Preparation method of malus hallings root extract, detection method and characteristic chromatogram construction method

Abstract

The present application relates to the field of traditional Chinese medicine analysis, and particularly relates to a preparation method, a detection method and a characteristic spectrum construction method of a Rosa roxburghii tratt root extract. The Rosa roxburghii tratt root extract is prepared by twice water decocting and filtering Rosa roxburghii tratt root decoction pieces to obtain a decoction, and then concentrating and drying the decoction. The extract yield of the Rosa roxburghii tratt root extract is 2.7-7.8%, which is relatively high. The mass content of ellagic acid in the Rosa roxburghii tratt root extract is 8.20-18.80 mg / g, and the transfer rate of ellagic acid is 20.38-98.32%. The detection method of the mass content of ellagic acid in the Rosa roxburghii tratt root extract and the characteristic spectrum construction method provided by the present application have good specificity, good repeatability and good stability, and provide a scientific basis for establishing the quality standards of the Rosa roxburghii tratt root extract and its preparation, and realize the overall quality control and effective supervision of traditional Chinese medicine formula granules.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine analysis, specifically to a method for preparing, detecting, and constructing a characteristic chromatogram of a Rosa rugosa root extract. Background Technology

[0002] The root of *Rosa roxburghii* Tratt.f.normalis Rehd.etWils., a plant in the Rosaceae family, is sour and astringent in taste, neutral in nature, and enters the spleen, stomach, liver, and kidney meridians. It has astringent and antidiarrheal properties, aids digestion and strengthens the spleen, and enhances the body's immunity. It is used in folk medicine to treat chronic gastritis, spleen deficiency dysentery, and abdominal pain due to food stagnation. Clinically, it is used to treat stomach pain, diarrhea, dysentery, seminal emission, metrorrhagia, leukorrhea, chronic cough, and hemorrhoids.

[0003] The clinical efficacy and pharmacological activity of *Rhizoma Rosa roxburghii* root are related to its active ingredients. Current research indicates that *Rhizoma Rosa roxburghii* root contains various components such as vitamin C, polyphenols, flavonoids, and organic acids. It also contains a variety of organic acids, including tartaric acid, malic acid, lactic acid, citric acid, and succinic acid, with tartaric acid and lactic acid being the most prevalent.

[0004] Extracts, also known as standard decoctions, are a traditional and widely used form of medicine in clinical practice. Extracts are prepared by following the theory of traditional Chinese medicine, decocting according to the standardized methods of clinical decoctions, separating solids and liquids, and then concentrating or drying them using appropriate methods. They serve as a standard reference for measuring whether the granules of traditional Chinese medicine formulas are basically consistent with the clinical decoctions.

[0005] Since extracts serve as a "bridge" connecting traditional Chinese medicine decoction pieces and modern Chinese medicine preparations, they provide a reference for controlling the quality of Chinese medicine end products, for standardizing different forms of Chinese medicine use, ensuring the uniformity of quality and the consistency of efficacy, and for evaluating the consistency of product quality among different manufacturers. Therefore, the formulation of quality standards for Chinese medicine extracts will provide a basis for the formulation of quality standards for all end products derived from decoction pieces.

[0006] Currently, there are no literature reports on the preparation methods, detection methods, and characteristic spectrum construction methods of Rosa rugosa root extract. Summary of the Invention

[0007] The technical problem to be solved by the present invention is to provide a method for preparing, detecting and constructing characteristic chromatograms of prickly pear root extract, which can provide quality control means for the quality evaluation of prickly pear root extract and its preparations.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] This application provides a method for preparing a prickly pear root extract, which includes decocting prickly pear root slices twice with water and filtering to obtain a decoction, and concentrating and drying the decoction to obtain the prickly pear root extract.

[0010] In some embodiments of this application, the first decoction is made with 6 to 10 times the amount of water, preferably 8 times the amount of water; the second decoction is made with 5 to 7 times the amount of water, preferably 6 times the amount of water.

[0011] In some embodiments of this application, the concentration temperature is 60-70°C; preferably, the concentration is carried out until the density of the concentrate is 1.01-1.03 g / mL.

[0012] In some embodiments of this application, the drying is freeze-drying, which is divided into three stages: a. pre-freezing: the pre-freezing temperature is -50 to -40°C; b. primary drying: the drying temperature is -30 to 0°C; c. secondary drying for analysis: the drying temperature is 5 to 25°C.

[0013] Preferably, the pre-freezing time is 3-5 hours, more preferably 4 hours; the first drying time is 12-16 hours, more preferably 14 hours; and the second drying time is 6-8 hours, more preferably 7 hours.

[0014] More preferably, the vacuum degree of the primary drying is 0 to 0.2 mbar;

[0015] More preferably, the vacuum degree of the secondary drying is 0 to 0.2 mbar.

[0016] In some embodiments of the present invention, filtration is performed using a 100-300 mesh sieve.

[0017] In some embodiments of this application, the yield of the prickly pear root extract is 2.7-7.8%.

[0018] This application also provides a prickly pear root extract, which is prepared by the above-described method.

[0019] This application also provides a method for detecting the ellagic acid content in the above-mentioned prickly pear root extract, which includes the following steps:

[0020] (1) Preparation of reference solution

[0021] Weigh out ellagic acid reference standard and add it to methanol to prepare a reference solution;

[0022] (2) Preparation of the test solution

[0023] Weigh out the root extract of Rosa rugosa and add it to the first solvent for extraction;

[0024] (3) High performance liquid chromatography analysis

[0025] The mass content of ellagic acid in the prickly pear root extract was obtained by high performance liquid chromatography analysis of the reference solution and the test solution.

[0026] This application also provides a method for detecting the transfer rate of ellagic acid in the above-mentioned Rosa rugosa root extract, which includes the following steps:

[0027] (1) Preparation of reference solution

[0028] Weigh out ellagic acid reference standard and add it to methanol to prepare a reference solution;

[0029] (2) Preparation of the test solution

[0030] Weigh out the root extract of Rosa rugosa and add it to the first solvent for extraction;

[0031] (3) Preparation of control solution of prickly pear root slices

[0032] Weigh out the prickly pear root slices and add the first solvent for extraction;

[0033] (4) High performance liquid chromatography analysis

[0034] The transfer rate of ellagic acid in the prickly pear root extract was obtained by high performance liquid chromatography analysis of the reference solution, the test solution, and the prickly pear root slices control solution.

[0035] In some embodiments of this application, the first solvent is selected from one or more of 25% ethanol, 50% ethanol, 75% ethanol, 100% ethanol, 25% methanol, 50% methanol, 75% methanol, 100% methanol and water, preferably water;

[0036] Preferably, the volume-to-mass ratio of the first solvent to the prickly pear root extract is 100-2000 mL / g, more preferably 500-1500 mL / g.

[0037] In some embodiments of this application, the extraction is performed using either reflux or ultrasound, preferably reflux extraction;

[0038] Preferably, the extraction time is 0.5-2 hours, and more preferably 1-2 hours.

[0039] In some embodiments of this application, the chromatographic conditions for the high-performance liquid chromatography analysis are as follows: using octadecylsilane-bonded silica gel as the packing material, and using mobile phase A and mobile phase B for gradient elution; preferably, methanol is used as mobile phase A, and 0.08-0.12% phosphoric acid solution is used as mobile phase B, more preferably 0.1% phosphoric acid solution is used as mobile phase B;

[0040] More preferably, the gradient elution conditions are as follows: from 0 to 15 min, mobile phase A is 15 vol% → 38 vol%, mobile phase B is 85 vol% → 62 vol%, and from 15 to 40 min, mobile phase A is 38 vol%, mobile phase B is 62 vol%.

[0041] In some embodiments of this application, the ellagic acid content in the prickly pear root extract, calculated on a dried basis, is 8.20-18.80 mg / g, and more preferably, the ellagic acid transfer rate is 20.38-98.32%.

[0042] This application also provides a method for constructing the characteristic spectrum of the above-mentioned Rosa rugosa root extract, which includes the following steps:

[0043] (1) Preparation of reference solution

[0044] Weigh out ellagic acid reference standard and add it to methanol to prepare a reference solution;

[0045] (2) Preparation of the test solution

[0046] Weigh out the root extract of Rosa rugosa and add a second solvent for extraction;

[0047] (3) High performance liquid chromatography analysis

[0048] The characteristic chromatograms of the prickly pear root extract were obtained by high performance liquid chromatography analysis of the reference solution and the test solution.

[0049] In some embodiments of this application, the second solvent is selected from one or more of 25% ethanol, 50% methanol, 75% ethanol, 100% ethanol, 25% methanol, 50% methanol, 75% methanol, 100% methanol and water, preferably water;

[0050] Preferably, the volume-to-mass ratio of the second solvent to the prickly pear root extract is 100-2000 mL / g, more preferably 500-1500 mL / g.

[0051] In some embodiments of this application, the extraction in step (2) is performed by either reflux or ultrasound, preferably reflux extraction;

[0052] Preferably, the extraction time in step (2) is 0.5-2 hours, and more preferably 1-2 hours.

[0053] In some embodiments of this application, the chromatographic conditions for high performance liquid chromatography analysis in step (3) are as follows: using octadecylsilane-bonded silica gel as the packing material, and using mobile phase A and mobile phase B for gradient elution; preferably, mobile phase A is methanol or acetonitrile, preferably methanol; mobile phase B is water, 0.1% phosphoric acid solution, 0.1% formic acid solution or 0.1% acetic acid solution, preferably 0.1% phosphoric acid solution;

[0054] More preferably, the gradient elution conditions are as follows: from 0 to 15 min, mobile phase A is 15 vol% → 38 vol%, mobile phase B is 85 vol% → 62 vol%, and from 15 to 40 min, mobile phase A is 38 vol%, mobile phase B is 62 vol%.

[0055] In some embodiments of this application, the characteristic spectrum of the prickly pear root extract contains seven characteristic peaks. Preferably, the characteristic peaks are those at retention times of 0.14-0.16, 0.67-0.71, 0.83-0.87, 0.90-0.92, 0.96-0.98, 1.00, and 1.19-1.24.

[0056] This application also provides the use of the above-mentioned prickly pear root extract in the preparation of health food and traditional Chinese medicine; preferably, the traditional Chinese medicine is a standard decoction, capsule, tablet, granule or formula granule.

[0057] This application also provides a standard decoction prepared from the above-mentioned prickly pear root extract.

[0058] This application also provides a prickly pear root formulation granule containing the above-mentioned prickly pear root extract. Preferably, the prickly pear root formulation granule also contains excipients. More preferably, the excipients are selected from one or more of maltodextrin, silicon dioxide, or magnesium stearate.

[0059] This application also provides the application of the above-mentioned detection method or the above-mentioned method for constructing the characteristic spectrum in the root extract of Rosa rugosa and its preparations.

[0060] The beneficial effects of this invention are:

[0061] 1. This invention uses the root of *Rhizophora pilosa* to prepare *Rhizophora pilosa* root extract. The extract has a high yield of 2.7-7.8%, and the ellagic acid content in the extract is 8.20-18.80 mg / g, with a transfer rate of 20.38-98.32%.

[0062] 2. The detection method for Rosa rugosa root extract established by this invention using ellagic acid content as an indicator component has good specificity, good repeatability, and good stability. It provides a scientific basis for establishing quality standards for Rosa rugosa root extract and its preparations, and realizes the overall quality control and effective supervision of traditional Chinese medicine formula granules.

[0063] 3. The characteristic chromatographic method established in this invention uses ultra-high performance liquid chromatography (UHPLC), which is simple, stable, precise, and reproducible. Moreover, the fingerprint chromatogram of the obtained extract has many peaks, good peak shape, is easy to identify, and is accurate and reliable. Furthermore, the method for constructing this characteristic chromatogram is time-saving and environmentally friendly, requiring only 30 minutes to analyze a batch of preparations. This can greatly shorten the detection and analysis time, improve production efficiency, and make it possible to conduct large-sample sampling and real-time monitoring of product quality. Attached Figure Description

[0064] Figure 1 This is a schematic diagram of the specificity investigation spectrum in Example 2.

[0065] Figure 2 This is a schematic diagram of the chromatographic peak purity in Example 2.

[0066] Figure 3 This is a schematic diagram of the linear relationship in Example 2.

[0067] Figure 4 The absorption spectrum of the prickly pear root extract in Example 4 is shown in the range of 190-400 nm.

[0068] Figure 5 These are characteristic spectra of different mobile phases A in Example 4.

[0069] Figure 6 These are characteristic spectra of different mobile phases B in Example 4.

[0070] Figure 7 The image shows the characteristic spectra of different batches of Rosa rugosa root extract in Example 4.

[0071] Figure 8 This is a schematic diagram of the common peak in Example 4.

[0072] Figure 9 The characteristic spectra are for methanol, 75% methanol and 50% methanol as solvents in Example 4.

[0073] Figure 10 The characteristic spectra are those of 25% methanol, anhydrous ethanol, and 75% ethanol in Example 4.

[0074] Figure 11 The characteristic spectra are those of solvents 50% ethanol, 25% ethanol and water in Example 4.

[0075] Figure 12This is a schematic diagram of the specificity investigation spectrum in Example 4.

[0076] Figure 13 This is a schematic diagram of the overall examination pattern in Example 4. Detailed Implementation

[0077] To make the objectives, technical solutions, and technical effects of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are described clearly and completely. The embodiments described below are some embodiments of the present invention, but not all embodiments. In conjunction with the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0078] In one specific embodiment of the present invention, the present invention provides a method for preparing a prickly pear root extract, wherein the preparation method includes:

[0079] The root slices of prickly pear were decocted twice and filtered to obtain the decoction.

[0080] The decoction was concentrated and dried to obtain the prickly pear root extract.

[0081] In this application, no restrictions are placed on the processing of prickly pear root slices. They can be prepared in accordance with conventional methods in the field. For example, the raw medicinal material can be taken, tender branches and leaves can be selected, residual roots and impurities can be removed, the leaves can be removed first, sieved and stored separately; the stems can be washed, soaked in water, cut into sections, dried in the sun, and then mixed with the leaves, or soaked in water, cut into sections, and dried, etc.

[0082] In this application, no restrictions are placed on the utensils used for decoction. Utensils in the field can be used for decoction. For example, the provisions on decoction containers in the "Management Standards for Traditional Chinese Medicine Decoction Rooms in Medical Institutions" (Document No. 3 of 2009, issued by the State Administration of Traditional Chinese Medicine) shall be used (decoction containers should preferably be made of ceramic, stainless steel, copper, etc.). In this application, a 5L automatic ceramic decoction pot is used as the decoction equipment for extracts.

[0083] In this application, no restrictions are placed on the dosage of prickly pear root slices. Conventional dosages in the field can be used. For example, according to the recommendations under "Standard Decoction Preparation" in the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Granules," the amount of slices used per decoction is generally not less than 100g, with the amount for flowers, leaves, etc., potentially reduced. In this application, the dosage of prickly pear root slices is 100g.

[0084] The extract described in this application can standardize the quality control and standardization research of traditional Chinese medicine formula granules, realize the overall quality control and effective supervision of traditional Chinese medicine formula granules, and provide a reference for the quality control of prickly pear root formula granules.

[0085] In some embodiments of this application, the first decoction is made with 6 to 10 times the weight of the prickly pear root slices of water, preferably 8 times the weight of water. Preferably, the second decoction is made with 5 to 7 times the weight of the prickly pear root slices of water, preferably 6 times the weight of water.

[0086] In this application, for the first decoction, 6-10 times the amount of water is added, preferably 8 times the amount of water. More preferably, for the second decoction, 5-7 times the amount of water is added, preferably 6 times the amount of water. This is based on the recommendations under "Standard Decoction Preparation" in the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Granules." The amount of water added is generally sufficient to cover the herbs by 2-5 cm. For flower and herbaceous medicinal materials or medicinal materials requiring a longer decoction time, the amount of water can be adjusted accordingly. For example, 6, 7, 8, 9, or 10 times the amount of water is added for the first decoction, and 5, 6, or 7 times the amount of water is added for the second decoction.

[0087] In this application, after adding water, soaking is required before decoction. This application does not impose any restrictions on the soaking time; the soaking time can be designed according to conventional methods in the field. For example, it is recommended under the "Standard Decoction Preparation" section of the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Granules" that the decoction pieces should be soaked first, with a soaking time generally not less than 30 minutes. Based on the "Technical Requirements," the soaking time for the prickly pear root extract is determined to be 30 minutes.

[0088] In this application, no restrictions are placed on the decoction time. The decoction time can be designed according to conventional methods in the field. For example, according to the recommendations under "Standard Decoction Preparation" in the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Granules," each dose of medicine is generally decocted twice. Generally, after boiling, it is decocted for 30 minutes. For diaphoretic, heat-clearing, and aromatic medicines, prolonged decoction is not advisable; boiling for 20 minutes is appropriate. For harder medicinal herbs, the decoction time can be appropriately extended. For tonifying medicinal herbs, first bring to a boil over high heat, then simmer over low heat for about 60 minutes. The second decoction time can be appropriately shortened. Therefore, according to the "Technical Requirements," the prickly pear root extract is decocted twice: the first decoction is boiled for 60 minutes after boiling, and the second decoction is boiled for 40 minutes after boiling.

[0089] In some implementations, filtration is performed using a 100-300 mesh sieve.

[0090] For example, filtration can be performed using 100-mesh, 200-mesh, or 300-mesh sieves. This is designed according to the recommendation in the "Standard Decoction Preparation" section of the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules" that the mesh size of the filter material for solid-liquid separation should be above 100 mesh. 100-mesh, 200-mesh, and 300-mesh sieves all facilitate filtration, but the filtrate obtained from 100-mesh and 200-mesh sieves has poorer clarity and produces more sediment after settling. The filtrate obtained from filtration through a 300-mesh sieve has better clarity and produces less sediment after settling. Therefore, a 300-mesh sieve is used for solid-liquid separation of Rosa rugosa root extract.

[0091] In this application, no restrictions are placed on the concentration method; concentration can be carried out according to conventional methods in the field. For example, it can be concentrated at low temperature to a specified amount of extract using the vacuum concentration method recommended under the "Preparation of Standard Decoctions" section of the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules". A suitable concentration temperature was determined by comparing different concentration temperatures, and a concentration ratio of 1:1 was tentatively set. The concentration state was observed by comparing and measuring the density, yield, and state of the concentrated liquid at different concentration temperatures. The results showed that the yield was basically consistent at different concentration temperatures, and there was no significant difference between concentration temperatures of 50℃ and 65℃. Considering all factors, 65℃ was chosen as the concentration temperature. When the concentration ratio was 1:1, the extract had moderate viscosity, good fluidity, and was easy to transfer, with a density between 1.01-1.03 g / ml. Therefore, a concentration ratio of 1:1 was determined, and other concentration ratios were not considered for the time being.

[0092] Therefore, for concentration, a rotary evaporator can be used to concentrate under reduced pressure (temperature: 65℃; vacuum degree: -0.080~-0.090MPa) to 100ml.

[0093] In some embodiments, the drying is freeze-drying, which includes pre-freezing, primary drying, and secondary drying. Preferably, the pre-freezing is maintaining the temperature at -50°C to -40°C for 3-5 hours, and more preferably maintaining the temperature at -45°C for 4 hours.

[0094] Preferably, the primary drying is carried out at -30°C to 0°C for 12-16 hours;

[0095] Preferably, the secondary drying is carried out at 5-25°C for 6-8 hours.

[0096] In this application, the freeze-drying method is chosen based on the recommendation in the "Preparation of Standard Decoctions" section of the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules" that freeze-drying is generally preferred for the drying of extracts to ensure their quality stability, ease of dissolution, and elimination of excipients. Therefore, freeze-drying is selected as the drying method for the Rosa rugosa root extract.

[0097] In this application, since the eutectic point test result of the vacuum freeze dryer for prickly pear root is -43.6℃, ​​the pre-freezing temperature is -45℃, the first drying temperature is -30~0℃, the second drying temperature is 5~25℃, and the total drying time is 28 hours, the prickly pear root extract has low moisture content under these freeze-drying conditions and is easy to preserve.

[0098] In this application, primary drying refers to sublimation drying, and secondary drying refers to desorption drying.

[0099] In this application, for a single drying process, the drying time can be maintained at -30°C for 420 min, -30°C for 120 min, -10°C for 120 min, or 0°C for 180 min.

[0100] In this application, for secondary drying, the drying process can be maintained at 5°C for 120 min, at 15°C for 120 min, or at 25°C for 180 min.

[0101] In some embodiments, the yield of the concentrate is 2.7-7.8%.

[0102] For example, the yield of the concentrate can be 2.7%, 3%, 4%, 5%, 6%, 7%, 7.8%, etc.

[0103] In this application, the solid content of the filtrate was also determined. In this application, no restrictions are placed on the method for determining the solid content of the filtrate. It can be determined according to conventional methods in the art. For example, it can be determined by referring to the "hot extraction method" in the General Chapter 2201 of the 2020 edition of the Chinese Pharmacopoeia, Part IV, Determination of Extracts. 25 mL of the extract is accurately pipetted into an evaporating dish that has been pre-weighed, evaporated to dryness on a water bath, dried at 105°C for 3 hours, cooled in a desiccator for 30 minutes, and the weight is quickly and accurately determined to obtain the solid content.

[0104] In this application, no restrictions are placed on the method for determining the yield of the concentrate. It can be determined according to conventional methods in the field. For example, the solid content of the concentrate can be determined first, and then the yield of the concentrate can be calculated according to the formula. For example, the determination of extract content by hot extraction method in General Chapter 2201 Extract Determination Method of Part IV of the 2020 edition of the Chinese Pharmacopoeia can be referred to: accurately weigh 10g of concentrate, place it in a pre-weighed evaporating dish, evaporate it to dryness on a water bath, dry it at 105℃ for 3 hours, cool it in a desiccator for 30 minutes, quickly and accurately weigh it, calculate the solid content of the concentrate, and then calculate the yield of the concentrate according to the following formula: Yield of concentrate % = weight of concentrate * solid content of concentrate / amount of medicinal slices * 100%.

[0105] This application also provides a prickly pear root extract prepared by the above method.

[0106] Understandably, this prickly pear root extract can be used directly as a standard decoction, or it can be mixed with other excipients to make formula granules.

[0107] This application also provides a prickly pear root formula granule, which contains the above-mentioned prickly pear root extract. Preferably, the prickly pear root formula granule also contains excipients. More preferably, the excipients are selected from one or more of maltodextrin, silicon dioxide, or magnesium stearate.

[0108] In some embodiments of this application, the prickly pear root extract is prepared by the following steps: boiling prickly pear root slices twice with water and filtering to obtain a decoction; concentrating and drying the decoction to obtain the prickly pear root extract.

[0109] The production of traditional Chinese medicine (TCM) preparations often involves the drying process of extracts. Currently, the main drying methods for TCM extracts include vacuum drying (drying oven), spray drying, microwave drying, and infrared drying. In recent years, with the development of TCM pharmaceutical equipment, vacuum drying (reduced pressure drying, low-temperature pulse drying, belt drying) and spray drying technologies have been widely used in the drying of TCM extracts. In particular, spray drying is widely used in TCM formulation granules. Its principle is to use an atomizer to disperse the liquid material into fine droplets, and then rapidly evaporate the solvent in a hot drying medium to form a dry powder product. Its main characteristics are: fast drying, short time, and the ability to directly dry solutions, emulsions, suspensions, and extracts into powder, granules, hollow spheres, or agglomerates. It can eliminate the need for evaporation, separation, and pulverization processes, retains better properties, and causes less damage to the effective components. Therefore, based on the effective components contained in the root of *Rhizoma Rosa rugosa* and considering the existing production equipment, a spray drying method was selected from various production equipment based on different principles. This method is characterized by high drying efficiency, fast drying speed, short drying time, minimal damage to effective components, uniform and fine dry powder, low moisture content, good solubility, and suitability for pilot-scale and large-scale commercial production. Therefore, in some embodiments of this application, the drying is spray drying. Preferably, the spray drying process parameters are: inlet air temperature: 165℃~195℃, outlet air temperature: 100~110℃.

[0110] In some embodiments of this application, in order to prevent foreign matter mixed in the extract from clogging the pipes, the extract is further heated to 60-100°C, and an appropriate amount of excipients is added to fully dissolve the extract. Preferably, the excipient is maltodextrin, and more preferably, the amount of maltodextrin added is 1.5-4.5%.

[0111] This application also provides a method for detecting the ellagic acid content in Rosa rugosa root extract, comprising:

[0112] (1) Preparation of reference solution

[0113] Weigh out ellagic acid reference standard and add it to methanol to prepare a reference solution;

[0114] (2) Preparation of the test solution

[0115] Weigh out the root extract of Rosa rugosa and add it to the first solvent for extraction;

[0116] (3) High performance liquid chromatography analysis

[0117] The mass content of ellagic acid in the prickly pear root extract was obtained by high performance liquid chromatography analysis of the reference solution and the test solution.

[0118] In some embodiments of this application, the first solvent is selected from one or more of 25% ethanol, 50% ethanol, 75% ethanol, 100% ethanol, 25% methanol, 50% methanol, 75% methanol, 100% methanol and water, preferably water;

[0119] Preferably, the volume-to-mass ratio of the first solvent to the prickly pear root extract is 100-2000 mL / g, more preferably 500-1500 mL / g. For example, based on the mass of the prickly pear extract being 0.1 g, the volume of the solvent can be 10 mL, 20 mL, 30 mL, 40 mL, 60 mL, 80 mL, 100 mL, 150 mL, etc.

[0120] In some embodiments of this application, the extraction in step (2) is performed by either reflux or ultrasound, preferably reflux extraction;

[0121] Preferably, the extraction time in step (2) is 0.5-2 hours, and more preferably 1-2 hours.

[0122] The methods described above can provide quality control for the quality evaluation of standard decoctions or granules of prickly pear root.

[0123] In some embodiments of this application, the chromatographic conditions for high performance liquid chromatography analysis in step (3) are as follows: using octadecylsilane-bonded silica gel as the packing material, and using mobile phase A and mobile phase B for gradient elution; preferably, mobile phase A is methanol or acetonitrile, preferably methanol; mobile phase B is water, 0.1% phosphoric acid solution, 0.1% formic acid solution or 0.1% acetic acid solution, preferably 0.1% phosphoric acid solution;

[0124] More preferably, the gradient elution conditions are as follows: from 0 to 15 min, mobile phase A is 15 vol% → 38 vol%, mobile phase B is 85 vol% → 62 vol%, and from 15 to 40 min, mobile phase A is 38 vol%, mobile phase B is 62 vol%.

[0125] In some embodiments, the flow rate of the mobile phase is 0.2-0.3 ml / min;

[0126] Preferably, the column temperature is 30-40℃;

[0127] Preferably, the detection wavelength is 250-260nm, and more preferably 254nm;

[0128] Preferably, the injection volume is 10-20 μL.

[0129] For example, the flow rate of the mobile phase can be 0.2 ml / min, 0.25 ml / min, 0.3 ml / min, etc.;

[0130] The column temperature can be 30℃, 31℃, 32℃, 33℃, 34℃, 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, etc.;

[0131] Wavelengths can be 250nm, 251nm, 252nm, 253nm, 254nm, 255nm, 256nm, 257nm, 258nm, 259nm, 260nm, etc.

[0132] The injection volume can be 10 μL, 11 μL, 12 μL, 13 μL, 14 μL, 15 μL, 16 μL, 17 μL, 18 μL, 19 μL, 20 μL, etc.

[0133] In some embodiments, the content of ellagic acid in the prickly pear root extract is 8.00-20.00 mg / g, preferably 8.20-18.80 mg / g.

[0134] For example, the content of ellagic acid in the prickly pear root extract can be 8 mg / g, 8.20 mg / g, 9 mg / g, 10 mg / g, 11 mg / g, 12 mg / g, 13 mg / g, 14 mg / g, 15 mg / g, 16 mg / g, 17 mg / g, 18 mg / g, 19 mg / g, 20 mg / g, etc.

[0135] In this application, the chromatographic column is a chromatographic column with octadecylsilane-bonded silica gel as the packing material (column length 150 mm, inner diameter 2.1 mm, particle size 1.8 μm).

[0136] This application also provides a method for detecting the ellagic acid transfer rate in Rosa rugosa root extract, comprising:

[0137] (1) Preparation of reference solution

[0138] Weigh out ellagic acid reference standard and add it to methanol to prepare a reference solution;

[0139] (2) Preparation of the test solution

[0140] Weigh out the root extract of Rosa rugosa and add it to the first solvent for extraction;

[0141] (3) Preparation of control solution of prickly pear root slices

[0142] Weigh out the prickly pear root slices and add the first solvent for extraction;

[0143] (4) High performance liquid chromatography analysis

[0144] The transfer rate of ellagic acid in the prickly pear root extract was obtained by high performance liquid chromatography analysis of the reference solution, the test solution, and the prickly pear root slices control solution.

[0145] In some embodiments of this application, the first solvent is selected from one or more of 25% ethanol, 50% ethanol, 75% ethanol, 100% ethanol, 25% methanol, 50% methanol, 75% methanol, 100% methanol and water, preferably water;

[0146] Preferably, the volume-to-mass ratio of the first solvent to the prickly pear root extract is 100-2000 mL / g, more preferably 500-1500 mL / g. For example, based on the mass of the prickly pear extract being 0.1 g, the volume of the solvent can be 10 mL, 20 mL, 30 mL, 40 mL, 60 mL, 80 mL, 100 mL, 150 mL, etc.

[0147] In some embodiments of this application, the extraction is performed using either reflux or ultrasound, preferably reflux extraction;

[0148] Preferably, the extraction time is 0.5-2 hours, and more preferably 1-2 hours.

[0149] It should be noted that the extraction conditions in step (3) are the same as those in step (2). For example, the root slices of prickly pear can be added to water and heated and refluxed for extraction.

[0150] In some embodiments of this application, the chromatographic conditions for high performance liquid chromatography analysis in step (4) are as follows: using octadecylsilane-bonded silica gel as the packing material, and using mobile phase A and mobile phase B for gradient elution; preferably, mobile phase A is methanol or acetonitrile, preferably methanol; mobile phase B is water, 0.1% phosphoric acid solution, 0.1% formic acid solution or 0.1% acetic acid solution, preferably 0.1% phosphoric acid solution;

[0151] More preferably, the gradient elution conditions are as follows: from 0 to 15 min, mobile phase A is 15 vol% → 38 vol%, mobile phase B is 85 vol% → 62 vol%, and from 15 to 40 min, mobile phase A is 38 vol%, mobile phase B is 62 vol%.

[0152] In some embodiments, the flow rate of the mobile phase is 0.2-0.3 ml / min;

[0153] Preferably, the column temperature is 30-40℃;

[0154] Preferably, the detection wavelength is 250-260 nm, and more preferably 254 nm;

[0155] Preferably, the injection volume is 10-20 μL.

[0156] For example, the flow rate of the mobile phase can be 0.2 ml / min, 0.25 ml / min, 0.3 ml / min, etc.;

[0157] The column temperature can be 30℃, 31℃, 32℃, 33℃, 34℃, 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, etc.;

[0158] Wavelengths can be 250nm, 251nm, 252nm, 253nm, 254nm, 255nm, 256nm, 257nm, 258nm, 259nm, 260nm, etc.

[0159] The injection volume can be 10 μL, 11 μL, 12 μL, 13 μL, 14 μL, 15 μL, 16 μL, 17 μL, 18 μL, 19 μL, 20 μL, etc.

[0160] In some embodiments, the transfer rate of ellagic acid in the prickly pear root extract is 20.00-99.00%, preferably 20.38-98.32%.

[0161] For example, the transfer rate of ellagic acid in the prickly pear root extract can be 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, etc.

[0162] In this application, the chromatographic column is a chromatographic column with octadecylsilane-bonded silica gel as the packing material (column length 150 mm, inner diameter 2.1 mm, particle size 1.8 μm).

[0163] This application also provides a method for constructing a characteristic map of a Rosa rugosa root extract, comprising the following steps:

[0164] (1) Preparation of reference solution

[0165] Weigh out ellagic acid reference standard and add it to methanol to prepare a reference solution;

[0166] (2) Preparation of the test solution

[0167] Weigh out the root extract of Rosa rugosa and add a second solvent for extraction;

[0168] (3) High performance liquid chromatography analysis

[0169] The characteristic chromatograms of the prickly pear root extract were obtained by high performance liquid chromatography analysis of the reference solution and the test solution.

[0170] In some embodiments of this application, the second solvent is selected from one or more of 25% ethanol, 50% ethanol, 75% ethanol, 100% ethanol, 25% methanol, 50% methanol, 75% methanol, 100% methanol and water, preferably water; preferably, the solvent in step (2) is selected from one or more of 25% ethanol, 50% ethanol, 75% ethanol, 100% ethanol, 25% methanol, 50% methanol, 75% methanol, 100% methanol and water, preferably water;

[0171] Preferably, the volume-to-mass ratio of the second solvent to the prickly pear root extract is 100-2000 mL / g, more preferably 500-1500 mL / g. For example, based on the mass of the prickly pear extract being 0.1 g, the volume of the solvent can be 10 mL, 20 mL, 30 mL, 40 mL, 60 mL, 80 mL, 100 mL, 150 mL, etc.

[0172] In some embodiments of this application, the extraction in step (2) is performed by either reflux or ultrasound, preferably reflux extraction;

[0173] Preferably, the extraction time in step (2) is 0.5-2 hours, and more preferably 1-2 hours.

[0174] In some embodiments of this application, the chromatographic conditions for high performance liquid chromatography analysis in step (3) are as follows: using octadecylsilane-bonded silica gel as the packing material, and using mobile phase A and mobile phase B for gradient elution; preferably, mobile phase A is methanol or acetonitrile, preferably methanol; mobile phase B is water, 0.1% phosphoric acid solution, 0.1% formic acid solution or 0.1% acetic acid solution, preferably 0.1% phosphoric acid solution;

[0175] More preferably, the gradient elution conditions are as follows: from 0 to 15 min, mobile phase A is 15 vol% → 38 vol%, mobile phase B is 85 vol% → 62 vol%, and from 15 to 40 min, mobile phase A is 38 vol%, mobile phase B is 62 vol%.

[0176] In some embodiments, the flow rate of the mobile phase is 0.2-0.3 ml / min;

[0177] Preferably, the column temperature is 30-40℃;

[0178] Preferably, the detection wavelength is 250-260 nm, and more preferably 254 nm;

[0179] Preferably, the injection volume is 10-20 μL.

[0180] For example, the flow rate of the mobile phase can be 0.2 ml / min, 0.25 ml / min, 0.3 ml / min, etc.;

[0181] The column temperature can be 30℃, 31℃, 32℃, 33℃, 34℃, 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, etc.;

[0182] Wavelengths can be 250nm, 251nm, 252nm, 253nm, 254nm, 255nm, 256nm, 257nm, 258nm, 259nm, 260nm, etc.

[0183] The injection volume can be 10 μL, 11 μL, 12 μL, 13 μL, 14 μL, 15 μL, 16 μL, 17 μL, 18 μL, 19 μL, 20 μL, etc.

[0184] In this application, the chromatographic column is a chromatographic column with octadecylsilane-bonded silica gel as the packing material (column length 150 mm, inner diameter 2.1 mm, particle size 1.8 μm).

[0185] In some embodiments, the characteristic spectrum of the prickly pear root extract contains seven characteristic peaks. Preferably, the characteristic peaks are those at retention times of 0.14-0.16, 0.67-0.71, 0.83-0.87, 0.90-0.92, 0.96-0.98, 1.00, and 1.19-1.24, and more preferably, those at retention times of 0.15, 0.69, 0.84, 0.91, 0.97, 1.0, and 1.21.

[0186] The beneficial effects of the present invention will be further illustrated below through specific embodiments.

[0187] All raw materials or reagents used in this invention are purchased from mainstream manufacturers on the market. Those without specified manufacturers or concentrations are all analytical grade raw materials or reagents that can be obtained routinely. There are no special restrictions as long as they can achieve the expected effect.

[0188] Unless otherwise specified in this embodiment, the techniques or conditions described in the literature in this field or in accordance with the product manual shall apply.

[0189] The present invention will now be described in more detail using examples and comparative examples, but the scope of the invention is not limited to these examples. It should be noted that, unless otherwise specified, all percentages, parts, and ratios used in the present invention are based on mass. Information on the raw materials and testing equipment used in the examples and comparative examples is shown in Tables 1 and 2, respectively.

[0190] Table 1. Information on raw materials used in this invention.

[0191] Raw material name Purity / Batch Number Origin prickly pear root YP220316-1 Huaxi District, Guiyang City, Guizhou Province prickly pear root YP220316-2 Wantanhe Town, Longli County, Guiyang City, Guizhou Province prickly pear root YP220316-3 Shihuo Town, Renhuai City, Zunyi City, Guizhou Province prickly pear root YP220316-4 Bulang Township, Puding County, Anshun City, Guizhou Province prickly pear root YP220316-5 Shuangliu Town, Kaiyang County, Guiyang City, Guizhou Province prickly pear root YP220316-6 Xiuwen County, Guiyang City, Guizhou Province prickly pear root YP220316-7 Shihuo Town, Renhuai City, Zunyi City, Guizhou Province prickly pear root YP220316-8 Wantanhe Town, Longli County, Guiyang City, Guizhou Province prickly pear root YP220316-9 Liuchang Town, Qingzhen City, Guiyang City, Guizhou Province prickly pear root YP220316-10 Shuangliu Town, Kaiyang County, Guiyang City, Guizhou Province prickly pear root YP220316-11 Jiangkou County, Tongren City, Guizhou Province prickly pear root YP220316-12 Wantanhe Town, Longli County, Guiyang City, Guizhou Province prickly pear root YP220316-13 Liuchang Town, Qingzhen City, Guiyang City, Guizhou Province prickly pear root YP220316-14 Liuchang Town, Qingzhen City, Guiyang City, Guizhou Province prickly pear root YP220316-15 Wantanhe Town, Longli County, Guiyang City, Guizhou Province Ellagic acid 111959-201903 China National Institutes for Food and Drug Control

[0192] Table 2 Information on the experimental equipment used in this invention

[0193]

[0194]

[0195] The solids content of the concentrate in the following examples was determined using the following method:

[0196] According to the "Hot Extraction Method" of General Chapter 2201 Extraction Determination Method in Part IV of the 2020 Chinese Pharmacopoeia: accurately weigh 10g of concentrated solution, place it in a pre-weighed evaporating dish, evaporate it to dryness on a water bath, dry it at 105℃ for 3 hours, cool it in a desiccator for 30 minutes, quickly and accurately weigh it, and calculate the solid content of the concentrated solution.

[0197] Example 1: Preparation of the extract

[0198] Optimization of the preparation process:

[0199] (1) Processing of medicinal slices: Take the raw medicinal material, remove impurities, and cut it into irregular slices of 3-5cm.

[0200] (2) Investigation on dosage of medicinal slices

[0201] According to the recommendations under "Standard Decoction Preparation" in the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules," the amount of medicinal slices used per decoction should generally not be less than 100g, with the amount for flowers, leaves, and other medicinal slices potentially reduced. Based on preliminary test results, 100g of Rosa rugosa root slices is convenient to handle, and the resulting dry extract is sufficient for experimental research on the extract; therefore, the amount of medicinal slices used in Rosa rugosa root extract is set at 100g.

[0202] (3) Water addition assessment

[0203] According to the recommendations under "Extract Preparation" in the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules," the amount of water added should generally be 2-5 cm above the surface of the herbs. For flower and herbaceous herbs or herbs requiring a longer decoction time, the amount of water added can be adjusted accordingly. When adding 8 times the weight of the herbs in water for the first decoction to meet the solvent requirement of "2-5 cm above the surface of the herbs," and adding 6 times the weight of the herbs in water for the second decoction, the water level exceeds 2 cm. Therefore, the final water addition is determined to be 8 times the weight of the herbs in water for the first decoction and 6 times the weight of the herbs in water for the second decoction.

[0204] (4) Investigation of soaking time

[0205] According to the "Preparation of Standard Decoctions" section of the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Granules," the medicinal slices to be decocted should be soaked first, with a soaking time generally not less than 30 minutes. Based on the "Technical Requirements," the soaking time for Rosa rugosa root extract is determined to be 30 minutes.

[0206] (5) Examination of cooking time

[0207] According to the recommendations under "Preparation of Standard Decoctions" in the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules," each dose of medicine is generally decocted twice. Generally, after boiling, it should be decocted for 30 minutes. For diaphoretic, heat-clearing, and aromatic herbs, prolonged decoction is not advisable; 20 minutes after boiling is sufficient. For harder herbs, the decoction time can be appropriately extended. For tonifying herbs, first bring to a boil over high heat, then simmer over low heat for about 60 minutes. The second decoction time can be appropriately shortened. Therefore, based on the "Technical Requirements," the extract of *Smilax china* is decocted twice: the first decoction is boiled for 60 minutes after boiling, and the second decoction is boiled for 40 minutes after boiling.

[0208] (6) Investigation of solid-liquid separation conditions

[0209] According to the "Standard Decoction Preparation" section of the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules," the mesh size of the filter material for solid-liquid separation should be above 100 mesh. To obtain a decoction consistent with traditional decoctions using modern solid-liquid separation methods, the filtration effects of 100-mesh, 200-mesh, and 300-mesh sieves on the decoction of *Rhizoma Rosa rugosa* root were mainly investigated. The results showed that filtration was relatively easy using 100-mesh, 200-mesh, and 300-mesh sieves. However, the filtrates obtained through filtration using 100-mesh and 200-mesh sieves had poor clarity and a larger amount of sediment after settling. The filtrate obtained through filtration using a 300-mesh sieve had better clarity and a smaller amount of sediment after settling. Therefore, a 300-mesh sieve was used for solid-liquid separation of *Rhizoma Rosa rugosa* root extract.

[0210] (7) Study on concentration process parameters

[0211] According to the "Preparation of Standard Decoctions" section of the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules," it is recommended to use a vacuum concentration method for low-temperature concentration to obtain the specified amount of extract. By comparing different concentration temperatures, a suitable concentration temperature is determined, and a concentration ratio of 1:1 is tentatively set. The concentration state is then observed.

[0212] The density, yield, and state of the concentrated extract of *Rhizoma Rosa rugosa* root extract were compared and measured at different concentration temperatures. The results showed that the yield was basically consistent across different concentration temperatures, with no significant difference between 50℃ and 65℃. Considering all factors, 65℃ was chosen as the optimal concentration temperature. When the concentration ratio was 1:1, the extract exhibited moderate viscosity, good fluidity, and ease of transfer, with densities ranging from 1.01 to 1.03 g / ml. Therefore, a concentration ratio of 1:1 was determined, and other concentration ratios were not considered.

[0213] (8) Study on freeze-drying process parameters

[0214] According to the "Preparation of Standard Decoctions" section of the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules," freeze-drying is generally recommended for the drying of standard decoctions to ensure quality stability, ease of dissolution, and the elimination of excipients. Therefore, freeze-drying was initially chosen as the drying method for Rosa rugosa root extract.

[0215] Under magnetic stirring, the extract was dispensed into 5ml brown vials, each with a volume of 2ml. The vials were partially capped, and after dispensing, they were transferred to a vacuum freeze dryer for freeze-drying. The vials were then removed, capped with aluminum, and the final product was obtained. The eutectic point of the *Rhizophora stylosa* root under vacuum freeze-drying was -43.6℃, ​​the pre-freezing temperature was -45℃, the sublimation drying temperature was -30℃ to 0℃, the desorption drying temperature was 5℃ to 25℃, and the total drying time was 28 hours. The results indicate that under these freeze-drying conditions, the standard *Rhizophora stylosa* root decoction has low moisture content and is easy to preserve. Therefore, the freeze-drying process parameters were determined as follows: Under magnetic stirring, the product was dispensed into 10ml brown vials, each with a volume of 2ml, half-stopped, and then transferred to a vacuum freeze dryer for freeze-drying. The freeze-drying parameters were as follows: the pre-freezing temperature of the prickly pear root extract concentrate was -45℃, the pre-freezing time was 240 minutes, the sublimation drying temperature was -30℃~0℃, the sublimation drying time was 1185 minutes, and the vacuum degree was -0.2mbar; the desorption drying temperature was 5℃~25℃, the desorption drying time was 445 minutes, and the vacuum degree was -0.2mbar.

[0216] The optimized preparation method of the extract is as follows:

[0217] Take 100g of prickly pear root slices, place them in an electric ceramic kettle, add water and decoct twice. For the first decoction, add 8 times the amount of water, soak for 30 minutes, bring to a boil over high heat (500W), then simmer over low heat (200W) for 60 minutes. Filter the decoction while hot through a 300-mesh sieve, and record the mass of the decoction after cooling to room temperature. For the second decoction, add 6 times the amount of water, heat to a boil over high heat, then simmer over low heat for 40 minutes. Filter the decoction while hot through a 300-mesh sieve, and record the mass of the decoction after cooling to room temperature. Combine the two decoctions. Transfer the decoction to a 2000ml round-bottom flask and concentrate under reduced pressure at low temperature using a rotary evaporator (temperature: 65℃; vacuum: -0.080~).

[0218] Extracts ranging from -0.090 MPa to 100 ml were prepared. Under magnetic stirring, the extract was dispensed into 5 ml brown vials, each with a volume of 2 ml. The vials were partially capped and then transferred to a vacuum freeze dryer for lyophilization. The lyophilization parameters were as follows: pre-freezing temperature of the prickly pear root concentrate was -45℃, pre-freezing time was 240 minutes; sublimation drying temperature was -30℃~0℃, sublimation drying time was 1185 minutes, and vacuum degree was -0.2 mbar; desorption drying temperature was 5℃~25℃, desorption drying time was 445 minutes, and vacuum degree was -0.2 mbar.

[0219] The preparation process was then validated.

[0220] Three batches of Rosa rugosa root slices (batch numbers: YP220316-1, YP220316-2, YP220316-3) were taken, with three replicates per batch and 100g per replicate. Rosa rugosa root extract was prepared according to the optimized preparation method described above. The yield of the extract from the three batches was determined to evaluate the stability and rationality of the extract preparation process. The results are shown in Table 3 below.

[0221] Table 3. Validation results of the preparation method of Rosa rugosa root extract

[0222]

[0223]

[0224] The experimental results show that the specific gravity, solid content and yield of the three parallel batches of prickly pear root extract are basically the same, indicating that the process is stable and reproducible and can be used as a preparation process for prickly pear root extract.

[0225] In addition, 15 batches of *Rhizophora stylosa* root extract were prepared according to the optimized preparation method described in the examples, and the verification results are shown in Table 4.

[0226] Table 4. Validation results of the preparation method of 15 batches of Rosa rugosa root extract

[0227]

[0228] The yield of prickly pear root extract from 15 batches varied from 4.03% to 6.48%, with a mean of 5.26% and a standard deviation (SD) of 0.85%. The range of 70% to 130% of the mean yield was 3.68% to 6.84%, and the ±3 SD range was 2.71% to 7.81%. Therefore, the optimal yield range for prickly pear root extract was determined to be 2.71% to 7.81% of the mean yield ±3 SD, approximately rounded to 2.7% to 7.8%.

[0229] Example 2: Method for determining the ellagic acid content in prickly pear root extract

[0230] 1. Determination of chromatographic conditions

[0231] Chromatographic conditions and system suitability tests were performed using octadecylsilane-bonded silica gel as the packing material (column length 150 mm, inner diameter 2.1 mm, particle size 1.8 μm); methanol as mobile phase A and 0.1% phosphoric acid solution as mobile phase B, with gradient elution as specified in Table 5; flow rate 0.20 mL / min; column temperature 35 °C; detection wavelength 254 nm. The theoretical plate number, calculated based on the ellagic acid peak, should be no less than 3000.

[0232] Table 5 Gradient elution conditions

[0233] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0～15 15→38 85→62 15～40 38→38 62→62

[0234] 2. Preparation of reference solution

[0235] Take an appropriate amount of ellagic acid reference standard, accurately weigh it, and add methanol to prepare a solution containing 18 μg of ellagic acid per 1 ml, which will be used as the reference solution.

[0236] 3. Optimization of preparation conditions for the test solution:

[0237] (1) Investigation of different extraction solvents

[0238] Take an appropriate amount of Rosa roxburghii root extract BT0316-1, a total of 9 portions (approximately 0.1g per portion), with 2 parallel samples per portion. Place each sample in a stoppered conical flask, and accurately add 20ml of 100% methanol, 75% methanol, 50% methanol, 25% methanol, 100% ethanol, 75% ethanol, 50% ethanol, 25% ethanol, and water sequentially. Weigh the samples, sonicate (500W power, 40kHz frequency) for 30 minutes, cool, weigh again, replenish the lost weight with the appropriate solvent, shake well, filter, and collect the filtrate to obtain the test solutions. Under the above chromatographic conditions, accurately inject 1μl each of the reference solution and the test solution into an ultra-high performance liquid chromatograph for determination. Calculate the effect of different extraction solvents on the ellagic acid content, and determine the optimal extraction solvent. The experimental results are calculated using the external standard one-point method. Detailed results are shown in Table 6.

[0239] Table 6. Effects of different extraction solvents on the content determination results of Rosa rugosa root extract.

[0240]

[0241] The experimental results show that different solvents have a significant impact on the content of ellagic acid in the root extract of Rosa rugosa. Taking into account the differences in content, water was selected as the best extraction solvent for further research.

[0242] (2) Examination of extraction methods

[0243] Two portions (approximately 0.1 g each) of Rosa rugosa root extract BT0316-1 were prepared, with two parallel samples per portion. Each sample was placed in a stoppered conical container, and 20 ml of water was precisely added sequentially. The samples were weighed, and each was ultrasonicated or refluxed for 30 min. After cooling, the samples were weighed again, and the lost weight was replenished with water. The mixture was shaken well, filtered, and the filtrate was collected to obtain the test solutions. Under the chromatographic conditions described above, 1 μl of each of the reference solution and the test solution were precisely injected into an ultra-high performance liquid chromatograph for determination. The content of ellagic acid was calculated based on the effect of different extraction methods, and the optimal extraction method was determined. The experimental results were calculated using the external standard one-point method. Detailed results are shown in Table 7.

[0244] Table 7. Effects of different extraction methods on the ellagic acid content in Rosa rugosa root extract.

[0245]

[0246] The experimental results show that different extraction methods have a significant impact on the content of ellagic acid in the root extract of Rosa rugosa. Considering the differences in content, and taking into account the experimental conditions of our laboratory and the ease of operation in practice, heating reflux extraction was selected as the extraction method for further research.

[0247] (3) Investigation of the volume of the extraction solvent

[0248] Take an appropriate amount of Rosa roxburghii root extract BT0316-1, a total of 8 portions (approximately 0.1g per portion), with 2 parallel samples per portion. Place each sample in a stoppered conical container, and accurately add 10ml, 20ml, 30ml, 40ml, 60ml, 80ml, 100ml, and 150ml of water sequentially. Weigh the samples and sonicate them (500W power, 40kHz frequency) for 30 minutes. After cooling, weigh them again and replenish the lost weight with water. Take an appropriate amount of the filtrate and filter it through a 0.22μm microporous membrane. Place the filtered solution in a high-performance liquid sample bottle to obtain the test solutions. Under the above chromatographic conditions, accurately inject 1μl each of the reference solution and the test solution into an ultra-high performance liquid chromatograph for determination. Calculate the effect of different extraction times on the ellagic acid content to determine the optimal extraction time. The experimental results are calculated using the external standard one-point method. Detailed results are shown in Table 8.

[0249] Table 8. Effect of different extraction solvent volumes on the ellagic acid content in Rosa rugosa root extract.

[0250]

[0251]

[0252] The experimental results show that the volume of different extraction solvents has a significant effect on the ellagic acid content in the prickly pear root extract. Considering the cost of the solvent and the difference in its content, an extraction volume of 100 ml was selected for further research.

[0253] (4) Examination of different extraction times

[0254] Take an appropriate amount of Rosa roxburghii root extract BT0316-1, a total of 4 portions (approximately 0.1g per portion), with 2 parallel samples per portion. Place each sample in a stoppered conical container, and accurately add 100ml of water sequentially. Weigh the samples, and heat under reflux for 0.5 hours, 1 hour, 1.5 hours, and 2 hours respectively. After cooling, weigh the samples again, and replenish the lost weight with water. Take an appropriate amount of the filtrate and filter it through a 0.22μm microporous membrane. Place the filtered solution in a high-performance liquid sample bottle to obtain the test solution for later use. Under the above chromatographic conditions, accurately pipette 1μl each of the reference solution and the test solution, and inject them into the ultra-high performance liquid chromatograph for determination. Calculate the effect of different extraction times on the ellagic acid content, and determine the optimal extraction time. The experimental results are calculated using the external standard one-point method. Detailed results are shown in Table 9.

[0255] Table 9. Effects of different extraction times on the ellagic acid content in Rosa rugosa root extract.

[0256]

[0257] The experimental results show that different extraction times have a significant effect on the ellagic acid content in the prickly pear root extract. Considering both time cost and content differences, reflux for 2 hours was chosen for further research.

[0258] (5) Determination of the preparation method of the test solution

[0259] Based on the results of the sample pretreatment experiment, the preparation method of the test sample can be determined as follows: Take about 0.1g of prickly pear root extract, accurately weigh it, place it in a stoppered conical flask, accurately add 100ml of water, weigh it, heat under reflux for 2h, cool it, weigh it again, replenish the lost weight with water, shake it well, filter it, and take the filtrate to obtain the test sample.

[0260] 4. Methodological Validation

[0261] (1) Specificity examination

[0262] Accurately pipette 1 μl each of the BT0316-1 test solution of prickly pear root extract, the ellagic acid reference solution, and the negative sample (using water as a blank solvent). Perform chromatographic analysis under the conditions described above and record the chromatograms. Results are as follows: Figure 1 As shown.

[0263] Depend on Figure 1 It can be seen that this analytical method is specific for the determination of ellagic acid content in prickly pear root extract.

[0264] (2) Peak purity

[0265] Accurately pipette 1 μl each of the test solution and reference solution of prickly pear root extract (BT0316-1) into the ultra-high performance liquid chromatograph and determine the peak purity of the target peak. The results are as follows: Figure 2 As shown in Table 10.

[0266] Table 10 Matching values ​​for target peaks and peak purity

[0267] Target Peak Ellagic acid Peak purity matching value 999.96

[0268] The experimental results show that the peak purity matching value of the components in this index is greater than 960, which indicates that the peak purity meets the analytical requirements.

[0269] (3) Examination of linear relationships

[0270] Accurately weigh an appropriate amount of ellagic acid reference standard and place it in a numbered 50 ml volumetric flask. Add methanol to prepare a solution containing 55.55328 μg of ellagic acid per ml. Shake well to obtain the ellagic acid reference standard stock solution, which should be stored in a refrigerator for later use. Dilute the ellagic acid reference standard stock solution by 1.25, 1.56, 1.95, 2.44, 3.05, 6.10, 12.21, 24.41, 48.83, and 97.66 times to obtain ellagic acid reference standard solutions of different concentrations. Perform chromatographic analysis under the above conditions. Plot the concentration on the x-axis and the peak area on the y-axis to investigate the linear range of ellagic acid. The linearity results are shown in Table 11 and [Table data missing]. Figure 3 As shown.

[0271] Table 11 Linearity Study of Ellagic Acid

[0272] serial number Ellagic acid concentration (μg / ml) Peak area (mAU) Ellagic acid-1 55.5533 3738.414 Ellagic acid-2 44.4426 2913.344 Ellagic acid-3 35.5541 2447.610 Ellagic acid-4 28.4433 1906.486 Ellagic acid-5 22.7546 1512.932 Ellagic acid-6 18.2037 1232.147 Ellagic acid-7 9.1018 635.890 Ellagic acid-8 4.5509 301.366 Ellagic acid-9 2.2755 158.205 Ellagic acid-10 1.1377 74.703 Ellagic acid-11 0.5689 35.212

[0273] Experimental results showed that ellagic acid concentration and peak area exhibited a good linear relationship within the concentration range of 0.5689 μg / ml to 55.5533 μg / ml, with a correlation coefficient R0. 2 =0.9994, the linear regression equation is: y = 66.923x + 5.4681.

[0274] (4) Precision test

[0275] Accurately pipette 1 μl each of the test solution of prickly pear root extract (batch number: BT0316-1) and the ellagic acid reference solution into an ultra-high performance liquid chromatograph and determine the results. The RSD (%) value of the target peak was calculated using the external standard one-point method, with ellagic acid as the criterion. The specific results are shown in Table 12.

[0276] Table 12 Precision test results of the method for determining the content of Rosa rugosa root extract

[0277]

[0278] The results show that the RSD (%) of the target peak ellagic acid is 0.20% < 1.5%, indicating that the method has good precision.

[0279] (5) Repeatability test

[0280] Accurately weigh approximately 0.1 g of the same batch of Rosa rugosa root extract (BT0316-1), prepare six parallel solutions according to the above-described method for preparing test solutions, and set them aside. Perform chromatographic analysis under the above-described conditions. Calculate the RSD (%) of the target peak content using the external standard single-point method, based on ellagic acid content in the test solutions. The specific results are shown in Table 13.

[0281] Table 13 Results of repeatability tests for the determination of the content of Rosa rugosa root extract.

[0282]

[0283] The experimental results show that the RSD (%) of the target peak ellagic acid content is 1.26% < 1.5%, indicating that the method has good repeatability.

[0284] (6) Intermediate precision

[0285] Other analysts in this project team operated on different dates and under different chromatographs, taking approximately 0.1 g of the same batch of Rosa rugosa root extract (BT0316-1), accurately weighed, and prepared the test solution according to the above-mentioned test solution preparation method. The test solution was then analyzed under the above-mentioned chromatographic conditions. The RSD (%) value of the target peak content was calculated using the external standard one-point method, based on ellagic acid in the test solution. The specific results are shown in Table 14.

[0286] Table 14 Results of intermediate precision test for the determination of content in Rosa rugosa root extract

[0287]

[0288]

[0289] The experimental results show that the intermediate precision RSD (%) value of the target peak ellagic acid content is 1.78% < 3%, indicating that the method has good precision.

[0290] (7) Accuracy test

[0291] Six portions of 0.1 g of *Rhizoma Rosa roxburghii* root extract (batch number BT0316-1, ellagic acid content 16.09 mg / g) with known content were accurately weighed. 9.2 ml of ellagic acid reference solution (0.086802 mg / ml) prepared in methanol was accurately added, and the mixture was evaporated to dryness. 100 ml of water was then accurately added. The weights were determined, and the test solution was prepared according to the method described above. The solution was then analyzed by chromatographic analysis under the conditions described above, with 1 μl injected into each sample. The chromatograms were recorded. The content of the target peak was calculated using the external standard single-point method, and the recovery rate and relative standard deviation were calculated according to the following formulas. The results are shown in Table 15.

[0292]

[0293] Table 15. Determination of content in Rosa rugosa root extract; Results of ellagic acid recovery experiment.

[0294]

[0295] The experimental results show that the recovery rate of ellagic acid in the root extract of Rosa rugosa is within the range of 98%-101%, and the RSD% (0.92%) is less than 1.5%, indicating that the accuracy of the assay method is good.

[0296] (8) Stability test

[0297] The test solution of prickly pear root extract (batch number BT0316-1) was prepared according to the preparation method of the test solution. The sample was injected at 0, 2, 4, 8, 12, 16, 20 and 24 hours according to the above chromatographic conditions, with an injection volume of 1 μl. The RSD (%) of the target peak content was calculated using the external standard one-point method based on ellagic acid. The specific results are shown in Table 16.

[0298] Table 16. Stability test results of the method for determining the content of Rosa roxburghii root extract.

[0299]

[0300] Experiments showed that the RSD (%) of the target peak ellagic acid content within 24 hours was 0.79% < 1.5%, indicating that the solution had good stability within 24 hours.

[0301] (9) Durability test

[0302] 1) Investigation of different chromatographic columns

[0303] The effects of three chromatographic columns—Endeavorsil C18 (2.1*150mm, 1.8μm), ACQUITY UPLCHSS T3C18 (2.1*150mm, 1.8μm), and Infintylab Poroshell 120Aq-C18 (2.1*150mm, 2.7μm)—on the peak shape and resolution of ellagic acid in Rosa rugosa root extract were compared. A test solution of Rosa rugosa root extract (batch number BT0316-1) was prepared, and the chromatographic data were recorded as ellagic acid, calculated using the above chromatographic conditions. The experimental results are shown in Table 17.

[0304] Table 17 Effect of different chromatographic columns on the determination results of the content of prickly pear root extract.

[0305]

[0306]

[0307] Experimental results show that the resolution and theoretical plate number of the Endeavorsil C18 (2.1*150mm, 1.8μm) column are superior to those of the ACQUITY UPLCHSS T3 C18 (2.1*150mm, 1.8μm) and Inertsil ODS-3 (2.1×150mm, 2μm) columns. Therefore, the Endeavorsil C18 (2.1*150mm, 1.8μm) column was selected as the preferred column for this experiment.

[0308] 2) Investigation using different chromatographs

[0309] Based on the existing equipment in the laboratory, Thermo Fisher Ultra High Performance Liquid Chromatography (Water, ACQUITY H class) and Agilent High Performance Liquid Chromatography (Agilent, 1290 Infinity II) were selected to compare the effects of the two chromatographic instruments on the peak shape and resolution of ellagic acid in Rosa rugosa root extract.

[0310] A test solution was prepared from the root extract of Rosa roxburghii (batch number (BT0316-1)). The chromatographic data were determined under the conditions described above, and the result was calculated as ellagic acid. The experimental results are shown in Table 18.

[0311] Table 18 Results of the investigation using different chromatographs

[0312]

[0313] Experimental results show that this analytical method exhibits good durability with different chromatographs. Variations in the chromatograph can meet the system adaptability requirements.

[0314] 3) Investigation at different column temperatures

[0315] The effects of different column temperatures (20℃, 25℃, and 30℃) on the peak shape of ellagic acid in *Rhizoma Rosa rugosa* root extract were compared. A test solution was prepared from *Rhizoma Rosa rugosa* root extract and analyzed under the chromatographic conditions described above. The chromatographic data were recorded as ellagic acid. The experimental results are shown in Table 19.

[0316] Table 19 Results of the determination of Rosa rugosa root extract at different column temperatures

[0317] Column temperature Indicator Components Resolution / R Theoretical number of plates 30℃ Ellagic acid 6.70 99644 35℃ Ellagic acid 2.80 92435 40℃ Ellagic acid 1.74 116005

[0318] The results showed that the peak shape and separation were good at all three column temperatures. At 35℃, the baseline of the chromatogram showed no drift, and the retention time was not significantly different compared to the other two temperatures. Considering the column's tolerance and the analysis time required, a column temperature of 35℃ was chosen.

[0319] 4) Investigation of different flow velocities

[0320] The effects of different flow rates of 0.20 ml / min, 0.25 ml / min, and 0.30 ml / min on the peak shape and resolution of ellagic acid in Rosa rugosa root extract were compared.

[0321] A test solution was prepared from the root extract of Rosa roxburghii (batch number: BT0316-1). The chromatographic data were determined under the chromatographic conditions described above and recorded as ellagic acid. The experimental results are shown in Table 20.

[0322] Table 20 Results of the determination of Rosa rugosa root extract by different flow rates

[0323] Flow rate Indicator Components Resolution / R Theoretical number of plates 0.20 ml / min Ellagic acid 2.80 92435 0.25ml / min Ellagic acid 2.22 106134 0.30ml / min Ellagic acid 1.79 116156

[0324] The results showed that the peak shape and separation were good at all three flow rates. The target component showed good separation with no baseline drift at a flow rate of 0.20 ml / min. Therefore, a flow rate of 0.20 ml / min was selected for this experiment.

[0325] Example 3: Determination of ellagic acid content and transfer rate in prickly pear root extract

[0326] (1) Preparation of reference solution

[0327] Take an appropriate amount of ellagic acid reference standard, accurately weigh it, and add methanol to prepare a solution containing 18 μg of ellagic acid per 1 ml, which will be used as the reference solution.

[0328] (2) Preparation of the test solution

[0329] Take about 0.1g of prickly pear root extract, accurately weigh it, place it in a stoppered conical flask, accurately add 100ml of water, weigh it, heat under reflux for 2 hours, cool it, weigh it again, add water to make up the weight loss, shake well, filter it, and take the filtrate to obtain the extract.

[0330] (3) Preparation of test solution of prickly pear root slices

[0331] Take about 1g of prickly pear root slices, weigh them accurately, place them in a stoppered conical flask, add 100ml of water accurately, weigh them, heat under reflux for 2 hours, cool them, weigh them again, add water to make up the weight loss, shake well, filter, and collect the filtrate to obtain the product.

[0332] (4) High performance liquid chromatography analysis

[0333] The column was packed with octadecylsilane-bonded silica gel (Endeavorsil C18, 150 mm in length, 2.1 mm in inner diameter, and 1.8 μm in particle size); methanol was used as mobile phase A, and 0.1% phosphoric acid solution was used as mobile phase B, with gradient elution performed according to the specifications in Table 5; the flow rate was 0.20 mL / min; the column temperature was 35 °C; and the detection wavelength was 254 nm. The theoretical plate number, calculated based on the ellagic acid peak, should be no less than 3000.

[0334] (5) The mass content and transfer rate of ellagic acid are calculated according to the following formula:

[0335] in,

[0336] w 鞣花酸 =A 供 ×C 对 ×V 供 ×10-3 / (m 供 ×A 对 )

[0337] w 饮 =A 饮供 ×C 对 ×V 饮供 ×10 -3 / (m 饮供 ×A 对 )

[0338] Ellagic acid transfer rate = (m 提 ×w 对羟基苯乙酮 ) / (m 饮 ×w 饮 )

[0339] In the formula:

[0340] w 鞣花酸 —The mass content of ellagic acid in the root extract of Rosa rugosa, mg / g;

[0341] w 饮 —The ellagic acid content in the prickly pear root slices, mg / g;

[0342] A 对 —The absorption peak area of ​​ellagic acid in the high performance liquid chromatography spectrum of the reference solution;

[0343] A 供 —The absorption peak area of ​​ellagic acid in the high performance liquid chromatography spectrum of the test solution;

[0344] A 饮供 —The absorption peak area of ​​ellagic acid in the high performance liquid chromatography spectrum of the sample solution;

[0345] C 对 —The concentration of the reference solution, in μg / mL;

[0346] m 供 —The oven-dry mass of the prickly pear root extract in the test solution, in g;

[0347] m 饮供 —The absolute dry mass (g) of the prickly pear root slices in the test solution;

[0348] V 供 —Volume of the test solution, ml;

[0349] V 饮供 —Volume of the test solution of the medicinal slices, in ml;

[0350] m 提 —The absolute dry weight of the obtained prickly pear root extract, in g;

[0351] m 饮 —Absolutely dry weight of raw prickly pear root slices, in g.

[0352] The content and transfer rate of ellagic acid in 15 batches of prickly pear extract prepared in Example 1 were determined, and the results are shown in Table 21.

[0353] Table 21 Results of content and transfer rate of prickly pear root extract from different batches

[0354]

[0355]

[0356] Based on the determination results of ellagic acid content in 15 batches of prickly pear root slices and prickly pear root extract, the mean and range of ellagic acid content and transfer rate in prickly pear root extract are calculated and shown in Table 22.

[0357] Table 22 Analysis of Ellagic Acid Content and Transfer Rate in 15 Batches of Rosa rugosa Root Extract

[0358]

[0359] As shown in Table 22, the average ellagic acid content in the prickly pear root extract is 13.50 mg / g. Based on the average ellagic acid content of 70% to 130%, its fluctuation range is calculated to be 9.45 mg / g to 17.55 mg / g. Based on ±2 times SD of the average ellagic acid content in the prickly pear root extract, its fluctuation range is calculated to be 9.97 mg / g to 17.04 mg / g. Based on ±3 times SD of the average ellagic acid content in the prickly pear root extract, its fluctuation range is calculated to be 8.20 mg / g to 18.80 mg / g.

[0360] As shown in Table 22, the average transfer rate of ellagic acid in the prickly pear root extract was 59.35%. Based on the average transfer rate of ellagic acid in the prickly pear root extract being 70% to 130%, its fluctuation range was 41.54% to 77.15%. Based on the average transfer rate of ellagic acid in the prickly pear root extract being ±3 times SD, its fluctuation range was 20.38% to 98.32%.

[0361] Example 4: Method for constructing the characteristic map of prickly pear root extract

[0362] 1. Preparation of reference solution

[0363] Take an appropriate amount of ellagic acid reference standard, accurately weigh it, and add methanol to prepare a solution containing 18 μg of ellagic acid per 1 ml, which will be used as the reference solution.

[0364] 2. Preparation method of control herbal solution

[0365] Take about 1g of prickly pear root slices, weigh them accurately, place them in a stoppered conical flask, add 100ml of water accurately, weigh them, heat under reflux for 2 hours, cool them, weigh them again, add water to make up the weight loss, shake well, filter, and collect the filtrate to obtain the product.

[0366] 3. Preparation of the test solution

[0367] Take about 0.1g of prickly pear root extract, accurately weigh it, place it in a stoppered conical flask, accurately add 100ml of water, weigh it, heat under reflux for 2 hours, cool it, weigh it again, add water to make up the weight loss, shake well, filter it, and take the filtrate to obtain the extract.

[0368] 4. Validation of chromatographic conditions

[0369] (1) Determination of detection wavelength

[0370] Inject the test sample solution of prickly pear root extract into the sample for analysis, and record the absorption spectrum in the range of 190–400 nm. Figure 4 As shown.

[0371] Depend on Figure 4 It can be seen that at a wavelength of 254 nm, the test solution of prickly pear root extract has more detectable chromatographic peak information and less baseline noise interference. Therefore, 254 nm was selected as the detection wavelength.

[0372] (2) Optimization of the mobile phase

[0373] ① Considering phase A as organic and phase B as aqueous, the elution gradient is shown in Table 5, and the results are as follows: Figure 5 As shown,

[0374] Depend on Figure 5 It is known that the elution ability of methanol-0.1% phosphoric acid solution is stronger than that of acetonitrile-0.1% phosphoric acid solution. Therefore, methanol-0.1% phosphoric acid solution was chosen to explore the conditions, and the various acids used in the mobile phase will be investigated later.

[0375] ② Examine the cases with and without acid.

[0376] Using methanol as mobile phase A, the effects of mobile phase B without acid and with different acids on the detection results were investigated. The results are as follows: Figure 6 As shown,

[0377] Depend on Figure 6 It can be seen that adding acid results in a better peak shape than not adding acid, and the peak shape is optimal when the mobile phase is a methanol-0.1% phosphoric acid solution system. Therefore, methanol-0.1% phosphoric acid solution is selected for elution.

[0378] (3) Determination of chromatographic conditions

[0379] The column was packed with octadecylsilane-bonded silica gel (150 mm column length, 2.1 mm inner diameter, 1.8 μm particle size); methanol was used as mobile phase A, and 0.1% phosphoric acid solution was used as mobile phase B, with gradient elution performed according to the specifications in Table 5; the flow rate was 0.20 mL / min; the column temperature was 35 °C; and the detection wavelength was 254 nm. The theoretical plate number, calculated based on the ellagic acid peak, should be no less than 3000.

[0380] 5. Optimization of test solution preparation conditions

[0381] First, the UPLC chromatograms of different batches of Rosa rugosa root extract samples were determined. The results were analyzed using the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine (2012 Edition)" recommended by the National Pharmacopoeia Commission. Common peaks were selected, and the results are as follows: Figure 7 As shown. By Figure 7 It can be seen that there are 7 relatively obvious common peaks in the spectrum of the prickly pear root extract, where R is the reference material and S1-S15 are different batches of prickly pear root extract.

[0382] Using peak 6 as a reference peak, a characteristic spectrum of the prickly pear root extract was established, such as... Figure 8 As shown, peak 6 (S peak) is ellagic acid.

[0383] from Figure 8 As can be seen, the characteristic chromatogram of the test sample should show 7 characteristic peaks, corresponding to the retention times of the 7 characteristic peaks in the chromatogram of the reference medicinal material. Among them, peak 6 should be consistent with the retention time of the ellagic acid reference peak. Taking the peak corresponding to the ellagic acid reference peak as the S peak, the relative retention times of each characteristic peak and the S peak are calculated. The relative retention times should be within ±10% of the specified values, which are: 0.15 (peak 1), 0.69 (peak 2), 0.84 (peak 3), 0.91 (peak 4), 0.97 (peak 5), and 1.21 (peak 7).

[0384] The effects of different extraction solvents on the characteristic chromatograms of Rosa rugosa root extract were then investigated. 100% methanol, 75% methanol, 50% methanol, 25% methanol, anhydrous ethanol, 75% ethanol, 50% ethanol, 25% ethanol, and water were selected as extraction solvents. The characteristic chromatograms of different extraction solvents were compared by using the total peak area / sample weight of seven tentatively determined chromatographic peaks and the chromatograms.

[0385] Take an appropriate amount of Rosa rugosa root extract (BT0316-1), approximately 0.1 g, accurately weigh it, and place it in a stoppered conical flask. Accurately add 100 ml each of 100% methanol, 75% methanol, 50% methanol, 25% methanol, anhydrous ethanol, 75% ethanol, 50% ethanol, 25% ethanol, and water. Weigh the flask, heat under reflux for 2 hours, cool, and weigh again. Make up the lost weight with water, shake well, filter, and collect the filtrate. Inject the extract under the determined chromatographic conditions and record the chromatogram. The results of the investigation of different extraction solvents for the characteristic chromatogram of Rosa rugosa root extract are as follows: Figure 9-11 As shown.

[0386] Figure 9-11 It is known that water is the extraction solvent with higher extraction efficiency, therefore water is chosen as the extraction solvent.

[0387] Based on the above experimental results, the sample pretreatment method for the characteristic chromatogram of Rosa rugosa root extract can be determined as follows:

[0388] Take about 0.1g of prickly pear root extract (BT0316-1), accurately weigh it, place it in a stoppered conical flask, accurately add 100ml of water, weigh it, heat under reflux for 2 hours, cool it, weigh it again, make up the lost weight with water, shake well, filter it, and take the filtrate to obtain the product.

[0389] 5. Methodological Validation

[0390] (1) Specificity examination

[0391] Accurately pipette 1 μl each of the test solution and blank solvent of *Rhizoma Rosa rugosa* root extract (BT0316-1) into the liquid chromatograph and determine the chromatographic conditions as specified above. The results are as follows: Figure 12 As shown.

[0392] Depend on Figure 12 It can be seen that the solvent does not interfere with the characteristic peaks in the characteristic spectrum of the prickly pear root extract.

[0393] (2) Holistic Examination

[0394] The test solution of Rosa rugosa root extract (BT0316-1) was injected into the liquid chromatograph. The elution time was extended by one time at the mobile phase ratio at the gradient endpoint, and the characteristic chromatograms were analyzed. The results are as follows: Figure 13 As shown.

[0395] Depend on Figure 13 It can be seen that no obvious chromatographic peak was observed after doubling the elution time under these chromatographic conditions, indicating that these chromatographic conditions basically meet the principle of maximizing information content.

[0396] (3) Precision test

[0397] The test solution of Rosa roxburghii root extract (BT0316-1) was injected 6 times, with an injection volume of 1 μl. Seven characteristic peaks were temporarily identified, and peak 6 was used as the reference peak. The relative retention times were calculated, and the experimental results are shown in Table 23.

[0398] Table 23 Precision results of characteristic chromatograms of Rosa rugosa root extract (relative retention time)

[0399] Serial Number Peak 1 Peak 2 Peak 3 Peak 4 Peak 5 Peak 6 Peak 7 1 0.15 0.69 0.83 0.90 0.96 1.00 1.22 2 0.15 0.69 0.83 0.90 0.96 1.00 1.22 3 0.15 0.69 0.83 0.90 0.96 1.00 1.22 4 0.15 0.69 0.83 0.90 0.96 1.00 1.22 5 0.15 0.69 0.83 0.90 0.96 1.00 1.22 6 0.15 0.69 0.84 0.91 0.97 1.00 1.22 average value 0.15 0.69 0.83 0.91 0.96 1.00 1.22 RSD (%) 0.24 0.17 0.16 0.05 0.05 0.00 0.10

[0400] As shown in Table 23, the RSD of the relative retention time of each chromatographic peak is less than 3.0%, indicating that the instrument has good precision.

[0401] (4) Stability test

[0402] The test solution of Rosa roxburghii root extract (BT0316-1) was injected at 0, 2, 4, 8, 12, 16, 20, and 24 hours, with an injection volume of 1 μl. Seven characteristic peaks were tentatively identified, and peak 6 was used as the reference peak. The relative retention times were calculated. The results are shown in Table 24.

[0403] Table 24. Stability results of characteristic spectra of Rosa roxburghii root extract (relative retention time)

[0404] sample Peak 1 Peak 2 Peak 3 Peak 4 Peak 5 Peak 6 (S) Peak 7 0h 0.15 0.69 0.84 0.91 0.97 1.00 1.22 2h 0.15 0.69 0.84 0.91 0.96 1.00 1.22 4h 0.15 0.69 0.83 0.91 0.96 1.00 1.22 8h 0.15 0.69 0.83 0.90 0.96 1.00 1.22 12h 0.15 0.69 0.84 0.91 0.97 1.00 1.22 16h 0.15 0.69 0.83 0.91 0.96 1.00 1.21 20h 0.15 0.69 0.84 0.91 0.97 1.00 1.21 24h 0.15 0.69 0.84 0.91 0.97 1.00 1.21 average value 0.15 0.69 0.84 0.91 0.97 1.00 1.22 RSD (%) 1.05 0.40 0.42 0.15 0.15 0.00 0.27

[0405] As shown in Table 24, the RSD of the relative retention time of the chromatographic peaks is in the range of 0.00% to 1.05%, and the RSD is <3.0%, indicating that the sample solution is relatively stable.

[0406] (5) Repeated examination

[0407] Approximately 0.1 g of the same batch of Rosa rugosa root extract (BT0316-1) was accurately weighed and prepared in six parallel batches. The sample solutions were prepared according to the pretreatment method for the characteristic chromatogram of Rosa rugosa root extract determined above, and 1 μl was injected into each sample. Seven characteristic peaks were tentatively identified, with peak 6 as the reference peak, and the relative retention times were calculated. The results are shown in Table 25.

[0408] Table 25. Repeatability results of characteristic chromatograms of Rosa rugosa root extract (relative retention time)

[0409]

[0410]

[0411] As shown in Table 25, the relative retention times (RSDs) of each chromatographic peak are between 0.00% and 0.10%, and the RSD values ​​are all <3.0%, indicating that the method has good repeatability.

[0412] (6) Durability test

[0413] ① Column analysis

[0414] The effects of different chromatographic columns (Endeavorsil C18 (2.1*150mm, 1.8μm), ACQUITY UPLCHSS T3C18 (2.1*150mm, 1.8μm), and Infintylab Poroshell 120Aq-C18 (2.1*150mm, 2.7μm)) on the characteristic peaks of Rosa rugosa root extract were investigated. The results are shown in Table 26.

[0415] Table 26. Chromatographic column analysis results of characteristic chromatograms of Rosa rugosa root extract (relative retention times)

[0416]

[0417] Table 26 shows that different chromatographic columns have a certain impact on the peak elution. The elution using an Endeavorsil C18 (2.1×150mm, 1.8μm) ultra-high performance liquid chromatography column resulted in better peak shapes and the best separation effect. Therefore, it is recommended that this method use an Endeavorsil C18 (2.1×150mm, 1.8μm) ultra-high performance liquid chromatography column.

[0418] ② Column temperature investigation

[0419] The effects of different column temperatures (25℃, 30℃, and 35℃) on the peak shapes of the characteristic chromatographic peaks of Rosa rugosa root extract were compared. The results are shown in Table 27.

[0420] Table 27. Characteristic spectrum of Rosa rugosa root extract: column temperature results (relative retention time)

[0421] Column temperature Peak 1 Peak 2 Peak 3 Peak 4 Peak 5 Peak 6 (S) Peak 7 25℃ 0.14 0.67 0.83 0.90 0.96 1.00 1.24 30℃ 0.15 0.69 0.84 0.91 0.97 1.00 1.22 35℃ 0.16 0.71 0.87 0.92 0.98 1.00 1.19

[0422] As shown in Table 27, column temperature has a certain impact on peak elution, but the separation effect is best and the peak shape is better when the column temperature is 35℃. Therefore, it is recommended to use a column temperature of 35℃ for the determination.

[0423] ③ Flow velocity investigation

[0424] The effects of different flow rates (0.20 ml / min, 0.25 ml / min, and 0.30 ml / min) on the peak shape of the characteristic chromatographic peaks of Rosa rugosa root extract were compared. The results are shown in Table 28.

[0425] Table 28. Flow rate analysis results (relative retention time) of characteristic spectra of Rosa roxburghii root extract.

[0426] Flow rate Peak 1 Peak 2 Peak 3 Peak 4 Peak 5 Peak 6 (S) Peak 7 0.20 ml / min 0.15 0.69 0.84 0.91 0.97 1.00 1.22 0.25ml / min 0.13 0.69 0.85 0.92 0.97 1.00 1.21 0.30ml / min 0.12 0.69 0.86 0.93 0.98 1.00 1.20

[0427] As shown in Table 28, the flow rate has a certain impact on the peak shape, but the peak shape is optimal when the flow rate is 0.20 ml / min. Therefore, it is recommended to use 0.20 ml / min as the measurement flow rate.

[0428] Example 5: Preparation of Prickly Pear Root Formula Granules

[0429] 1. Preparation of extract

[0430] The extract was prepared according to the optimized preparation method in Example 1. 16 kg of prickly pear root slices (YP220316-1) were placed in an extraction tank and decocted twice with water. Before decoction, the root slices were soaked for 30 minutes. For the first decoction, 10 times the amount of water was added, and the mixture was heated to boiling, maintained at a gentle boil for 90 minutes, and then filtered (200 mesh). For the second decoction, 8 times the amount of water was added, and the mixture was heated to boiling, maintained at a gentle boil for 60 minutes, and then filtered (200 mesh). Concentration was performed under reduced pressure at a set temperature of 65°C and a vacuum degree of -0.080 to -0.090 MPa. The filtrate was concentrated under reduced pressure to a relative density of 1.04–…

[0431] The extract was obtained at 1.06 g / mL (45±5℃). The yield was measured according to the method in Example 1 and compared with the yield of the prickly pear root extract in Example 1. The results are shown in Table 29.

[0432] Table 29 Comparison of extract yield between extract and granular extract concentrated extract

[0433] Sample Name batch number Extract yield (%) Prickly pear root granule extract 230619 6.01 Example 1 Extract BT0316-1 5.87

[0434] The results in the table above show that the yield of extract from the concentration process of prickly pear root is basically consistent with that of extract prepared from the same batch of medicinal slices, and both are within the yield range of prickly pear root extract, indicating that the selected concentration method and concentration process parameters are relatively reasonable.

[0435] 2. Drying of the extract

[0436] (1) Drying method

[0437] The production of traditional Chinese medicine (TCM) preparations often involves the drying process of extracts. Currently, the main drying methods for TCM extracts include vacuum drying (drying oven), spray drying, microwave drying, and infrared drying. In recent years, with the development of TCM pharmaceutical equipment, vacuum drying (reduced pressure drying, low-temperature pulse drying, belt drying) and spray drying technologies have been widely used in the drying of TCM extracts. In particular, spray drying is widely used in the production of TCM granules. Its principle is to use an atomizer to disperse the liquid material into fine droplets, and then rapidly evaporate the solvent in a hot drying medium to form a dry powder product. Its main characteristics are: fast drying, short drying time, and the ability to directly dry solutions, emulsions, suspensions, and extracts into powder, granules, hollow spheres, or agglomerates. It can eliminate the need for evaporation, separation, and pulverization processes, retains better properties, and causes less damage to the active ingredients. Therefore, based on the effective components contained in the root of Rosa rugosa and the existing production equipment, this study selected a spray drying method from various production equipment based on different principles. This method is characterized by high drying efficiency, fast drying speed, short drying time, minimal damage to effective components, uniform and fine dry powder, low water content, good solubility, and suitability for pilot-scale and large-scale commercial production. The extract samples used in the experiment were prepared using the above method.

[0438] (2) Drying of extract samples

[0439] To prevent foreign matter from clogging the pipes, the extract was properly heated and boiled to fully dissolve it. An appropriate amount of excipients (maltodextrin was added in this study) were added, mixed well, and then passed through a No. 6 sieve (100 mesh). The mixture was placed in a mixing tank, and the material temperature was maintained between 60 and 100°C. The mixture was continuously stirred for later use.

[0440] (3) Screening of excipient types and dosages

[0441] Selection of excipients: Pharmaceutical excipients are the general term for all components in a formulation other than the active ingredient. Their main functions include excipients, stabilizers, and improving patient compliance. Because aqueous extracts and pastes of traditional Chinese medicine are rich in polysaccharides, tannins, mucilage, and other medicinal components, they are prone to sticking to the drying walls during spray drying. Besides improving equipment performance, the ideal method to prevent sticking is to add appropriate excipients as fillers that are effective against various sticky substances, thus promoting the spray drying of the extracts and pastes. Considering the quality requirements for granule solubility, drug compliance, and the needs of the target population, this experiment prioritizes maltodextrin as a commonly used excipient during drying due to its low sweetness, low calorie content, high nutritional value, good solubility, strong heat resistance, low hygroscopicity, resistance to clumping, and economic safety.

[0442] Screening of excipient dosage: Referring to the requirements of the production process of Chinese herbal medicine formula granules, unless otherwise specified, the ratio of excipients to intermediates (based on dried product) should generally not exceed 1:1. The dosage of excipients used in the preliminary study was investigated. The evaluation was based on the moisture content of the intermediates, the yield of the dry extract, the drying state, and the ease of drying. The specific method was to take an appropriate amount of the above-mentioned extracted and concentrated extract, divide it into 4 equal parts, each part weighing 1.8 kg (each part is equivalent to 4 kg of medicinal slices). According to the design scheme in the table below, reasonable spray drying process parameters (inlet air temperature: 165℃~195℃, outlet air temperature: 100~110℃) were selected to conduct an experiment on the amount of spray drying excipients added. The experimental results are shown in Table 30.

[0443] Table 30 Results of the investigation on the dosage of excipients for spray drying concentrated prickly pear root extract

[0444]

[0445] Note: The yield of dry extract powder is the yield of dry extract powder including excipients, and the dry extract powder yield refers to the yield of dry extract powder without excipients.

[0446] The results in the table above show that without excipients, the powder adheres to the drying wall and is difficult to dry. Adding 1.5%–4.5% maltodextrin as an excipient to the powder facilitates drying, resulting in lower moisture content and better drying performance. This indicates that adding a certain amount of maltodextrin to the concentrated extract of *Rhizoma prickly pear* root (single-petaled silk flower) significantly improves the spray drying effect and makes drying easier. Based on the observations of different excipient addition ratios, adding maltodextrin at 1.5%–4.5% of the powder weight in the drying process is more reasonable.

[0447] The optimized preparation method for the dry extract powder of prickly pear root is as follows: Take 16 kg of prickly pear root slices (YP220316-1) into an extraction tank, add water and decoct twice. Soak for 30 minutes before decoction. For the first decoction, add 10 times the amount of water, heat to boiling, maintain a gentle boil for 90 minutes, and filter (200 mesh). For the second decoction, add 8 times the amount of water, heat to boiling, maintain a gentle boil for 60 minutes, and filter (200 mesh). The concentration method is vacuum concentration, with the concentration set at 65℃ and vacuum degree: -0.080~-0.090MPa. The filtrate was concentrated under reduced pressure to a relative density of 1.04–1.06 (45±5℃) to obtain an extract. Then, 1.5% maltodextrin was added, and finally, the extract was dried by spray drying. The inlet air temperature was set to 165–195℃ and the outlet air temperature to 100–110℃. After the system reached the temperature, purified water was used for a test spray. After stabilization, the system was switched to extract feeding for spray drying, collecting powder while drying. After the extract feeding was completed, the system was switched back to water spray until the powder collection was finished, resulting in prickly pear root formula granules dry extract powder.

[0448] (4) Comparative study on the quality consistency between prickly pear root extract and spray-dried dry powder

[0449] Using the yield and characteristic chromatogram as evaluation indicators, the consistency of quality between the Rosa roxburghii root extract and the dry extract powder from the same batch of processed Rosa roxburghii in Example 1 was compared. The experimental results are shown in Table 31. The relative retention times of the characteristic chromatograms of the Rosa roxburghii root extract and the dry extract powder from the same batch of processed Rosa roxburghii in Example 1 are shown in Table 32.

[0450] Table 31 Comparison of the yield of prickly pear root extract and formulated granules / powder.

[0451]

[0452] Table 32 Relative retention times of characteristic spectra of Rosa rotundus root extract and formulated granules / powder

[0453]

[0454] The comparative study results show that the yield of the dried extract of prickly pear root granules meets the requirements of the yield range of prickly pear root extract. Characteristic peaks are present in the chromatograms of the dried extract, which should show seven characteristic peaks, corresponding to the retention times of the seven characteristic peaks in the chromatogram of prickly pear root extract in Example 4. Peak number 6, corresponding to the prickly pear root extract, is peak S. This indicates that the material basis of the dried extract of prickly pear root granules is consistent with that of the prickly pear root extract. The selected drying method and drying process parameters are preliminarily confirmed to be reasonable, and the drying process is basically feasible.

[0455] 3. Molding process research

[0456] 1) Ingredients are sieved and mixed.

[0457] ① Selection of granulation method and types of excipients

[0458] Granulation Method Selection: Dry granulation refers to granulation without wetting agents or liquid binders. It involves uniformly mixing the drug and excipients, compressing them into large flakes or strips, and then pulverizing them into particles of the desired size. This method relies on compression force to create bonding between particles. Preparation methods include heavy compression and rolling. Compared to traditional wet granulation, it eliminates the processes of preparing the soft material, drying, and granulation, simplifying the process and overcoming the disadvantage of large excipient usage, thus significantly increasing drug loading. However, relevant regulations for traditional Chinese medicine (TCM) formula granules require that the finished product contain as few or no excipients as possible. Therefore, this embodiment selects dry granulation as the granulation method for TCM formula granules and uses this method for screening excipient usage and preparing granule samples.

[0459] Selection of excipients: Since dry granulation excipients should have good flowability and compressibility, i.e., good drying and binding properties, different types and proportions of excipients such as maltodextrin, magnesium stearate, silica, talc, and microcrystalline cellulose can be considered. These can alter the powder properties, directly converting the powder raw material into granular products that meet the requirements. Granulation significantly increases the bulk density, achieving both pollution control and reduced powder waste, improved material appearance and flowability, and easier storage and transportation. It also allows for control of solubility, porosity, and specific surface area. This experiment selected inexpensive and readily available magnesium stearate and silica as excipients for granulation.

[0460] ② Ingredients are sieved and mixed.

[0461] According to the experimental design, add appropriate amounts of silica and magnesium stearate to the dry powder of prickly pear root formula granules, pass it through an 80-mesh sieve, and mix it evenly (usually for 30 minutes) to investigate the granulation process and prepare prickly pear root formula granules.

[0462] 2) Investigation of granulation molding process

[0463] ① Screening of auxiliary material dosage and investigation of granulation process

[0464] Given that a certain proportion of maltodextrin has been added to the prickly pear root formula granules, and considering the impact of the flowability of the prickly pear root formula powder on the efficiency of dry granulation and the influence of the resulting formula granules on the filling volume difference, the commonly used flow aids in the process of traditional Chinese medicine preparation are mainly silica and magnesium stearate. Therefore, this experiment uses the prickly pear root formula powder prepared above to investigate the dosage of excipients and granulation process. Each time, 1.0 kg of prickly pear root formula powder is taken, and the excipients are added according to the experimental design ratio, mixed thoroughly, and then placed in a GZL100-30L dry granulator to investigate the effects of process parameters such as the dosage of excipients, roller speed, and pressure on the granulation effect. The sieve mesh size is selected as 12-40 mesh for granulation. The dosage of excipients and granulation process are evaluated based on indicators such as particle size, flowability, first-pass yield, bulk density, solubility, and fine powder rate. The experimental results of the prickly pear root granulation process parameters are shown in Table 33, and the excipient dosage screening is shown in Table 34.

[0465] Table 33. Production Equipment Parameters for Granulation Molding Process Investigation

[0466]

[0467] Table 33 lists the parameters for commonly used equipment in screening the dosage of excipients for dry granulation of prickly pear root and investigating the granulation process. Simultaneously, the angle of repose was measured to determine the flowability of the prickly pear root formulation granules prepared by screening the dosage of excipients; the results are shown in Table 34.

[0468] Table 34: Evaluation of Granulation Process Parameters for Prickly Pear Root Formula Granules

[0469]

[0470]

[0471] The data in Tables 33 and 34 show that the feeding speed of the prickly pear root granule dry extract powder should not be too fast. Too fast a feeding speed will affect the yield of finished products and the strength of the granules. Adding a certain amount of flow aid before dry granulation of prickly pear root dry extract powder can significantly improve the flowability of the granules.

[0472] Based on the data in Tables 33 and 34, the dosage of excipients and the granulation process were evaluated using particle size, flowability, yield per batch, bulk density, and solubility as evaluation indicators. The results showed that the dosage of excipients (silica and magnesium stearate) in dry granulation should be ≤0.3wt% (based on the weight of the raw medicinal slices). The granulation process parameters are those in Table 33.

[0473] 3) Particle quality control requirements

[0474] The prepared prickly pear root formulation granules should have a uniform color and meet the particle size requirements.

[0475] ①Particle total blend

[0476] The prepared qualified particles were mixed evenly, and the total mixing time was initially set at 30 minutes.

[0477] ② Evaluation of molding process

[0478] Based on the results of the screening and investigation of excipient types, dosages and granulation processes in the early stage, and taking into account the characteristics of Chinese herbal dry powder and large-scale production efficiency, combined with the process parameters screened in the above studies, the content and transfer rate of index components, characteristic chromatograms and relative retention time were used as evaluation indicators to compare the consistency of the quality of the standard decoction and the experimental verification granules. The results of the comparative study on transfer rate and relative retention time of characteristic chromatograms are shown in Tables 35 and 36, respectively.

[0479] Table 35 Comparison of the content and transfer rate of indicative components between Rosa rugosa root extract and formulated granules.

[0480]

[0481] As shown in Table 35, the particle transfer rate is slightly lower than that of the standard decoction prepared from the same batch of medicinal slices, but is generally consistent with that of the standard decoction.

[0482] Table 36 Relative retention times of characteristic chromatograms of standard decoctions and formula granules

[0483]

[0484] The comparative study results above show that the chromatogram of the prickly pear root formula granules has 7 characteristic peaks, which correspond to the retention times of the 7 characteristic peaks in the chromatogram of the standard decoction; peak number 6, corresponding to the standard decoction, is peak S. This indicates that the material basis of the prickly pear root formula granules is consistent with that of the standard prickly pear root decoction, suggesting that the selected granulation method and granulation process parameters are reasonable and the granulation process is basically feasible.

[0485] In this embodiment, the determination methods for the content and transfer rate of the index component (ellagic acid) of the formulation granule extract and formulation granules are performed according to the method in Example 3; the detection method for the characteristic spectrum is performed according to the method in Example 4.

[0486] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for constructing a characteristic spectrum of a Rosa rugosa root extract, characterized in that, Includes the following steps: (1) Preparation of reference solution Weigh out ellagic acid reference standard and add it to methanol to prepare a reference standard solution; (2) Preparation of control herbal solution The control herbal solution was obtained by extracting the root slices of Rosa roxburghii with water. (3) Preparation of the test solution The root slices of prickly pear were decocted twice and filtered to obtain the decoction. The decoction was concentrated and dried to obtain the prickly pear root extract. The prickly pear root extract was weighed and added to water for reflux extraction to obtain the test solution. (4) High performance liquid chromatography analysis The characteristic chromatograms of the *Rhizoma Rosa roxburghii* root extract were obtained by high-performance liquid chromatography (HPLC) analysis of the reference solution and the test solution. The characteristic chromatograms of the *Rhizoma Rosa roxburghii* root extract contained seven characteristic peaks, which were located at retention times of 0.14-0.16, 0.67-0.71, 0.83-0.87, 0.90-0.92, 0.96-0.98, 1.00, and 1.19-1.

24. The characteristic peak at 1.00 was identified as ellagic acid. The chromatographic conditions for high-performance liquid chromatography (HPLC) analysis in step (4) were as follows: octadecylsilane-bonded silica gel was used as the packing material; the column length was 150 mm, the inner diameter was 2.1 mm, and the particle size was 1.8 μm; gradient elution was performed using mobile phase A and mobile phase B; mobile phase A was methanol; mobile phase B was 0.1% phosphoric acid solution; the gradient elution conditions were as follows: from 0 to 15 min, mobile phase A was 15 vol% → 38 vol%, and mobile phase B was 85 vol% → 62 vol%; from 15 to 40 min, mobile phase A was 38 vol%, and mobile phase B was 62 vol%. The flow rate was 0.20 ml per minute; the column temperature was 35℃; and the detection wavelength was 254 nm.

2. The construction method according to claim 1, characterized in that, The volume-to-mass ratio of water to the prickly pear root extract is 100-2000 mL / g.

3. The construction method according to claim 1, characterized in that, The reflux extraction time is 0.5-2 hours.

4. The construction method according to claim 2, characterized in that, The volume-to-mass ratio of water to the prickly pear root extract is 500-1500 mL / g.

5. The construction method according to claim 3, characterized in that, The reflux extraction time is 1-2 hours.

6. The method for constructing the characteristic spectrum according to any one of claims 1-5 is used as a quality control measure in the quality evaluation of Rosa rugosa root extract and its preparations.

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