Quality standard of Rungia pectinata medicinal material and detection method thereof

Abstract

The invention provides a quality standard of a rungia pectinata medicinal material and a detection method thereof, and relates to the technical field of traditional Chinese medicines. The invention provides a quality standard detection method for Rungia pectinata medicinal materials, which detects the quality of Rungia pectinata from the aspects of medicinal material sources, character identification, microscopic identification, thin-layer identification, moisture, total ash content, acid-insoluble ash content and extracts, and establishes a thin-layer chromatography identification method with strong specificity, stability and reliability. Therefore, the authenticity of the rungia pectinata medicinal material is identified, the quality of the rungia pectinata medicinal material is evaluated, and the medication safety is ensured. According to the quality standard of the Rungia pectinata medicinal material established by the method, the identification result is stable and reliable, and the accuracy is high.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and in particular to a quality standard and testing method for a herbal medicine for children. Background Technology

[0002] Child's grass, a commonly used folk remedy in Guangdong for treating infantile malnutrition, is a traditional Chinese medicine with a long history of application. It comes from the plant Child's grass (Acanthaceae family). Rungia pectinata The dried whole herb of (L.) Nees, also known as "blue grass", "malnutrition grass" or "wasp grass", grows in grassy areas along fields, slopes and villages. It is a common wild weed in grasslands and is widely used in Guangdong, Guangxi and other places to treat malnutrition, indigestion, colds and fever in children.

[0003] Its original plant morphology is as follows: an annual slender herb; branches cylindrical, yellow when dry. Leaves thin and papery, lower leaves oblong-ovate, up to 6 cm long, usually about 4 cm long, apex obtuse, base gradually narrowed or sometimes nearly acute, both surfaces covered with appressed sparse soft hairs; lateral veins 5 on each side, usually not very obvious; petiole 3-4 mm long or longer. The spike-like inflorescences are densely flowered, terminal and axillary, 1–3 cm long; bracts are in 4 rows, only 2 rows bearing flowers; the flowering bracts are nearly orbicular or broadly ovate, about 4 mm long, with long soft hairs on the back, and a membranous margin about 0.5 mm wide, ciliate; the unflowering bracts are oblong-lanceolate, about 6.5 mm long, with a hard pointed apex, and a narrow membranous margin and ciliate margin on one or sometimes both sides; the bracteoles are slightly smaller; the calyx lobes are linear, equal in size, about 3 mm long; the corolla is pale blue or white, about 5 mm long, glabrous except for the lower lip, the upper lip abruptly narrowed at the apex, and the lower lip lobes nearly triangular. The capsule is about 3 mm long, glabrous. Flowering occurs in early spring.

[0004] Currently, *Hedyotis diffusa* is only recorded in books such as *Guangdong Materia Medica*, *National Compendium of Chinese Herbal Medicine*, *Dictionary of Chinese Materia Medica*, *Chinese Materia Medica*, and *Lingnan Herbal Collection*. *Guangdong Materia Medica* records that *Hedyotis diffusa* has the effects of clearing liver heat and treating infantile malnutrition, promoting diuresis and relieving indigestion. It is used for infantile malnutrition with fever, infantile food stagnation, damp-heat diarrhea, and dysentery; recently, it has also been used for acute infectious hepatitis, acute conjunctivitis, epidemic keratitis (pink eye), and cervical lymph node tuberculosis. *National Compendium of Chinese Herbal Medicine* records that *Hedyotis diffusa* has the effects of clearing heat and dampness, relieving indigestion and promoting digestion. It is used for infantile malnutrition, indigestion, hepatitis, enteritis, colds, sore throat, conjunctivitis, cervical lymph node tuberculosis, and boils. *Dictionary of Chinese Materia Medica* and *Chinese Materia Medica* both record that *Hedyotis diffusa* has the effects of relieving indigestion, purging liver fire, and clearing damp-heat. It is used to treat indigestion in children, red and swollen eyes, diarrhea due to damp heat, hepatitis, scrofula, carbuncles, and snake bites. The *Lingnan Caiyao Lu* records that it "eliminates indigestion in children and clears liver fire," hence its nickname "Children's Grass." The *Guangdong Zhongyao Zhi* describes its inflorescence as: "Short spike-like inflorescences, bluish-green, flattened, 1.5–2.5 cm long, terminal or axillary, resembling a cockroach."

[0005] As an important medicinal plant in the traditional folk medicine system in South China, Rungia pectinata has a long history of clinical application and has long been widely used in folk remedies and traditional Chinese medicine hospital formulations. Although it shows significant clinical demand in regional medical practice, the current systematic research on Rungia pectinata is still in its initial stage, and there is a lack of authoritative research data at home and abroad. Moreover, Rungia pectinata has not been included in the current version of the Chinese Pharmacopoeia and the local standards of various provinces and cities, lacking a unified standard, which makes it impossible to effectively evaluate the authenticity, quality and superiority of Rungia pectinata药材, and also restricts its transformation from folk empirical medicine to standardized clinical practice. At the same time, there are many common adulterants of Rungia pectinata, such as Rungia chinensis Rungiachinensis Benth. (also known as Ming'e Cao), etc. The alias of Rungia pectinata is "Ganji Cao", and in folk, the dried whole herbs of Rostellularia procumbens Rostellularia procumbens (L.) Nees of Acanthaceae and Striga asiatica Striga asiatica (L.) O. Kuntze of Scrophulariaceae are also used as "Ganji Cao", which are actually different plants with the same name, etc. The lack of a systematic and effective quality control method for Rungia pectinata will affect the internal quality and medication quality of the药材, and even cause problems with medication safety. To ensure the efficacy and medication safety of Rungia pectinata药材, it is crucial to establish a complete quality standard detection method.

[0006] In view of this, the present invention is specifically proposed. Summary of the Invention

[0007] The first object of the present invention is to provide a quality standard detection method for Rungia pectinata药材 to solve the above technical problems.

[0008] The second object of the present invention is to provide a quality standard for Rungia pectinata药材.

[0009] To achieve the above objects, the following technical solutions are specifically adopted: In the first aspect, the present invention provides a quality standard detection method for Rungia pectinata药材, including药材 source, character identification, transverse section microscopic identification, surface microscopic identification, powder microscopic identification and thin layer chromatography identification; The thin layer chromatography identification includes the following steps: a. The test药材 and the control药材 of Rungia pectinata are respectively extracted with an ethanol solution to obtain extraction solutions, and after concentration of the extraction solutions, test solution and control solution are prepared; b. The test solution and the control solution are spotted on a thin layer plate, and then developed with a developing agent and sprayed with a coloring agent for coloring; The developing agent is a mixed solution of petroleum ether, chloroform and ethyl acetate, and the volume ratio of petroleum ether, chloroform and ethyl acetate is 5:10:0.8; The coloring agent is a 5 vol% - 15 vol% sulfuric acid ethanol solution.

[0010] As a further technical solution, the concentration of ethanol in the ethanol solution is 70 vol%-100 vol%. The extraction material-to-liquid ratio is 1 g: 5 ml - 1 g: 30 ml; The extraction was performed under ultrasonic conditions for 5-20 minutes.

[0011] As a further technical solution, the thin-layer plate is a silicone G thin-layer plate; The volume of the spotting sample is 2-6 μl; The unfolding temperature is 10-23℃; The humidity level during operation is 32%-88%. The color development temperature is 100℃~180℃.

[0012] As a further technical solution, it also includes moisture determination, total ash determination, and acid-insoluble ash determination.

[0013] As a further technical solution, the moisture content is determined according to the method described in General Chapter 0832, Method II of the 2020 edition of the Chinese Pharmacopoeia; The determination of total ash and acid-insoluble ash was performed according to the method described in General Chapter 2302 of the 2020 edition of the Chinese Pharmacopoeia.

[0014] As a further technical solution, hydrothermal determination of leachate is also included; As a further technical solution, the hydrothermal immersion method is determined according to the method described in General Chapter 2201 of the 2020 edition of the Chinese Pharmacopoeia.

[0015] Secondly, this invention provides a quality standard for herbal medicine for children, and the test results obtained by using the aforementioned quality standard testing method meet the following requirements: Source of medicinal materials: Herba Amaranthus, a plant of the Acanthaceae family Rungia pectinata Dried whole herb of (L.) Nees; The morphological identification results are as follows: The whole herb is 20-60 cm long; the root is slender and cylindrical, curved, and grayish-yellow; the stem is slender and slightly hard, branched, bluish-green, grayish-brown, or grayish-brown, 0.7-3.5 mm in diameter, with slightly raised longitudinal ridges on the surface, covered with fine soft hairs, the nodes are obvious and slightly swollen, with fibrous roots growing from the nodes near the base, brittle and easily broken, the cross-section is yellowish-white, the pith is hollow and needle-like or white; the leaves are opposite, bluish-green or yellowish-green, thin and papery, mostly wrinkled or broken, when intact they are flattened they are narrowly lanceolate or oblong-ovate, 1.2-7 cm long, entire, sparsely soft hairs on both sides, with short petioles; the spike-like inflorescence is short, terminal or axillary, yellowish-white to bluish-green or grayish-brown, flattened, and shaped like a cockroach; Microscopic identification of cross-sections revealed the following: the stems were mostly obtuse hexagonal; the epidermis consisted of a single layer of square-shaped cells with crystal cells containing cystoliths between the cells; stomata, glandular scales, and non-glandular hairs were visible, with non-glandular hairs being more common at the corners; the cortex was narrow, composed of 2–6 layers of parenchyma cells; there were 2–4 layers of collenchyma at the corners, and some corners had a large parenchyma cell below the collenchyma, 110–210 μm long; the endodermis was distinct, with occasional small columnar crystals in some cells; the pericycle fibers were visible on the inner side, arranged discontinuously in a ring, with a wall thickness of 2–5 μm; the vascular bundles were collateral; the phloem was narrow; the cambium was indistinct; the xylem consisted of vessels and fibers connected in a ring; the vessels were mostly scattered singly or arranged in several radially, with a diameter of 7–35 μm; the pith parenchyma cells were relatively large. Microscopic identification of leaf cross sections revealed the following: upper epidermal cells were square or rectangular, while lower epidermal cells were smaller; both upper and lower epidermis contained crystal cells with cystoliths, and non-glandular hairs and glandular scales were visible; the lower epidermis contained more stomata; the palisade tissue consisted of a single layer of cells passing above the midrib; the spongy tissue consisted of 2–4 loosely arranged layers of cells; the midrib protruded upwards and downwards, the vascular bundles were collateral, and the phloem was narrow; the inner surface of both upper and lower epidermis contained 1–2 layers of collenchyma cells. Microscopic identification of the leaf surface revealed the following: the anticlinal walls of the upper epidermal cells were relatively straight or slightly curved, while those of the lower epidermal cells were wavy; the lower epidermis had densely distributed stomata, which were straight-axis type, and the subsidiary cells varied greatly in size; both the upper and lower epidermis contained numerous crystal cells of cystoliths, which were oblong or rod-shaped, slightly curved, tapering or blunt at both ends, 100–500 μm long and 15–55 μm in diameter; non-glandular hairs with 1–5 cells were visible, with warty protuberances on the surface, mostly located on the leaf veins and leaf margins; the heads of glandular scales were mostly 4-celled, with a diameter of 25–40 μm. Microscopic identification of the powder revealed the following: the powder was yellowish-green; the anticlinal walls of the upper epidermal cells were relatively straight or slightly curved; the lower epidermal cells were polygonal with wavy anticlinal walls, and the stomata were straight-axis; non-glandular hairs were mostly broken, with the intact ones having pointed tips and often showing warty protrusions. Glandular scales typically had 4-celled heads with short, unicellular stalks; cystoliths were oblong or club-shaped, slightly curved, with acuminate or obtuse ends; mesophyll cells contained calcium oxalate crystals; the anticlinal walls of the persistent calyx epidermal cells were deeply wavy; the vessels were mostly spiral vessels or bordered pitted vessels; pollen grains were elliptical, suborbicular, or subtriangular, with a diameter of 10–30 μm. Wood fibers were slender, mostly in bundles, with distinct cell cavities. The thin-layer chromatography identification results were as follows: In the chromatogram of the test medicinal material, fluorescent main spots of the same color appeared at the corresponding positions as in the chromatogram of the reference medicinal material of *Hedyotis diffusa*.

[0016] As a further technical solution, the following must also be met: The moisture content test results are as follows: the moisture content must not exceed 13.0%. The total ash content determination result is: the total ash content shall not exceed 20.0%; The results of the acid-insoluble ash determination are as follows: the acid-insoluble ash content shall not exceed 6.0%.

[0017] As a further technical solution, the following must also be met: Results of hydrothermal extraction of leachate: The leachate content shall not be less than 15.0%.

[0018] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a quality standard testing method for *Hedyotis diffusa* herbal medicine. It tests the quality of *Hedyotis diffusa* from multiple aspects, including the source of the medicinal material, morphological identification, microscopic identification, thin-layer chromatography identification, moisture, total ash, acid-insoluble ash, and extractives. It also establishes a highly specific, stable, and reliable thin-layer chromatography identification method to facilitate the identification of the authenticity of *Hedyotis diffusa* herbal medicine, evaluate the quality of *Hedyotis diffusa* herbal medicine, and ensure the safety of medication.

[0019] The quality standard for children's herbal medicine established by the method of this invention has stable, reliable, and highly accurate identification results. Attached Figure Description

[0020] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0021] Figure 1 : Phyllanthus urinaria sample characteristics (No. 12); Figure 2 Cross-section of the stem of *Heliotropium indicum* (A: detailed view; B: enlarged view); Note: 1 is epidermis, 2 is stoma, 3 is collenchyma, 4 is non-glandular hair, 5 is crystal cell containing cystoliths, 6 is glandular scale, 7 is cortex, 8 is endodermis, 9 is phloem, 10 is xylem, 11 is pith, 12 is pericycle fiber, 13 is large parenchyma cell, 14 is small columnar crystal; Figure 3 Detailed cross-section of a leaf of *Heliotropium indicum*; Note: 1 is collenchyma cell, 2 is crystal cell containing cystolith, 3 is upper epidermis, 4 is palisade tissue, 5 is xylem, 6 is phloem, 7 is spongy tissue, 8 is lower epidermis, 9 is stoma, 10 is non-glandular hair. Figure 4 Detailed view of the leaf surface of *Heliotropium indicum*; Note: 1 is upper surface cell, 2 is lower surface cell, 3 is stoma, 4 is crystal cell containing cystoliths, 5 is non-glandular hair, 6 is glandular scale; Figure 5Microscopic identification of powder from *Heliotropium indicum*; Note: 1 is upper leaf epidermal cell, 2 is lower leaf epidermal cell, 3 is non-glandular hair, 4 is glandular scale, 5 is cystolith, 6 is mesophyll cell, 7 is persistent calyx epidermal cell, 8 is vascular bundle, 9 is pollen grain, 10 is wood fiber. Figure 6 Chromatograms of different extraction solvents for *Heliotropium indicum*; Note: 1 is methanol solvent, 2 is 70% methanol solvent, 3 is 70% ethanol solvent, and 4 is ethanol solvent. Figure 7 Chromatograms of different extraction solvent volumes for *Heliotropium indicum*; Note: 1 is 5 ml of ethanol, 2 is 10 ml of ethanol, 3 is 15 ml of ethanol, 4 is 20 ml of ethanol, 5 is 25 ml of ethanol, and 6 is 30 ml of ethanol. Figure 8 Chromatograms of different ultrasonic extraction times of *Hedyotis diffusa* for identification; Note: 1 represents extraction time of 5 min, 2 represents extraction time of 10 min, 3 represents extraction time of 15 min, and 4 represents extraction time of 20 min. Figure 9 Chromatograms of different spotting amounts of *Heliotropium indicum*; Note: 1 represents a spotting amount of 1 μl, 2 represents a spotting amount of 2 μl, 3 represents a spotting amount of 3 μl, 4 represents a spotting amount of 4 μl, 5 represents a spotting amount of 5 μl, and 6 represents a spotting amount of 6 μl. Figure 10 : Thin-layer chromatograms of 16 batches of *Hedyotis diffusa* for identification; Note: Lane D is β-sitosterol reference standard, 1 is *Hedyotis diffusa* working reference material (batch 20240501), and 3-16 are samples 3-16 in Table 1; Figure 11 Thin-layer chromatography (TLC) development effects of different manufacturers' plates on *Heliotropium indicum* (A. Merck G TLC plate, Germany; B. Silica G TLC plate, Qingdao Ocean Chemical Co., Ltd.); Note: Lane D is β-sitosterol reference standard, 1 is *Heliotropium indicum* working reference material (batch 20240501), and 2, 3, 5, 7, 10, and 12 are samples 2, 3, 5, 7, 10, and 12 in Table 1. Figure 12 Thin-layer chromatographic development of *Hedysarum heterotropoides* at different temperatures (A. 24℃; B. 10℃); Note: Lane D is β-sitosterol reference standard, 1 is *Hedysarum heterotropoides* working reference material (batch 20240501), and 2, 3, 5, 7, 10, and 12 are samples 2, 3, 5, 7, 10, and 12 in Table 1. Figure 13Thin-layer chromatographic development of *Heliotropium indicum* under different humidity conditions (A. 32% humidity; B. 88% humidity); Note: Lane D is β-sitosterol reference standard, 1 is *Heliotropium indicum* working reference material (batch 20240501), and 2, 3, 5, 7, 10, and 12 are samples 2, 3, 5, 7, 10, and 12 in Table 1. Figure 14 Thin-layer chromatographic development of Andrographis paniculata and Hedyotis diffusa; Note: Lane C is the reference material of Andrographis paniculata, 1 is the working reference material of Hedyotis diffusa (batch 20240501), and 8 is Hedyotis diffusa 20240602. Figure 15 Thin-layer chromatograms of different developing solvents composed of two reagents (A. chloroform-ethyl acetate; B. petroleum ether (60-90 ℃)-chloroform; C. petroleum ether (60-90 ℃)-ethyl acetate); Note: Lane D is β-sitosterol reference standard, 1 is *Hedysarum heterotropoides* working reference material (batch 20240501), and 8 is *Hedysarum heterotropoides* 20240602; Figure 16 Thin-layer chromatograms of developing solvents composed of three reagents in different ratios (in the AD chromatogram, the volume ratios of petroleum ether, chloroform, and ethyl acetate are 5:10:5, 5:10:0.5, 5:15:1, and 5:10:0.8, respectively); Note: Lane D represents β. Sitosterol reference standard, 1 is the working reference herb of *Hedyotis diffusa* (batch 20240501), 8 is *Hedyotis diffusa* 20240602. Detailed Implementation

[0022] The embodiments and examples of the present invention will be described in detail below. However, those skilled in the art will understand that the following embodiments and examples are for illustrative purposes only and should not be considered as limiting the scope of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention. Unless otherwise specified, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0023] In a first aspect, the present invention provides a quality standard detection method for children's medicinal materials, including identification of the source of medicinal materials, identification of physical characteristics, identification of cross-section microscopy, identification of surface microscopy, identification of powder microscopy and identification by thin layer chromatography. The thin-layer chromatography identification includes the following steps: a. Extract the test herb and the reference herb, *Heliotropium indicum*, with ethanol solution to obtain extracts. Concentrate the extracts to prepare the test solution and the reference solution. b. Spot the test solution and the reference solution onto a thin-layer plate, then develop the solution with the developing solvent, and finally spray the developing agent to develop the color. The developing solvent is a mixed solution of petroleum ether, chloroform and ethyl acetate, with a volume ratio of 5:10:0.8. The colorimetric reagent is a 5 vol% to 15 vol% sulfuric acid ethanol solution. The concentration of the sulfuric acid ethanol solution can be, for example, but is not limited to, 5 vol%, 10 vol% or 15 vol%.

[0024] This invention provides a quality standard testing method for *Hedyotis diffusa* herbal medicine. The method tests the quality of *Hedyotis diffusa* from multiple aspects, including the source of the medicinal material, morphological identification, microscopic identification, thin-layer chromatography, moisture, total ash, acid-insoluble ash, and extractives. This method is used to identify the authenticity of *Hedyotis diffusa* herbal medicine, evaluate its quality, and ensure safe use.

[0025] In some alternative embodiments, the concentration of ethanol in the ethanol solution may be, for example, but not limited to, 70 vol%, 80 vol%, or 100 vol%. The extraction liquid-to-material ratio can be, for example, but not limited to, 1 g: 5 ml, 1 g: 20 ml, or 1 g: 30 ml; The extraction is performed under ultrasonic conditions, and the ultrasonic time can be, for example, but not limited to, 5 min, 10 min or 5-20 min.

[0026] In some alternative embodiments, the thin film is a silicone G thin film; The volume of the spotting can be, for example, but not limited to, 2 μl, 4 μl or 6 μl; The unfolding temperature can be, for example, but is not limited to, 10°C, 15°C, or 23°C; The humidity level can be, for example, but is not limited to, 32%, 50%, or 88%; The color development temperature can be, for example, but is not limited to, 100℃, 140℃ or 180℃.

[0027] The present invention will be further illustrated below with specific embodiments. However, it should be understood that these embodiments are merely for the purpose of more detailed illustration and should not be construed as limiting the present invention in any way.

[0028] Example 1 1. Sample Collection Sixteen representative samples of *Heliotropium indicum* were collected from different production areas. Detailed sample information is shown in Table 1.

[0029] Table 1. Sample information of *Heliotropium indicum* from different origins.

[0030] 2. Source of medicinal materials Each batch of samples was identified by Liu Maogui, Chief Pharmacist of the Quality Management Department of Kangmei Pharmaceutical Co., Ltd., as *Hymenochloa crus-galli*, a plant belonging to the Acanthaceae family. Rungia pectinata The dried whole herb of (L.) Nees, including batches 20240501 and 20240606, was also identified by the Institute of Microbiology, Guangdong Academy of Sciences, as *L. Nees*, a plant in the Acanthaceae family. Rungia pectinata The dried whole herb of (L.)Nees. Based on the above identification results, the source is revised to: This product is *Hedyotis diffusa*, a plant belonging to the Acanthaceae family. Rungia pectinata The dried whole herb of (L.) Nees. Harvest when the flowers are just in full bloom, remove impurities, and dry.

[0031] 3. Identification of morphological characteristics Based on observations of the characteristics of the aforementioned 16 batches of *Heliotropium indicum*, the following revisions were made to their characteristics: [Characteristics] The whole herb is 20–60 cm long. The root is slender, cylindrical, curved, and grayish-yellow. The stem is slender and slightly stiff, branched, bluish-green, grayish-brown, or grayish-brown, 0.7–3.5 mm in diameter, with slightly raised longitudinal ridges on the surface, covered with fine soft hairs, with distinct, slightly swollen nodes, and fibrous roots growing from the nodes near the base. It is brittle and easily broken, with a yellowish-white cross-section, and the pith is hollow and needle-like or white. The leaves are opposite, bluish-green or yellowish-green, thin and papery, mostly wrinkled or broken; when intact and flattened, they are narrowly lanceolate or oblong-ovate, 1.2–7 cm long, entire, sparsely pubescent on both sides, and have short petioles. The spike-like inflorescence is short, terminal or axillary, yellowish-white to bluish-green or grayish-brown, flattened, and shaped like a cockroach. (See the characteristics of the herbal medicine for children.) Figure 1 ).

[0032] 4. Microscopic identification A systematic study was conducted on the microscopic tissue structure characteristics of *Heliotropium indicum* from three parts: root, stem, and leaf. The specific methods and results are as follows: 4.1 Instruments and Reagents Instruments: The experiment was conducted using an Olympus BX43 biological microscope (Olympus Corporation).

[0033] Reagents: The chloral hydrate (Shanghai Maclean Biochemical Technology Co., Ltd., batch number: C16759433), anhydrous ethanol (Xilong Scientific Co., Ltd.), glycerol and glacial acetic acid (Tianjin Damao Chemical Reagent Factory) used in the experiment were all analytical grade.

[0034] 4.2 Methods Cross-section preparation: Take the parts of the *Heliotropium indicum* sample to be observed (roots, stems, and leaves), soften them, and then cut them into thin sections of 10–20 μm using the freehand sectioning method. Place a flat thin section on a glass slide, add 1–2 drops of glycerol-acetic acid test solution, and cover with a coverslip. Separately, place another flat thin section on a glass slide, add chloral hydrate test solution, heat to permeate, add glycerol-ethanol test solution, and cover with a coverslip.

[0035] Surface preparation: After moistening and softening the leaves of the *Heliotropium indicum* sample, tear off approximately 4 mm of the epidermis from the area to be observed. 2 Place one on a glass slide and the other on a glass slide. Add chloral hydrate solution, heat to allow it to permeate, and then add glycerol ethanol solution. Cover with a coverslip.

[0036] Powder preparation: Pass the *Hedysarum heterotropoides* sample powder through a No. 4 sieve, take a small amount and place it on a glass slide, add chloral hydrate test solution, heat to thaw, and then add glycerol ethanol test solution. Cover with a coverslip. Take another small amount and place it on a glass slide, add 1-2 drops of glycerol acetic acid test solution, and cover with a coverslip.

[0037] 4.3 Results and Analysis The prepared microscopic specimens were observed under a microscope. Based on the observed microscopic features, the microscopic structural characteristics of *Heliotropium indicum* were determined, and the microscopic identification was revised as follows:

Identification

[0038] Leaf cross-section: Upper epidermal cells are square or rectangular, lower epidermal cells are smaller (i.e., smaller than epidermal cells); both upper and lower epidermis contain crystal cells with cystoliths, and non-glandular trichomes and glandular scales are visible; stomata are more numerous on the lower epidermis (compared to the upper epidermis). Palisade tissue consists of a single layer of cells, passing above the midrib; spongy tissue consists of 2-4 loosely arranged layers of cells. The midrib protrudes upwards and downwards, the vascular bundles are collateral, and the phloem is narrow; the inner part of the upper and lower epidermis consists of 1-2 layers of collenchyma cells (see the histological characteristics of the leaf cross-section of *Hemiberlesia lataniae* for details). Figure 3 ).

[0039] Leaf surface view: The anticlinal walls of the upper epidermal cells are relatively straight or slightly curved, while those of the lower epidermal cells are wavy. The lower epidermis has densely distributed stomata, which are straight-axis type, and the subsidiary cells vary greatly in size. Both the upper and lower epidermis contain numerous crystal cells of cystoliths, which are oblong or rod-shaped, slightly curved, tapering or obtuse at both ends, 100–500 μm long and 15–55 μm in diameter. Non-glandular trichomes with 1–5 cells and warty protuberances on the surface are visible, mostly located on the leaf veins and leaf margins. The glandular scale heads are mostly 4-celled, 25–40 μm in diameter (see the histological characteristics of the leaf surface view of *Hemiberlesia lataniae* for more information). Figure 4 ).

[0040] The powder is yellowish-green. The anticlinal walls of the upper epidermal cells are relatively straight or slightly curved; the lower epidermal cells are polygonal with wavy anticlinal walls, and the stomata are straight-axis type. Non-glandular hairs are mostly broken, with the intact ones having pointed tips and often showing warty protrusions. Glandular scales usually have a 4-celled head, a short, unicellular stalk. Cystomas are oblong or club-shaped, slightly curved, with acuminate or obtuse ends. Mesophyll cells contain calcium oxalate crystals. The anticlinal walls of the persistent calyx epidermal cells are deeply wavy. Vessels are mostly spiral vessels or bordered pitted vessels. Pollen grains are elliptical, suborbicular, or subtriangular, 10–30 μm in diameter. Wood fibers are slender, mostly in bundles, with distinct cell cavities (see Microscopic identification of *Heliotropium indicum* powder for details). Figure 5 ).

[0041] 5. Thin-layer chromatography identification Based on the information on its chemical composition, a corresponding thin-layer chromatography identification method was established.

[0042] 5.1 Instruments and Materials Instrument: Disek DD70 thin-layer imaging system.

[0043] Reagents: Silica gel G thin-layer plate (Merck, Germany, batch number: Hx31898026, size: 10×20cm), silica gel G thin-layer plate (Qingdao Ocean Chemical Co., Ltd., batch number: 2022113, size: 10×20cm); methanol, petroleum ether (60~90 ℃), chloroform, sulfuric acid, and ethanol (all from Xilong Scientific Co., Ltd.) were all analytical grade; ethyl acetate (Guinness High Purity Chemicals, USA) was chromatographic grade.

[0044] Control material: The sample of *Hedyotis diffusa* (batch number: 20240501) identified by the Institute of Microbiology, Guangdong Academy of Sciences, was used as the working control material.

[0045] Reference standard: β-sitosterol reference standard (China National Institutes for Food and Drug Control, batch number: 110851-201909).

[0046] 5.2 Investigation of the preparation method of the test sample (1) Preliminary experimental method Take 1 g of sample powder (sample No. 8), add 20 ml of methanol, extract by ultrasonication for 15 min, filter, and concentrate the filtrate to 1 ml as the test solution. Take 1 g of *Hedyotis diffusa* working reference material and prepare a reference material solution using the same method. Take β-sitosterol reference standard, add methanol to prepare a solution containing 1 mg per ml as the reference solution. Perform thin-layer chromatography, apply 2 µl of each of the above three solutions to the same silica gel G thin-layer plate, develop using petroleum ether (60–90 °C)-chloroform-ethyl acetate (5:10:0.8) as the developing solvent, remove, air dry, spray with 10 vol% sulfuric acid ethanol solution, heat at 105 °C until the spots are clearly visible, and examine under ultraviolet light (365 nm).

[0047] Results: In the chromatogram of the test sample, fluorescent spots of the same color appeared at the corresponding positions as in the chromatograms of the working control medicinal material and the reference substance.

[0048] (2) Investigation of extraction solvent Different extraction solvents were investigated: methanol, 70 vol% methanol, ethanol, and 70 vol% ethanol.

[0049] Take 1 g of sample powder (sample number 8), add 20 ml of each of the above-mentioned extraction solvents, and extract ultrasonically for 15 min. Filter, concentrate the filtrate to 1 ml, and use this as the test solution. Spot 2 μl of each test solution, and develop and visualize using the same method. Results: The thin-layer chromatographic bands in the results extracted with 70 vol% methanol became lighter, while the thin-layer chromatographic bands in the results extracted with ethanol and 70 vol% ethanol were relatively stable. Ethanol was ultimately chosen as the extraction solvent (see results below). Figure 6 ).

[0050] (3) Investigation of the volume of extraction solvent Take 1 g of sample powder (sample number 8), add 5 ml, 10 ml, 15 ml, 20 ml, 25 ml, and 30 ml of ethanol respectively, and extract by ultrasonication for 15 min. Filter, concentrate the filtrate to 1 ml, and use as the test solution. Take 2 μl of each test solution and spot it, develop and develop the color using the same method. Results: With the increase of extraction volume, the band clarity increases. When the volume reaches 20 ml, the band clarity and resolution are almost the same. Therefore, a 20 ml extraction volume was selected for subsequent experiments (see results). Figure 7 ).

[0051] (4) Investigation of ultrasonic extraction time Take 1 g of sample powder (sample number 8), add 20 ml of ethanol, and extract by ultrasonication for 5 min, 10 min, 15 min, and 20 min respectively. Filter the solution, concentrate the filtrate to 1 ml, and use this as the test solution. Spot 2 μl of each test solution, and develop and visualize using the same method. Results: The thin-layer chromatography showed the best band separation and a higher number of bands in the 15 min ultrasonic extraction time. Therefore, 15 min was selected as the extraction time (see results below). Figure 8 ).

[0052] 5.3 Investigation of sample size Spotting 1 μl, 2 μl, 3 μl, 4 μl, 5 μl, and 6 μl of the test solution were performed, and the same development and colorimetric methods were used. Results: The clarity of the bands increased with increasing spotting volume. However, when the spotting volume reached 4 μl, the band clarity became essentially uniform. Therefore, 4 μl was ultimately selected as the spotting volume (see results). Figure 9 ).

[0053] 5.4 Identification of *Heliotropium indicum* by Thin-Layer Chromatography Based on the above-mentioned conditions, a thin-layer chromatography identification method for *Heliotropium indicum* was finally established and revised as follows: Take 1 g of the powder, add 20 ml of ethanol, extract by sonication for 15 min, filter, and concentrate the filtrate to 1 ml to obtain the test solution. Take 1 g of *Hedyotis diffusa* reference material and prepare a reference material solution using the same method. Take β-sitosterol reference standard, add ethanol to prepare a solution containing 1 mg per ml to obtain the reference standard solution. Perform thin-layer chromatography (General Chapter 0502, 2020 Edition of the Chinese Pharmacopoeia). Apply 4 µl of each of the above three solutions to the same silica gel G thin-layer plate. Develop using petroleum ether (60–90 °C)-chloroform-ethyl acetate (5:10:0.8) as the developing solvent. Remove, air dry, spray with 10 vol% sulfuric acid ethanol solution, and heat at 105 °C until the spots are clearly visible. Examine under ultraviolet light (365 nm). In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference standard.

[0054] Sixteen batches of *Hedyotis diffusa* samples were analyzed using the above method. Results showed that the test sample chromatogram exhibited fluorescent spots of the same color at the corresponding positions as the reference medicinal material and reference standard chromatograms. (See the thin-layer chromatograms for the 16 batches of *Hedyotis diffusa* for details.) Figure 10 The unfolding temperature was 23℃ and the humidity was 60%.

[0055] 5.5 Methodological Examination (1) Evaluation of Thin-Layer Plates from Different Manufacturers: To verify the feasibility of this method, 6 batches were randomly selected from 16 batches of *Heliotropium indicum* for experimentation. The influence of thin-layer plates from different manufacturers on the development effect was investigated. Silica gel G thin-layer plates from Merck (Germany) and Qingdao Ocean Chemical Co., Ltd. were selected. Results: Both types of thin-layer plates met the identification requirements. The Merck silica gel pre-prepared thin-layer plate showed more concentrated spots, better separation, and superior chromatographic effect. (Comparison of the thin-layer chromatographic development effects of different manufacturers on *Heliotropium indicum* can be found in...) Figure 11 ).

[0056] (2) Comparison of different developing temperatures: To investigate the effect of different developing temperatures, development was performed at 10℃ and 23℃ respectively. Results: The spots in the chromatograms at different temperatures were clear, indicating that this method has good adaptability to different temperatures. (For a comparison of the thin-layer chromatographic development effects of different temperatures on *Heliotropium indicum*, see...) Figure 12 (The humidity is 60%).

[0057] (3) Comparison of different developing humidity levels: To investigate the effect of relative humidity on this method, the humidity was adjusted to 32% and 88% using different concentrations of sulfuric acid, and the developing effect under different humidity conditions was examined. Results: The spots in the chromatograms were clear under different humidity levels, indicating that this method is well adapted to different humidity environments. (For a comparison of the thin-layer chromatographic developing effect of different temperatures on *Heliotropium indicum*, see...) Figure 13 ).

[0058] 5.6 Comparison with the thin-layer chromatography method in the Andrographis paniculata pharmacopoeia The thin-layer chromatography method for Andrographis paniculata according to the 2025 edition of the Chinese Pharmacopoeia is as follows: Take 0.5 g of Andrographis paniculata reference material, add 25 ml of 40 vol% methanol, sonicate for 30 minutes, filter, and use the filtrate as the reference material solution. Perform the thin-layer chromatography test (General Rule 0502), taking 10 μl each of the reference solution, the test solution, and the above-mentioned reference material solution from the "Assay" section, and spot them separately on the same silica gel G thin-layer plate. Use chloroform-toluene-methanol (8:1:1) as the developing solvent, develop, remove, air dry, spray with 10 vol% sulfuric acid ethanol solution, heat at 105℃ until the spots are clearly visible, and examine under ultraviolet light (365 nm). In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatograms of the reference material and the reference solution.

[0059] Sample No. 8 of *Hypericum edulis*, *Hypericum edulis* working control material, and *Andrographis paniculata* control material (i.e., *Andrographis paniculata*, a plant of the Acanthaceae family) will be taken. Andrographis paniculata[Dried aerial parts of (Burm. f.) Nees] The experiment was conducted according to the above-described thin-layer chromatography method for Andrographis paniculata. Results: The Andrographis paniculata reference material showed clear fluorescent spots, but the *Heliotropium indicum* sample and the *Heliotropium indicum* working reference material did not show clear fluorescent spots; it can be seen that the thin-layer chromatography method for Andrographis paniculata in the 2025 edition of the Chinese Pharmacopoeia is applicable to the specific identification of Andrographis paniculata, but not to the specific identification of *Heliotropium indicum*, a species in the same family and genus. (Comparison of the thin-layer chromatographic development effects of Andrographis paniculata and *Heliotropium indicum* can be found in the figure below.) Figure 14 ).

[0060] 5.7 Comparison of thin-layer chromatograms with different developing solvents (1) Different developing conditions using two reagents were investigated, including chloroform-ethyl acetate (10:0.8), petroleum ether (60–90 °C)-chloroform (5:10), and petroleum ether (60–90 °C)-ethyl acetate (5:0.8). After development, the samples were removed, dried, sprayed with 10 vol% sulfuric acid ethanol solution, and heated at 105 °C until the spots were clearly visible. The samples were then examined under a UV lamp (365 nm). Results: In the developed images of all three developing solvents, some spots accumulated and could not be clearly separated, resulting in some main spots not being separated (see...). Figure 15 ).

[0061] (2) The developing conditions of different ratios of petroleum ether (60-90 ℃)-chloroform-ethyl acetate were investigated. The volume ratios included (5:10:5), (5:10:0.5), (5:15:1), and (5:10:0.8). After development, the samples were removed, dried, sprayed with 10 vol% sulfuric acid ethanol solution, and heated at 105 ℃ until the spots were clearly visible. The samples were then examined under a UV lamp (365 nm). Results: In the thin-layer chromatograms developed with different ratios of petroleum ether (60-90 ℃)-chloroform-ethyl acetate, the spots of the sample developed with a volume ratio of 5:10:5 accumulated at the leading edge (see...). Figure 16 In the thin-layer chromatography image developed with a volume ratio of 5:10:0.5 (A in the image), the spots below the corresponding positions of the β-sitosterol reference standard are indistinguishable (see [reference]). Figure 16 In the thin-layer chromatography image developed with a volume ratio of 5:15:1, spots below the corresponding position of the β-sitosterol reference were separated that were too close to the corresponding position of the reference, and the sample chromatogram showed too many non-primary spots, affecting the interpretation (see B). Figure 16 (C) Compared with the thin-layer chromatograms of other developing solvent ratios, the developing solvent with a volume ratio of 5:10:0.8 produced the best thin-layer chromatogram, with clear separation of the main spot and an Rf value of 0.35 for the reference standard, which meets the requirements of the 2025 edition of the Chinese Pharmacopoeia, Part IV, General Chapter 0502, Thin-Layer Chromatography (see C). Figure 16Therefore, the optimal development conditions are petroleum ether (60-90℃)-chloroform-ethyl acetate (5:10:0.8).

[0062] 6. Quality control of inspection items 6.1 Moisture Moisture content was determined according to the moisture determination method (General Chapter 0832, Method II, Chinese Pharmacopoeia 2020 Edition). Sixteen batches of samples were tested, with results ranging from 10% to 11.4%, and an average value of 10.7%. The limit was set at 20% above the average value (12.84%), therefore the moisture content of this product should not exceed 13.0%. The test results are shown in Table 2.

[0063] Table 216 shows the moisture content determination results of *Heliotropium indicum*.

[0064] 6.2 Total Ash Content The ash content was determined according to the method for ash determination (General Chapter 2302, Chinese Pharmacopoeia 2020). Sixteen batches of samples were tested, with results ranging from 11.8% to 21.6%, and an average value of 16.0%. The total ash content of this product is limited to 20.0%, with a margin of 20% (19.2%) above the average value. The test results are shown in Table 3.

[0065] Table 3. Results of total ash content determination for 16 batches of *Heliotropium indicum*

[0066] 6.3 Acid-insoluble ash The ash content was determined according to the method for determination of ash content (General Chapter 2302, Chinese Pharmacopoeia 2020). Sixteen batches of samples were tested, with results ranging from 1.1% to 9.5%, and an average value of 3.5%. The limit is 20% (4.2%) above the average value, therefore the acid-insoluble ash content of this product must not exceed 6.0%. The test results are shown in Table 4.

[0067] Table 416 Results of Acid-Insoluble Ash Determination for Herba Herba 416

[0068] 7Extracts The extractives of sample (number 10) were determined using water and dilute ethanol as solvents, respectively, according to the hot and cold maceration methods under the water-soluble extractives determination method (General Chapter 2201 of the Chinese Pharmacopoeia 2020 Edition) and the hot maceration method under the alcohol-soluble extractives determination method (General Chapter 2201 of the Chinese Pharmacopoeia 2020 Edition). The results showed that the hydrothermal maceration method yielded the highest result (see Table 5), therefore, the hydrothermal maceration method was selected for determining the extractive content. The extractive content of 16 more batches of samples was then determined, with results ranging from 17.2% to 28.3%, and an average value of 23.05%. A 20% reduction (18.44%) below the average value was deemed acceptable, and the extractive content of this product was specified to be no less than 15.0%. The extractive content determination results are shown in Table 6.

[0069] Table 5. Results of solvent analysis of extracts from 516 batches of *Heliotropium indicum*.

[0070] Table 6. Results of extract determination of 16 batches of *Heliotropium indicum*

[0071] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A quality standard testing method for children's herbal medicine, characterized in that, This includes identification of medicinal material sources, morphological characteristics, cross-sectional microscopic identification, surface microscopic identification, powder microscopic identification, and thin-layer chromatography identification. The thin-layer chromatography identification includes the following steps: a. Extract the test herb and the reference herb, *Heliotropium indicum*, with ethanol solution to obtain extracts. Concentrate the extracts to prepare the test solution and the reference solution. b. Spot the test solution and the reference solution onto a thin-layer plate, then develop the solution with the developing solvent, and finally spray the developing agent to develop the color. The developing solvent is a mixed solution of petroleum ether, chloroform and ethyl acetate, with a volume ratio of 5:10:0.

8. The colorimetric reagent is a 5 vol% to 15 vol% sulfuric acid ethanol solution.

2. The method for quality standard testing of herbal medicine for children according to claim 1, characterized in that, The concentration of ethanol in the ethanol solution is 70 vol%-100 vol%. The extraction material-to-liquid ratio is 1 g: 5 ml - 1 g: 30 ml; The extraction was performed under ultrasonic conditions for 5-20 minutes.

3. The method for quality standard testing of herbal medicine for children according to claim 1, characterized in that, The thin-layer plate is a silicone G thin-layer plate; The volume of the spotting sample is 2-6 μl; The unfolding temperature is 10-23℃; The humidity level during operation is 32%-88%. The color development temperature is 100℃~180℃.

4. The method for quality standard testing of herbal medicine for children according to claim 1, characterized in that, It also includes moisture determination, total ash determination, and acid-insoluble ash determination.

5. The method for quality standard testing of herbal medicine for children according to claim 4, characterized in that, The moisture content was determined according to the method described in General Chapter 0832, Method II of the 2020 edition of the Chinese Pharmacopoeia. The determination of total ash and acid-insoluble ash was performed according to the method described in General Chapter 2302 of the 2020 edition of the Chinese Pharmacopoeia.

6. The method for quality standard testing of herbal medicine for children according to claim 1, characterized in that, It also includes the hydrothermal method for determining leachate.

7. The method for quality standard testing of herbal medicine for children according to claim 6, characterized in that, The hydrothermal immersion method was determined according to the method described in General Chapter 2201 of the 2020 edition of the Chinese Pharmacopoeia.

8. A quality standard for a herbal medicine for children, characterized in that, The quality standard testing method according to any one of claims 1-7 is used for determination, and the test results meet the following requirements: Source of medicinal material: Herba Acanthaceae, plant *Hedyotis diffusa* Rungia pectinata Dried whole herb of (L.) Nees; The morphological identification results are as follows: The whole herb is 20-60 cm long; the root is slender and cylindrical, curved, and grayish-yellow; the stem is slender and slightly hard, branched, bluish-green, grayish-brown, or grayish-brown, 0.7-3.5 mm in diameter, with slightly raised longitudinal ridges on the surface, covered with fine soft hairs, the nodes are obvious and slightly swollen, with fibrous roots growing from the nodes near the base, brittle and easily broken, the cross-section is yellowish-white, the pith is hollow and needle-like or white; the leaves are opposite, bluish-green or yellowish-green, thin and papery, mostly wrinkled or broken, when intact they are flattened they are narrowly lanceolate or oblong-ovate, 1.2-7 cm long, entire, sparsely soft hairs on both sides, with short petioles; the spike-like inflorescence is short, terminal or axillary, yellowish-white to bluish-green or grayish-brown, flattened, and shaped like a cockroach; Microscopic identification of cross-sections revealed the following: the stems were mostly obtuse hexagonal; the epidermis consisted of a single layer of square-shaped cells with crystal cells containing cystoliths between the cells; stomata, glandular scales, and non-glandular hairs were visible, with non-glandular hairs being more common at the corners; the cortex was narrow, composed of 2–6 layers of parenchyma cells; there were 2–4 layers of collenchyma at the corners, and some corners had a large parenchyma cell below the collenchyma, 110–210 μm long; the endodermis was distinct, with occasional small columnar crystals in some cells; the pericycle fibers were visible on the inner side, arranged discontinuously in a ring, with a wall thickness of 2–5 μm; the vascular bundles were collateral; the phloem was narrow; the cambium was indistinct; the xylem consisted of vessels and fibers connected in a ring; the vessels were mostly scattered singly or arranged in several radially, with a diameter of 7–35 μm; the pith parenchyma cells were relatively large. Microscopic identification of leaf cross sections revealed the following: upper epidermal cells were square or rectangular, while lower epidermal cells were smaller; both upper and lower epidermis contained crystal cells with cystoliths, and non-glandular hairs and glandular scales were visible; the lower epidermis contained more stomata; the palisade tissue consisted of a single layer of cells passing above the midrib; the spongy tissue consisted of 2–4 loosely arranged layers of cells; the midrib protruded upwards and downwards, the vascular bundles were collateral, and the phloem was narrow; the inner surface of both upper and lower epidermis contained 1–2 layers of collenchyma cells. Microscopic identification of the leaf surface revealed the following: the anticlinal walls of the upper epidermal cells were relatively straight or slightly curved, while those of the lower epidermal cells were wavy; the lower epidermis had densely distributed stomata, which were straight-axis type, and the subsidiary cells varied greatly in size; both the upper and lower epidermis contained numerous crystal cells of cystoliths, which were oblong or rod-shaped, slightly curved, tapering or blunt at both ends, 100–500 μm long and 15–55 μm in diameter; non-glandular hairs with 1–5 cells were visible, with warty protuberances on the surface, mostly located on the leaf veins and leaf margins; the heads of glandular scales were mostly 4-celled, with a diameter of 25–40 μm. Microscopic identification results of the powder were as follows: the powder was yellowish-green; the anticlinal walls of the upper epidermal cells were relatively straight or slightly curved; the lower epidermal cells were polygonal with wavy anticlinal walls and straight stomata; non-glandular hairs were mostly broken, with the intact ones having pointed tips and many warty protrusions; the glandular scales had mostly 4-celled heads with short, single-celled stalks; the cystoliths were oblong or rod-shaped, slightly curved, with acuminate or obtuse ends; the mesophyll cells contained calcium oxalate crystals; the anticlinal walls of the persistent calyx epidermal cells were deeply wavy; the vessels were mostly spiral vessels or bordered pitted vessels; the pollen grains were elliptical, suborbicular, or subtriangular, with a diameter of 10–30 μm; the wood fibers were slender, mostly in bundles, and had distinct cell cavities. The thin-layer chromatography identification results were as follows: In the chromatogram of the test medicinal material, fluorescent main spots of the same color appeared at the corresponding positions as in the chromatogram of the reference medicinal material of *Hedyotis diffusa*.

9. The quality standard for children's herbal medicine according to claim 8, characterized in that, It also needs to meet the following requirements: The moisture content test results are as follows: the moisture content must not exceed 13.0%. The total ash content determination result is: the total ash content shall not exceed 20.0%; The results of the acid-insoluble ash determination are as follows: the acid-insoluble ash content shall not exceed 6.0%.

10. The quality standard for children's herbal medicine according to claim 8, characterized in that, It also needs to meet the following requirements: Results of hydrothermal extraction of leachate: The leachate content shall not be less than 15.0%.

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