratios: American ginseng 9.6-14.4 parts, Astragalus membranaceus 17.6-26.4 parts, Atractylodes macrocephala 10.4-15.6 parts, Poria cocos 10.4-15.6 parts, Glycyrrhiza uralensis 8-12.5 parts, Angelica sinensis 9.6-14.4 parts, Picrorhiza scrophulariiflora 8-12.5 parts, Lonicera japonica 12-18 parts, Papaver somniferum husk 8.8-13.2 parts, Catechu 8-12 parts, Dioscorea opposita 10.4-15.6 parts, Rehmannia glutinosa 10.4-15.6 parts, Gentiana macrophylla 10.4-15.6 parts, Alpinia oxyphylla 10.4-15.6 parts, Corydalis yanhusuo 17.6-26.4 parts.

2. The traditional Chinese medicine composition for drug addiction treatment according to claim 1, characterized in that: It is prepared from the following raw materials in the indicated weight ratios: American ginseng 12 parts, astragalus 22 parts, atractylodes macrocephala 13 parts, poria cocos 13 parts, licorice 10 parts, angelica sinensis 12 parts, coptis chinensis 10 parts, honeysuckle 15 parts, corn husk 11 parts, catechu 10 parts, dula 13 parts, rehmannia glutinosa 13 parts, gentiana macrophylla 13 parts, alpinia oxyphylla 13 parts, corydalis 22 parts.

3. The traditional Chinese medicine composition for drug addiction treatment according to claim 1 or 2, characterized in that: It is prepared into a commonly used oral formulation by adding pharmaceutically acceptable excipients or auxiliary ingredients, using the original drug powder, water or organic solvent extract as the active ingredient.

4. The traditional Chinese medicine composition for drug addiction treatment according to claim 3, characterized in that: The oral preparations mentioned are tablets, capsules, pills, granules, and oral liquids.

5. The traditional Chinese medicine composition for drug addiction treatment according to claim 4, characterized in that: The pills are honey pills containing at least 0.019 mg / g of tarazine; at least 0.082 mg / g of cryptochlorogenic acid, at least 0.565 mg / g of catechin, at least 0.291 mg / g of berberine II, at least 0.085 mg / g of isochlorogenic acid C, at least 0.110 mg / g of berberine I, and a total content of at least 1.134 mg / g of cryptochlorogenic acid, catechin, berberine II, isochlorogenic acid C, and berberine I.

6. A method for preparing the traditional Chinese medicine composition for drug addiction treatment according to any one of claims 1-5, characterized in that: It includes the following steps: a. Weigh the raw materials according to their respective weight ratios; b. Take Astragalus membranaceus, Angelica sinensis, Lonicera japonica, Picrorhiza scrophulariiflora, Rehmannia glutinosa, Gentiana macrophylla, and Corydalis yanhusuo, add water and decoct to concentrate; c. Grind the remaining raw materials into powder, add the concentrate from step b, and then add pharmaceutically acceptable excipients or auxiliary ingredients to prepare a commonly used oral pharmaceutical preparation.

7. Use of the traditional Chinese medicine combination according to any one of claims 1-5 in the preparation of a drug for drug addiction treatment.

8. The use according to claim 7, characterized in that: The aforementioned medication is used to treat the coexistence of deficiency of vital energy and excess of pathogenic toxins during addiction withdrawal; or to treat extreme weakness of qi, blood, yin and yang caused by withdrawal symptoms from opioid drugs.

9. A method for detecting the traditional Chinese medicine composition for drug addiction treatment according to any one of claims 1-5, characterized in that: It uses high-performance liquid chromatography (HPLC) to detect tarazine in the composition. The chromatographic conditions are as follows: Octadecylsilane-bonded silica gel was used as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile was used as mobile phase A, and 0.1% triethylamine solution was used as mobile phase B, with gradient elution performed according to the specifications in the table below; the flow rate was 1.0 ml per minute; the column temperature was 35 °C; the detection wavelength was 235 nm; the theoretical plate number, calculated based on the Tarrasque peak, should not be less than 5000.

10. The method for detecting the traditional Chinese medicine composition for drug addiction treatment according to claim 9, characterized in that: It includes the following steps: a. Preparation of reference solution Prepare a reference solution by adding methanol to tarassamine reference standard. b. Preparation of the test solution Take an appropriate amount of the test sample, add ammonia solution and ether, seal tightly and soak for 24 hours, then sonicate, cool, shake well, filter, and take the filtrate to obtain the sample. c. Detection under the chromatographic conditions described in claim 9.

11. A method for detecting the traditional Chinese medicine composition for drug addiction treatment according to any one of claims 1-5, characterized in that: The contents of cryptochlorogenic acid, catechin, berberine II, isochlorogenic acid C, and berberine I were determined by high performance liquid chromatography (HPLC). The chromatographic conditions were as follows: Using octadecylsilane-bonded silica gel as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); using acetonitrile as mobile phase A and 0.1% formic acid solution (containing 1.62 g ammonium formate per 1000 ml) as mobile phase B, gradient elution was performed according to the specifications in the table below; the flow rate was 1.0 ml per minute; the column temperature was 30 °C; the detection wavelength was 280 nm; the theoretical plate number calculated based on the cryptochlorogenic acid peak should not be less than 5000; 12. The method for detecting the traditional Chinese medicine composition for drug addiction treatment according to claim 11, characterized in that: It includes the following steps: a. Preparation of reference solution Prepare a reference solution by adding methanol to cryptochlorogenic acid reference standard, catechin reference standard, berberine II, isochlorogenic acid C, and berberine I reference standards. b. Preparation of the test solution Take the sample to be tested, add 70% methanol, and sonicate for 30 minutes to obtain the sample. c. Detection under the chromatographic conditions described in claim 11.

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