Traditional Chinese medicine composition for treating turbid phlegm and blood stasis type erectile dysfunction and preparation method thereof

Abstract

The invention provides a traditional Chinese medicine composition for treating turbid phlegm and blood stasis type erectile dysfunction and a preparation method thereof. The composition is prepared from various traditional Chinese medicinal materials such as roses, purslane, carrot seeds, turnip seeds, anchusa, muskmelon seeds, basil seeds, cornflower and limonium sinense, and the traditional Chinese medicinal materials are combined according to a specific weight ratio and have the effects of eliminating phlegm and heat, dissipating blood stasis, activating blood circulation, tonifying the body and strengthening yang. The preparation method comprises the steps of medicinal material treatment, extraction, concentration, drying and forming, an extraction solvent is preferably water, and the extraction process is optimized, so that efficient extraction and stability of effective components can be ensured. The composition is suitable for being prepared into various dosage forms such as tablets, granules, capsules, oral liquid, ointment, gel or spray, and has a remarkable curative effect and a good application prospect.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to the preparation and application of a Chinese medicine composition, and in particular to a Chinese medicine composition for treating phlegm-turbidity and blood-stasis type erectile dysfunction and a preparation method thereof. Background Art

[0002] Erectile dysfunction (ED) refers to the inability of the penis to achieve or maintain an erection sufficient for a satisfactory sexual life. It is one of the common sexual dysfunctions in adult men. With the increase of social pressure and the change of lifestyle, the incidence of ED is increasing and becoming younger. Research data shows that the number of ED patients worldwide is increasing year by year, which has a serious impact on men's reproductive health and quality of life.

[0003] Existing ED treatments mainly include oral phosphodiesterase-5 (PDE-5) inhibitors, hormone replacement therapy, device-assisted therapy, and psychological counseling. However, these methods have certain limitations in practical application, such as obvious side effects, limited efficacy, strong dependence, and contraindications for some patients, making these treatments unsuitable.

[0004] In traditional Chinese medicine theory, ED is classified as "impotence" and "impotence". Its etiology and pathogenesis are complex and are often related to factors such as kidney deficiency, phlegm turbidity, and blood stasis. Among them, phlegm turbidity and blood stasis type ED is a common syndrome in clinical practice. Its characteristics are phlegm dampness obstruction and blood stasis, which leads to the loss of nourishment to the penis meridians and dysfunction. For the treatment of phlegm turbidity and blood stasis type ED, traditional Chinese medicine often adopts the treatment methods of clearing phlegm heat, dispersing blood stasis and activating blood circulation, and nourishing the body and strengthening yang.

[0005] In recent years, Chinese medicine has shown unique advantages in the treatment of ED, with characteristics such as multiple targets, overall regulation, and low toxicity and side effects. However, existing Chinese medicine preparations still have deficiencies in terms of compatibility, extraction technology, and quality control, making it difficult to fully exert the efficacy of Chinese medicine. Especially in the specific treatment of phlegm-turbidity and blood-stasis type ED, there is a lack of Chinese medicine compositions that have been systematically studied and clinically verified.

[0006] Therefore, there is an urgent need to develop a Chinese medicine composition for erectile dysfunction of phlegm-turbidity and blood stasis type, which has a clear compatibility composition, an optimized extraction process and a stable quality control method, so as to improve the clinical efficacy and meet the treatment needs of patients. Summary of the invention

[0007] The purpose of the present invention is to provide a Chinese medicine composition for phlegm turbidity and blood stasis type erectile dysfunction and its preparation method. Through the carefully selected combination of Chinese medicinal materials and the optimized extraction process, it aims to effectively improve the symptoms of patients with this type of erectile dysfunction, has significant efficacy and good safety, is suitable for long-term use, and meets the needs of clinical treatment.

[0008] To achieve the above-mentioned purpose, the present invention provides a Chinese medicine composition, which is composed of the following raw medicinal materials in parts by weight: rose: 1 to 15 parts; purslane: 1 to 15 parts; carrot seeds: 1 to 15 parts; turnip seeds: 1 to 15 parts; bugloss: 1 to 15 parts; melon seeds: 1 to 15 parts; basil seeds: 1 to 15 parts; cornflower: 1 to 12 parts; and limonium herb: 1 to 12 parts.

[0009] Preferably, the weight portions of each raw medicinal material are: rose: 6 to 15 parts; purslane: 6 to 15 parts; carrot seeds: 6 to 15 parts; turnip seeds: 6 to 15 parts; bugloss: 6 to 15 parts; melon seeds: 6 to 15 parts; basil seeds: 6 to 15 parts; cornflower: 5 to 12 parts; limonium: 5 to 12 parts.

[0010] More preferably, the weight proportions of each raw medicinal material are: rose: 15 parts; purslane: 15 parts; carrot seeds: 15 parts; turnip seeds: 15 parts; bugloss: 15 parts; melon seeds: 15 parts; basil seeds: 15 parts; cornflower: 12 parts; limonium: 12 parts.

[0011] The components of the above-mentioned Chinese medicine composition have the following effects: Rose flower: regulating qi and relieving depression, promoting blood circulation and removing blood stasis; Purslane: clearing away heat and detoxifying, dispersing blood stasis and reducing swelling; Carrot seed: promoting diuresis and drying dampness, strengthening the spleen and kidney; Turnip seed: drying dampness and resolving phlegm, eliminating accumulation and regulating qi; Herba Lycopodii: clearing away heat and detoxifying, promoting dampness and reducing swelling; Melon seed: nourishing kidney and strengthening yang, drying dampness and consolidating essence; Basil seed: strengthening spleen and stomach, dispelling wind and regulating qi; Cornflower: nourishing liver and kidney, enhancing physical strength; Blood-replenishing Grass: nourishing blood and replenishing qi, regulating qi and blood. These medicinal materials are scientifically matched to synergistically play the effects of clearing phlegm and heat, dispersing blood stasis and promoting blood circulation, and nourishing the body and strengthening yang, specifically targeting the pathological characteristics of erectile dysfunction of phlegm turbidity and blood stasis type, and fundamentally regulating the patient's body functions.

[0012] Selection of extraction solvent

[0013] The present invention compares the extraction effects of whole powder, water extract, and different concentrations of ethanol extracts (10%, 30%, 50%, 70%, 95%) on the effective components of the Chinese medicine composition through systematic experimental research. The results of behavioral and biochemical index screening in animal experiments show that the water extract with water as the extraction solvent has the best effect in improving the biological signs, blood lipid levels, blood rheology indicators and hormone levels of rats with erectile dysfunction model of phlegm turbidity and blood stasis. Therefore, water is determined to be the best extraction solvent for the Chinese medicine composition of the present invention.

[0014] Preparation process

[0015] The present invention optimizes the key process parameters such as the number of extractions, extraction time, and water addition times in the preparation method of the Chinese medicine composition by using the response surface design method. The extraction rate of the main active ingredients and the extract yield are used as evaluation indicators to determine the optimal extraction process conditions, so that the active ingredients of the Chinese medicine composition are extracted to the maximum extent, ensuring the quality stability and consistency of the efficacy of the product.

[0016] The preparation method of the Chinese medicine composition of the present invention comprises the following steps:

[0017] 1) Raw material processing: weigh each raw medicinal material according to the above proportion, remove impurities, clean it, and dry it for later use.

[0018] 2) Extraction step: Mix the treated medicinal materials evenly, add 8 to 15 times the amount of water, and soak for 0.5 to 2 hours. Then heat and reflux at 80 to 100°C for 1 to 3 hours, and extract 2 to 4 times. Filter after each extraction and combine the extracts. The preferred extraction conditions are: add 12 times the amount of water, soak for 1 hour, heat and reflux at 85°C for 1.5 hours, and extract 3 times.

[0019] 3) Concentration step: The combined extract is concentrated under reduced pressure to a thick paste with a relative density of 1.15 to 1.25 (measured at 60° C.) The preferred concentration condition is: concentrated under reduced pressure to a relative density of 1.20.

[0020] 4) Drying step: The concentrated paste is dried by spray drying or vacuum drying at a temperature of 50 to 90° C. to obtain a dry powder of the Chinese medicine composition. The preferred drying conditions are: spray drying at a drying temperature of 70° C.

[0021] 5) Formulation: The obtained dry powder is prepared into different dosage forms as required, including but not limited to tablets, granules, capsules, oral liquids, ointments, etc., supplemented with pharmaceutically acceptable excipients such as adhesives, fillers, disintegrants, lubricants, etc.

[0022] The preparation of the preparation can be carried out according to conventional processes, for example:

[0023] Tablets: Mix the dry powder with an appropriate amount of binder and disintegrant and press into tablets;

[0024] Capsules: Put the dry powder directly into capsules;

[0025] Granules: Mix the dry powder with the auxiliary materials, granulate and dry to obtain granules;

[0026] Oral solution: dissolve or suspend the dry powder in a suitable amount of solvent, add appropriate stabilizers, preservatives, etc. to obtain the oral solution.

[0027] The Chinese medicine composition of the present invention can be prepared into a variety of dosage forms to meet the needs of different patients and clinical application scenarios. In addition to oral preparations, it can also be prepared into external preparations, such as ointments, gels, sprays, etc., for local administration, to increase the concentration of the drug in the affected area and improve the efficacy. In addition, the Chinese medicine composition of the present invention can also be applied to other male reproductive health problems related to phlegm turbidity and blood stasis, such as loss of libido, infertility, etc., and has broad application prospects.

[0028] The novelty of the invention

[0029] Compared with the prior art, the present invention has significant innovations and advantages in the following aspects:

[0030] 1) Unique combination of medicinal materials: Based on traditional Uyghur medical theory and combined with modern pharmacological research, nine medicinal materials such as rose, purslane, carrot seeds, etc. are combined in a specific proportion for the first time to achieve a comprehensive multi-target therapeutic effect for erectile dysfunction of phlegm and blood stasis type.

[0031] 2) Optimization of extraction solvent: Through systematic experimental research, water was determined to be the best extraction solvent, which not only ensures the extraction efficiency of effective ingredients, but also meets the requirements of green environmental protection, reducing production costs and environmental pressure.

[0032] 3) Optimization of preparation process: The response surface design method was used to optimize the key process parameters to ensure the stability of product quality and consistency of efficacy. The process is simple and suitable for industrial production.

[0033] 4) Broad application prospects: The Chinese medicine composition of the present invention can be prepared into a variety of dosage forms, suitable for different application scenarios. It not only has significant therapeutic effects in the treatment of erectile dysfunction of phlegm and blood stasis type, but can also be used for the prevention and treatment of other related diseases.

[0034] In summary, the Chinese medicine composition and the preparation method thereof provided by the present invention have the advantages of significant efficacy, high safety, and suitability for industrial production through scientific combination of medicinal materials and optimized preparation process, filling the technical gap in this field and having important clinical application value and broad market prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 Quantitative results of biological signs of rats in the normal group and the modeling group after 8 weeks of modeling

[0036] Figure 2 Results of the APO erectile function and sexual behavior function experiment in rats in the normal group and the modeling group after 8 weeks of modeling Figure 3Results of erectile function and sexual behavior function of APO rats in each group after drug intervention for 0-3 weeks

[0037] Figure 4 ICP / MAP test results of rats in each group after drug intervention

[0038] Figure 5 Results of blood lipid level test in rats

[0039] Figure 6 Results of blood rheology test in rats

[0040] Figure 7 Results of hormone level test in rats

[0041] Figure 8 Masson staining results of rat penis tissue

[0042] Fig. 9 HPLC chart of specificity test for detection of Chinese medicine composition, where a is test sample, b is mixed reference sample, and c is blank control;

[0043] 1. Caffeic acid; 2. Rutin; 3. Rosmarinic acid; 4. Quercetin; 5. Luteolin;

[0044] Fig.10 Design space diagram of extraction process of traditional Chinese medicine composition DETAILED DESCRIPTION

[0045] The present invention is described in detail below by specific embodiments to better understand the technical solutions and advantages of the present invention. However, the specific embodiments described below are only illustrative contents and do not limit the scope of protection of the present invention. The specific conditions not specified in the present invention are operated according to the conventional conditions in the art or the conditions recommended by the manufacturer, and the raw materials, reagents or equipment used are all commercially available conventional products unless otherwise specified. Unless otherwise defined, the terms and expressions in the present invention should be consistent with the meanings generally understood by those skilled in the art.

[0046] Example 1 Establishment and evaluation of rat model of erectile dysfunction with phlegm turbidity and blood stasis

[0047] 1. Model building

[0048] 160 6-week-old male SD rats with a body weight of 200±10g were selected. Ten rats were randomly selected as the normal control group (N group), and the remaining 150 rats were used as the phlegm-turbidity and blood-stasis type erectile dysfunction modeling group (M group). The rats in group N were fed with ordinary feed, and the rats in group M were fed with a high-fat feed with a 60% energy supply ratio. The modeling time was 8 weeks. During the modeling period, the biological signs of the rats were observed regularly every day, including hair, tongue coating, urination and defecation status, and quantitative indicators such as body weight, core temperature, body surface temperature, diet, and water intake. The erectile function and sexual behavior ability of the modeling rats were evaluated using the APO penile erection experiment and sexual behavior experiment.

[0049] 2. Experimental results

[0050] (1) Biological signs

[0051] During the 8-week modeling period, the rats in group N showed normal biological characteristics, with neat and shiny hair, light red tongue coating, clear urine, formed stool, and stable body weight, core temperature, body surface temperature, diet and water intake. However, the rats in group M gradually showed pathological characteristics of phlegm turbidity and blood stasis. At 4 weeks after modeling, the hair of the rats in group M was malnourished, the tongue coating gradually thickened, and the color changed from light red to dark purple, accompanied by symptoms such as dry stool and turbid urine. By 8 weeks after modeling, the weight of the rats in group M increased significantly, the core temperature increased significantly, the body surface temperature decreased significantly, and the diet and water intake decreased, showing typical pathophysiological characteristics of phlegm turbidity and blood stasis ( Figure 1 shown).

[0052] (2) Results of the APO penis erection experiment

[0053] In the APO penile erection experiment, the erection latency of rats in group N was shorter and the number of erections was more frequent, showing normal erectile function. In contrast, the erection latency of rats in group M was significantly prolonged and the number of erections decreased 4 weeks after modeling. By 8 weeks, the erectile function was significantly reduced, and some rats had no erectile response ( Figure 2 shown).

[0054] (3) Results of sexual behavior experiments

[0055] In the sexual behavior experiment, the rats in group N showed normal sexual behavior, including shorter latency periods of climbing, insertion, and ejaculation. However, the sexual behavior of the rats in group M decreased significantly after 4 weeks of modeling, and the number of insertions decreased significantly (P<0.01). At 8 weeks, the sexual behavior of the rats in group M was severely inhibited ( Figure 2 shown).

[0056] (4) Model evaluation

[0057] The results of biological signs, APO penile erection experiment and sexual behavior experiment confirmed that the M group rats successfully established the phlegm turbidity and blood stasis type erectile dysfunction model. According to the experimental results after modeling, the M group rats were further divided into the phlegm turbidity and blood stasis syndrome group (S group) and the erectile dysfunction group (ED group). The S group rats showed the characteristics of phlegm turbidity and blood stasis syndrome, while the ED group rats had no erection and sexual behavior response in the APO penile erection experiment and sexual behavior experiment, successfully verifying the phlegm turbidity and blood stasis type erectile dysfunction model.

[0058] Example 2: Preparation of whole powder

[0059] Take 6.61g of rose, 6.61g of purslane, 6.61g of carrot seeds, 6.61g of turnip seeds, 6.61g of bugloss, 6.61g of melon seeds, 6.61g of basil seeds, 5.29g of cornflower, and 5.29g of limonium herb, mix all the Chinese medicinal materials, grind them and pass through a 24-mesh standard medicine sieve to obtain drug powder within 850±29μm.

[0060] Example 3: Preparation of water extract

[0061] Take 13.22g of rose, 13.22g of purslane, 13.22g of carrot seed, 13.22g of turnip seed, 13.22g of bugloss, 13.22g of melon seed, 13.22g of basil seed, 10.57g of cornflower and 10.57g of limonium, add appropriate amount of pure water, heat and reflux for extraction, filter, combine the filtrate and concentrate under reduced pressure to a thick paste, and vacuum dry at 55°C to obtain a dry extract.

[0062] Example 4: Preparation of 10% ethanol extract

[0063] Take 13.22g of rose, 13.22g of purslane, 13.22g of carrot seeds, 13.22g of turnip seeds, 13.22g of bugloss, 13.22g of melon seeds, 13.22g of basil seeds, 10.57g of cornflower, and 10.57g of limonium herb, add appropriate amount of 10% ethanol solution, heat and reflux to extract, filter and combine the filtrate, concentrate under reduced pressure to a thick paste, and dry in a vacuum at 55°C to obtain a dry extract.

[0064] Example 5: Preparation of 30% ethanol extract

[0065] Take 13.22g of rose, 13.22g of purslane, 13.22g of carrot seeds, 13.22g of turnip seeds, 13.22g of bugloss, 13.22g of melon seeds, 13.22g of basil seeds, 10.57g of cornflower, and 10.57g of limonium herb, add appropriate amount of 30% ethanol solution, heat and reflux to extract, filter and combine the filtrate, concentrate under reduced pressure to a thick paste, and dry in a vacuum at 55°C to obtain a dry extract.

[0066] Example 6: Preparation of 50% ethanol extract

[0067] Take 13.22g of rose, 13.22g of purslane, 13.22g of carrot seeds, 13.22g of turnip seeds, 13.22g of bugloss, 13.22g of melon seeds, 13.22g of basil seeds, 10.57g of cornflower, and 10.57g of limonium herb, add appropriate amount of 50% ethanol solution, heat and reflux to extract, filter and combine the filtrate, concentrate under reduced pressure to a thick paste, and dry in a vacuum at 55°C to obtain a dry extract.

[0068] Example 7: Preparation of 70% ethanol extract

[0069] Take 13.22g of rose, 13.22g of purslane, 13.22g of carrot seeds, 13.22g of turnip seeds, 13.22g of bugloss, 13.22g of melon seeds, 13.22g of basil seeds, 10.57g of cornflower, and 10.57g of limonium herb, add appropriate amount of 70% ethanol solution, heat and reflux to extract, filter and combine the filtrate, concentrate under reduced pressure to a thick paste, and dry in a vacuum at 55°C to obtain a dry extract.

[0070] Example 8: Preparation of 95% ethanol extract

[0071] Take 13.22g of rose, 13.22g of purslane, 13.22g of carrot seeds, 13.22g of turnip seeds, 13.22g of bugloss, 13.22g of melon seeds, 13.22g of basil seeds, 10.57g of cornflower, and 10.57g of limonium herb, add appropriate amount of 95% ethanol solution, heat and reflux to extract, filter and combine the filtrate, concentrate under reduced pressure to a thick paste, and dry in a vacuum at 55°C to obtain a dry extract.

[0072] Example 9: Effects of the Chinese herbal composition on clearing phlegm and heat, dispersing blood stasis and promoting blood circulation, and strengthening the body and yang in rats with erectile dysfunction of phlegm turbidity and blood stasis type

[0073] The experiment selected the rat model of phlegm turbidity and blood stasis type erectile dysfunction successfully established in Example 1. Rats were divided into a normal control group (N group), a phlegm turbidity and blood stasis syndrome group (S group), and a phlegm turbidity and blood stasis type erectile dysfunction group (ED group). The experimental drugs included different extracts of the Chinese medicine composition of the present invention and the positive control drug Imusak tablets, which were specifically divided into original drug powder group (O group), water extract group (W group), 10% ethanol extract group (10% group), 30% ethanol extract group (30% group), 50% ethanol extract group (50% group), 70% ethanol extract group (70% group) and 95% ethanol extract group (95% group), and Imusak tablets (Y group) were used as positive control drugs.

[0074] Biological sign evaluation results

[0075] During the whole experiment, the rats in group N maintained a good physiological state, with smooth hair, light red tongue coating, clear urine, dark brown stool and moderate water content. Compared with group N, the rats in groups S and ED showed symptoms such as dry hair, thick purple tongue coating, turbid urine, and constipation. After 2 weeks of intervention, the hair glossiness of the rats in group W increased, the color of the tongue coating gradually became normal, the urine color improved, and the stool hardness decreased ( Figure 3 shown).

[0076] In terms of quantitative physical signs, the weight of rats in the ED group increased significantly, the core temperature increased, and the body surface temperature, water intake, and urine and feces volume decreased significantly. Groups W, Y, and O showed significant regulatory effects after 2 weeks of intervention, especially Group W, which performed best in weight control, water intake, and urine and feces volume ( Figure 4 shown).

[0077] Results of APO penis erection experiment and sexual behavior experiment

[0078] After 2 weeks of administration, the erection latency of rats in group W was significantly shortened, the number of erections increased, and the sexual activity was significantly improved. The effect of group W in improving the erectile function and sexual behavior function of rats was better than that of other extract groups and positive drugs ( Figure 3 shown).

[0079] ICP / MAP results

[0080] The ICP / MAP ratio of rats in the ED group was significantly lower than that in the N group, indicating that their erectile function was severely impaired. After intervention, the ICP / MAP of rats in the W group was significantly improved, indicating that their erectile function recovery was better than that of the other extract groups and the positive control drug ( Figure 4 shown).

[0081] Lipid Level Results

[0082] The levels of triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in the ED group were significantly higher than those in the N group, and the phlegm and turbidity signs were significant. After intervention in the W group, the levels of TG and LDL-C were significantly reduced and returned to near normal levels, which was better than that in the other extract groups ( Figure 5 shown).

[0083] Blood flow changes

[0084] Blood rheology indexes showed that the whole blood viscosity (WBV) and hematocrit (Hct) of rats in the ED group increased significantly, showing typical blood stasis characteristics. The W group rats had significant effects in regulating WBV and Hct, indicating that it effectively improved blood circulation ( Figure 6 shown).

[0085] Hormone Level Results

[0086] The levels of prolactin (PRL) and estradiol (E2) in rats in the ED group increased significantly, while the level of testosterone (T) decreased significantly. After 3 weeks of intervention, the levels of PRL and E2 in rats in the W group decreased significantly, while the level of T increased significantly, indicating that the W group had a significant regulatory effect on hormone levels ( Figure 7 shown).

[0087] Histopathological results

[0088] HE staining results showed that the corpus cavernosum sinus cavity of the ED group rats was enlarged and the tissue structure was significantly damaged. After intervention in the W group, the pathological damage of the corpus cavernosum tissue was reduced, the sinus cavity was reduced, and the tissue structure was repaired, which was better than the other extract groups and the positive control drug.

[0089] The results of Masson staining showed that the collagen fibers in the penis tissue of the rats in the ED group increased significantly, while the smooth muscle fibers decreased. After intervention, the collagen fibers in the rats in the W group decreased significantly, the proportion of smooth muscle fibers increased, and the degree of tissue fibrosis was significantly reduced, with significant effects ( Figure 8 shown).

[0090] In summary, the experimental results show that the Chinese medicine composition of the present invention, especially the water extract (W group), shows significant effects in improving the erectile function, sexual behavior function, blood lipid level, blood circulation, hormone level and repairing tissue pathology of rats with phlegm turbidity and blood stasis type erectile dysfunction. The W group is superior to other extracts and positive drug groups in many biological and pathological indicators, showing its potential clinical application value in the treatment of phlegm turbidity and blood stasis type erectile dysfunction.

[0091] Example 10: HPLC Detection of Main Components of Traditional Chinese Medicine Composition

[0092] This experiment uses a Waters 2695-2998 high performance liquid chromatograph and a ZORBAX StableBond Aq chromatographic column (4.6 x 250 mm, 5 μm). The test drugs include the various components of the Chinese medicine composition of the present invention, such as rose, purslane, carrot seeds, etc. All medicinal materials meet the standards of the "Chinese Pharmacopoeia". Reagents such as caffeic acid, rutin, rosmarinic acid and other reference substances are provided by Shanghai Yuanye Biotechnology Co., Ltd. to ensure that all materials and reagents used in the experiment meet the precision and stability requirements of the experiment.

[0093] Preparation of reference and test solutions and chromatographic conditions

[0094] According to the requirements of chromatographic analysis, mixed reference solution and test solution were prepared respectively. The mixed reference included caffeic acid, rutin, rosmarinic acid, quercetin and luteolin, which were dissolved and fixed with methanol. The test sample was extracted from the Chinese medicine composition, prepared by ultrasonic reflux method and filtered through a microporous membrane. A ZORBAX StableBond Aq chromatographic column (4.6x 250mm, 5μm) was used, with a gradient elution mobile phase and a detection wavelength of 330nm.

[0095] Specificity test

[0096] The chromatograms of the test sample and mixed reference solution were determined according to the above method. The results showed that the retention time of the chromatographic peaks of the test sample and the mixed reference solution was consistent, there was no interference from other components, and the chromatographic peaks were well separated, ensuring the specificity of the method ( Fig. 9 shown).

[0097] Linear relationship investigation

[0098] In order to study the linear relationship of the detection method, reference solutions of different concentrations were accurately aspirated, their peak areas were recorded, and standard curves were drawn. The results showed that caffeic acid, rutin, rosmarinic acid, quercetin and luteolin had good linear relationships, and the correlation coefficient reached 0.999, indicating that the method had a good linear range.

[0099] Precision test

[0100] To test the precision of the instrument, the sample was injected 6 times continuously and the peak area of ​​each index component was recorded. The relative standard deviation (RSD) of the peak area of ​​each component was within the allowable range, and the RSD of caffeic acid, rutin, rosmarinic acid, quercetin, and luteolin were 0.890%, 2.748%, 0.328%, 3.799%, and 2.489%, respectively, indicating that the chromatographic method has good precision.

[0101] Stability test

[0102] The test solution was placed at room temperature for 0 to 36 hours, and samples were taken regularly for chromatographic determination. The results showed that the test sample was relatively stable within 24 hours, and the RSD of the peak area of ​​each component within 36 hours was small, indicating that the test solution had good stability at room temperature.

[0103] Repeatability test

[0104] In order to verify the repeatability of the method, 6 test solutions were prepared using the same preparation method and chromatographic determination was performed. The results showed that the RSDs of caffeic acid, rutin, rosmarinic acid, quercetin and luteolin were 0.649%, 0.397%, 0.161%, 3.10% and 3.016%, respectively, indicating that the method had good repeatability.

[0105] Sample recovery test

[0106] In order to test the accuracy of the detection method, a known amount of reference solution was added to the test sample, and the peak area of ​​each component was measured and the recovery rate was calculated. The results showed that the recovery rates of caffeic acid, rutin, rosmarinic acid, quercetin, and luteolin were 99.618%, 99.729%, 98.893%, 96.349%, and 97.430%, respectively. The RSD of each component was within the allowable range, indicating that the detection method was accurate and reliable.

[0107] The experimental results of this example verify the specificity, precision, stability, repeatability and accuracy of the high performance liquid chromatography detection method for the main components of the Chinese medicine composition of the present invention, ensuring that the method can effectively and accurately detect the key components in the Chinese medicine composition of the present invention, providing a reliable basis for subsequent quality control and further research.

[0108] Example 11 Optimization of extraction process of traditional Chinese medicine composition based on Box-Behnken response surface design

[0109] Box-Behnken Experimental Design

[0110] In this experiment, the Box-Behnken response surface design combined with failure mode and effect analysis (FMEA) was used to determine the number of extractions, extraction time and water addition as the main factors affecting the extraction process. By investigating the effects of these factors on the extraction process of the Chinese medicine composition, the extract yield (Y1), caffeic acid (Y2), rutin (Y3), rosmarinic acid (Y4), quercetin (Y5) and luteolin (Y6) were used as evaluation indicators. The experimental design included 3 factors and 3 levels, with a total of 17 experimental points designed, and the center point was repeated 5 times. Table 1 shows the Box-Behnken experimental design and results.

[0111] Table 1 Box-Behnken design and results

[0112]

[0113]

[0114] Model establishment and evaluation

[0115] According to the principle of selecting the highest-order polynomial model when multiple fitting model results are significant, the quadratic polynomial regression model was used to fit the six evaluation indicators of caffeic acid, rutin, rosmarinic acid, quercetin and luteolin content and extract yield and the corresponding three influencing factors of water addition, extraction time and extraction times. The results are shown in Table 2.

[0116] Table 2 Box-Behnken design optimization model parameters and statistical results for the extraction process of traditional Chinese medicine compound

[0117]

[0118]

[0119] Control space development

[0120] To further optimize the extraction process conditions, the optimization ranges of the six evaluation indicators were set at different extraction times. Taking 80% of the maximum extraction amount of each component as the optimization target, the control space of extraction times of 1, 2 and 3 was determined (see Fig.10 ). The results showed that when the number of extractions was 3 times, the extraction time was 1.5 hours, and the amount of water added was 12 times, all indicators could reach the optimization target. Under this process condition, the content of caffeic acid, rutin, rosmarinic acid, quercetin, luteolin and the yield of extract all reached the optimal balance point, and it had high economy and operability. Therefore, the optimal extraction process conditions finally determined were: 3 extractions, 1.5 hours of extraction time, and 12 times of water added.

[0121] Validation of the optimal extraction process

[0122] In order to verify the reliability and stability of the optimal extraction process, this study further adopted the optimal extraction process conditions: extraction times 3 times, extraction time 1.5h, water addition 12 times, and 6 verification samples were prepared in parallel. The verification results are shown in Table 3. After extraction according to the optimal extraction process, the results can meet the preset target: caffeic acid content > 1.40μg·mL -1 , rutin content>10.952μg·mL -1 , rosmarinic acid content>24.160μg·mL -1 , quercetin content>1.552μg·mL -1 , luteolin content>0.584μg·mL -1 , the extract yield was >23.064%, and the RSD values ​​of each CQAs were less than 5%. The verification results showed that the optimal extraction process conditions were stable and reliable, and could lay the foundation for the next step of production process amplification and preparation process standardization.

[0123] Table 3 Optimal extraction process verification results

[0124]

Claims

1. A Chinese medicine composition for treating phlegm-turbidity and blood-stasis type erectile dysfunction, characterized in that: The invention is composed of the following raw medicinal materials in parts by weight: 1-15 parts of rose, 1-15 parts of purslane, 1-15 parts of carrot seeds, 1-15 parts of turnip seeds, 1-15 parts of bugloss, 1-15 parts of melon seeds, 1-15 parts of basil seeds, 1-12 parts of cornflower and 1-12 parts of limonium herb.

2. The Chinese medicine composition according to claim 1, characterized in that: The weight proportions of the raw medicinal materials are: 6-15 parts of rose, 6-15 parts of purslane, 6-15 parts of carrot seeds, 6-15 parts of turnip seeds, 6-15 parts of bugloss, 6-15 parts of melon seeds, 6-15 parts of basil seeds, 5-12 parts of cornflower, and 5-12 parts of limonium.

3. The Chinese medicine composition according to claim 1 or 2, characterized in that: The weight proportions of the raw medicinal materials are as follows: 15 parts of rose, 15 parts of purslane, 15 parts of carrot seeds, 15 parts of turnip seeds, 15 parts of bugloss, 15 parts of melon seeds, 15 parts of basil seeds, 12 parts of cornflower, and 12 parts of limonium.

4. The Chinese medicine composition according to any one of claims 1 to 3, characterized in that: The traditional Chinese medicine composition uses water as an extraction solvent.

5. The Chinese medicine composition according to claim 4, characterized in that: The Chinese medicine composition can be prepared into one or more of the following dosage forms: tablets, granules, capsules, oral liquids, pastes, gels or sprays.

6. A method for preparing the Chinese medicine composition according to any one of claims 1 to 5, characterized in that: The following steps are involved: (1) Weigh each raw medicinal material according to the proportion, clean it, and dry it for later use; (2) After mixing the medicinal materials, add 8 to 15 times the amount of water and soak for 0.5 to 2 hours; (3) heating and refluxing at 80 to 100° C. for 1 to 3 hours, extracting 2 to 4 times, combining the extracts, filtering and removing the residue; (4) concentrating the extract under reduced pressure to a thick paste having a relative density of 1.15 to 1.25 (measured at 60° C.); (5) using a spray drying or vacuum drying method at a drying temperature of 50 to 90° C. to obtain a dry powder; (6) preparing the desired dosage form with the dry powder and adding pharmaceutically acceptable excipients.

7. The preparation method according to claim 6, characterized in that: In step (2), preferably 12 times of water is added and soaked for 1 hour; in step (3), heating and reflux extraction are performed at 85° C. for 1.5 hours and extraction is performed 3 times.

8. The preparation method according to claim 6 or 7, characterized in that: In step (4), the extract is preferably concentrated under reduced pressure to a relative density of 1.20 (measured at 60°C).

9. The preparation method according to any one of claims 6 to 8, characterized in that: In step (5), spray drying is preferably used, and the drying temperature is 70°C.

10. Use of the Chinese medicine composition according to any one of claims 1 to 5 in the preparation of a drug for treating erectile dysfunction of phlegm turbidity and blood stasis type.

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