Peach and safflower four-component decoction compound preparation as well as preparation method and quality control method thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicine preparation, and particularly relates to a peach kernel and safflower four-ingredient decoction compound preparation and a preparation method and a quality control method thereof. The invention provides a preparation method of a peach kernel and safflower four-component decoction compound preparation by screening and optimizing an enrichment method of volatile oil in extraction, concentration, inclusion and drying processes on the basis of an ancient water decoction process. According to the peach kernel and safflower four-drug decoction compound preparation prepared by the invention, the quality consistency with a reference sample is realized, including the consistency of each content index in the compound preparation with the reference sample and the consistency of a fingerprint spectrum with the reference sample; especially, characteristic peaks of volatile components-ligustilide, senkyunolide I and senkyunolide A are presented in the fingerprint spectrum, and the consistency of the content of the volatile oil is determined. Moreover, the quality control method provided by the invention is expected to be used for quality control of industrial large-scale production of a classic famous prescription, namely the peach kernel and safflower four-ingredient decoction compound preparation, and has a good application prospect.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine preparation, and particularly relates to a Taohong Siwu Decoction compound preparation and a preparation method and a quality control method thereof. Background Art

[0002] The classic formula, Taohong Siwu Decoction, first appeared in the "Yi Zong Jin Jian" (Medical Ancestral Mirror). This recipe is a combination of Siwu Decoction with peach kernel and safflower added to promote blood circulation. Modern research demonstrates that Taohong Siwu Decoction has vasodilatory, anti-inflammatory, anti-fatigue, anti-shock, immune regulation, lipid-lowering, trace element supplementation, and anti-allergic effects. Taohong Siwu Decoction focuses on removing blood stasis, supplemented by nourishing blood and promoting qi. The formula primarily features peach kernel and safflower, which are potent blood-dissolving herbs, to promote blood circulation and dissipate stasis. The sweet and warming ingredients Rehmannia glutinosa and Angelica sinensis nourish yin and tonify the liver, nourishing blood and regulating menstruation. Peony root nourishes blood and harmonizes nutrients, enhancing blood tonification. Chuanxiong promotes blood circulation and qi circulation, regulating qi and blood, and aiding blood circulation. The formula's well-balanced composition is notable for its ability to dissipate blood stasis, generate new blood, and promote the smooth flow of qi. Taohong Siwu Decoction is widely used in TCM to treat dozens of conditions, including gynecology, orthopedics, cardiology, and dermatology, making it a widely used formula in clinical practice.

[0003] Angelica sinensis and Chuanxiong rhizome are often used as a herbal pair in Traditional Chinese Medicine (TCM) to nourish and replenish blood, and to activate blood circulation and remove blood stasis. First mentioned in the Song Dynasty's "Taiping Huimin Hejijufang," the herbal pair is known for its ability to nourish and invigorate blood, regulate menstruation, and relieve pain. It is not only a key herbal remedy for regulating menstruation in gynecology but also an excellent blood-tonifying herbal remedy in internal medicine, earning it the nickname "the holy blood-tonifying herb." The chemical composition of TCM is fundamental to its efficacy, and its volatile components are a key and essential component in the research of classic prescriptions. The angelica sinensis and Chuanxiong rhizome pair in the classic prescription, Taohong Siwu Tang, nourishes and invigorates blood, regulates menstruation, and relieves pain, with well-established pharmacological effects. Correlation analysis between the efficacy of the angelica sinensis and Chuanxiong rhizome pair revealed that chemical components such as ligustilide, ferulic acid, and ligustilide significantly contribute to its menstrual regulation and analgesic effects. Clinical practice demonstrates that the synergistic combination of angelica sinensis and Chuanxiong can enhance its blood-nourishing and blood-activating effects. Therefore, in the large-scale production of the classic formula - Taohong Siwu Decoction compound preparation, effectively controlling the quantitative transfer of volatile components and ensuring consistency with the benchmark sample are key projects and important contents in the research and development process.

[0004] With the continuous development of the modernization of traditional Chinese medicine, new dosage forms are constantly emerging, and some large and medium-sized traditional Chinese medicine companies are competing to develop them. Since the overall research ideas of classic formula compound preparations are different from other traditional Chinese medicine new drug research models, for the research and development of classic formula compound preparations, the preferred dosage form is granules. According to the traditional production process of granules, they must go through decoction, concentration, drying, granulation and other processes. Each link will affect the effective retention of volatile components, especially in the process of reduced pressure concentration, the volatile components are almost lost.

[0005] The first core requirement for research on compound preparations of classic formulas is the use of traditional water-decoction techniques. Relevant regulations and guidelines emphasize that extraction routes must be consistent with ancient texts. However, the traditional method of extracting a combination of herbs using a decoction of multiple herbs prevents the individual extraction of volatile oils from individual herbs, significantly limiting the development of processes for extracting volatile oils from classic formulas. While decoction extraction of herbs containing light oils typically yields high concentrations of aromatic water, it is difficult to obtain volatile oils or high concentrations of aromatic water from herbs containing heavy oils. Because heavy oils have a higher boiling point and a greater relative density than water, they are difficult to evaporate with water vapor through distillation, making oil-water separation even more challenging. Therefore, the enrichment of heavy oils in the research of compound preparations of classic formulas has always been a key technical barrier. Given the unique characteristics of the volatile oils contained in the herbs Angelica sinensis and Chuanxiong, organic solvent extraction or supercritical extraction is generally used, making this nearly impossible using traditional water extraction techniques. Classical Chinese medicine formulas use traditional decoction methods for a relatively short period of time, with no more than two decoctions. This presents challenges in the large-scale production of compound preparations from these formulas. The classic formula, Taohong Siwu Decoction, contains a significant proportion of the medicinal ingredients Angelica sinensis and Chuanxiong. The volatile component, ligustilide, has a high boiling point, high density, and low solubility. During the extraction process, when the volatile oil dissolution reaches equilibrium, only a small portion is carried away by the steam, while the majority remains in the extract and the medicinal residue. Consequently, achieving high concentrations of volatile oil using traditional decoction extraction techniques is difficult. Currently, no successful case has been found of the successful extraction of high concentrations of volatile oil using traditional decoction techniques in the large-scale production of compound preparations from the classic Chinese medicine formula, Taohong Siwu Decoction.

[0006] The second core requirement for research on compound preparations of classic formulas is that product quality should embody the qualities of the ancient "bowl of soup." This means that the quality of mass-produced compound preparations must be essentially consistent with that of the benchmark sample, with all content indicators ranging from 70% to 130% of the average of 15 batches of benchmark samples, fingerprints consistent with those of the benchmark samples, and a paste yield within ±10% of the average of the benchmark samples. Conventional processes used in mass production of compound preparations of the classic formula, Taohong Siwu Decoction, have struggled to achieve consistency in all product parameters with the benchmark sample, primarily with regard to the retention of the volatile components, ligustilide and ligustilide. Currently, despite multiple registration applications for compound preparations of the classic formula, Taohong Siwu Decoction, none have successfully passed the technical review of the Center for Drug Evaluation. The fundamental reason is that the consistency of volatile components in mass-produced compound preparations with the benchmark sample has not been adequately addressed.

[0007] Therefore, a method for enriching volatile oil in the Taohong Siwu Decoction compound preparation was developed so that the quality of the Taohong Siwu Decoction compound preparation product can restore the quality attributes of the ancient "bowl of soup" based on the ancient water decoction process. Summary of the Invention

[0008] In response to the defects of the existing technology, the present invention provides a Taohong Siwu Decoction compound preparation and its preparation method and quality control method, aiming to adopt the ancient water decoction process to achieve the product quality of the Taohong Siwu Decoction compound preparation and restore the quality attributes of the ancient "bowl of soup".

[0009] The present invention provides a Taohong Siwu Decoction compound preparation, which is prepared according to the following steps:

[0010] Step 1: Take the medicinal materials of Taohong Siwu Decoction or its slices, add water and decoct them by double extraction method, perform primary oil-water separation and secondary oil-water separation in reflux, collect and combine aromatic water, and combine the two extracts, wherein the temperature of the secondary oil-water separation is 40-60° C.;

[0011] Step 2, adding an adsorbent to the extract for adsorption, and concentrating under reduced pressure to obtain a concentrated solution;

[0012] Step 3, adding an adsorbent to the aromatic water for inclusion to prepare an inclusion liquid;

[0013] Step 4: mixing the concentrated solution with the inclusion solution, spray drying, and obtaining a dry powder.

[0014] Preferably, in step 1, the double extraction step includes: adding 10-15 times the mass of the medicinal slices of water for the first time, decocting and refluxing for 1-1.5 hours, and adding 8-12 times the mass of the medicinal slices of water for the second time, decocting and refluxing for 50-60 minutes; and / or, in step 1, the temperature of the primary oil-water separation is 70-90°C.

[0015] Preferably, after combining the two extracts in step 1, a residue is obtained, and the residue is extracted again by steam distillation for 1-3 hours or refluxing the residue for 1-3 hours, and the distilled condensate or the reflux condensate is subjected to primary oil-water separation and secondary oil-water separation, and the aromatic water is collected and combined, and then combined with the aromatic water described in step 1.

[0016] Preferably, in steps 2 and 3, the adsorbent is independently selected from at least one of modified starch, porous starch, porous silica, beta-cyclodextrin, and hydroxypropyl-beta-cyclodextrin.

[0017] Preferably, in steps 2 and 3, the adsorbent is independently selected from at least one of modified starch, beta-cyclodextrin, and hydroxypropyl-beta-cyclodextrin.

[0018] Preferably, in steps 2 and 3, the amount of the adsorbent is 2-5% of the mass of the extract or aromatic water; and / or, in steps 2 and 3, the temperature of the adsorption or inclusion is 50-60°C; and / or, in steps 2 and 3, the time of the adsorption or inclusion is 1-3 hours; and / or, in steps 2 and 3, the adsorption or inclusion method is stirring or colloid mill inclusion adsorption, and the stirring rate is 200-500r / min.

[0019] Preferably, in step 2, the temperature of the reduced pressure concentration is 60-70°C; and / or, in step 2, the reduced pressure concentration is performed to a relative density of the extract of 1.04-1.10 g / ml; and / or, in step 4, the spray drying conditions include: an inlet air temperature of 165-175°C and an outlet air temperature of 90-100°C; and / or, dry granulation is performed after the dry powder is obtained; and / or, the primary oil-water separation and the secondary oil-water separation are performed in an oil-water separator.

[0020] The present invention provides a method for preparing any of the above-mentioned Taohong Siwu Decoction compound preparations, comprising the following steps:

[0021] Step 1: Take the medicinal materials of Taohong Siwu Decoction or its slices, add water and decoct them by double extraction method, perform primary oil-water separation and secondary oil-water separation in reflux, collect and combine aromatic water, and combine the two extracts, wherein the temperature of the secondary oil-water separation is 40-60° C.;

[0022] Step 2, adding an adsorbent to the extract for adsorption, and concentrating under reduced pressure to obtain a concentrated solution;

[0023] Step 3, adding an adsorbent to the aromatic water for inclusion to prepare an inclusion liquid;

[0024] Step 4: mixing the concentrated solution with the inclusion solution, spray drying, and obtaining a dry powder.

[0025] The present invention provides a quality control method for a Taohong Siwu Decoction compound preparation, which comprises the following steps:

[0026] High performance liquid chromatography is used to perform quality control of fingerprints and content indicators on the medicinal materials of Taohong Siwu Decoction or the extract, concentrate, aromatic water, inclusion liquid or dry powder prepared by the preparation method according to claim 7;

[0027] The index component is selected from at least one of ligustilide, amygdalin, ferulic acid, hydroxysafflor yellow A, rehmannia glutinosa D, and paeoniflorin.

[0028] Preferably, the specific contents of the quality control of the fingerprint and index component content of the medicinal materials include: the fingerprint of the medicinal taste of Angelica sinensis contains characteristic peaks of ligustilide I, ligustilide H, coniferyl ferulate and ligustilide, the fingerprint of the medicinal taste of Chuanxiong contains characteristic peaks of ligustilide A, ligustilide I, ligustilide H and ligustilide, and the total content of ligustilide in the prescription ratio of the Angelica sinensis-Ligustilide drug pair is 0.9-1.8%;

[0029] And / or, the specific contents of the quality control of the fingerprint and index component content of the extract include: the fingerprint of the extract contains characteristic peaks of amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, ligustilide, ligustilide A, and ligustilide I, the content of amygdalin in the extract is 18-40 mg / half prescription, the content of ferulic acid is 3-7 mg / half prescription, and the content of ligustilide is 20-35 mg / half prescription;

[0030] And / or, the specific contents of the quality control of the fingerprint and index component content of the concentrated solution include: the fingerprint of the concentrated solution contains characteristic peaks of amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, ligustilide, ligustilide A and ligustilide I, the content of amygdalin in the concentrated solution is 18-40 mg / halo prescription, the content of ferulic acid is 3-7 mg / halo prescription, and the content of ligustilide is 15-30 mg / halo prescription;

[0031] And / or, the specific contents of the quality control of the fingerprint spectrum and index component content of the aromatic water include: the fingerprint spectrum of the aromatic water contains characteristic peaks of ligustilide and ligustilide A, the content of ligustilide in the aromatic water>

[0032] 5mg / single prescription.

[0033] And / or, the specific contents of the quality control of the fingerprint spectrum and the content of the index components of the inclusion liquid include: the fingerprint spectrum of the inclusion liquid contains characteristic peaks of ligustilide and ligustilide A, the content of ligustilide in the inclusion liquid is greater than 3.5 mg / single prescription, and the inclusion rate of ligustilide is greater than 70%;

[0034] And / or, the specific contents of the quality control of the fingerprint and index component content of the dry powder include: the fingerprint of the dry powder contains characteristic peaks of amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, ligustilide, ligustilide A and ligustilide I, the content of amygdalin in the dry powder is 10-25 mg / single prescription, the content of ferulic acid is 3-7 mg / single prescription, the content of rehmannia glutinosa D is 2.5-5.5 mg / single prescription, the content of paeoniflorin is 30-60 mg / single prescription, and the content of ligustilide is 20-35 mg / single prescription;

[0035] Preferably, the chromatographic conditions of the high performance liquid chromatography method include:

[0036] Octadecylsilane bonded silica gel is used as the filler, acetonitrile or methanol is used as the mobile phase A, and 0.05-0.15% phosphoric acid solution is used as the mobile phase B, and gradient elution is performed.

[0037] Preferably, the chromatographic conditions for quality control of fingerprints using high performance liquid chromatography include: acetonitrile as mobile phase A; the gradient elution process includes: 0-25 min, 0-15% mobile phase A; 25-50 min, 15-20% mobile phase A; 50-80 min, 20-75% mobile phase A; 80-90 min, 75-90% mobile phase A; and / or, the chromatographic conditions of the high performance liquid chromatography also include: flow rate 0.8-1.2 ml / min, column temperature 30-35°C, and detection wavelength of 220±5 nm.

[0038] Preferably, the chromatographic conditions for quality control of content indicators using high performance liquid chromatography include: using methanol as mobile phase A; the gradient elution process includes: 0-35 min, 15-25% mobile phase A; 35-60 min, 25% mobile phase A; 60-62 min, 25-80% mobile phase A; 62-75 min, 80% mobile phase A; and / or, the chromatographic conditions of the high performance liquid chromatography also include: flow rate 0.8-1.2 ml / min, column temperature 25-30°C, detection wavelengths: amygdalin and rehmannia glutinosa D detection wavelength is 205±5 nm, paeoniflorin detection wavelength is 230±5 nm, ferulic acid and ligustilide detection wavelength is 321±5 nm, hydroxysafflor yellow A detection wavelength is 403±5 nm.

[0039] The "fragrant water" refers to a nearly saturated or saturated aqueous solution of a volatile substance, wherein the main component of the volatile substance is volatile oil.

[0040] The present invention establishes an effective method and control technology for the enrichment of volatile oils in a Taohong Siwu Decoction compound preparation that can be industrialized and mass-produced. Based on the technical requirements for the development of classic prescriptions, the present invention achieves quality consistency between the Taohong Siwu Decoction compound preparation and a reference sample, ensuring consistency of various content indicators in the compound preparation with those of the reference sample and consistency of the fingerprint with those of the reference sample. Furthermore, the fingerprint presents characteristic peaks of the volatile components ligustilide, ligustilide I, and ligustilide A, particularly clarifying the consistency of volatile oil content. Furthermore, the quality control method provided by the present invention is expected to be used for quality control of the industrialized mass production of the classic prescription Taohong Siwu Decoction compound preparation, and has good application prospects.

[0041] Conventional extraction processes for volatile oils from traditional Chinese medicines typically separate the volatile medicinal flavors through distillation to collect aromatic water or volatile oils. However, the specific extraction requirements of classic Chinese medicine formulas limit the development of volatile oil processes. Extracting volatile oils using a co-decoction extraction method presents significant challenges, requiring consideration of both other medicinal components in the medicinal solution and the consistency of the volatile components. In this extraction method for classic Chinese medicine formulas, the primary consideration is that the overall quality attributes of the extract should be able to replicate the qualities of the ancient "bowl of soup." Ancient decoctions were taken, resulting in relatively little loss of active ingredients. Modern compound formulations typically undergo concentration and drying stages, which can alter the volatile components and heat-sensitive substances. Therefore, in the development of compound formulations based on classic Chinese medicine formulas, it is necessary to consider developing multiple process pathways based on the co-decoction extraction process, breaking away from conventional methods for extracting and enriching volatile oils from traditional Chinese medicines, maximizing the retention of volatile oil active substances, and ensuring that the overall quality of compound formulations from classic Chinese medicine formulas is consistent with that of a "bowl of soup."

[0042] At present, there is no industrial production of the classic formula - Taohong Siwu Decoction compound preparation of volatile oil enrichment method and control technology implementation. Patent publication number CN202211617476.0 discloses a characteristic spectrum construction method, quality control method and preparation method of a Taohong Siwu Decoction benchmark sample, which is limited to the quality control of the benchmark sample and cannot be used for industrial production; CN105535237A discloses a preparation method and quality control method of Taohong Siwu Decoction formula granules. According to current relevant regulations, the preparation of formula granules should be the extraction of single medicinal materials, which cannot be included in the category of formula granules, and of course does not belong to the research field of classic prescriptions; Application publication number CN112656866A discloses a Taohong Siwu granule and its preparation method. The application also mentions that the ligusticum lactone is almost completely lost during the concentration process. There is no effective measure to retain the volatile components in the application. Therefore, a sufficient amount of volatile oil is not obtained, and the fingerprint spectrum of the volatile components of the benchmark sample is not provided. The research data in the method cannot effectively prove the consistency with the material basis of the benchmark sample, and does not meet the basic requirements of the classic prescription research. Research has shown that due to The reflux extraction time is short, and the amount of ligustilide contained in the collected aromatic water is far from meeting the content requirements of the benchmark sample. It can be determined that it does not belong to the development of the classic prescription - Taohong Siwu Decoction compound preparation. Its efficacy needs to be further verified in clinical trials and does not meet the technical review requirements of classic prescriptions; Application Publication No. CN118903023A is a method for preparing Taohong Siwu granules. Although there are fingerprints of benchmark samples and embodiments, it can be seen from the fingerprints that the characteristic peak of ligustilide is significantly smaller, indicating that the retention rate of ligustilide in the application method is extremely low. As a study of classic prescription compound preparations, the process has major defects. There are fewer characteristic peaks of volatile components in the fingerprint, and there are no volatile substances such as ligustilide A and ligustilide I. There is insufficient evidence for consistency with the benchmark sample. The amount of cyclodextrin used has exceeded the safe dosage in pharmaceutical medicine. The efficacy and safety of the product are questionable and do not meet the research technical requirements of classic prescriptions. This patent application cannot support the large-scale production of the classic prescription - Taohong Siwu Decoction.

[0043] Obviously, based on the above contents of the present invention, according to common technical knowledge and customary means in this field, without departing from the above basic technical ideas of the present invention, other various forms of modifications, replacements or changes can be made.

[0044] The following further describes the above content of the present invention in detail through specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention fall within the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1This is the fingerprint of the reference sample of Taohong Siwu Decoction. Among the 10 common peaks, peak 4 is amygdalin, peak 5 is hydroxysafflor yellow A, peak 6 is paeoniflorin, peak 7 is ferulic acid, peak 8 is ligustilide I, peak 9 is ligustilide A, and peak 10 is ligustilide.

[0046] Figure 2 This is a positioning map of chemical active ingredient reference substances, which includes, from bottom to top, amygdalin, hydroxysafflor yellow A, ferulic acid, verbascoside, ligustilide I, ligustilide H, ligustilide A, the chromatogram of the ligustilide reference substance, and the fingerprint of the Taohong Siwu Decoction benchmark sample;

[0047] Figure 3 The middle left picture shows a photograph of the volatile oils or aromatic waters enriched during the extraction process; Figure 3 The middle right picture shows the volatile oil or aromatic water enriched in the medicinal residue;

[0048] Figure 4 The fingerprints of Chinese Angelica sinensis, Chinese Ligusticum chuanxiong and their chemical active ingredients are as follows: ligustrazine (S1), chlorogenic acid (S2), ferulic acid (S3), ligustrin lactone I (S4), ligustrin lactone H (S5), coniferyl ferulate (S6), ligustrin lactone A (S7), ligustilide (S8), Chinese Ligusticum chuanxiong (S9), Chinese Angelica sinensis (S10).

[0049] Figure 5 The volatile components identification diagram of the extract, aromatic water and concentrated liquid, from bottom to top are the fingerprints of the extract, aromatic water and concentrated liquid;

[0050] Figure 6 1 is a comparison chart of the similarity of the fingerprints of the extracts of Comparative Example 1 and Examples 1-3;

[0051] Figure 7 It is the fingerprint of the essential oils in the collected aromatic waters;

[0052] Figure 8 is the fingerprint of the inclusion compound;

[0053] Figure 9 The fingerprint comparison diagram of the extract, concentrate and spray-dried powder, from bottom to top are the fingerprints of the extract, concentrate and spray-dried powder;

[0054] Figure 10 This is the fingerprint of the spray-dried powder prepared in Comparative Example 1;

[0055] Figure 11 These are the fingerprints of the spray-dried powders prepared in Examples 1-3, wherein from top to bottom are the fingerprints of the spray-dried powders of Examples 1, 2, and 3. DETAILED DESCRIPTION

[0056] In the following examples and experimental examples, reagents and materials not otherwise specified are commercially available.

[0057] 1. The reference sample prescription of Taohong Siwu Decoction in the present invention is: 3.78g of stewed peach kernel, 3.73g of safflower with wine, 11.19g of rehmannia root with wine, 14.92g of angelica root with wine, 5.60g of white peony root with wine, and 3.73g of Chuanxiong.

[0058] The preparation of the present invention adopts the decoction pieces processed from Chinese medicinal materials.

[0059] 2. Quality Control Method for Taohong Siwu Decoction Reference Sample

[0060] 1. Fingerprint control of reference samples

[0061] The classic prescription regulations require the preparation of 15 batches of medicinal materials from a fixed origin into slices, followed by the preparation of 15 batches of reference samples. The key to this material basis research is the chemical composition of the reference samples. Fifteen batches of reference samples were prepared using the traditional decoction process, and their chemical composition was analyzed. Fingerprinting is a key evaluation indicator for effectively demonstrating chemical composition consistency and is an essential component of research for classic prescription compound preparations. Through fingerprint analysis of the Taohong Siwu Decoction reference sample, characteristic peaks in the fingerprint were identified, originating from each of the medicinal ingredients in the prescription, with a particular focus on the volatile components derived from Angelica sinensis and Chuanxiong. The establishment of the reference sample fingerprint, through the identification and attribution of shared peaks, can more effectively illustrate the primary chemical basis of the "bowl of soup," providing strong support for the development of large-scale compound preparations and providing a judgment standard and reference for consistency research in compound preparation quality.

[0062] Fingerprint chromatographic conditions: acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, gradient elution as specified in the table below; flow rate: 1.0 ml / min, column temperature: 35°C, detection wavelength: 250 nm. The theoretical plate number calculated based on the ligustilide peak should be no less than 10,000.

[0063] Table 1 Gradient elution conditions of the fingerprint of the reference sample

[0064]

[0065]

[0066] The reference sample fingerprint is as follows Figure 1 As shown. By using the reference sample positioning method, the common characteristic peaks in the reference samples are assigned, such as Figure 2 shown.

[0067] The main active indicators of each medicinal material in the prescription were attributed by the reference sample positioning method, including: the active ingredient of peach kernel - amygdalin (peak 4), the active ingredient of safflower - hydroxysafflor yellow A (peak 5), the active ingredient of white peony - paeoniflorin (peak 6), the active ingredient of angelica - ferulic acid (peak 7) and ligustilide (peak 10), the active ingredient of Chuanxiong - ligustilide I (peak 8), ligustilide A (peak 9) and ligustilide (peak 10), etc. These chemical substances are all common characteristic peaks in the fingerprint of the reference samples.

[0068] According to the polarity of the common peaks in the fingerprint spectrum, the main low-level volatile components include ligustilide, ligustilide A and ligustilide I. The volatile components in the reference sample correspond to the volatile components in the medicinal materials.

[0069] 2. Control of reference sample content index

[0070] Prescription of benchmark sample: 3.78g of stewed peach kernel, 3.73g of safflower with wine, 11.19g of Rehmannia root with wine, 14.92g of Angelica sinensis with wine, 5.60g of white peony root with wine, and 3.73g of Chuanxiong.

[0071] Based on the above prescription, 15 batches of benchmark samples were prepared in accordance with the "Technical Guidelines for Pharmaceutical Research of Traditional Chinese Medicine Compound Preparations Managed by the Catalog of Ancient Classic Prescriptions", and the chemical composition of the 15 batches of benchmark samples were analyzed. The control ranges of the quality indicators of different medicinal flavors from the prescriptions were determined, providing a basis for judging the quality consistency between subsequent compound preparations and benchmark samples. During the process development of compound preparations, everything is based on the control limits of the indicator components and cannot deviate from the control range.

[0072] Determination was performed by HPLC under the following conditions: octadecylsilane bonded silica gel as the filler; methanol as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, with gradient elution as specified in the table below; flow rate 1.0 ml / min; column temperature 25°C; multi-wavelength detection of each index component: amygdalin and rehmannia glutinin D at 205 nm, paeoniflorin at 230 nm, ferulic acid and ligustilide at 321 nm, and hydroxysafflor yellow A at 403 nm. The number of theoretical plates, calculated based on the amygdalin peak, should be no less than 3000.

[0073] Table 2 Gradient elution conditions of the index components of the reference sample

[0074]

[0075] Each indicator component should comply with the requirements in Table 3 below.

[0076] Table 3 Quality control standards for each index component of the benchmark sample

[0077]

[0078]

[0079] The core of the research on compound preparations of classic prescriptions lies in the use of ancient water decoction technology to ensure that the product quality attributes are consistent with the benchmark sample "a bowl of soup". The benchmark samples are prepared in accordance with the technical requirements of classic prescriptions and the "Management Standards for Chinese Medicine Decoction Rooms in Medical Institutions", and the chemical composition of the benchmark samples is analyzed to determine the control range of the index components.

[0080] Example 1 Preparation Method of Taohong Siwu Decoction Compound Preparation and Volatile Oil Enrichment Method

[0081] This embodiment provides a preparation method of Taohong Siwu Decoction compound preparation, which includes extraction, concentration, inclusion, drying, and granulation. The specific steps are as follows:

[0082] Step 1: Prepare the material according to the 1200 times reference sample prescription, put it into a 1T extraction tank, add 12 times the amount of water for the first time, reflux extraction for 1 hour, add 10 times the amount of water for the second time, reflux extraction for 50 minutes, filter them separately, and combine the two extracts to about 1050kg. The extraction process is carried out in an oil-water separator (the first oil-water separation temperature is 70-80℃, the second oil-water separation temperature is 50-60℃), and 25.3kg of milky white high-concentration aromatic water (such as Figure 3 ).

[0083] Step 2: Add hydroxypropyl-beta-cyclodextrin to the extract, the amount of hydroxypropyl-beta-cyclodextrin is 3% of the mass of the extract, stir and adsorb at 50°C to 60°C for 2 hours, the stirring rate is 250r / min, and concentrate and reduce pressure at 65°C using a 500L single-effect concentrator to a relative density of about 1.05 (g / ml).

[0084] Step 3: The medicinal residue is directly distilled by steam for 1.5 hours. During the distillation and condensation process, the distilled condensate is separated by oil and water in an oil-water separator (the first oil-water separation temperature is 70-80°C, and the second oil-water separation temperature is 50-60°C) to enrich 23.6 kg of milky white high-concentration aromatic water (such as Figure 3 ), and then combined with the aromatic water of step 1, beta-cyclodextrin is added to the combined aromatic water, the amount of beta-cyclodextrin is 5% of the quality of the aromatic water, and the inclusion is stirred at a temperature of 50°C to 60°C for 2 hours at a stirring rate of 250r / min.

[0085] Step 4: The concentrate and the inclusion liquid are mixed and spray-dried at an inlet air temperature of 170°C and an outlet air temperature of 95°C.

[0086] Step 5: Spray dry powder for dry granulation and pack in aluminum-plastic composite film bags.

[0087] In other embodiments, the stirring rate in step 2 and step 3 can be adjusted within the range of 200-500 r / min.

[0088] Table 4 Determination results of single prescription index components at each stage of Example 1

[0089]

[0090] Analysis of the results of Example 1: The content of ligustilide in the extract is relatively high, and nearly 70% of ligustilide is retained in the concentrate. The process of enriching volatile oil in the extract is stable. The collected high-concentration aromatic water is milky white and has a strong aroma. The high concentration of volatile oil is conducive to inclusion. The loss of volatile components in the spray drying stage is very small, and the volatile components in each stage are also effectively retained. All indicators of the spray-dried powder fully meet the limit requirements of the benchmark sample.

[0091] Example 2 Preparation Method of Taohong Siwu Decoction Compound Preparation and Volatile Oil Enrichment Method

[0092] This embodiment provides a preparation method of Taohong Siwu Decoction compound preparation, which includes extraction, concentration, inclusion, drying, and granulation. The specific steps are as follows:

[0093] Step 1: Prepare materials according to the 3000 times benchmark sample prescription, put them into a 3T extraction tank, add 12 times the amount of water for the first time, extract for 60 minutes, add 10 times the amount of water for the second time, extract for 50 minutes, filter them separately, and combine the two extracts of about 2630 kg; during the extraction process, perform secondary oil-water separation in an oil-water separator (the primary oil-water separation temperature is 70-80°C, and the secondary oil-water separation temperature is 50-60°C), and enrich and combine 70.8 kg of milky white high-concentration aromatic water.

[0094] Step 2: Add modified starch to the extract at a rate of 3% of the mass of the extract, stir and adsorb at 50°C to 60°C for 2 hours at a stirring rate of 250 r / min, and concentrate under reduced pressure at 65°C using a 1T single-effect concentrator to a relative density of approximately 1.05 (g / ml).

[0095] Step 3: Add 5 times the amount of water to the medicinal residue and continue heating and reflux extraction for 1 hour. The reflux condensate in the reflux process passes through an oil-water separator (the first-level oil-water separation temperature is 70-80°C, and the second-level oil-water separation temperature is 50-60°C) to enrich 30.5 kg of milky white high-concentration aromatic water, and then merge it with the aromatic water in step 1. Beta-cyclodextrin is added to the combined aromatic water. The amount of beta-cyclodextrin is 5% of the mass of the aromatic water. Stir and include at 50°C ~ 60°C for 2 hours, and the stirring rate is 250r / min.

[0096] Step 4: The concentrate is mixed with the inclusion liquid and spray-dried at an inlet air temperature of 170°C and an outlet air temperature of 90°C.

[0097] Step 5: Spray dry powder for dry granulation and pack in aluminum-plastic composite film bags.

[0098] Table 5: Determination results of haploid index components at each stage of Example 2

[0099]

[0100] Analysis of the results of Example 2: The content of ligustilide in the extract is relatively high, and nearly 70% of ligustilide is retained in the concentrate. The process of enriching volatile oil in the extract is stable, the collected aromatic water is milky white and has a strong aroma, and the high concentration is conducive to inclusion. The loss of volatile components in the spray drying stage is less than 10%, and the volatile components in each stage are also effectively retained. All indicators of the spray-dried powder fully meet the limit requirements of the benchmark sample.

[0101] Example 3 Preparation Method of Taohong Siwu Decoction Compound Preparation and Volatile Oil Enrichment Method

[0102] This embodiment provides a preparation method of Taohong Siwu Decoction compound preparation, which includes extraction, concentration, inclusion, drying, and granulation. The specific steps are as follows:

[0103] Step 1: Prepare materials according to the prescription of 600 times the benchmark sample, put them into a 0.5T extraction tank, add 12 times the amount of water for the first time, decoct and extract for 1 hour, add 10 times the amount of water for the second time, decoct and extract for 50 minutes, filter them separately, and combine the two extracts of about 520kg; during the extraction process, perform secondary oil-water separation in an oil-water separator (the primary oil-water separation temperature is 70-80℃, and the secondary oil-water separation temperature is 50-60℃), and enrich 12.5kg of milky white high-concentration aromatic water.

[0104] Step 2: Add cyclodextrin to the extract at a rate of 3% of the extract mass, stir and adsorb at 50°C to 60°C for 2 hours at a stirring rate of 250 r / min, and concentrate under reduced pressure at 65°C in a 500L single-effect concentrator to a relative density of approximately 1.05 (g / ml).

[0105] Step 3: Add 5% beta-cyclodextrin to the high-concentration aromatic water, with the amount being 5% of the mass of the aromatic water, and stir the inclusion complex at 50°C to 60°C for 2 hours at a stirring rate of 250 r / min.

[0106] Step 4: The concentrate is mixed with the inclusion liquid and spray-dried at an inlet air temperature of 175°C and an outlet air temperature of 95°C.

[0107] Step 5: Spray dry powder for dry granulation and pack in aluminum-plastic composite film bags.

[0108] Table 6 Determination results of prescription index components at each stage of Example 3

[0109]

[0110] Analysis of the results of Example 3: The content of ligustilide in the extract is relatively high, and nearly 70% of ligustilide is retained in the concentrate. The process of enriching volatile oil in the extract is stable, the collected aromatic water is milky white and has a strong aroma, and the high concentration is conducive to inclusion. The loss of volatile components in the spray drying stage is less than 10%, and the volatile components in each stage are also effectively retained. All indicators of the spray-dried powder fully meet the limit requirements of the benchmark sample.

[0111] Example 4 Quality Control Method for a Taohong Siwu Decoction Compound Preparation

[0112] Effectively controlling the quality of intermediates during large-scale production is a key factor in ensuring that compound preparations are consistent with benchmark samples. A full-process quality control method must be established during the development of Taohong Siwu Decoction compound preparations. By effectively controlling the indicator components of each link, the quality deviation of intermediate products and the quality difference of final products can be reduced, thereby ensuring the production of products with uniform quality and making the quality of compound preparations consistent with that of benchmark samples.

[0113] Effectively controlling the quality of the extract during large-scale production is the primary step in ensuring that the compound preparation is consistent with the benchmark sample. During the development of the Taohong Siwu Decoction compound preparation, control standards for the extract (intermediate) must be established. By controlling the index components, the quality deviation of the production process can be effectively reduced, ensuring the consistency of the compound preparation with the benchmark sample.

[0114] 1. Control method of feeding medicinal materials

[0115] 1. Fingerprint control of feeding medicinal materials

[0116] Fingerprint of Chinese Angelica sinensis Figure 4 ) should include the characteristic peaks of volatile components such as ligustilide I, ligustilide H, coniferyl ferulate and ligustilide.

[0117] Ligusticum chuanxiong medicinal material fingerprint Figure 4 ) should include characteristic peaks of volatile components such as ligustilide A, ligustilide I, ligustilide H and ligustilide.

[0118] The shared mass attributes of the volatile components of the two medicinal materials were determined by the chemical reference substance positioning method, providing a basis for the extraction of volatile oils. The main volatile components contained in the Chinese angelica and Chuanxiong medicinal materials used in this embodiment are ligustilide and ligustilide, and the volatile components are basically the same. Therefore, the fingerprint of the Taohong Siwu Decoction compound preparation of the Chinese angelica and Chuanxiong medicinal materials should contain the chromatographic peaks of ligustilide and ligustilide (ligustilide I and ligustilide H).

[0119] Quality control of the source of medicinal materials is the basic guarantee for obtaining qualified finished preparations. The volatile components of the source of medicinal materials can be effectively controlled through fingerprint mapping.

[0120] 2. Control of the content of volatile components in the raw medicinal materials

[0121] In order to ensure that the volatile oil is effectively extracted and enriched, the total amount of the volatile component in the medicinal materials, ligustilide, is controlled. If the content of the medicinal materials is too low, it will be difficult to enrich the volatile components in the medicinal solution, and if the content of the medicinal materials is too high, the procurement cost will increase.

[0122] (1) Chromatographic conditions: Octadecylsilane bonded silica gel as the filler; methanol-water (52:48) as the mobile phase; detection wavelength for ligustilide at 330 nm; column temperature at 35°C. The theoretical plate number calculated for ligustilide should be no less than 5000.

[0123] (2) Preparation of reference solution: Take an appropriate amount of ligustilide reference substance, weigh accurately, and add methanol to make a solution containing 80 μg per 1 ml.

[0124] (3) Preparation of test solution: Take about 0.2 g of Angelica sinensis-Chuanxiong mixed slice powder (passed through No. 4 sieve), where the ratio of Angelica sinensis-Chuanxiong slices is the same as that of Angelica sinensis to Chuanxiong in the reference sample of Taohong Siwu Decoction, accurately weigh it, place it in a stoppered conical flask, accurately add 25 ml of methanol, weigh it, ultrasonically treat it (300 W, 40 kHz) for 30 minutes, let it cool, weigh it again, make up the lost weight with methanol, shake it well, filter it, take the filtrate, and place it in a brown bottle.

[0125] (4) Determination method: Accurately aspirate 10 μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0126] (5) Calculation: The content of ligustilide in the mixed raw materials was calculated by weighted average according to the proportion of Angelica sinensis-Ligusticum chuanxiong slices in the raw materials prescription.

[0127] According to the content and average transfer rate of ligustilide in 15 batches of slices used in the preparation of the benchmark sample, the content of ligustilide in the Angelica sinensis-Chuanxiong mixed slices used in the Taohong Siwu Decoction compound preparation should be controlled at 0.9-1.8% to ensure the consistency of the ligustilide content in the preparation with that of the benchmark sample.

[0128] 2. Fingerprint control of volatile components of extracts, aromatic waters and concentrates

[0129] According to the reference sample fingerprint method established in the present invention, the characteristic peaks of the volatile components - ligustilide and ligustilide - were confirmed in the extract, aromatic water, and concentrated solution. In this example, the extract, aromatic water, inclusion solution, and concentrated solution were all prepared according to the method of Example 1.

[0130] like Figure 5 The extraction, concentration and volatile oil enrichment steps of the present invention can effectively retain the volatile components - ligustilide and ligustilide, which are consistent with the volatile components in the reference sample.

[0131] 3. Control Methods for Active Ingredients in the Extraction Process

[0132] 1. Extract content index control: According to the content method of the reference sample of the present invention, the content of each index component in the extract is basically consistent with that of the reference sample. The content control range of the active ingredient in the extract is formulated based on the range of ±30% of the mean value of the content of 15 batches of reference samples (decoctions).

[0133] Table 7 Quality control standards for the extract

[0134] Indicator components Control standard (equivalent to the content in a single prescription) Amygdalin 18.0mg~40.0mg Ferulic acid 3.0mg~7.0mg Ligustilide 20mg–35mg

[0135] 2. Fingerprint control of the extract: According to the fingerprint method of the reference sample of the present invention, the fingerprint of the extract should show the characteristic peaks of amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, ligustilide, ligustilide A and ligustilide I. According to the similarity evaluation system of traditional Chinese medicine chromatographic fingerprint, the similarity of the fingerprint should be greater than 0.9 ( Figure 6 ).

[0136] 4. Active ingredient control method during concentration process

[0137] 1. Concentrate Content Control: The contents of all target components in the concentrate, except ligustilide, should be essentially consistent with those in the reference sample, as determined using the same method as described for the reference sample. The transfer rate of ligustilide during the concentration process is approximately 70%, while the transfer rates of the other target components are 90% to 110%. Limits are established based on the transfer rates of the target components during the concentration process.

[0138] Table 8 Quality control standards for concentrate

[0139] Indicator components Control standard (equivalent to the content in a single prescription) Amygdalin 18.0mg~40.0mg Ferulic acid 3.0mg~7.0mg Ligustilide 15.0mg~30.0mg

[0140] 2. Fingerprint control of the concentrate: According to the fingerprint method of the benchmark sample of the present invention, the fingerprint of the concentrate should show characteristic peaks such as amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, and ligustilide. The characteristic peaks of the volatile components - ligustilide A and ligustilide I cannot be missing. According to the traditional Chinese medicine chromatographic fingerprint similarity evaluation system, the similarity of the fingerprint is greater than 0.9.

[0141] 5. Methods for controlling volatile components in the process of enriching volatile oils in aromatic water

[0142] 1. Volatile oil content index control: According to the method for determining the content of ligustilide in the benchmark sample of the present invention, the ligustilide content in the single-dose prescription volatile oil should be greater than 5 mg.

[0143] 2. Volatile oil fingerprint control: According to the fingerprint conditions of the benchmark sample of the present invention, the volatile oil or high-concentration aromatic water fingerprint should show the characteristic peaks of ligustilide and ligustilide A ( Figure 7 ).

[0144] 6. Methods for controlling volatile components in the inclusion process

[0145] 1. Control of inclusion compound content index: According to the HPLC method of the present invention, the content of ligustilide in the standard sample ligustilide should be greater than 3.5 mg, and the inclusion rate of ligustilide should be above 70%.

[0146] 2. Inclusion compound fingerprint control: According to the HPLC method of the present invention, the fingerprint of the reference sample should show the characteristic peaks of ligustilide and ligustilide A. The inclusion compound control fingerprint is as follows: Figure 8 shown.

[0147] 7. Active ingredient control method during drying process

[0148] 1. Control of spray-dried powder content index: According to the content method of the reference sample of the present invention, the content of each index component in the single-prescription spray-dried powder should be basically consistent with that of the reference sample.

[0149] Table 9 Quality control standards for spray-dried powder

[0150] Indicator components Control standard (equivalent to the content in a single prescription) Amygdalin 18.0mg~40.0mg Hydroxysafflor Yellow A 10.0mg~25.0mg Ferulic acid 3.0mg~7.0mg Rehmannia glutinosa D 2.5mg–5.5mg Paeoniflorin 30.0mg~60.0mg Ligustilide 20.0mg~35.0mg

[0151] 2. Spray-dried powder fingerprint control: Determined according to the benchmark sample fingerprint method, the spray-dried powder fingerprint should show characteristic peaks of amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, ligustilide and ligustilide A and I. Calculated according to the Chinese medicine chromatographic fingerprint similarity evaluation system, the fingerprint similarity should be greater than 0.9.

[0152] The range of the quality control standard in this example was determined based on 15 batches of benchmark samples and the transfer rate of the concentration process.

[0153] Fingerprints of extracts, concentrates and spray-dried powders are as follows: Figure 9 The three stages of extraction, concentration, and drying in the present invention effectively retain the volatile components, ligustilide and ligustilide. Through quality control and quantitative transfer analysis of the aforementioned processes, the classic formula, Taohong Siwu Decoction, can achieve consistency in quality with a reference sample, while ensuring quantitative transfer of the material basis of each process step and the enrichment of volatile components in the product.

[0154] Comparative Example 1

[0155] A preparation method of Taohong Siwu Decoction compound preparation comprises extraction, concentration, inclusion, drying, and granulation, and the specific steps are as follows:

[0156] Step 1: Prepare materials according to 600 times the prescription amount of the benchmark sample, put them into a 0.5T extraction tank, use the double extraction method, add water and decoct and reflux extraction twice, add 12 times the amount of water in the first time, decoct and extract for 1 hour, add 10 times the amount of water in the second time, decoct and extract for 50 minutes, filter the extracts separately, and combine the two extracts of 510kg; during the extraction process, 13.6kg of clear aromatic water is collected in the first-level oil-water separator (the first-level oil-water separation temperature is 70-80℃).

[0157] Step 2: Use a 500L single-effect concentrator to directly concentrate the extract at 65°C under reduced pressure to a relative density of about 1.05 (g / ml).

[0158] Step 3: Add beta-cyclodextrin to the aromatic water, the amount of beta-cyclodextrin is 5% of the mass of the aromatic water, stir and include at 50℃~60℃ for 2 hours, the stirring rate is 250r / min, refrigerate overnight, filter, and dry the inclusion complex under reduced pressure at 50℃.

[0159] Step 4: The concentrate is spray-dried at an inlet air temperature of 170°C and an outlet air temperature of 95°C.

[0160] Step 5: Mix the spray-dried powder with the inclusion compound, add appropriate amount of auxiliary materials for dry granulation, and package in aluminum-plastic composite film bags.

[0161] Table 10 Determination results of single prescription index components at each stage of comparative example 1

[0162]

[0163] Analysis of the results of Comparative Example 1 based on traditional technology: The content of ligustilide in the extract is relatively high, but the content of ligustilide in the concentrate is negligible. The collected aromatic water is clear and has a strong aroma. The concentration of ligustilide is relatively low, which is not conducive to the inclusion of ligustilide. The volatile components cannot be completely transferred to the granules. The small amount of ligustilide in the granules is mainly obtained through the aromatic water inclusion complex, which is far from meeting the benchmark sample limit requirements.

[0164] The technical solution of the present invention is further illustrated by experiments below.

[0165] Experimental Example 1 Effect of different preparation methods on the content of index components

[0166] 1. Experimental Methods

[0167] Taohong Siwu Decoction compound preparations (granules) were prepared according to the methods of Examples 1-3 and Comparative Example 1, and the contents and fingerprints of the haploid prescriptions of the index components in the granules were determined.

[0168] 2. Experimental Results

[0169] The results of the index component determination are shown in Table 11. The volatile component ligustilide in Comparative Example 1 differs significantly from the standard requirements of the benchmark sample in Example 4, while the other index components are basically consistent and are almost completely lost during the concentration process. The traditional process retains very little of the volatile components in the classic prescription - Taohong Siwu Decoction, and cannot restore the basic quality attributes of "a bowl of soup". It does not meet the basic quality consistency requirements of the classic prescription, and clinical efficacy cannot be guaranteed. The content of each index component in Examples 1 to 3 is consistent with the control standards of the benchmark samples, and the volatile components are also effectively retained, meeting the technical requirements for the development of compound preparations of classic prescriptions.

[0170] Table 11 Determination results of single prescription index components in Taohong Siwu Decoction compound preparation

[0171]

[0172] Fingerprint results are as follows Figure 10 、 11 As shown. The characteristic peaks of the volatile components - ligustilide and ligustilide A are obviously missing in the fingerprint of Comparative Example 1, which is quite different from the fingerprint of the benchmark sample. The traditional extraction process cannot produce a qualified classic prescription - Taohong Siwu Decoction compound preparation. In the fingerprints of Examples 1 to 3, the volatile components - ligustilide and ligustilide A are effectively retained, and the active substances in each medicinal flavor are all present, which is consistent with the reference fingerprint of the benchmark sample. It can restore the quality attributes of "a bowl of soup", ensure the efficacy of the product, and meet the technical requirements for the research and development of classic prescription compound preparations.

[0173] Experimental Example 2 Effect of adsorption conditions on the adsorption effect of volatile oil

[0174] 1. Adsorbent type selection test

[0175] Prepare materials according to the 120-fold benchmark sample prescription, put them into a 100L extraction tank, add 12-fold water for the first time, reflux and extract for 1 hour, add 10-fold water for the second time, reflux and extract for 50 minutes, combine the extracts, weigh them, and determine the content of ligustilide in the extracts. Divide the extracts into four equal parts, add different types of adsorbents at a concentration of 4%, keep warm at 60°C with stirring for adsorption for 2 hours, and concentrate under reduced pressure to a relative density of about 1.05. Determine the content of ligustilide in the concentrates and calculate the content equivalent to that in the single-fold prescription.

[0176] The experimental results are shown in Table 12. Under the same conditions, the adsorption rates of different adsorbent types vary to a certain extent. Among them, hydroxypropyl-β-cyclodextrin has the best adsorption effect, while modified starch and β-cyclodextrin have relatively high cost performance, which is suitable for cost saving in industrial large-scale production.

[0177] Table 12 Adsorbent type selection test results

[0178]

[0179] 2. Adsorbent concentration selection test

[0180] Prepare the extract according to the 120-fold reference sample recipe and place it in a 100-L extraction tank. Add 12-fold the amount of water for the first extraction, and reflux for 1 hour. Add 10-fold the amount of water for the second extraction, and reflux for 50 minutes. Combine the extracts. Weigh and determine the ligustilide content in the extract. Divide the extract into four equal portions, add modified starch at concentrations of 2%, 3%, 4%, and 5%, respectively, and heat and stir at 60°C for 2 hours. Concentrate under reduced pressure to a relative density of approximately 1.05. Determine the ligustilide content in each concentrate and calculate the equivalent to the single-fold recipe.

[0181] The experimental results are shown in Table 13. Under the same conditions, the adsorption rate increases with the increase of adsorbent concentration, but at concentrations of 3% and 4%, the adsorption effect is not significantly increased. Adding 3% modified starch can meet the process requirements.

[0182] Table 13 Adsorbent concentration selection test results

[0183] Modified starch addition amount 2％ 3％ 4％ 5％ Ligustilide content in single dose prescription 12.5mg 18.8mg 20.5mg 22.5mg Adsorption rate 45.8％ 68.8％ 75.1％ 82.4％

[0184] 3. Adsorption time selection test

[0185] (1) Adsorption time selection test when the adsorbent is modified starch

[0186] Prepare the material according to the 120-fold reference sample recipe and place it in a 100-L extraction tank. Add 12-fold the amount of water for the first extraction, and reflux for 1 hour. Add 10-fold the amount of water for the second extraction, and reflux for 50 minutes. Combine the extracts. Weigh and determine the ligustilide content in the extract. Divide the extract into three equal portions, add 3% modified starch, and adsorb with stirring at 60°C for 1, 2, and 3 hours, respectively. Concentrate under reduced pressure to a relative density of approximately 1.05. Determine the ligustilide content in each concentrate and calculate the equivalent to the single-fold recipe.

[0187] The experimental results are shown in Table 14. Under the same conditions, the adsorption rate increased with the increase of adsorption time, but the increase in adsorption effect after 3 hours was not large, and the adsorption time of 2 hours could meet the process requirements.

[0188] Table 14 Adsorption time selection test results when the adsorbent is modified starch

[0189] Adsorption time 1 hour 2 hours 3 hours Ligustilide content in single dose prescription 15.6mg 20.4mg 22.5mg Adsorption rate 55.7％ 72.8％ 80.4％

[0190] (2) Adsorption time selection test when the adsorbent is beta-cyclodextrin

[0191] Prepare the extract according to the 120x reference sample recipe and place it in a 100L extraction tank. Add 12x the amount of water for the first extraction and reflux for 1 hour. Add 10x the amount of water for the second extraction and reflux for 50 minutes. Combine the extracts. Weigh and determine the ligustilide content in the extract. Divide the extract into three equal portions, add 3% beta-cyclodextrin, and adsorb with stirring at 60°C for 1, 2, and 3 hours, respectively. Concentrate under reduced pressure to a relative density of approximately 1.05. Determine the ligustilide content in each concentrate and calculate the equivalent to the single-dose recipe.

[0192] The experimental results are shown in Table 15. Under the same conditions, the adsorption rate increased with the increase of adsorption time, but the increase in adsorption effect after 3 hours was not large, and the adsorption time of 2 hours could meet the process requirements.

[0193] Table 15 Adsorption time selection test results when the adsorbent is beta-cyclodextrin

[0194] Adsorption time 1 hour 2 hours 3 hours Ligustilide content in single dose prescription 14.2mg 18.5mg 21.3mg Adsorption rate 54.6％ 71.2％ 81.9％

[0195] The results of this experimental example show that the type, dosage, and adsorption time of the adsorbent have a significant impact on the adsorption of volatile oils. The best adsorption effect is achieved when the adsorbent is hydroxypropyl-β-cyclodextrin. However, from the perspective of large-scale industrial production, it is necessary to balance the effect and cost (time cost and economic cost). The preferred adsorbents are β-cyclodextrin and modified starch, the preferred adsorbent dosage is 3%, and the preferred adsorption time is 2 hours.

[0196] Experimental Example 3 Effect of different temperatures on oil-water separation

[0197] Traditional extraction of volatile oils from traditional Chinese medicines (TCMs) typically relies on steam distillation. Volatile components pass through a condenser and then into an oil-water separator, where the oil-water mixture separates into separate layers. The separated oil is collected, and the separated water is discharged or returned to the extraction tank, completing the extraction process. The operating temperature of the oil-water separator is generally controlled between 40°C and 60°C. However, temperature control is often lacking during the oil-water separation process. Traditional distillation methods for volatile oil extraction only measure the yield of volatile oils, often yielding only a small amount of clear aromatic water. Vigorous boiling of the medicinal solution is necessary to remove a small amount of heavy oil along with the steam. The aromatic water at the outlet of the shell-and-tube condenser is typically around 80°C, entering the oil-water separator directly. Due to the high temperature, the surface tension of the oil is high, while the aromatic water content is low, preventing oil molecules from condensing. Under these conditions, the small density difference between oil and water makes it difficult for the aromatic water to separate through natural sedimentation or centrifugation in the oil-water separator. This is the fundamental reason why many companies only produce low-concentration aromatic water when extracting volatile oils. To this end, the present invention adopts a temperature control method to increase the concentration of volatile oil after oil-water separation.

[0198] 1. Separation effect test of primary oil-water separator

[0199] Prepare the material according to the 120-fold benchmark sample recipe and place it in a 100L extraction tank. Add 12-fold the amount of water for the first extraction, and reflux extract for 1 hour. Add 10-fold the amount of water for the second extraction, and reflux extract for 50 minutes. Turn on the cooling water to control the flow rate and minimize the temperature of the aromatic water, ensuring that the aromatic water enters only the primary oil-water separator. The separated water returns to the tank. Extract the two tanks, lowering the temperature of the aromatic water in each oil-water separator to the predetermined value. Collect the separated volatile oil or aromatic water separately, weigh them, take samples, and determine the ligustilide content. Calculate the ligustilide content in the single-fold recipe. Set the temperature of the primary oil-water separator to 80°C-90°C and 70°C-80°C, respectively.

[0200] The experimental results are shown in Table 16. When only one-stage oil-water separation is performed, the volatile oil enrichment effect is still poor even if temperature control is implemented.

[0201] Table 16 Results of the first-stage oil-water separation test

[0202]

[0203] 2. Secondary oil-water separator separation effect test

[0204] Prepare the ingredients according to a 120x base sample recipe and place them in a 100L extraction tank. Add 12x the amount of water for the first extraction and reflux for 1 hour. Add 10x the amount of water for the second extraction and reflux for 50 minutes. Turn on the cooling water, allowing the aromatic water to enter the primary oil-water separator first. After further cooling, it enters the secondary oil-water separator, and the separated water returns to the tank. Perform three extractions, lowering the secondary oil-water separator to the desired temperature. Collect the separated volatile oil or aromatic water from each tank, weigh them, take samples, and determine the ligustilide content. Calculate the ligustilide content in the single-dose recipe. Set the primary oil-water separator to 70°C–80°C, and the secondary oil-water separators to 60°C–70°C, 50°C–60°C, and 40°C–50°C, respectively.

[0205] The experimental results are shown in Table 17. After the extract was subjected to secondary oil-water separation, the enrichment effect of volatile oil was significantly improved. In particular, when the temperature of the secondary oil-water separator was 50℃~60℃, the enrichment effect was the best.

[0206] Table 17 Secondary oil-water separation test results

[0207]

[0208]

[0209] The results of this experiment demonstrate that secondary temperature control of the oil-water separator significantly improves volatile oil enrichment, meeting the requirements for volatile oil enrichment and addition in classic formulas, enabling the compound preparation to replicate the quality attributes of the classic "bowl of soup" formula. Specifically, the optimal volatile oil enrichment effect was achieved when the secondary oil-water separator temperature was between 50°C and 60°C.

[0210] Experimental Example 4 Secondary extraction test of residual volatile oil

[0211] Since the solubility of volatile components is low, a large amount of residual oil remains in the medicinal residue after extraction. Two extraction methods were used to study the residual volatile oil.

[0212] 1. Investigation on the amount of aromatic water collected during the reflux time of medicinal residue

[0213] Prepare materials according to the 3000 times benchmark sample prescription, put them into a 3T extraction tank, add 12 times the amount of water for the first time, reflux extraction for 1 hour, add 10 times the amount of water for the second time, reflux extraction for 50 minutes, and the distilled aromatic water enters the first oil-water separator, and then enters the second oil-water separator after cooling. The separated water returns to the extraction tank to collect high-concentration volatile oil or aromatic water.

[0214] Add 5 times the amount of water to the medicinal residue in the extraction tank, continue heating and reflux for 3 hours to enrich the high-concentration aromatic water separately, take samples at different time points of 1, 2, and 3 hours, weigh them, determine the content, and calculate the amount of ligustilide in the single prescription.

[0215] The experimental results are shown in Table 18. The longer the reflux time, the greater the amount of volatile oil / aromatic water, and the higher the content of ligustilide in the single-fold formulation. Among them, the amount of volatile oil enriched by reflux extraction of the medicinal residue for 1 to 2 hours can fully meet the loss during the concentration process.

[0216] Table 18 Effect of reflux time of medicinal residue on the enrichment of volatile oil

[0217]

[0218] 2. Investigation on the time of direct steaming of medicinal residue and the amount of aromatic water collected

[0219] Prepare materials according to the 1200 times benchmark sample prescription, put into a 1T extraction tank, add 12 times the amount of water for the first time, reflux extraction for 1 hour, add 10 times the amount of water for the second time, reflux extraction for 50 minutes, turn on the cooling water, and let the aromatic water flow into the first-level oil-water separator for preliminary oil-water separation. After cooling, it enters the second-level oil-water separator. The separated water returns to the tank, and the separated high-concentration aromatic water is collected.

[0220] Steam was continued to be introduced into the residue in the extraction tank from the bottom for 3 hours to separately enrich the volatile oil. Samples were taken at different time periods of 0.5, 1, 1.5, 2, 2.5, and 3 hours, and the weight and content were measured to calculate the amount of ligustilide in the single-fold prescription.

[0221] The experimental results are shown in Table 19. The longer the direct steaming time of the medicinal residue, the more volatile oil / aromatic water was collected separately, and the higher the content of ligustilide in the single-dose formula. Among them, the amount of volatile oil enriched by direct steaming of the medicinal residue for 1 to 1.5 hours can fully meet the loss during the concentration process.

[0222] Table 19 Effect of direct steaming time of medicinal residue on the enrichment of volatile oil

[0223]

[0224]

[0225] The results of this experimental example show that after the medicinal residues are subjected to reflux or direct steam treatment, the residual volatile oil in the medicinal residues is effectively extracted; considering the application of large-scale industrial production, the amount of volatile oil enriched by reflux extraction of the medicinal residues for 1 to 2 hours, or direct steam extraction for 1 to 1.5 hours, can meet the loss amount during the concentration process.

[0226] Through the above embodiments and experimental examples, it can be seen that the core of the present invention is:

[0227] (1) A process for enriching volatile components from the extract of Taohong Siwu Decoction, a classic prescription that can be industrially produced. Porous adsorption excipients with good safety are added to the extract, including porous starch, modified starch, hydroxypropyl-hexyl cyclodextrin, porous silica, etc. The amount of the porous adsorption excipient is 2% to 5% of the total amount of the extract, which meets the amount of pharmaceutical excipients added. The adsorption temperature of the extract is 60°C to 80°C, the adsorption method is stirring adsorption or colloid milling inclusion adsorption, and the adsorption time is 1 to 3 hours. After the extract is concentrated under reduced pressure and spray-dried, the retention rate of the volatile component - ligustilide in the spray-dried powder is about 70%, which is close to the lower limit of the average content of ligustilide in the reference sample. The fingerprint of the spray-dried powder is basically consistent with the fingerprint of the reference sample. The common peaks in the fingerprint should include volatile components such as ligustilide, ligustilide A and ligustilide I. According to the Chinese medicine chromatographic fingerprint similarity evaluation system, the similarity of the fingerprint is greater than 0.9.

[0228] (2) A classic prescription that can be industrially produced - Taohong Siwu Decoction compound preparation adopts a double extraction process. While extracting and refluxing, the distilled aromatic water is separated into oil and water twice. The temperature of the first oil-water separator should be in the range of 70-90℃, and the temperature of the second oil-water separator should be lower than 60℃. The volatile oil or aromatic water with high concentration of ligustilide is enriched by the oil-water separator. The concentration of ligustilide can reach 10 times the concentration of ligustilide in the extract, which is more conducive to inclusion, reduces the amount of inclusion compound, and improves the quality and inclusion efficiency of the inclusion compound. After spray drying, the fingerprint of the inclusion compound shows characteristic peaks of ligustilide and ligustilide A. The content of volatile component - ligustilide in the spray-dried powder is approximately equivalent to 30% of the average content of the benchmark sample. The volatile oil inclusion compound can also be effectively retained after spray drying with the concentrate. The content of volatile component - ligustilide in the spray-dried powder is equivalent to more than 90% of the average content of the benchmark sample. The enriched volatile oil makes up for the loss during the concentration process and meets the consistency requirements of volatile components.

[0229] (3) Extraction process of residual volatile oils in prescription medicines. The volatile components of the classic prescription - Taohong Siwu Decoction compound preparation can be enriched by collecting the remaining volatile components in the medicinal residue by distillation, directly steam distilling or adding water to reflux the medicinal residue for 1 to 3 hours, collecting a certain amount of aromatic water for replenishment, and enriching high-concentration aromatic water or volatile oil by oil-water separation. The volatile oil is included, and the total amount of volatile component - Ligusticum chuanxiong lactone in the inclusion complex can reach at least 30% of the average content of the benchmark sample. The enriched volatile oil also makes up for the loss during the concentration process, meeting the consistency requirements of the volatile components.

[0230] By effectively combining three approaches to enrich volatile components, we can achieve large-scale industrial production of the classic prescription - Taohong Siwu Decoction compound preparation, which not only achieves consistency with the quality of the benchmark sample, but also saves production time and the amount of excipients.

[0231] The present invention also provides a technology for controlling volatile components of a compound preparation of Taohong Siwu Decoction, a classic prescription that can be industrially produced. The core of the technology lies in controlling the volatile components enriched in the extract, the volatile components enriched in the aromatic water, and the volatile components of the inclusion compound. According to the production process of the compound preparation of Taohong Siwu Decoction, a content determination method and a fingerprint control method are established respectively. The consistency comparison with the benchmark sample is carried out at each stage, so that all links of the compound preparation in large-scale production are fully controlled. An overall quality control method is established for the extract, concentrate, aromatic water, inclusion compound and spray-dried powder respectively, providing a strong technical guarantee for the large-scale production of the compound preparation of Taohong Siwu Decoction.

[0232] The present invention provides a Taohong Siwu Decoction compound preparation and its preparation method and quality control method. Through preliminary screening experiments, the present invention establishes an effective enrichment method and control technology for volatile oil in the Taohong Siwu Decoction compound preparation that can be industrially mass-produced. Based on the technical requirements for the research and development of classic prescriptions, the quality consistency of the Taohong Siwu Decoction compound preparation and a reference sample is achieved, ensuring that the various content indicators in the compound preparation are consistent with those of the reference sample and that the fingerprint spectrum is consistent with that of the reference sample. The fingerprint spectrum also shows characteristic peaks of the volatile components ligustilide, ligustilide I, and ligustilide A, particularly clarifying the consistency of volatile oil content. The overall quality control technology is used as a control method for the industrial mass production of the classic prescription Taohong Siwu Decoction compound preparation. The preparation method and quality control method of the present invention have good application prospects in the industrial mass production and quality control of classic prescription compound preparations.

Claims

1. A Taohong Siwu Decoction compound preparation, characterized in that: Prepared according to the following steps: Step 1: Take the medicinal materials of Taohong Siwu Decoction or its slices, add water and decoct them by double extraction method, perform primary oil-water separation and secondary oil-water separation in reflux, collect and combine aromatic water, and combine the two extracts, wherein the temperature of the secondary oil-water separation is 40-60° C.; Step 2, adding an adsorbent to the extract for adsorption, and concentrating under reduced pressure to obtain a concentrated solution; Step 3, adding an adsorbent to the aromatic water for inclusion to prepare an inclusion liquid; Step 4: mixing the concentrated solution with the inclusion solution, spray drying, and obtaining a dry powder.

2. The Taohong Siwu Decoction compound preparation according to claim 1, characterized in that: In step 1, the double extraction step includes: adding 10-15 times the mass of the medicinal slices of water for the first time, decocting and refluxing for 1-1.5 hours, and adding 8-12 times the mass of the medicinal slices of water for the second time, decocting and refluxing for 50-60 minutes; and / or, in step 1, the temperature of the primary oil-water separation is 70-90°C.

3. The Taohong Siwu Decoction compound preparation according to claim 1, characterized in that: After combining the two extracts in step 1, the residue is obtained, and the residue is extracted again by steam distillation for 1-3 hours or refluxing the residue for 1-3 hours. The distilled condensate or the reflux condensate is subjected to primary oil-water separation and secondary oil-water separation, and the aromatic water is collected and combined, and then combined with the aromatic water described in step 1.

4. The Taohong Siwu Decoction compound preparation according to claim 1, characterized in that: In steps 2 and 3, the adsorbent is independently selected from at least one of modified starch, porous starch, porous silica, beta-cyclodextrin, and hydroxypropyl-beta-cyclodextrin.

5. The Taohong Siwu Decoction compound preparation according to claim 1, characterized in that: In steps 2 and 3, the amount of the adsorbent is 2-5% of the mass of the extract or aromatic water; and / or, in steps 2 and 3, the adsorption or inclusion temperature is 50-60°C; and / or, in steps 2 and 3, the adsorption or inclusion time is 1-3 hours; and / or, in steps 2 and 3, the adsorption or inclusion method is stirring or colloid mill inclusion adsorption, and the stirring rate is 200-500r / min.

6. The Taohong Siwu Decoction compound preparation according to claim 1, characterized in that: In step 2, the temperature of the reduced pressure concentration is 60-70°C; and / or, in step 2, the reduced pressure concentration is performed to a relative density of the extract of 1.04-1.10 g / ml; and / or, in step 4, the spray drying conditions include: an inlet air temperature of 165-175°C and an outlet air temperature of 90-100°C; and / or, dry granulation is performed after the dry powder is obtained; and / or, the primary oil-water separation and the secondary oil-water separation are performed in an oil-water separator.

7. The method for preparing the Taohong Siwu Decoction compound preparation according to any one of claims 1 to 6, characterized in that: The steps include: Step 1: Take the medicinal materials of Taohong Siwu Decoction or its slices, add water and decoct them by double extraction method, perform primary oil-water separation and secondary oil-water separation in reflux, collect and combine aromatic water, and combine the two extracts, wherein the temperature of the secondary oil-water separation is 40-60° C.; Step 2, adding an adsorbent to the extract for adsorption, and concentrating under reduced pressure to obtain a concentrated solution; Step 3, adding an adsorbent to the aromatic water for inclusion to prepare an inclusion liquid; Step 4: mixing the concentrated solution with the inclusion solution, spray drying, and obtaining a dry powder.

8. A quality control method for Taohong Siwu Decoction compound preparation, characterized in that: The following steps are involved: High performance liquid chromatography is used to perform quality control of fingerprints and content indicators on the medicinal materials of Taohong Siwu Decoction or the extract, concentrate, aromatic water, inclusion liquid or dry powder prepared by the preparation method according to claim 7; The index component is selected from at least one of ligustilide, amygdalin, ferulic acid, hydroxysafflor yellow A, rehmannia glutinosa D, and paeoniflorin.

9. The method for quality control of fingerprints according to claim 8, characterized in that: The specific contents of the quality control of the fingerprint and index component content of the medicinal materials include: the fingerprint of the medicinal flavor of Angelica sinensis contains characteristic peaks of ligustilide I, ligustilide H, coniferyl ferulate and ligustilide; the fingerprint of the medicinal flavor of Chuanxiong contains characteristic peaks of ligustilide A, ligustilide I, ligustilide H and ligustilide; the total content of ligustilide in the prescription ratio of the Angelica sinensis-Ligustilide medicinal pair is 0.9-1.8%; And / or, the specific contents of the quality control of the fingerprint and index component content of the extract include: the fingerprint of the extract contains characteristic peaks of amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, ligustilide, ligustilide A, and ligustilide I, the content of amygdalin in the extract is 18-40 mg / half prescription, the content of ferulic acid is 3-7 mg / half prescription, and the content of ligustilide is 20-35 mg / half prescription; And / or, the specific contents of the quality control of the fingerprint and index component content of the concentrated solution include: the fingerprint of the concentrated solution contains characteristic peaks of amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, ligustilide, ligustilide A and ligustilide I, the content of amygdalin in the concentrated solution is 18-40 mg / halo prescription, the content of ferulic acid is 3-7 mg / halo prescription, and the content of ligustilide is 15-30 mg / halo prescription; And / or, the specific contents of the quality control of the fingerprint spectrum and index component content of the aromatic water include: the fingerprint spectrum of the aromatic water contains the characteristic peaks of ligustilide and ligustilide A, and the content of ligustilide in the aromatic water is >5mg / single prescription. And / or, the specific contents of the quality control of the fingerprint spectrum and the content of the index components of the inclusion liquid include: the fingerprint spectrum of the inclusion liquid contains characteristic peaks of ligustilide and ligustilide A, the content of ligustilide in the inclusion liquid is greater than 3.5 mg / single prescription, and the inclusion rate of ligustilide is greater than 70%; And / or, the specific contents of the quality control of the fingerprint and index component content of the dry powder include: the fingerprint of the dry powder contains characteristic peaks of amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, ligustilide, ligustilide A and ligustilide I, the content of amygdalin in the dry powder is 10-25 mg / half prescription, the content of ferulic acid is 3-7 mg / half prescription, the content of rehmannia glutinosa D is 2.5-5.5 mg / half prescription, the content of paeoniflorin is 30-60 mg / half prescription, and the content of ligustilide is 20-35 mg / half prescription.

10. The fingerprint quality control method according to claim 8, characterized in that: The chromatographic conditions of the high performance liquid chromatography method include: Octadecylsilane bonded silica gel is used as the filler, acetonitrile or methanol is used as the mobile phase A, and 0.05-0.15% phosphoric acid solution is used as the mobile phase B, and gradient elution is performed.

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