Solid beverage with sleep promoting effect and identification method thereof

Abstract

The invention provides a solid beverage with a sleep promoting effect and an identification method thereof, and relates to the technical field of large health food. The beverage is prepared from medicinal and edible traditional Chinese medicinal materials such as radix puerariae, longan aril, mulberries, rhizoma polygonati, jujubes, lilies, wheat, chrysanthemums, red roses with double petals, spina date seeds and liquorice through scientific compatibility, the beverage is prepared by adopting modern pharmaceutical processes such as supercritical CO2 extraction and low-temperature and high-pressure homogeneous extraction, and the quality of a product is controlled and identified through a liquid-phase fingerprint technology. The invention provides a safe and effective product with homology of medicine and food for promoting sleep. By adopting extraction preparation technologies such as supercritical CO2 extraction and low-temperature high-pressure homogenization, the purity and activity of the components are ensured, and the modern pharmaceutical process is advanced; fingerprint analysis shows that the consistency of the relative retention time and the relative peak area of each component is good, and the similarity of chromatograms of all batches of products is more than 0.9998, so that the stability and the consistency of the products are ensured.

Description

Technical field

[0001] The present invention relates to the technical field of health foods, and in particular to a solid beverage having a sleep-promoting effect and an identification method thereof. [Background Technology]

[0002] Perimenopausal insomnia (PMI) refers to insomnia symptoms that occur during perimenopause (the transition period between menopause and postmenopause). Perimenopausal insomnia, which typically occurs between the ages of 45 and 55, is a natural phase in a woman's menstrual cycle that culminates in the cessation of menstruation, known as menopause. During this period, hormone levels in a woman's body fluctuate, particularly with a decrease in estrogen levels. These changes can affect sleep patterns and lead to insomnia. The Traditional Chinese Medicine book "Suwen: Shanggu Tianzhen Lun" states: "At the age of 49, a woman's Ren meridian becomes weak, her Taichong meridian weakens, her Tiangui (energy essence) is exhausted, and her Didao (disease channel) is blocked, resulting in a weakened body and childlessness." Women around the age of 49 often experience symptoms such as restlessness, insomnia, palpitations, irritability, hot flashes, sweating, dizziness, tinnitus, and back pain. This stage is also known as perimenopausal insomnia.

[0003] Coupled with the accelerated pace of modern life and increased work pressure, perimenopausal women face even more severe insomnia issues due to both physiological and psychological changes. Perimenopause insomnia not only impacts women's daily lives and work but can also lead to a range of health problems, including memory loss, weakened immune function, mood swings, and even serious illnesses like hypertension and coronary heart disease.

[0004] Currently, treatments for insomnia in perimenopausal women primarily include pharmacological and non-pharmacological therapies. Pharmacological treatments typically include sedatives, hypnotics, and antidepressants. While these medications can provide short-term relief, long-term use can lead to dependence, tolerance, and other side effects. Non-pharmacological treatments, such as cognitive behavioral therapy, sleep hygiene education, relaxation techniques, acupuncture, and massage, have fewer side effects, but their effectiveness is inconsistent, the treatment process is time-consuming, and adherence is poor.

[0005] In view of the shortcomings of the current treatment methods for insomnia in perimenopausal women, it is necessary to propose a new solution. Under the guidance of traditional Chinese medicine theory, a health food solid beverage with sleep-promoting effect specifically for insomnia in perimenopausal women is developed. After scientific formulation, modern preparation methods are combined with modern fingerprint technology to conduct comprehensive, objective and scientific evaluation and control of product quality to ensure the efficacy and safety of the product. [Summary of the invention]

[0006] The purpose of the present invention is to: address the problems raised in the background technology, provide a solid beverage with a sleep-promoting effect and an identification method thereof, wherein the solid beverage can effectively improve the insomnia symptoms of perimenopausal women through scientific formulation; explore advanced preparation technology to obtain a product with stable quality and easy use; and at the same time establish liquid fingerprint technology to achieve comprehensive, scientific and accurate identification and quality control of multiple ingredients in the solid beverage.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0008] A solid beverage with a sleep-promoting effect comprises the following raw materials in parts by weight: 10-30 parts of kudzu root, 10-30 parts of longan pulp, 8-24 parts of mulberry, 6-20 parts of polygonatum, 6-20 parts of jujube, 6-20 parts of lily, 5-15 parts of wheat, 5-15 parts of chrysanthemum, 3-11 parts of double red rose, 3-11 parts of sour jujube kernel, and 1.5-4.5 parts of liquorice.

[0009] The optimized solution is composed of the following raw materials in the following weight ratios: 20 parts of kudzu root, 20 parts of longan pulp, 17 parts of mulberry, 13 parts of polygonatum, 13 parts of jujube, 13 parts of lily, 10 parts of wheat, 10 parts of chrysanthemum, 7 parts of double red rose, 7 parts of sour jujube kernel, and 3 parts of licorice.

[0010] A method for preparing a solid beverage having a sleep-promoting effect comprises the following steps:

[0011] A. First, crush the Chinese date seeds by weight and pass them through a 20-40 mesh sieve. Then, take chrysanthemum and double red rose, and extract the above three materials through supercritical CO2 to obtain a compound extract and a compound extract residue;

[0012] B. Take longan meat, mulberry, jujube, lily, wheat, licorice by weight, and then take the compound extract residue of step A, crush the above materials, use low temperature and high pressure homogenization extraction, take the extract centrifugation, microfiltration, membrane concentration to prepare a concentrate, mix with auxiliary material A, spray dry and slurry, and then spray dry to prepare the compound spray-dried powder ①;

[0013] C. Weigh kudzu root and polygonatum by weight, crush and extract using low-temperature high-efficiency extraction, take the extract and centrifuge, microfilter, membrane concentrate, and vacuum concentrate to make a concentrated solution, which is then mixed with auxiliary material A, spray-dried and slurried, and then spray-dried to make the compound spray-dried powder ②;

[0014] D. The compound extract, compound spray-dried powder ①, and compound spray-dried powder ② are pre-mixed by granulation and then granulated in a one-step process to prepare a granular product, thereby obtaining a solid beverage having a sleep-promoting effect.

[0015] Optimized, the carbon dioxide flow rate of supercritical CO2 extraction in step A is 300-400 L / h, the extraction temperature is 40-50°C, and the extraction pressure is 20-30 MPa; the temperature of separation kettle I is 35-40°C, and the temperature of separation kettle II is 30-35°C; the pressure of separation kettle I is 9-11 MPa, and the pressure of separation kettle II is 5.5-6.5 MPa; the extraction time is 1.0-3.0 h.

[0016] Optimized, in step B, the temperature of the low-temperature and high-pressure homogenization extraction is lower than 25° C., the extraction pressure is 25-40 MPa, the extraction solvent amount is 10-30 times, and the microfiltration pore size is 200-500 nm.

[0017] Optimized, auxiliary material A is maltodextrin.

[0018] Optimized, Pueraria root and Polygonatum sibiricum are processed before being crushed, and the processing method is: steaming for 20min-120min at a temperature of 100-121℃.

[0019] A method for identifying a solid beverage having a sleep-promoting effect comprises the following steps:

[0020] a. Preparation of test solution: Take solid beverage, add water, dissolve and shake well, filter with a filter membrane to obtain the test solution;

[0021] b. Preparation of reference solution: Weigh the puerarin reference substance and dissolve it in an organic solvent to obtain a reference solution;

[0022] c. Solution determination: Accurately aspirate the test sample solution and the reference sample solution into a high performance liquid chromatograph for determination to obtain the test sample fingerprint and the reference sample fingerprint. The chromatographic conditions of the high performance liquid chromatograph are as follows: the chromatographic column stationary phase is filled with octadecylsilane bonded silica gel, acetonitrile is used as mobile phase A, and 0.01-0.15% by volume of an inorganic acid aqueous solution is used as mobile phase B for gradient elution. The gradient elution is performed according to the following procedure: by volume percentage,

[0023] From 0 to 10 minutes, mobile phase A was maintained at 2% and mobile phase B was maintained at 98%;

[0024] 10-15 minutes, mobile phase A increased from 2% to 7%, and mobile phase B decreased from 98% to 93%;

[0025] From 15 to 60 minutes, the mobile phase A concentration was increased from 7% to 25%, and the mobile phase B concentration was decreased from 93% to 75%;

[0026] From 60 to 80 minutes, the mobile phase A concentration was increased from 25% to 60%, and the mobile phase B concentration was decreased from 75% to 40%;

[0027] d. Similarity evaluation: The fingerprints of multiple batches of test samples and reference samples obtained according to steps a to c are imported into the Chinese medicine chromatographic fingerprint similarity evaluation system for analysis to obtain the fingerprint of the product composition. The fingerprint of the reference sample is then compared with the fingerprints of different batches of products in production for similarity, thereby evaluating the quality of the current batch of products.

[0028] For optimization, the chromatographic column was Inertsil ODS-3, the detection wavelength was 265 nm, the flow rate was 1 ml / min, the column temperature was 35 °C, and the mobile phase B was 0.1% phosphoric acid aqueous solution.

[0029] The optimized method for obtaining the control fingerprint spectrum is to prepare ≥9 batches of composition samples, and after obtaining the sample fingerprint spectrum, perform a similarity evaluation. The similarity evaluation method is: import the fingerprint spectrum into the similarity evaluation software for similarity evaluation analysis, determine the similarity ≥0.999, and determine 16 common peaks, numbered 1 to 16, among which peak 7 is puerarin and serves as a reference peak for the control fingerprint spectrum.

[0030] Research based on Traditional Chinese Medicine (TCM) theories on perimenopausal insomnia has revealed that women in their 79th year of life, known as the "seven-seventh month," experience a unique phase characterized by the loss of yin energy at the age of 40. During this period, the balance of kidney yin and yang is disrupted, with the gradual depletion of Tiangui (Essence of Heavenly Roots), leading to Chong and Ren meridian deficiency and liver and kidney deficiency. Since women are frequently damaged by blood loss during menstruation, pregnancy, childbirth, and lactation, kidney yin deficiency is often seen during this period. This insufficiency of kidney yin leads to a loss of heart fire, causing heat to disturb the mind. Since Yi and Gui share the same origin, water cannot contain wood, leading to hyperactivity of liver yang. This manifests as a series of clinical symptoms: heart-kidney disharmony, liver-kidney yin deficiency, and restlessness. Therefore, perimenopausal insomnia is often associated with kidney yin deficiency and heart-kidney disharmony.

[0031] Based on the above research, the formulation principle of the present invention is mainly based on nourishing the kidney and liver, and calming the mind and tranquilizing the nerves.

[0032] Sovereign medicine: Pueraria root, mulberry fruit, and longan pulp, all three are sovereign medicines;

[0033] Pueraria root is sweet, pungent and cool, and enters the spleen, stomach and lung meridians. It relaxes muscles and tendons, and relieves chest and diaphragm heat. Pueraria root contains chemical components such as protein, amino acids, and isoflavones, which regulate endocrine function and improve symptoms of dryness, heat and excessive sweating.

[0034] Mulberry, sweet and cold in nature, enters the liver and kidney meridians, nourishes yin and replenishes blood, and treats symptoms such as dizziness, back pain, tinnitus, premature graying of hair, insomnia and dreaminess caused by liver and kidney deficiency and blood and essence deficiency.

[0035] Longan meat is sweet and warm, and enters the heart and spleen meridians. It replenishes qi and calms the mind. It is mild and not greasy.

[0036] Ministerial drugs: Polygonatum sibiricum, lily, spinach seed, liquorice, wheat, jujube, the above 6 ingredients are ministerial drugs;

[0037] Polygonatum sibiricum, nourishes qi and yin, and benefits the kidneys. After steaming, its spleen-tonifying, lung-moistening, and kidney-benefiting functions are enhanced, and the numb taste can be removed to avoid irritation to the throat.

[0038] Lily, soothes the mind and nourishes yin and moistens the lungs;

[0039] Ziziphus jujuba seed, soothes the stomach, nourishes the heart and calms the mind, relieves boredom, and helps with sleep. Modern pharmacological research shows that ziziphus jujuba seed saponins can be sedative and hypnotic.

[0040] Licorice, wheat (floating wheat), and jujube work together synergistically to nourish the heart and produce blood, strengthen the spleen and bring peace of mind, thereby improving sleep quality.

[0041] Adjuvants: double red rose and chrysanthemum, the above two are adjuvants;

[0042] Double-petaled red roses, with their aromatic fragrance promoting qi circulation and bitter taste promoting dispersing qi, enter the liver meridian and have the effects of soothing the liver and harmonizing the stomach, promoting qi circulation and relieving depression. They can be used to treat chest, flank, and abdominal distension and pain caused by liver-stomach disharmony.

[0043] Chrysanthemum, clears away heat and detoxifies, clears the head;

[0044] Assistant drug: Licorice harmonizes the stomach, relieves urgency, and harmonizes all flavors. It is an assistant drug.

[0045] The solid beverage of the present invention utilizes a scientific combination of multiple Chinese medicinal herbs that are both medicinal and edible, to comprehensively improve insomnia symptoms in perimenopausal women by regulating liver and kidney function, nourishing the heart and calming the mind, harmonizing the spleen and stomach, and soothing the liver and relieving depression. This formulation not only has a solid foundation in traditional Chinese medicine theory but has also been validated in modern pharmacological research, ensuring the product's high efficiency, safety, and innovation, and possessing significant market value and promotional prospects.

[0046] In summary, due to the adoption of the above technical solution, the beneficial effects of the present invention are:

[0047] 1. The principle of the prescription of the present invention is centered on "nourishing the kidney and liver, calming the mind and soothing the nerves". By scientifically combining a variety of Chinese medicinal materials with the same origin as medicine and food, combined with traditional Chinese medicine theory and modern pharmacological research, the physiological and psychological states of perimenopausal women are comprehensively regulated. Due to the imbalance of kidney yin and yang in perimenopausal women, the celestial essence is gradually exhausted and the Chong and Ren meridians are deficient, which manifests as liver and kidney deficiency and heart-kidney disharmony, thereby causing a series of insomnia symptoms. The present invention uses Pueraria root, mulberry fruit and longan meat to nourish the liver and kidney, restore the balance of yin and yang, regulate endocrine system, and improve symptoms such as dryness and sweating; uses Polygonatum sibiricum, lily, and Ziziphus jujuba seed to calm the mind and soothe the nerves, nourish the heart and spleen, relieve restlessness, insomnia, anxiety and irritability, and improve sleep quality; and supplemented with double-petal red roses and chrysanthemums to soothe the liver and relieve depression, promote qi circulation and regulate emotions, and alleviate mood swings and insomnia caused by liver qi stagnation. In addition, the present invention uses licorice to harmonize the various medicines, enhance the harmony of the overall compatibility, and achieve synergistic effects while each component plays its own role.

[0048] 2. The solid beverage of the present invention realizes an efficient, safe and stable product preparation process by comprehensively applying a variety of advanced technologies in the preparation method. In the preparation process of the present invention, supercritical CO2 extraction technology is used to treat double-petal red roses and Chinese jujube kernels. The rose essential oil contained in double-petal red roses and Chinese jujube kernel oil in Chinese jujube kernels both have significant sleep-inducing effects; extraction is carried out at a relatively low temperature and in an organic solvent-free environment, thereby avoiding the destruction and contamination of active ingredients by high temperature and organic solvents, and ensuring the high purity and safety of the extract. The residue after extraction is not wasted. The present invention innovatively extracts these extracted residues together with the remaining materials, further ensuring the full extraction and utilization of the effective ingredients, and maximizing the efficiency of resource utilization.

[0049] When processing other main ingredients such as longan pulp, mulberry, jujube, lily, wheat, and licorice, the present invention adopts low-temperature, high-pressure homogenization extraction technology. This technology applies high pressure under low-temperature conditions to break the plant cell walls and release more active ingredients. Low-temperature, high-pressure homogenization extraction technology not only maintains the high activity and high stability of the extract, but also significantly improves the extraction efficiency and purity of the extract. The application of this technology avoids the damage to heat-sensitive components caused by traditional high-temperature extraction, ensuring the quality and effectiveness of the product. The application of this technology combination in the preparation of traditional Chinese medicine is a creative attempt, which significantly improves the efficacy of the product.

[0050] 3. During the processing of the extract, the present invention employs a series of sophisticated processing steps, including centrifugation, microfiltration, and membrane concentration, to further purify and concentrate the extract. The extract is then mixed with excipient A (e.g., maltodextrin) and spray-dried to produce the compound spray-dried powder. Spray-drying technology rapidly converts the liquid extract into a powder, maintaining the stability and uniform distribution of the active ingredients while ensuring the uniformity of the ingredients and the overall effectiveness of the compound. Through this modern pharmaceutical process, the present invention significantly improves the quality and stability of the product, giving it a pleasant taste and ease of portability.

[0051] 4. During the processing of the extract, the present invention employs a series of sophisticated processing steps, including centrifugation, microfiltration, and membrane concentration, to further purify and concentrate the extract. The extract is then mixed with excipient A (e.g., maltodextrin) and spray-dried to produce the compound spray-dried powder. Spray-drying technology rapidly converts the liquid extract into a powder, maintaining the stability and uniform distribution of the active ingredients while ensuring the uniformity of the ingredients and the overall effectiveness of the compound. Through this modern pharmaceutical process, the present invention significantly improves the quality and stability of the product, giving it a pleasant taste and ease of portability.

[0052] 5. The present invention's innovative fingerprint identification technology is demonstrated by its efficient identification and quality control of multiple complex ingredients. During the research process, the present invention fully considered the diverse medicinal ingredients contained in the solid beverage, including kudzu root, mulberry fruit, longan pulp, polygonatum, lily, jujube seed, licorice, wheat, jujube, double-petaled red rose, and chrysanthemum. To ensure that each ingredient has a clear and highly resolved peak in the chromatogram, the present invention conducted extensive experiments and optimization to identify the most suitable high-performance liquid chromatography (HPLC) conditions.

[0053] After much deliberation, we ultimately chose an Inertsil ODS-3 column, with a detection wavelength of 265 nm, a flow rate of 1 ml / min, and a column temperature of 35°C. Through repeated experiments, we determined that mobile phase A was acetonitrile and mobile phase B was a 0.1% aqueous phosphoric acid solution, and designed the following gradient elution program:

[0054] From 0 to 10 minutes, mobile phase A was maintained at 2% and mobile phase B was maintained at 98%;

[0055] 10-15 minutes, mobile phase A increased from 2% to 7%, and mobile phase B decreased from 98% to 93%;

[0056] From 15 to 60 minutes, the mobile phase A concentration was increased from 7% to 25%, and the mobile phase B concentration was decreased from 93% to 75%;

[0057] From 60 to 80 minutes, the concentration of mobile phase A increased from 25% to 60%, and that of mobile phase B decreased from 75% to 40%.

[0058] This chromatographic condition cannot be determined through conventional means or simple experiments, but is obtained through systematic research on various parameters such as chromatographic column type, mobile phase ratio, gradient program and detection wavelength, as well as continuous thinking based on the previous experiment, thinking on this basis and considering the next experiment, and the result obtained after precise optimization through an indefinite number of experiments.

[0059] By optimizing the chromatographic conditions described above, the present invention successfully obtained an HPLC fingerprint of the solid beverage. This fingerprint clearly separated and identified multiple components, including kudzu root, mulberry fruit, longan pulp, polygonatum, lily bulb, jujube seed, licorice root, wheat, jujube, double-petal red rose, and chrysanthemum, ensuring that each component had a unique and stable peak representation in the fingerprint.

[0060] 6. By analyzing the fingerprint spectra of multiple batches of test products, we identified 16 common peaks and clearly defined Peak 7 as the unique characteristic component of Pueraria lobata. This result can only be ensured through in-depth analysis and comparison of multiple batches of data to ensure the quality and efficacy of each batch of products.

Brief Description of the Drawings

[0061] Figure 1 A comparison chart of rat sleep latency

[0062] Figure 2 A comparison of rat sleep time

[0063] Figure 3 PSQI score comparison Comparison Chart

[0064] Figure 4 It is the acetonitrile-water 3D fingerprint

[0065] Figure 5 It is the methanol-water 3D fingerprint

[0066] Figure 6 Acetonitrile-0.1% phosphoric acid water 3D fingerprint

[0067] Figure 7 Comparison of chromatograms at different wavelengths

[0068] Figure 8 Comparison of different mobile phase gradient spectra

[0069] Figure 9 Precision test fingerprint comparison

[0070] Figure 10 Comparison of fingerprints of repeated tests

[0071] Figure 11 Comparison of stability test fingerprints

[0072] Figure 12 It is the common peak of the product fingerprint

[0073] Figure 13 Comparison of the reference and product spectra

[0074] Figure 14 It is a superposition and comparison of the fingerprints of 9 batches of products [Specific implementation method]

[0075] The following is a clear and complete description of the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0076] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention pertains. The terms used in this specification of the present invention are for the purpose of describing specific embodiments only and are not intended to limit the present invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0077] Example 1

[0078] The following is a specific formulation of the solid beverage of the present invention and examples thereof, illustrating the proportions of the various ingredients. Furthermore, to further verify the rationality of the formulation of the present invention, six comparative examples are provided; a total of four examples and six comparative examples are provided in the proportions, by weight, shown in Table 1.

[0079] Table 1 Formulations of Examples and Comparative Examples

[0080]

[0081] Example 2

[0082] The formulations listed in Examples 1-3 and Comparative Examples 1-4 were respectively prepared by the following method.

[0083] The chrysanthemum, double red rose, and Chinese jujube kernel were weighed by weight, and the Chinese jujube kernel was crushed through a 20-mesh sieve. The three materials were extracted with supercritical CO2 to obtain a chrysanthemum compound extract and a chrysanthemum compound extract residue. The carbon dioxide flow rate was 350 L / h, the extraction temperature was set to 45° C., the extraction pressure was 25 MPa, the temperature of the separation kettle I was 35° C., the temperature of the separation kettle II was 30° C., the pressure of the separation kettle I was 10 MPa, the pressure of the separation kettle II was 6 MPa, the extraction time was 2 h, and after removing water, the compound extract and the compound extract residue were obtained;

[0084] Longan pulp, compound extract residue, mulberry, jujube, lily, wheat, and licorice were weighed by weight and subjected to pulverization, low-temperature and high-pressure homogenization extraction, centrifugation, microfiltration, and membrane concentration to produce a concentrated solution. This was then mixed with an appropriate amount of maltodextrin and spray-dried to produce a compound spray-dried powder. The low-temperature and high-pressure homogenization extraction temperature was 20°C, the extraction pressure was 35 MPa, the extraction solvent volume was 30 times, and the microfiltration pore size was 500 nm.

[0085] Weigh kudzu root and polygonatum by weight and steam, crush, extract at low temperature and high efficiency, centrifuge, microfilter, membrane concentrate, and vacuum concentrate to produce a concentrated solution. Then, add an appropriate amount of maltodextrin and spray dry to produce a compound spray-dried powder. The kudzu root and polygonatum are steamed at 100°C for 30 minutes.

[0086] The compound extract, compound spray-dried powder ① and compound spray-dried powder ② are prepared into granules through processes such as granulation premixing and one-step granulation. The granules are then put into storage after inner packaging, outer packaging and inspection to obtain a solid beverage product.

[0087] The solid beverage products are numbered as Example 1-3 samples and Comparative Example 1-4 samples respectively.

[0088] Example 3

[0089] According to the formulation of Example 3 of Example 1, the preparation was carried out according to the following method.

[0090] The chrysanthemum, double red rose and Chinese jujube kernel were weighed by weight, the Chinese jujube kernel was crushed through a 40-mesh sieve, and the three materials were extracted by supercritical CO2 to obtain a chrysanthemum compound extract and a chrysanthemum compound extract residue, the carbon dioxide flow rate was 400 L / h, the extraction temperature was set to 50° C., the extraction pressure was 30 MPa, the temperature of separation kettle I was 40° C., the temperature of separation kettle II was 35° C., the pressure of separation kettle I was 11 MPa, the pressure of separation kettle II was 6.5 MPa, and the extraction time was 3.0 h to obtain the compound extract and the compound extract residue;

[0091] Longan pulp, compound extract residue, mulberry, jujube, lily, wheat, and licorice were weighed by weight and subjected to pulverization, low-temperature and high-pressure homogenization extraction, centrifugation, microfiltration, and membrane concentration to produce a concentrated solution. This was then mixed with an appropriate amount of maltodextrin and spray-dried to produce a compound spray-dried powder. The low-temperature and high-pressure homogenization extraction temperature was 25°C, the extraction pressure was 25 MPa, the extraction solvent volume was 10 times, and the microfiltration pore size was 200 nm.

[0092] Weigh kudzu root and polygonatum by weight and steam, crush, extract at low temperature, centrifuge, microfilter, membrane concentrate, and vacuum concentrate to produce a concentrated solution. This solution is then mixed with an appropriate amount of maltodextrin and spray-dried to produce a compound spray-dried powder. The kudzu root and polygonatum are steamed at 121°C for 120 minutes.

[0093] The compound extract, compound spray-dried powder ① and compound spray-dried powder ② are prepared into granules through processes such as granulation premixing and one-step granulation. The granules are then put into storage after inner packaging, outer packaging and inspection to obtain a solid beverage product.

[0094] The solid beverage products are numbered as samples of Example 4.

[0095] Example 4

[0096] The products listed in the examples can also be prepared according to the following preparation methods.

[0097] The chrysanthemum, double red rose and Chinese jujube kernel were weighed by weight, the Chinese jujube kernel was crushed through a 20-mesh sieve, and the three materials were extracted by supercritical CO2 to obtain a chrysanthemum compound extract and a chrysanthemum compound extract residue, the carbon dioxide flow rate was 300 L / h, the extraction temperature was set to 40°C, the extraction pressure was 20 MPa, the temperature of separation kettle I was 37°C, the temperature of separation kettle II was 32°C, the pressure of separation kettle I was 9 MPa, the pressure of separation kettle II was 5.5 MPa, the extraction time was 1.0 h, and after removing water, the compound extract and the compound extract residue were obtained;

[0098] Longan pulp, compound extract residue, mulberry, jujube, lily, wheat, and licorice were weighed by weight and subjected to pulverization, low-temperature and high-pressure homogenization extraction, centrifugation, microfiltration, and membrane concentration to produce a concentrated solution. This was then mixed with an appropriate amount of maltodextrin and spray-dried to produce a compound spray-dried powder. The low-temperature and high-pressure homogenization extraction temperature was 22.5°C, the extraction pressure was 40 MPa, the extraction solvent volume was 20 times, and the microfiltration pore size was 300 nm.

[0099] Pueraria root and Polygonatum sibiricum were weighed by weight and steamed, crushed, extracted at low temperature, centrifuged, microfiltered, membrane concentrated, and vacuum concentrated to produce a concentrated solution. The solution was then mixed with an appropriate amount of maltodextrin and spray-dried to produce a compound spray-dried powder. The Pueraria root and Polygonatum sibiricum were steamed at 121°C for 20 minutes.

[0100] The compound extract, compound spray-dried powder ① and compound spray-dried powder ② are prepared into granules through processes such as granulation premixing and one-step granulation. The granules are then put into storage after inner packaging, outer packaging and inspection to obtain a solid beverage product.

[0101] Example 5

[0102] According to the formulation listed in Comparative Example 5, the preparation was carried out according to the following method.

[0103] Longan pulp, chrysanthemum, double-petal red rose, wild jujube kernel, mulberry, jujube, lily, wheat, and licorice were weighed by weight. These materials were crushed, subjected to low-temperature and high-pressure homogenization extraction, centrifuged, microfiltered, and membrane concentrated to obtain a concentrated solution. This was then mixed with an appropriate amount of maltodextrin and spray-dried to obtain a longan pulp compound spray-dried powder. The low-temperature and high-pressure homogenization extraction temperature was 25°C, the extraction pressure was 25 MPa, the extraction solvent volume was 10 times, and the microfiltration pore size was 200 nm.

[0104] Weigh kudzu root and polygonatum by weight, steam, crush, low-temperature high-efficiency extraction, centrifugation, microfiltration, membrane concentration, vacuum concentration and other processes to prepare steam concentrate, and then add appropriate amount of maltodextrin to spray dry slurry, spray dry and other processes to prepare kudzu root compound spray-dried powder; wherein, the kudzu root and polygonatum are steamed at a temperature of 121°C and a steaming time of 120 minutes;

[0105] The longan pulp compound spray-dried powder and the kudzu root compound spray-dried powder are prepared into granules through the processes of granulation premixing and one-step granulation; and the granules are then put into storage after inner packaging, outer packaging and inspection to obtain a solid beverage product.

[0106] The solid beverage products were numbered as comparative example 5 samples.

[0107] Example 6

[0108] According to the formulation listed in Comparative Example 6, the preparation was carried out according to the following method.

[0109] Weigh chrysanthemum, double red rose, and wild jujube kernel according to weight, crush the wild jujube kernel through a 40-mesh sieve, and extract the three materials through supercritical CO2 to obtain a chrysanthemum compound extract and a chrysanthemum compound extract residue. The carbon dioxide flow rate is 400 L / h, the extraction temperature is set to 50° C., the extraction pressure is 30 MPa, the temperature of separation kettle I is 40° C., the temperature of separation kettle II is 35° C., the pressure of separation kettle I is 11 MPa, the pressure of separation kettle II is 6.5 MPa, the extraction time is 3.0 h, and after removing water, the chrysanthemum compound extract is obtained;

[0110] Longan pulp, chrysanthemum compound extract residue, kudzu root, polygonatum, mulberry, jujube, lily, wheat, and licorice were weighed by weight and subjected to pulverization, low-temperature and high-pressure homogenization extraction, centrifugation, microfiltration, and membrane concentration to produce a concentrated solution. This was then mixed with an appropriate amount of maltodextrin and spray-dried to produce a longan pulp compound spray-dried powder. The low-temperature and high-pressure homogenization extraction temperature was 25°C, the extraction pressure was 25 MPa, the extraction solvent volume was 10 times, and the microfiltration pore size was 200 nm.

[0111] The chrysanthemum compound extract and longan pulp compound spray-dried powder are prepared into granules through granulation premixing, one-step granulation and other processes; and then the granules are put into storage after inner packaging, outer packaging and inspection to obtain a solid beverage product.

[0112] The solid beverage products were numbered as comparative example 6 samples.

[0113] Example 7

[0114] Implementation samples: The products obtained in Examples 2, 3, 5, and 6 were respectively: Example 1 sample, Example 2 sample, Example 3 sample, Example 4 sample, Comparative Example 1 sample, Comparative Example 2 sample, Comparative Example 3 sample, Comparative Example 4 sample, Comparative Example 5 sample, and Comparative Example 6 sample. The above samples were subjected to an efficacy test on promoting sleep in perimenopausal rats.

[0115] Rats were taken and bilateral ovariectomy and parachlorophenylalanine were intraperitoneally injected to establish a PI model. The success of the model was determined by vaginal exfoliative cell smear and righting reflex test. The modeling group was then randomly divided into sham-operated group, model group, positive control group, embodiment 1-4 group, comparative example 1-6 group, a total of 13 groups; 8 rats in each group. The sham-operated group was given 10mL / kg of normal saline, and the model group was given 10mg / kg of zolpidem. The embodiment 1-4 group and the comparative example 1-6 group were respectively administered at 630mg / kg standard. All administration methods were gavage, and the continuous administration was 8d. The body weight of the rats was weighed 8h after the last gavage, and the pentobarbital sodium 28mg / kg standard was used for intraperitoneal injection. The righting reflex test coordinated by pentobarbital sodium was performed to compare the sleep latency and sleep duration of each group of rats.

[0116] The results are shown in Table 1 and the statistical results are shown in Figure 1 、 Figure 2 .

[0117] Table 1 Results of sleep latency and sleep duration tests in rats

[0118]

[0119] The results showed that the model group had a significantly prolonged sleep latency and shorter sleep time than the sham operation group; the positive control group had a shorter sleep interval and longer sleep time than the model group (P<0.01), while the sleep interval of rats in each example group was shorter and the sleep time was longer than that in the model group (P<0.01). The sleep-promoting effect of Example 3 was equivalent to that of the positive control group.

[0120] Example 8

[0121] In order to further verify the sleep-promoting effect of the medicine-food formula of the present invention, the sleep-promoting effect after use was studied.

[0122] 1. Diagnostic criteria

[0123] 1.1 Western medicine diagnostic criteria: formulated with reference to the Chinese Classification and Diagnostic Criteria for Mental Disorders (CCMD-3) and Obstetrics and Gynecology edited by Le Jie. (1) Insomnia is the main symptom, including difficulty falling asleep (sleep time > 0.5h), shortened sleep time (< 5h / d), shallow sleep, frequent dreams, early awakening, or difficulty falling asleep again after waking up, fatigue and sleepiness after waking up, etc.; (2) Dissatisfaction with the quantity and quality of sleep, causing obvious distress or decreased mental activity efficiency, or impaired social function; (3) The above sleep disorders occur at least 3 times a week and last for more than 1 month; (4) Women aged 45 to 55 years old, with typical perimenopausal symptoms such as menstrual disorders, hot flashes and sweating, or accompanied by irritability, anxiety, palpitations, dizziness, memory loss, blood pressure fluctuations, muscle and joint pain, etc.

[0124] 1.2 Traditional Chinese Medicine Diagnostic Criteria: Developed with reference to the "Guiding Principles for Clinical Research of New Chinese Medicines" and "Traditional Chinese Medicine Gynecology." Primary symptoms: insomnia and frequent dreams; hot flashes and sweating; irritability or depression; emotional instability; Secondary symptoms: soreness of the waist and knees, menstrual irregularities / amenorrhea, headache, dizziness and forgetfulness, tinnitus, dry eyes, chest and flank pain, dry mouth and throat, decreased libido, genital dryness, hot palms and soles, palpitations, excessive sighing, dry skin, itching, and red cheeks; tongue and pulse: red tongue with minimal coating, or a thin or thin yellow coating; a thin, rapid, or stringy pulse. A diagnosis can be made based on the tongue and pulse if four or more of the primary and secondary symptoms are present.

[0125] 1.3 Inclusion criteria: (1) Those who meet both the above-mentioned Western medicine and traditional Chinese medicine diagnostic criteria; (2) Women aged 45 to 55 years; (3) Those who have not received perimenopausal insomnia-related treatment within 1 week before treatment; (4) Those who signed the informed consent form before treatment and were able to adhere to the treatment course and cooperate with the treatment and follow-up.

[0126] 1.4 Exclusion criteria:

[0127] (1) Patients with secondary insomnia and insomnia caused by external environmental interference factors; (2) Patients with primary diseases of the heart, brain, liver, kidney and hematopoietic system and mental illness; (3) Patients with unexplained irregular vaginal bleeding; (4) Patients with breast tumors, oophorectomy, organic ovarian lesions, premature ovarian failure, polycystic ovary syndrome, and hyperprolactinemia; (5) Patients with poor compliance, inability to determine the efficacy or incomplete data.

[0128] 2. Test Method

[0129] According to the above diagnostic criteria, volunteers were randomly divided into 11 groups, each with 20 participants. The treatment group had an age range of 46-54 years, with an average age of (50.3±2.6) years, and a disease course of 2-48 months, with an average course of (16.7±11.8) months. The control group was given estazolam tablets, 1.0 mg / tablet, one tablet / time, orally 30 minutes before bedtime every night for a course of 4 weeks; Example 1-4, Comparative Example 1-4, Group 5, and Group 6 were each given 3 g / bag of the product, one bag / time, twice a day for 4 weeks.

[0130] 3. Observation of therapeutic effects:

[0131] 3.1 Efficacy standards:

[0132] Cured: sleep returns to normal (more than 6 hours), daytime energy is abundant, and the PSQI score reduction rate is ≥80%; markedly effective: sleep time is extended (more than 3 hours), sleep efficiency is increased, night awakenings are reduced, and there is no obvious fatigue during the day, and the PSQI score reduction rate is ≥50%, <80%; effective: sleep time is extended but less than 3 hours, night awakenings are reduced, and daytime fatigue is relieved, and the PSQI score reduction rate is ≥25%, <50%; ineffective: sleep conditions do not improve, and the PSQI score reduction is <25%.

[0133] 3.2 Observation indicators: Observe and record the Pittsburgh Sleep Quality Index (PSQI) scores of the two groups before and after treatment (end of medication) and the clinical efficacy and adverse reactions of each group after treatment. Pittsburgh Sleep Quality Index (PSQI) score: Assess the patient's sleep quality, with a total score of 0 to 21 points. The higher the score, the worse the sleep quality. This embodiment uses the Pittsburgh Sleep Quality Index (PSQI) for scoring. The result data are shown in Table 2 and the comparison chart is shown in Figure 3 .

[0134] Table 2 Pittsburgh Sleep Quality Index (PSQI) scores

[0135]

[0136]

[0137] The results show that the effects of Examples 1-4 after treatment are significantly better than those before treatment, and are better than those of the positive control group and the comparative example groups.

[0138] 3.3 Statistical methods: All data were analyzed using SPSS 22.0 statistical software package. The data were expressed as normal distribution using t test, and those that did not meet the normal distribution were expressed as nonparametric rank sum test; the enumeration data were expressed as rate (%), and the comparison was performed using χ test. 2 test.

[0139] 4 Treatment results

[0140] The clinical efficacy of each group after treatment is shown in Table 3 below. In the positive control group, 3 cases of dizziness and 2 cases of gastrointestinal reactions occurred, while no adverse events occurred in the examples and comparative example groups.

[0141] Table 3 Comparison of clinical efficacy

[0142] Group get well Significant effect efficient invalid total control group 3 13 4 0 20 Example 1 6 11 3 0 20 Example 2 7 12 1 0 20 Example 3 8 11 1 0 20 Example 4 8 10 2 0 20 Comparative Example 1 1 6 11 2 20 Comparative Example 2 2 8 10 0 20 Comparative Example 3 1 7 10 2 20 Comparative Example 4 0 4 13 3 20 Comparative Example 5 3 10 6 0 19 (1 case dropped out midway) Comparative Example 6 4 8 7 0 19 (1 case dropped out midway)

[0143] Example 9

[0144] Experimental Materials

[0145] Equipment: Agilent 1100 high-performance liquid chromatograph (Agilent, USA), Inertsil ODS-3 (4.6 mm × 250 mm, 5 μm) & Diamonsi C18 (4.6 mm × 250 mm, 5 μm), AUW220D electronic balance (Shimadzu, Japan)

[0146] Reagents: acetonitrile (chromatographic grade), methanol (chromatographic grade), phosphoric acid (analytical grade), puerarin reference substance (Beijing Yihua Tongbiao Technology Co., Ltd., batch number: 191005), and first-grade water.

[0147] Sample: According to the formula of Example No. 3 in Example 1, a solid beverage product with a sleep-promoting effect was obtained. According to different production batch numbers, the following batches of products were obtained: 20230201, 20230202, 20230203, 20230204, 20230301, 20230302, 20230303, 20230304, and 20230305.

[0148] Preparation of test solution: Take 0.5 g of solid beverage product sample, accurately weigh it, place it in a 5 ml volumetric flask, add ultrapure water to the volume, ultrasonicate, filter through a 0.45 μm microporous filter membrane into a sample injection bottle to obtain the test solution.

[0149] Preparation of reference solution: Accurately weigh puerarin reference solution to prepare 1.0 mg / mL puerarin reference solution.

[0150] Selection of chromatographic conditions

[0151] 1. Selection of chromatographic column:

[0152] According to the same liquid phase conditions, the analysis was carried out using Inertsil ODS-3 (4.6 mm × 250 mm, 5 μm) & Diamonsi C18 (4.6 mm × 250 mm, 5 μm). The spectral results showed that the Inertsil ODS-3 chromatographic column was suitable for this experiment, with good separation and rich spectral information. Therefore, the Diamonsi C18 chromatographic column was used for sample analysis in the later experiments of the present invention.

[0153] 2. Selection of mobile phase system:

[0154] To ensure the comprehensiveness of the fingerprint of the solid beverage granule product of the present invention, we selected a suitable mobile phase system based on the raw materials of the product and carried out systematic screening and optimization.

[0155] Two common mobile phase systems were initially selected for the experiment: acetonitrile-water and methanol-water. Full wavelength scanning was used. Figure 4 and Figure 5 .

[0156] Discover Figure 4The spectral information is relatively richer, but the spectral information is still not comprehensive enough, so we further screened the acetonitrile-acid water system and selected a variety of organic acids, including formic acid, acetic acid and phosphoric acid, and conducted experiments at concentrations of 0.01%, 0.05%, 0.1%, and 0.15%. By comparing the chromatograms obtained under different organic acids and their concentrations, it was found that phosphoric acid had the best effect. Finally, it was found that the chromatographic experiment in the acetonitrile-0.1% phosphoric acid aqueous solution system gave the most comprehensive spectral information, which can fully reflect the chemical composition of solid beverage granule products. Figure 6 The present invention finally determined to use acetonitrile-0.1% phosphoric acid water as the mobile phase system.

[0157] 3. Selection of detection wavelength:

[0158] like Figure 6 The obtained spectrum detection wavelength was detected by DAD detector for three-dimensional spectrum analysis. Under the condition of avoiding strong absorption that interferes with the overall peak profile, the spectrum components at 220-320nm were selected with strong absorption, small interference, good separation and stable baseline. It is planned to further screen the detection wavelength within this range.

[0159] The wavelengths of 225, 235, 245, 255, 265, 275, 285, 295, 305, and 315 nm were further screened. Figure 7 As shown. Figure 7 It can be seen that the spectrum information of the solid beverage test sample is the richest at 265nm, which is the best scanning wavelength selected in this experiment. Therefore, 265nm is used as the detection wavelength in the later experiments of the present invention.

[0160] 4. Optimization of mobile phase elution gradient:

[0161] To ensure the comprehensiveness and accuracy of the fingerprint generated for the solid beverage granules, we conducted detailed experiments to create a mobile phase gradient system. By systematically optimizing the gradient conditions of the acetonitrile-0.1% phosphoric acid mobile phase, we determined the optimal gradient conditions, demonstrating the inventiveness of the present invention in its detection method.

[0162] Preliminary gradient condition test (NO.1)

[0163] We first used a relatively simple gradient condition for preliminary experiments:

[0164] Mobile phase: A: acetonitrile, B: 0.1% phosphoric acid

[0165] Gradient conditions: 0-90 min, 5%-95% acetonitrile, 95%-5% 0.1% phosphoric acid

[0166] Using this gradient condition, we obtained a preliminary chromatogram. Figure 8 The analysis results of S1 showed that due to the large variation in acetonitrile concentration, some components were not effectively separated, resulting in insufficient spectral information and difficulty in fully reflecting the full range of chemical components in the solid beverage. Therefore, we decided to further optimize the gradient conditions.

[0167] Gradient optimization test (NO.2)

[0168] In order to improve the separation effect, we optimized the gradient conditions and designed a new experimental plan:

[0169] Mobile phase: A: acetonitrile, B: 0.1% phosphoric acid

[0170] Gradient conditions:

[0171] 0-10min: 2%-2% acetonitrile, 98%-98% 0.1% phosphoric acid; 10-50min: 2%-30% acetonitrile, 98%-70% 0.1% phosphoric acid; 50-70min: 30%-50% acetonitrile, 70%-50% 0.1% phosphoric acid. For new gradient conditions, see Figure 8 The chromatogram of S2 showed improved separation of some components and increased spectral information, but the characteristic peaks were concentrated between 40 and 52 minutes, resulting in poor resolution. We decided to continue optimizing the gradient conditions to further improve separation and the richness of spectral information.

[0172] Gradient optimization test (NO.3)

[0173] Based on the experimental results of NO.2, we made more detailed adjustments to the gradient conditions:

[0174] Mobile phase: A: acetonitrile, B: 0.1% phosphoric acid

[0175] Gradient conditions:

[0176] 0-10 min: 2%-2% acetonitrile, 98%-98% 0.1% phosphoric acid; 10-15 min: 2%-5% acetonitrile, 98%-95% 0.1% phosphoric acid; 15-65 min: 5%-30% acetonitrile, 95%-70% 0.1% phosphoric acid; 50-75 min: 30%-50% acetonitrile, 70%-50% 0.1% phosphoric acid.

[0177] Under this gradient condition, see Figure 8 The chromatogram of S3 shows a significant improvement in separation, with more components being well separated. However, we found that some key components still overlap in the middle and later stages, indicating that further optimization is necessary.

[0178] Gradient optimization test (NO.4)

[0179] We continue to adjust the gradient conditions to improve the separation effect in the middle and later stages:

[0180] Mobile phase: A: acetonitrile, B: 0.1% phosphoric acid

[0181] Gradient conditions:

[0182] 0-10 min: 2%-2% acetonitrile, 98%-98% 0.1% phosphoric acid; 10-20 min: 2%-10% acetonitrile, 98%-90% 0.1% phosphoric acid; 20-60 min: 10%-25% acetonitrile, 90%-75% 0.1% phosphoric acid; 60-100 min: 25%-95% acetonitrile, 75%-5% 0.1% phosphoric acid.

[0183] Under the new gradient conditions, see Figure 8 The S4 chromatogram further improves separation and component separation becomes clearer. However, the peak interval becomes longer, the peak elution time is longer, the gradient condition time is longer, and the experimental efficiency is lower.

[0184] Gradient optimization test (NO.5)

[0185] To further improve experimental efficiency and separation effect, we further optimized the gradient conditions:

[0186] Mobile phase: A: acetonitrile, B: 0.1% phosphoric acid

[0187] Gradient conditions:

[0188] 0-10 min: 2%-2% acetonitrile, 98%-98% 0.1% phosphoric acid; 10-23 min: 2%-15% acetonitrile, 98%-85% 0.1% phosphoric acid; 23-60 min: 15%-25% acetonitrile, 85%-75% 0.1% phosphoric acid; 60-80 min: 25%-60% acetonitrile, 75%-40% 0.1% phosphoric acid.

[0189] Under this gradient condition, see Figure 8 The chromatogram of S5 shows that most components in the second half are well separated and the information is relatively rich. However, the separation effect of some complex components in 28-33 minutes is still not completely ideal.

[0190] Gradient optimization test (NO.6)

[0191] To further improve experimental efficiency and separation effect, we further optimized the gradient conditions:

[0192] Mobile phase: A: acetonitrile, B: 0.1% phosphoric acid

[0193] Gradient conditions:

[0194] 0-10 min: 2%-2% acetonitrile, 98%-98% 0.1% phosphoric acid; 10-15 min: 2%-7% acetonitrile, 98%-93% 0.1% phosphoric acid; 15-60 min: 7%-25% acetonitrile, 93%-75% 0.1% phosphoric acid; 60-80 min: 25%-60% acetonitrile, 75%-40% 0.1% phosphoric acid.

[0195] Under this optimal gradient condition, see Figure 8 The chromatogram of S6 shows that all components are fully separated, and the spectrum is rich in information, which can fully reflect the complete chemical composition of the solid beverage. This condition is used as the optimal gradient of the present invention, ensuring the comprehensiveness and accuracy of the product fingerprint, providing a reliable basis for quality control.

[0196] Through the aforementioned systematic gradient optimization experiments, the present invention not only selected the optimal acetonitrile-0.1% phosphoric acid mobile phase system but also determined the optimal gradient conditions through multiple optimization experiments. This optimization process not only systematically discovered the optimal conditions but also demonstrated that these conditions can effectively separate complex components, ensuring the comprehensiveness and accuracy of the fingerprint.

[0197] 5. Determination of chromatographic conditions:

[0198] According to the above experimental results, the chromatographic conditions for the subsequent experiments are determined as follows:

[0199] (1) Chromatographic column: Inertsil ODS-3 (250*4.6nm, 5μm)

[0200] (2) Detection wavelength: 265nm

[0201] (3) Injection volume: 20ul, flow rate: 1.0ml / min

[0202] (4) Column temperature: 35°C

[0203] (5) Mobile phase: A: acetonitrile, B: 0.1% phosphoric acid

[0204] (6) Mobile phase gradient conditions:

[0205] 0-10 min: 2%-2% acetonitrile, 98%-98% 0.1% phosphoric acid; 10-15 min: 2%-7% acetonitrile, 98%-93% 0.1% phosphoric acid; 15-60 min: 7%-25% acetonitrile, 93%-75% 0.1% phosphoric acid; 60-80 min: 25%-60% acetonitrile, 75%-40% 0.1% phosphoric acid.

[0206] Example 10 Methodological Investigation of Detection Methods

[0207] 1. Precision test

[0208] In order to evaluate the precision of the solid beverage detection method of the present invention, a precision investigation test was conducted. The specific steps are as follows:

[0209] Preparation of test solution: Take the newly prepared test solution of the same batch (batch number: 20230201) and prepare it according to the test solution preparation method under liquid chromatography conditions.

[0210] Injection test: Inject the prepared test solution 6 times continuously and record the chromatogram of each time, numbering S1-S6.

[0211] Data analysis: Calculate the relative retention time and relative peak area of ​​the common peaks in each chromatogram, and calculate the relative standard deviation (RSD).

[0212] Test results: The chromatogram similarity was analyzed using the "Chinese Herbal Medicine Chromatographic Fingerprint Similarity Evaluation System" (2012 version) software. Chromatographic peaks with good separation were selected. There were 16 common peaks in the HPLC chromatograms of the six samples, and the similarity of each chromatogram was above 0.9998, indicating that the instrument had good precision and met the technical requirements of the fingerprint spectrum. Figure 9 .

[0213] Relative retention time: Six consecutive injections were performed, and the relative retention time of each chromatogram was recorded and the RSD was calculated. The results showed that the RSD of the relative retention time was ≤ 0.3%, indicating good instrument precision. Specific data are shown in Table 4.

[0214] Table 4. Relative retention time of common peaks in precision test

[0215]

[0216] Relative peak area: Six consecutive injections were performed, and the relative peak area of ​​each chromatogram was recorded and the RSD was calculated. The results showed that the RSD of the relative peak area was ≤5%. Specific data are shown in Table 5.

[0217] Table 5. Relative peak areas of common peaks in precision test

[0218]

[0219]

[0220] 2. Precision test

[0221] To evaluate the repeatability of the solid beverage detection method presented herein, a detailed reproducibility study was conducted. Six samples of solid beverage batch 20230201 were prepared according to the test solution preparation conditions. Each of the six samples was injected sequentially under optimized liquid chromatography conditions, and the chromatograms for each were recorded, numbered S1-S6. The relative retention time and relative peak area of ​​the common peaks in each chromatogram were calculated, and the relative standard deviation (RSD) was calculated.

[0222] Six sample injections were performed, and the relative retention times of each chromatogram were recorded and RSDs were calculated. The results showed that the RSDs of the relative retention times were ≤ 0.7%, indicating that the method has good reproducibility. Specific data are shown in Table 6.

[0223] Table 6 Relative retention time of common peaks in repeatability test

[0224]

[0225] Six sample injections were performed, and the relative peak areas of each chromatogram were recorded and RSDs were calculated. The results showed that the RSDs of the relative peak areas were ≤ 5%. Specific data are shown in Table 7.

[0226] Table 7 Relative peak areas of the total peaks in the repeatability test

[0227]

[0228] S1 to S6, see Figure 10 The results showed that the similarity of each chromatogram was above 0.9998, which proved that the liquid chromatography detection method of the present invention had good repeatability and met the technical requirements of fingerprint.

[0229] 3. Stability test

[0230] In order to evaluate the stability of the solid beverage test solution of the present invention, a detailed stability test was conducted. Take a freshly prepared test solution of the same batch (batch number: 20230201) and prepare it according to the liquid chromatography conditions. Samples were injected and tested at 0, 2, 4, 6, 8, 12, 16, and 24 hours respectively, and the chromatograms of each time were recorded and numbered S1-S8. The relative retention time and relative peak area of ​​the common peaks in the chromatogram at each time point were calculated, and the relative standard deviation (RSD) was calculated.

[0231] The relative retention times of the chromatograms were recorded and RSDs were calculated by injection testing at different time points. The results showed that the RSDs of the relative retention times were ≤ 0.3%, indicating that the test solution had good stability within 24 hours. Specific data are shown in Table 8.

[0232] Table 8 Relative retention time of common peaks in stability test

[0233]

[0234]

[0235] Samples were injected at different time points, and the relative peak areas of each chromatogram were recorded and RSDs were calculated. The results showed that the RSDs of the relative peak areas were ≤ 5%. Specific data are shown in Table 9.

[0236] Table 9 Total peak relative peak area in stability test

[0237]

[0238] Compare the chromatograms at different time points and calculate their similarity. The results show that the similarity of each chromatogram is above 0.9998, indicating that the test solution has good stability within 24 hours and meets the technical requirements of fingerprint spectrum. Figure 11 .

[0239] Example 11 Establishment of fingerprint and identification of common peaks

[0240] Nine batches of solid beverage samples (batch numbers: 20230201, 20230202, 20230203, 20230204, 20230301, 20230302, 20230303, 20230304, and 20230305) were collected. Test solutions were prepared according to the test solution preparation method. Test solutions and reference solutions were injected and tested under optimized liquid chromatography conditions, and the chromatograms were recorded.

[0241] The chromatographic fingerprint data of 9 batches of samples were analyzed using the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine" (2012 Edition). Sample S1 was used as the reference spectrum, and the common pattern was generated by the average method as the reference spectrum. The time window width was 0.1s. The chromatograms of the 9 batches of samples all had 16 common peaks. After comparison with the reference substance and ultraviolet spectrum, peak 7 was confirmed to be puerarin. Figure 12 Among them, peak 7 has better peak shape and separation, so it is selected as the reference peak. The relative retention time and relative peak area of ​​each common peak and the reference peak are calculated. The results are as follows Figure 13 As shown, the comparison of the chromatograms of 9 batches of products is shown in Figure 14 .

[0242] The chromatograms of nine batches of products were analyzed and the relative retention times of the common peaks were calculated. The results showed that the RSD of the relative retention times was ≤ 0.6%. Specific data are shown in Table 10.

[0243] Table 109 Relative retention time of common peaks in fingerprints of batches of products

[0244]

[0245] The chromatograms of the nine batches of products were analyzed, and the relative peak areas of the common peaks were calculated. The results showed that the RSD of the relative peak areas was ≤ 5.0%. Specific data are shown in Table 11.

[0246] Table 11. Relative peak areas of common peaks in fingerprints of 9 batches of products

[0247]

[0248]

[0249] Using the "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" (2012 edition), we calculated the similarity of the chromatograms of nine batches of products. The results showed that the similarity of each chromatogram was above 0.9998, indicating good product quality consistency. Specific data are shown in Table 12.

[0250] Table 129 Calculation of similarity of fingerprints of batches of products

[0251]

[0252] Fingerprint analysis of 9 batches of solid beverage products showed that: RSD of relative retention time ≤ 0.6% (see Table 10); RSD of relative peak area ≤ 5.0% (see Table 11); similarity of each chromatogram was above 0.9998 (see Table 12). Figure 14 .

[0253] The results show that the process of this product is stable and the product quality is well controlled during the production process. The use of fingerprint technology can provide a fast and accurate quality standard for the quality control of this product.

[0254] The above description is a detailed description of the preferred embodiments of the present invention, but the embodiments are not intended to limit the scope of the patent application of the present invention. Any equivalent changes or modifications completed under the technical spirit suggested by the present invention should fall within the patent scope covered by the present invention.

Claims

1. A solid beverage having a sleep-promoting effect, characterized in that: The invention is composed of the following raw materials in parts by weight: 10-30 parts of kudzu root, 10-30 parts of longan pulp, 8-24 parts of mulberry, 6-20 parts of polygonatum, 6-20 parts of jujube, 6-20 parts of lily, 5-15 parts of wheat, 5-15 parts of chrysanthemum, 3-11 parts of double red rose, 3-11 parts of spiny jujube kernel and 1.5-4.5 parts of liquorice.

2. A solid beverage having a sleep-promoting effect according to claim 1, characterized in that: The invention is composed of the following raw materials in parts by weight: 20 parts of kudzu root, 20 parts of longan pulp, 17 parts of mulberry, 13 parts of polygonatum, 13 parts of jujube, 13 parts of lily, 10 parts of wheat, 10 parts of chrysanthemum, 7 parts of double red rose, 7 parts of sour jujube kernel and 3 parts of liquorice.

3. A method for preparing a solid beverage having a sleep-promoting effect according to claim 1 or 2, characterized in that: The following steps are involved: A. By weight, firstly crush the Chinese date kernels and pass them through a 20-40 mesh sieve, then take chrysanthemum and double red rose, and extract the above three materials by supercritical CO2 to obtain a compound extract and a compound residue; B. Take longan pulp, mulberry, jujube, lily, wheat, licorice by weight, and then take the compound extract residue of step A, crush the above materials, use low temperature and high pressure homogenization extraction, take the extract and centrifuge, microfilter, membrane concentration to make a concentrate, mix with auxiliary material A, spray dry and slurry, spray dry, and make a compound spray-dried powder①; C. Weigh kudzu root and polygonatum sibiricum by weight, crush and extract with low temperature and high efficiency, take the extract and make it into concentrated solution after centrifugation, microfiltration, membrane concentration and vacuum concentration, mix it with auxiliary material A, spray dry and prepare compound spray-dried powder after slurry preparation②; D. The compound extract, the compound spray-dried powder ① and the compound spray-dried powder ② are taken, pre-mixed by granulation and subjected to a one-step granulation process to prepare a granular product, thereby obtaining a flavored solid beverage having a sleep-promoting effect.

4. The method for preparing a solid beverage having a sleep-promoting effect according to claim 3, characterized in that: In step A, the carbon dioxide flow rate of supercritical CO2 extraction is 300-400 L / h, the extraction temperature is 40-50° C., and the extraction pressure is 20-30 MPa; the temperature of separation kettle I is 35-40° C., and the temperature of separation kettle II is 30-35° C.; the pressure of separation kettle I is 9-11 MPa, and the pressure of separation kettle II is 5.5-6.5 MPa; and the extraction time is 1.0-3.0 h.

5. The method for preparing a solid beverage having a sleep-promoting effect according to claim 3, characterized in that: In step B, the temperature of the low-temperature and high-pressure homogenization extraction is lower than 25° C., the extraction pressure is 25-40 MPa, the extraction solvent amount is 10-30 times, and the microfiltration pore size is 200-500 nm.

6. The method for preparing a solid beverage having a sleep-promoting effect according to claim 3, characterized in that: Excipient A is maltodextrin.

7. The method for preparing a solid beverage having a sleep-promoting effect according to claim 3, characterized in that: The kudzu root and polygonatum sibiricum are processed before being crushed, and the processing method is: steaming for 20min-120min at a temperature of 100-121°C.

8. A method for identifying the solid beverage having a sleep-promoting effect according to claim 1 or 2, characterized in that: The following steps are involved: a. Preparation of test solution: Take solid beverage, add water, dissolve and shake well, filter with a filter membrane to obtain the test solution; b. Preparation of reference solution: weigh puerarin reference substance, add organic solvent to dissolve it, and obtain reference solution; c. Solution determination: Accurately aspirate the test solution and the reference solution and inject them into the high performance liquid chromatograph for determination to obtain the fingerprint of the test sample and the reference solution; the chromatographic conditions of the high performance liquid chromatograph are as follows: the stationary phase of the chromatographic column is octadecylsilane bonded silica gel as filler, acetonitrile is used as mobile phase A, and 0.01-0.15% by volume of inorganic acid aqueous solution is used as mobile phase B for gradient elution; the gradient elution is carried out according to the following procedure: by volume percentage, 0-10 minutes, mobile phase A is maintained at 2%, and mobile phase B is maintained at 98%; 10-15 minutes, mobile phase A increased from 2% to 7%, and mobile phase B decreased from 98% to 93%; From 15 to 60 minutes, mobile phase A increased from 7% to 25%, and mobile phase B decreased from 93% to 75%; From 60 to 80 minutes, mobile phase A increased from 25% to 60%, and mobile phase B decreased from 75% to 40%; d. Similarity evaluation: Import the fingerprints of multiple batches of test samples and reference samples obtained according to steps a to c into the Chinese medicine chromatographic fingerprint similarity evaluation system for analysis to obtain the fingerprint of the product composition. Use the reference fingerprint to compare the similarity with the fingerprints of different batches of products in production, so as to evaluate the quality of the current batch of products.

9. The method for identifying a solid beverage having a sleep-promoting effect according to claim 8, characterized in that: The chromatographic column selected was Inertsil ODS-3, the detection wavelength was 265 nm, the flow rate was 1 ml / min, the column temperature was 35° C., and the mobile phase B was 0.1% phosphoric acid aqueous solution.

10. The method for identifying a solid beverage having a sleep-promoting effect according to claim 8, characterized in that: The method for obtaining the control fingerprint spectrum is to prepare ≥9 batches of composition samples, and after obtaining the sample fingerprint spectrum, perform a similarity evaluation. The similarity evaluation method is: import the fingerprint spectrum into the similarity evaluation software for similarity evaluation analysis, determine the similarity ≥0.999, and determine 16 common peaks, which are numbered 1 to 16, among which peak 7 is puerarin and is also used as a reference peak of the control fingerprint spectrum.

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