Method for detecting and distinguishing ardisia crenata and ardisia quinquegona in pharmaceutical preparation

Abstract

The invention discloses a method for detecting and distinguishing ardisia crenata and ardisia dolichos in a medicinal preparation, which adopts a high performance liquid chromatography to construct a fingerprint spectrum for detection, and the chromatographic conditions are as follows: a chromatographic column is a C18 chromatographic column; the detection wavelength is 275 nm; mobile phases: a mobile phase A is water, a mobile phase B is methanol, gradient elution is adopted, the mobile phase proportion is volume percent, and the elution procedure is as follows: 0-5 minutes, 92-85% of the mobile phase A and 8-15% of the mobile phase B; 5-15 min, 85%-70% of the mobile phase A and 15%-30% of the mobile phase B; 15-25 min, the mobile phase A accounts for 70%-40%, and the mobile phase B accounts for 30%-60%; 25-35 min, the mobile phase A accounts for 40%, and the mobile phase B accounts for 60%; 35.1-35 min, 40%-92% of the mobile phase A and 60%-8% of the mobile phase B; the mobile phase A accounts for 92% and the mobile phase B accounts for 8% in 35.1-45min. The method provided by the invention can more accurately distinguish the preparation, especially whether the swelling and pain relieving tincture contains the ardisia quinquegona medicinal material or the ardisia crenata medicinal material extract.

Description

Technical Field

[0001] The present invention relates to the field of a method for detecting and distinguishing radix cinnabarinii and radix scutellariae in a pharmaceutical preparation of a traditional Chinese medicine, and in particular to a method for detecting and distinguishing radix cinnabarinii and radix scutellariae in a pharmaceutical preparation. Background Art

[0002] Although cinnabar root and small umbrella are often used interchangeably in the Lingnan region, they are not identical medicinal sources. According to the Chinese Pharmacopoeia (Chinese Pharmacopoeia) and authoritative literature, cinnabar root (Ardisia crenata Sims) is the official source plant, while small umbrella usually refers to Ardisia punctata Lindl. or Ardisiacrenata var. bicolor, both of the Myrsinaceae family. These are closely related species or variants and are not officially included in the Chinese Pharmacopoeia. The two differ in their clinical applications and efficacy. In the treatment of sprains and bruises, cinnabar root is suitable for the acute phase of injury, rapidly relieving local redness, swelling, heat, and pain, and is particularly effective for joint sprains and infections. Small umbrella is more suitable for chronic injuries or those in the recovery phase, as its blood-activating components promote tissue repair and reduce scarring. Clinically, precise selection should be based on the specific syndrome, prioritizing cinnabar root to ensure efficacy and safety.

[0003] The prescription for analgesic and swelling-relieving tincture contains 21 herbs that relax muscles and tendons, reduce swelling, and relieve pain. It is suitable for treating injuries caused by falls, rheumatic bone pain, pain that is localized, resists pressure, has difficulty moving, or has localized swelling, bruises, or hematomas. The main herbs in the prescription are Da Luosan (Greater Rhizome) and Xiao Luosan (Small Rhizome), which promote blood circulation, remove blood stasis, and relieve swelling and pain. Da Luosan (Greater Rhizome) has the effects of reducing swelling and relieving pain, promoting blood circulation, and removing blood stasis; Xiao Luosan (Small Rhizome) has the effects of promoting blood circulation, removing blood stasis, and dispelling wind and dampness. Based on the prescription's main functions and explanation, Cinnabar Root is more suitable for this prescription than Xiao Luosan. To better distinguish preparations made from Cinnabar Root and Xiao Luosan and improve the accuracy of prescription use, it is necessary to develop a method for detecting and distinguishing Cinnabar Root and Xiao Luosan in pharmaceutical preparations. Summary of the Invention

[0004] The purpose of the present invention is to overcome the shortcomings of the existing technology and provide a method for detecting and distinguishing Cinnabar Root and Rhizoma Cinnamomi in pharmaceutical preparations. The method can more accurately distinguish whether the preparations, especially the anti-inflammatory and analgesic tinctures, contain Rhizoma Cinnabaromi medicinal materials or Cinnabar Root medicinal material extracts.

[0005] The technical solutions of the present invention are as follows: A method for detecting and distinguishing cinnabar root and Rhizoma Cibotii in the pharmaceutical preparation; High performance liquid chromatography was used to construct the fingerprint for detection, and the chromatographic conditions were as follows: Chromatographic column: C18 column; Detection wavelength: 275nm; Mobile phase: Mobile phase A is water, mobile phase B is methanol, gradient elution is used, the mobile phase ratios are all volume percentages, and the elution procedure is as follows: 0-5 min, mobile phase A 92-85%, mobile phase B 8-15%; 5-15 min, mobile phase A 85%→70%, mobile phase B 15%→30%; 15-25 min, mobile phase A 70%→40%, mobile phase B 30%→60%; 25-35 min, mobile phase A 40%, mobile phase B 60%; 35.1-35 min, mobile phase A 40%→92%, mobile phase B 60%→8%; 35.1-45min, mobile phase A is 92%, mobile phase B is 8%.

[0006] The C18 chromatographic column has specifications of 4.6×25 mm, 5 μm. The chromatographic conditions also include: column temperature of 40-50° C. and flow rate of 0.8-1.3 mL / min. The C18 chromatographic column is: SHIMADZU Shim-pack GIST C18 chromatographic column.

[0007] The method for detecting and distinguishing Cinnabar Root and Rhizoma Cibotii in the pharmaceutical preparation further comprises a thin layer chromatography identification method for detection, using Rhizoma Cibotii and Cinnabar Root as references, comprising the following steps: The test samples made from the medicinal materials of Xiaoluosan, Zhushagen, and preparations were respectively spotted on the same silica gel G thin layer plate, and chloroform-ethyl acetate-methanol was used as the developing solvent in a volume ratio of 4-6:3-5:1-3. The plate was developed, taken out, dried, and examined under a 365nm ultraviolet lamp. A fluorescent spot was detected in the chromatogram of the medicinal material of Zhushagen, and no fluorescent spot of the same color was detected at the corresponding position of the spot in the chromatogram of the medicinal material of Xiaoluosan. Based on whether the preparation test sample has the spot at that position, it can be distinguished whether the preparation contains Zhushagen.

[0008] The pharmaceutical preparation is a liquid preparation.

[0009] In the high performance liquid chromatography method for constructing the fingerprint, the method for preparing the test solution comprises the following steps: Preparations made from the medicinal materials of Cinnabar Root and Rhizoma Cibotii were taken respectively; If it is a solid preparation, grind the solid preparation containing 0.212-0.318g of cinnabar root or Radix Aconiti Lateralis Preparata and place it in a 50mL volumetric flask. Add 35-42mL of n-butanol and sonicate for 30-50 minutes. After cooling, dilute to the mark with n-butanol, shake, filter, accurately measure 25mL of the filtrate, evaporate to dryness, dissolve the residue with methanol and dilute to a 10mL volumetric flask. Filter through a 0.45μm microporous filter membrane to obtain the product. If it is a liquid preparation, 20-30 mL of the liquid preparation of the cinnabar root or the small umbrella medicinal material is measured, evaporated to dryness, the residue is dissolved with 30-41 mL of n-butanol, transferred to a 50 mL volumetric flask, ultrasonically treated for 30-50 minutes, cooled, diluted to the scale with n-butanol, shaken, filtered, accurately measured 25 mL of the filtrate, evaporated to dryness, the residue is dissolved with methanol and diluted to a 10 mL volumetric flask, filtered with a 0.45 μm microporous filter membrane, and the product is obtained.

[0010] In the thin layer chromatography identification method, the test solution preparation method is as follows: The preparation of the test solution of the medicinal material of Radix Cinnabarinii and the test solution of the medicinal material of Radix Cinnabarinii comprises the following steps: Take 0.2-0.6g of powder of Radix Angelicae Dahuricae and Radix Cinnabarifolia respectively, add 30-50mL of methanol respectively, and ultrasonically treat for 20-40 minutes. Let cool, filter, evaporate the filtrate to dryness, and dissolve the residue in 1-3mL of methanol respectively. The method for preparing the test solution of the preparation comprises the following steps: Take 20-30 mL of the liquid preparation made from the medicinal materials of Cinnabar Root and Small Parashorea chinensis, add 40-60 mL of methanol, ultrasonicate for 20-40 minutes, cool, filter, evaporate the filtrate to dryness, add 2-4 mL of methanol to dissolve the residue, centrifuge, and take the supernatant as the test solution.

[0011] Preferably, the pharmaceutical preparation is a swelling-reducing and analgesic tincture prepared using cinnabar root or Rhizoma Cinnabarinii as one of the raw materials; Prepared from the following raw materials: 40-100 parts of costus root, 40-100 parts of siler, 40-100 parts of schizonepeta, 40-100 parts of asarum, 40-100 parts of acanthopanax bark, 40-100 parts of cassia twig, 40-100 parts of cyperus rotundus, 40-100 parts of ligusticum chuanxiong, 40-100 parts of cyperus rotundus, 70-120 parts of angelica dahurica, 40-100 parts of zedoaria, 70-120 parts of red eucommia bark, 130-180 parts of datura, 130-180 parts of cinnabar root, 130-180 parts of zymophora japonica, 120-160 parts of yellow twig, 130-180 parts of gardenia, 70-120 parts of trigonosciadium japonicum, 30-60 parts of agarwood, 60-100 parts of camphor, and 60-100 parts of menthol; Or prepared from the following raw materials: 40-100 parts of costus root, 40-100 parts of siler, 40-100 parts of schizonepeta, 40-100 parts of asarum, 40-100 parts of acanthopanax bark, 40-100 parts of cassia twig, 40-100 parts of cyperus rotundus, 40-100 parts of ligusticum chuanxiong, 40-100 parts of cyperus rotundus, 70-120 parts of angelica dahurica, 40-100 parts of zedoaria, 70-120 parts of red eucommia, 130-180 parts of radix scutellariae, 130-180 parts of radix scutellariae, 130-180 parts of zizyphus chinensis, 120-160 parts of truncatum, 130-180 parts of gardenia, 70-120 parts of trigonosciadium japonicum, 30-60 parts of aquilariae, 60-100 parts of camphor, and 60-100 parts of menthol.

[0012] The following characteristic peaks were detected in the fingerprint of the swelling and analgesic tincture prepared with Xiaoluosan as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; auxiliary characteristic peaks: RT = 16.004 min, RT = 17.021 min, RT = 17.648 min, RT = 20.203 min, RT = 21.324 min; The following characteristic peaks were detected in the fingerprint of the anti-swelling and analgesic tincture prepared with cinnabar root as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; there is no characteristic peak at the position of the above-mentioned auxiliary characteristic peaks.

[0013] The beneficial effects of the present invention are as follows: The high performance liquid chromatography method of the present invention is used to test the fingerprints of preparations made from the medicinal materials of cinnabar root and small umbrella respectively, and can better distinguish the preparations made from the medicinal materials of small umbrella and cinnabar root. There are large differences in the fingerprints of the two preparations, which proves that the quality of the preparations made from the two medicinal materials is greatly different.

[0014] The thin layer chromatography method of the present invention is used to test the medicinal materials of Rhizoma Cibotii and Rhizoma Cinnamomi respectively, and can better distinguish the medicinal materials of Rhizoma Cibotii and Rhizoma Cinnamomi. The quality of the two medicinal materials has certain differences. The thin layer chromatography method can also better distinguish whether the medicinal materials of Rhizoma Cibotii or Rhizoma Cinnamomi are used in preparations, especially in the finished product of the swelling and analgesic tincture.

[0015] The present invention is based on thin-layer chromatography and high-performance liquid chromatography, and develops a detection method that can identify and differentiate preparations, especially the medicinal materials of Rhizoma Cibotii and Rhizoma Cinnabaris in the prescription of anti-inflammatory and analgesic tincture. By adopting the quality detection method, it is possible to accurately identify that the medicinal materials used in the anti-inflammatory and analgesic tincture preparation are Rhizoma Cibotii and Rhizoma Cinnabaris, thereby further improving the accuracy of the prescription of the anti-inflammatory and analgesic tincture. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The thin layer chromatography spectra of three batches of medicinal materials, cinnabar root and scutellaria baicalensis, in Example 4; Figure 2The thin layer chromatography spectra of 7 batches of medicinal materials Cinnabar Root and Rhizoma Cibotii of Example 4 are shown; Figure 3 The thin layer chromatography spectrum of the anti-swelling and analgesic tincture finished product (made from the medicinal material of cinnabar root) and the negative sample lacking small umbrella; Figure 4 The fingerprint of the swelling-reducing and analgesic tincture (made from the medicinal material of Radix Aspergillus niger) of Example 5; Figure 5 The fingerprint of the swelling-reducing and analgesic tincture (made from the medicinal material Cinnabar root) of Example 5; Figure 6 This is a comparison of the fingerprints of the two preparations in Example 5 (the upper one is the medicinal material preparation of Osmanthus fragrans, and the lower one is the medicinal material preparation of Cinnabar Root). DETAILED DESCRIPTION

[0017] Example 1

[0018] Methods for detecting and distinguishing Root of Cinnabarinum and Rhizoma Cibotii in pharmaceutical preparations, including high performance liquid chromatography fingerprint construction and thin layer chromatography identification; 1. High performance liquid chromatography fingerprint construction: (1) The method for preparing the test solution includes the following steps: The anti-swelling and analgesic composition gel preparations made from the medicinal materials of Cinnabar Root and Rhizoma Cibotii were respectively used as test samples; The anti-swelling and analgesic composition gel preparation made from the medicinal material cinnabar root is prepared from the following raw materials: 40g of costus root, 40g of siler, 40g of schizonepeta, 40g of asarum, 40g of acanthopanax bark, 40g of cinnamon twig, 40g of oxaline root, 40g of ligusticum chuanxiong, 40g of cyperus rotundus, 70g of angelica dahurica, 40g of zedoaria, 70g of red eucommia bark, 130g of radix dahuricae, 130g of cinnabar root, 130g of dianthus, 120g of yellow vine, 130g of gardenia, 70g of trifoliate lanceolate, 30g of agarwood, 60g of camphor, and 60g of menthol; The anti-swelling and analgesic composition gel preparation made from the medicinal material of Rhizoma Cibotii is prepared from the following raw materials: 40g of costus root, 40g of siler, 40g of schizonepeta, 40g of asarum, 40g of acanthopanax bark, 40g of cinnamon twig, 40g of chyranthes bidentata, 40g of ligusticum chuanxiong, 40g of cyperus rotundus, 70g of angelica dahurica, 40g of curcuma, 70g of red eucommia, 130g of large parasol, 130g of small parasol, 130g of lancea chinensis, 120g of yellow vine, 130g of gardenia, 70g of trillium, 30g of agarwood, 60g of camphor, and 60g of menthol.

[0019] In the high performance liquid chromatography method for constructing the fingerprint, the method for preparing the test solution comprises the following steps: A detumescent and analgesic composition gel preparation containing 0.212 g of cinnabar root or Radix Aconiti Lateralis Preparata was placed in a 50 mL volumetric flask, 35 mL of n-butanol was added, and ultrasonic treatment was performed for 3 minutes. After cooling, the mixture was diluted to the mark with n-butanol, shaken, and filtered. 25 mL of the filtrate was accurately measured and evaporated to dryness. The residue was dissolved in methanol and diluted to a 10 mL volumetric flask. The mixture was filtered through a 0.45 μm microporous membrane to obtain the product. (2) Chromatographic conditions are: Column: Agilent ZORBAX Eclipse XDB-C18; 4.6 × 25 mm, 5 μm; Detection wavelength: 275nm; Column temperature: 40°C; Flow rate: 0.8 mL / min; Mobile phase: Mobile phase A is water, mobile phase B is methanol, gradient elution is used, and the proportions of mobile phases are all volume percentages. The elution procedure is as follows: 0-5 min, mobile phase A 92-85%, mobile phase B 8-15%; 5-15 min, mobile phase A 85%→70%, mobile phase B 15%→30%; 15-25 min, mobile phase A 70%→40%, mobile phase B 30%→60%; 25-35 min, mobile phase A 40%, mobile phase B 60%; 35.1-35 min, mobile phase A 40%→92%, mobile phase B 60%→8%; 35.1-45min, mobile phase A is 92%, mobile phase B is 8%.

[0020] (3) Test results: The following characteristic peaks were detected in the fingerprint of the swelling and analgesic tincture prepared with Xiaoluosan as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; auxiliary characteristic peaks: RT = 16.004 min, RT = 17.021 min, RT = 17.648 min, RT = 20.203 min, RT = 21.324 min; The following characteristic peaks were detected in the fingerprint of the anti-swelling and analgesic tincture prepared with cinnabar root as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; there is no characteristic peak at the position of the above-mentioned auxiliary characteristic peaks.

[0021] 2. Thin layer chromatography identification method: Taking the medicinal materials of Xiao Luosan and Zhu Shagen as reference, the method includes the following steps: (1) The test solution is prepared as follows: The preparation of the test solution of the medicinal material of Radix Cinnabarinii and the test solution of the medicinal material of Radix Cinnabarinii comprises the following steps: Take 0.2 g of powder of Radix Angelicae Dahuricae and Radix Cinnabarifolia respectively, add 30 mL of methanol respectively, and ultrasonically treat for 20 minutes. Let cool, filter, evaporate the filtrate to dryness, and dissolve the residue in 1 mL of methanol respectively. The method for preparing the test solution of the preparation comprises the following steps: Take a detumescent and analgesic composition gel preparation equivalent to 0.212g of cinnabar root or small umbrella medicinal material, add 40mL of methanol, ultrasonicate for 20 minutes, let cool, filter, evaporate the filtrate to dryness, add 2mL of methanol to dissolve the residue, centrifuge, and take the supernatant as the test solution.

[0022] (2) Detection method: The test samples made from the medicinal materials of Xiaoluosan, Zhushagen, and preparations were respectively spotted on the same silica gel G thin layer plate, and chloroform-ethyl acetate-methanol = 4:3:1 by volume was used as the developing solvent. The plate was developed, taken out, dried, and examined under a 365nm ultraviolet lamp. A fluorescent spot was detected in the chromatogram of the medicinal material of Zhushagen, but no fluorescent spot of the same color was detected at the corresponding position of the spot in the chromatogram of the medicinal material of Xiaoluosan. Based on whether the preparation test sample has the spot at that position, it can be distinguished whether the preparation contains Zhushagen.

[0023] Example 2 Methods for detecting and distinguishing Root of Cinnabarinum and Rhizoma Cibotii in pharmaceutical preparations, including high performance liquid chromatography fingerprint construction and thin layer chromatography identification; 1. Construction of fingerprint by high performance liquid chromatography (1) The test solution preparation method includes the following steps: The anti-swelling and analgesic composition gel preparations made from the medicinal materials of Cinnabar Root and Rhizoma Cibotii were respectively used as test samples; The anti-swelling and analgesic composition gel preparation made from the medicinal material cinnabar root is prepared from the following raw materials: 100g of costus root, 100g of siler, 100g of schizonepeta, 100g of asarum, 100g of acanthopanax bark, 100g of cinnamon twig, 100g of cyperus rotundus, 100g of ligusticum, 100g of ligusticum chuanxiong, 100g of cyperus rotundus, 120g of angelica dahurica, 100g of zedoaria, 120g of red eucommia bark, 180g of radix cinnabaris, 180g of zizyphus chinensis, 160g of yellow vine, 180g of gardenia, 120g of trifoliate lanceolate, 60g of agarwood, 100g of camphor, and 100g of menthol; The anti-swelling and analgesic composition gel preparation made from the medicinal material of Rhizoma Cibotii is prepared from the following raw materials: 100g of costus root, 100g of siler, 100g of schizonepeta, 100g of asarum, 100g of acanthopanax bark, 100g of cinnamon twig, 100g of chyranthes bidentata, 100g of ligusticum chuanxiong, 100g of cyperus rotundus, 120g of angelica dahurica, 100g of zedoaria, 120g of red eucommia bark, 180g of large parasol, 180g of small parasol, 180g of zizyphus chinensis, 160g of yellow vine, 180g of gardenia, 120g of trillium, 60g of agarwood, 100g of camphor, and 100g of menthol.

[0024] In the high performance liquid chromatography method for constructing the fingerprint, the method for preparing the test solution comprises the following steps: A detumescent and analgesic composition gel preparation containing 0.318 g of cinnabar root or Radix Cinnabarinii was placed in a 50 mL volumetric flask, 42 ​​mL of n-butanol was added, and ultrasonic treatment was performed for 50 minutes. After cooling, the mixture was diluted to the mark with n-butanol, shaken, and filtered. 25 mL of the filtrate was accurately measured and evaporated to dryness. The residue was dissolved in methanol and diluted to a 10 mL volumetric flask. The mixture was filtered through a 0.45 μm microporous filter membrane to obtain the product. (2) Chromatographic conditions are: Column: Agilent ZORBAX SB-C18; 4.6 × 25 mm, 5 μm; Detection wavelength: 275nm; Column temperature: 50°C; Flow rate: 1.3 mL / min; Mobile phase: Mobile phase A is water, mobile phase B is methanol, gradient elution is used, the mobile phase ratios are all volume percentages, and the elution procedure is as follows: 0-5 min, mobile phase A 92-85%, mobile phase B 8-15%; 5-15 min, mobile phase A 85%→70%, mobile phase B 15%→30%; 15-25 min, mobile phase A 70%→40%, mobile phase B 30%→60%; 25-35 min, mobile phase A 40%, mobile phase B 60%; 35.1-35 min, mobile phase A 40%→92%, mobile phase B 60%→8%; 35.1-45min, mobile phase A is 92%, mobile phase B is 8%.

[0025] (3) Test results: The following characteristic peaks were detected in the fingerprint of the swelling and analgesic tincture prepared with Xiaoluosan as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; auxiliary characteristic peaks: RT = 16.004 min, RT = 17.021 min, RT = 17.648 min, RT = 20.203 min, RT = 21.324 min; The following characteristic peaks were detected in the fingerprint of the anti-swelling and analgesic tincture prepared with cinnabar root as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; there is no characteristic peak at the position of the above-mentioned auxiliary characteristic peaks.

[0026] 2. Thin layer chromatography identification method, using the medicinal materials of Rhizoma Cibotii and Radix Cinnabari as reference, includes the following steps: (1) The test solution is prepared as follows: The preparation of the test solution of the medicinal material of Radix Cinnabarinii and the test solution of the medicinal material of Radix Cinnabarinii comprises the following steps: Take 0.6 g of powder of Radix Angelicae Dahuricae and Radix Cinnabarifolia respectively, add 50 mL of methanol respectively, and ultrasonically treat for 40 minutes. Let cool, filter, evaporate the filtrate to dryness, and dissolve the residue in 3 mL of methanol respectively. The method for preparing the test solution of the preparation comprises the following steps: Take a detumescent and analgesic composition gel preparation equivalent to 0.318g of cinnabar root or small umbrella medicinal material, add 60mL of methanol, ultrasonicate for 40 minutes, let cool, filter, evaporate the filtrate to dryness, add 4mL of methanol to dissolve the residue, centrifuge, and take the supernatant as the test solution.

[0027] (2) Detection method: The test samples made from the medicinal materials of Xiaoluosan, Zhushagen, and preparations were respectively spotted on the same silica gel G thin layer plate, and chloroform-ethyl acetate-methanol = 6:5:3 by volume was used as the developing solvent. The plate was developed, taken out, dried, and examined under a 365nm ultraviolet lamp. A fluorescent spot was detected in the chromatogram of the medicinal material of Zhushagen, but no fluorescent spot of the same color was detected at the corresponding position of the spot in the chromatogram of the medicinal material of Xiaoluosan. Based on whether the preparation test sample has the spot at that position, it can be distinguished whether the preparation contains Zhushagen.

[0028] Example 3 Methods for detecting and distinguishing Root of Cinnabarinum and Rhizoma Cibotii in pharmaceutical preparations, including high performance liquid chromatography fingerprint construction and thin layer chromatography identification; 1. Construction of fingerprint by high performance liquid chromatography (1) The test solution preparation method includes the following steps: The anti-swelling and analgesic composition gel preparations made from the medicinal materials of Cinnabar Root and Rhizoma Cibotii were respectively used as test samples; The anti-swelling and analgesic composition gel preparation made from the medicinal material cinnabar root is prepared from the following raw materials: Costus root 71g, Saposhnikovia root 71g, Schizonepeta tenuifolia 71g, Asarum 71g, Acanthopanax acanthopanax bark 71g, Cinnamon twig 71g, Achyranthes bidentata 71g, Chuanxiong rhizome 71g, Cynanchum indica 71g, Angelica dahurica 106g, Curcuma 71g, Eucommia barbata 106g, Daluosan 152g, Cinnabar root 106g, Zanthoxylum bungeanum 152g, Phellodendron chinense 144g, Gardenia jasminoides 152g, Tripterygium wilfordii 106g, Aquilaria sinensis 49g, Camphor 83g, Menthol 83g; The anti-swelling and analgesic composition gel preparation made from the medicinal material of Rhizoma Cibotii is prepared from the following raw materials: Costus root 71g, Saposhnikovia root 71g, Schizonepeta tenuifolia 71g, Asarum 71g, Acanthopanax acanthopanax 71g, Cinnamon twig 71g, Achyranthes bidentata 71g, Ligusticum chuanxiong 71g, Cynanchum indicum 71g, Angelica dahurica 106g, Curcuma 71g, Eucommia bark 106g, Radix Angelicae Dahuricae 152g, Radix Angelicae Sinensis 106g, Zanthoxylum bungeana 106g, Zanthoxylum bungeana 152g, Radix Zanthoxylum bungeana 144g, Gardenia jasminoides 152g, Trigonosciadium japonicum 106g, Aquilaria sinensis 49g, Camphor 83g, Menthol 83g.

[0029] In the high performance liquid chromatography method for constructing the fingerprint, the method for preparing the test solution comprises the following steps: A detumescent and analgesic composition gel preparation containing 0.26 g of cinnabar root or Radix Cinnabarinii was placed in a 50 mL volumetric flask, 40 mL of n-butanol was added, and ultrasonic treatment was performed for 40 minutes. After cooling, the mixture was diluted to the mark with n-butanol, shaken, and filtered. 25 mL of the filtrate was accurately measured and evaporated to dryness. The residue was dissolved in methanol and diluted to a 10 mL volumetric flask. The mixture was filtered through a 0.45 μm microporous membrane to obtain the product. (2) Chromatographic conditions are: Column: SHIMADZU Shim-pack GIST C18; 4.6 × 25 mm, 5 μm; Detection wavelength: 275nm; Column temperature: 40-50°C; Flow rate: 0.8-1.3 mL / min; Mobile phase: Mobile phase A is water, mobile phase B is methanol, gradient elution is used, the mobile phase ratios are all volume percentages, and the elution procedure is as follows: 0-5 min, mobile phase A 92-85%, mobile phase B 8-15%; 5-15 min, mobile phase A 85%→70%, mobile phase B 15%→30%; 15-25 min, mobile phase A 70%→40%, mobile phase B 30%→60%; 25-35 min, mobile phase A 40%, mobile phase B 60%; 35.1-35 min, mobile phase A 40%→92%, mobile phase B 60%→8%; 35.1-45min, mobile phase A is 92%, mobile phase B is 8%.

[0030] (3) Test results: The following characteristic peaks were detected in the fingerprint of the swelling and analgesic tincture prepared with Xiaoluosan as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; auxiliary characteristic peaks: RT = 16.004 min, RT = 17.021 min, RT = 17.648 min, RT = 20.203 min, RT = 21.324 min; The following characteristic peaks were detected in the fingerprint of the anti-swelling and analgesic tincture prepared with cinnabar root as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; there is no characteristic peak at the position of the above-mentioned auxiliary characteristic peaks.

[0031] 2. Thin layer chromatography identification method, using the medicinal materials of Rhizoma Cibotii and Radix Cinnabari as reference, includes the following steps: (1) The test solution is prepared as follows: The preparation of the test solution of the medicinal material of Radix Cinnabarinii and the test solution of the medicinal material of Radix Cinnabarinii comprises the following steps: Take 0.3 g of powder of Radix Angelicae Dahuricae and Radix Cinnabarifoliae respectively, add 40 mL of methanol, ultrasonically treat for 30 minutes, cool, filter, The filtrate was evaporated to dryness, and 2 mL of methanol was added to dissolve the residue to obtain the obtained product. The method for preparing the test solution of the preparation comprises the following steps: Take a detumescent and analgesic composition gel preparation equivalent to 0.26g of cinnabar root or small umbrella medicinal material, add 60mL of methanol, ultrasonicate for 40 minutes, let cool, filter, evaporate the filtrate to dryness, add 4mL of methanol to dissolve the residue, centrifuge, and take the supernatant as the test solution.

[0032] (2) Detection method: The test samples made from the medicinal materials of Xiaoluosan, Zhushagen, and preparations were respectively spotted on the same silica gel G thin layer plate, and chloroform-ethyl acetate-methanol = 5:4:2 by volume was used as the developing solvent. The plate was developed, taken out, dried, and examined under a 365nm ultraviolet lamp. A fluorescent spot was detected in the chromatogram of the medicinal material of Zhushagen, but no fluorescent spot of the same color was detected at the corresponding position of the spot in the chromatogram of the medicinal material of Xiaoluosan. Whether the preparation contained Zhushagen was determined by whether the spot existed at that position in the test sample of the preparation.

[0033] Example 4 Methods for detecting and distinguishing Root of Cinnabarinum and Rhizoma Cibotii in pharmaceutical preparations, including high performance liquid chromatography fingerprint construction and thin layer chromatography identification; 1. Construction of fingerprint by high performance liquid chromatography (1) The test solution preparation method comprises the following steps: The Zhu Sharen and Xiaoluosan medicinal materials are respectively prepared into 20 mL of Xiaozhong Zhitong Tincture, and then respectively evaporated to dryness. The test solution preparation method in the high performance liquid chromatography fingerprint construction comprises the following steps: The Zhu Sharen and Xiaoluosan medicinal materials are respectively prepared into 20 mL of Xiaozhong Zhitong Tincture, and then respectively evaporated to dryness.

[0034] (2) The chromatographic conditions are as follows: The chromatographic column is SHIMADZU Shim-pack GIST C18; 4.6*25 mm, 5 μm. The detection wavelength is 275 nm. The column temperature is 40-50 DEG C. The flow rate is 0.8-1.3 mL / min. The mobile phase is water for mobile phase A and methanol for mobile phase B, and gradient elution is adopted, and the proportion of the mobile phase is all volume percent, and the elution program is as follows: 0-5 min, mobile phase A 92-85%, mobile phase B 8-15%; 5-15 min, mobile phase A 85%→70%, mobile phase B 15%→30%; 15-25 min, mobile phase A 70%→40%, mobile phase B 30%→60%; 25-35 min, mobile phase A 40%, mobile phase B 60%; 35.1-35 min, mobile phase A 40%→92%, mobile phase B 60%→8%; 35.1-45 min, mobile phase A is 92%, and mobile phase B is 8%.

[0035] (3) The detection results are as follows: The Xiaozhong Zhitong Tincture prepared by taking Xiaoluosan as one of the raw materials has the following characteristic peaks in the fingerprint spectrum: Main peak: RT=13.105±0.1 min; auxiliary characteristic peaks: RT=16.004 min, RT=17.021 min, RT=17.648 min, RT=20.203 min, RT=21.324 min; The following characteristic peaks were detected in the fingerprint of the anti-swelling and analgesic tincture prepared with cinnabar root as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; there is no characteristic peak at the position of the above-mentioned auxiliary characteristic peaks.

[0036] 2. Thin layer chromatography identification method, using the medicinal materials of Rhizoma Cibotii and Radix Cinnabari as reference, includes the following steps: (1) The test solution is prepared as follows: The preparation of the test solution of the medicinal material of Radix Cinnabarinii and the test solution of the medicinal material of Radix Cinnabarinii comprises the following steps: Take 0.3 g of powder of Radix Angelicae Dahuricae and Radix Cinnabarifolia respectively, add 40 mL of methanol respectively, and ultrasonically treat for 30 minutes. Let cool, filter, evaporate the filtrate to dryness, and dissolve the residue in 2 mL of methanol respectively. The method for preparing the test solution of the preparation comprises the following steps: The method for preparing the test solution of the preparation comprises the following steps: 30 mL of the finished product of the anti-swelling and analgesic tincture prepared from the medicinal materials of Rhizoma Cinnabarinii and Radix Cinnabarinii were measured respectively, and 60 mL of methanol was added respectively. The mixture was ultrasonicated for 40 minutes respectively, cooled, filtered, and the filtrate was evaporated to dryness. 4 mL of methanol was added to the residue to dissolve it, centrifuged, and the supernatant was used as the test solution.

[0037] (2) Detection method: The test samples made from the medicinal materials of Xiaoluosan, Zhushagen, and preparations were respectively spotted on the same silica gel G thin layer plate, and chloroform-ethyl acetate-methanol = 5:4:2 by volume was used as the developing solvent. The plate was developed, taken out, dried, and examined under a 365nm ultraviolet lamp. A fluorescent spot was detected in the chromatogram of the medicinal material of Zhushagen, but no fluorescent spot of the same color was detected at the corresponding position of the spot in the chromatogram of the medicinal material of Xiaoluosan. Whether the preparation contained Zhushagen was determined by whether the spot existed at that position in the test sample of the preparation.

[0038] Example 5 Methods for detecting and distinguishing Root of Cinnabarinum and Rhizoma Cibotii in pharmaceutical preparations, including high performance liquid chromatography fingerprint construction and thin layer chromatography identification; 1. High performance liquid chromatography fingerprint construction: (1) The method for preparing the test solution includes the following steps: The anti-swelling and analgesic tinctures made from the medicinal materials of Cinnabar Root and Rhizoma Cibotii were used as test samples; In the high performance liquid chromatography method for constructing the fingerprint, the method for preparing the test solution comprises the following steps: 30 mL of each of the anti-swelling and analgesic tinctures made from cinnabar root and radix scutellariae were measured and evaporated to dryness, and the residues were dissolved in 41 mL of n-butanol, transferred to different 50 mL volumetric flasks, and ultrasonically treated for 50 minutes. After cooling, n-butanol was added to dilute to the scale, shaken, and filtered. 25 mL of the filtrate was accurately measured and evaporated to dryness, and the residues were dissolved in methanol and diluted to different 10 mL volumetric flasks. After filtering with a 0.45 μm microporous filter membrane, the product was obtained.

[0039] (2) Chromatographic conditions are: Column: SHIMADZU Shim-pack GIST C18; 4.6 × 25 mm, 5 μm; Detection wavelength: 275nm; Column temperature: 40-50°C; Flow rate: 0.8-1.3 mL / min; Mobile phase: Mobile phase A is water, mobile phase B is methanol, gradient elution is used, the mobile phase ratios are all volume percentages, and the elution procedure is as follows: 0-5 min, mobile phase A 92-85%, mobile phase B 8-15%; 5-15 min, mobile phase A 85%→70%, mobile phase B 15%→30%; 15-25 min, mobile phase A 70%→40%, mobile phase B 30%→60%; 25-35 min, mobile phase A 40%, mobile phase B 60%; 35.1-35 min, mobile phase A 40%→92%, mobile phase B 60%→8%; 35.1-45min, mobile phase A is 92%, mobile phase B is 8%.

[0040] (3) Test results: The following characteristic peaks were detected in the fingerprint of the swelling and analgesic tincture prepared with Xiaoluosan as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; auxiliary characteristic peaks: RT = 16.004 min, RT = 17.021 min, RT = 17.648 min, RT = 20.203 min, RT = 21.324 min; The following characteristic peaks were detected in the fingerprint of the anti-swelling and analgesic tincture prepared with cinnabar root as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; there is no characteristic peak at the position of the above-mentioned auxiliary characteristic peaks.

[0041] 2. Thin layer chromatography identification method, using the medicinal materials of Rhizoma Cibotii and Radix Cinnabari as reference, includes the following steps: (1) The preparation method of the test solution of the sample is as follows: The preparation of the test solution of the sample of the Xialuozao medicinal material and the test solution of the sample of the Zhushagen medicinal material includes the following steps: 0.3g of the Xialuozao and Zhushagen medicinal material powder was respectively added with 40ml of methanol, and ultrasonic treatment was performed for 30 minutes, and then the mixture was cooled, filtered, and the filtrate was evaporated to dryness. The preparation method of the test solution of the sample of the preparation includes the following steps: 30ml of the prepared Xialuozao and Zhushagen medicinal material preparation was respectively added with 60ml of methanol, and ultrasonic treatment was performed for 40 minutes, and then the mixture was cooled, filtered, and the filtrate was evaporated to dryness.

[0042] (2) The detection method: The test solutions of the samples of the Xialuozao, Zhushagen medicinal material, and preparation were respectively spotted on the same silica gel G thin layer plate, and chloroform-ethyl acetate-methanol=5:4:2 was used as the developing agent, and then the mixture was developed, taken out, dried, and observed under a 365nm ultraviolet lamp.

[0043] Example 6 The detection and differentiation method of Zhushagen and Xialuozao in the pharmaceutical preparation includes the construction of a fingerprint by high performance liquid chromatography and thin layer chromatography identification. 1. The construction of a fingerprint by high performance liquid chromatography: (1) The preparation method of the test solution of the sample includes the following steps: (1) The preparation method of the test solution of the sample includes the following steps: The Zhushagen and Xialuozao medicinal material prepared into the anti-inflammatory analgesic tincture were used as the test samples. In the construction of a fingerprint by high performance liquid chromatography, the preparation method of the test solution of the sample includes the following steps: 25ml of the anti-inflammatory analgesic tincture prepared from Zhushagen and Xialuozao medicinal material was respectively evaporated to dryness, and the residue was dissolved in 35ml of n-butanol, and then transferred into different 50ml volumetric flasks, and ultrasonic treatment was performed for 40 minutes, and then n-butanol was added to dilute to the mark, and the mixture was shaken, filtered, and 25ml of the filtrate was precisely measured, evaporated to dryness, dissolved in methanol, and diluted to different 10ml volumetric flasks, and then filtered through a 0.45μm microporous filter to obtain the test solution of the sample.

[0044] (2) The chromatographic conditions are as follows: Column: SHIMADZU Shim-pack GIST C18; 4.6 × 25 mm, 5 μm; Detection wavelength: 275nm; Column temperature: 40-50°C; Flow rate: 0.8-1.3 mL / min; Mobile phase: Mobile phase A is water, mobile phase B is methanol, gradient elution is used, the mobile phase ratios are all volume percentages, and the elution procedure is as follows: 0-5 min, mobile phase A 92-85%, mobile phase B 8-15%; 5-15 min, mobile phase A 85%→70%, mobile phase B 15%→30%; 15-25 min, mobile phase A 70%→40%, mobile phase B 30%→60%; 25-35 min, mobile phase A 40%, mobile phase B 60%; 35.1-35 min, mobile phase A 40%→92%, mobile phase B 60%→8%; 35.1-45min, mobile phase A is 92%, mobile phase B is 8%.

[0045] (3) Test results: The following characteristic peaks were detected in the fingerprint of the swelling and analgesic tincture prepared with Xiaoluosan as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; auxiliary characteristic peaks: RT = 16.004 min, RT = 17.021 min, RT = 17.648 min, RT = 20.203 min, RT = 21.324 min; The following characteristic peaks were detected in the fingerprint of the anti-swelling and analgesic tincture prepared with cinnabar root as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; there is no characteristic peak at the position of the above-mentioned auxiliary characteristic peaks.

[0046] 2. Thin layer chromatography identification method, using the medicinal materials of Rhizoma Cibotii and Radix Cinnabari as reference, includes the following steps: (1) The test solution is prepared as follows: The preparation of the test solution of the medicinal material of Radix Cinnabarinii and the test solution of the medicinal material of Radix Cinnabarinii comprises the following steps: Take 0.3 g of powder of Radix Angelicae Dahuricae and Radix Cinnabarifolia respectively, add 40 mL of methanol respectively, and ultrasonically treat for 30 minutes. Let cool, filter, evaporate the filtrate to dryness, and dissolve the residue in 2 mL of methanol respectively. The method for preparing the test solution of the preparation comprises the following steps: The method for preparing the test solution of the preparation comprises the following steps: 30 mL of the finished product of the anti-swelling and analgesic tincture prepared from the medicinal materials of Rhizoma Cinnabarinii and Radix Cinnabarinii were measured respectively, and 60 mL of methanol was added respectively. The mixture was ultrasonicated for 40 minutes respectively, cooled, filtered, and the filtrate was evaporated to dryness. 4 mL of methanol was added to the residue to dissolve it, centrifuged, and the supernatant was used as the test solution.

[0047] (2) Detection method: The test samples made from the medicinal materials of Xiaoluosan, Zhushagen, and preparations were respectively spotted on the same silica gel G thin layer plate, and chloroform-ethyl acetate-methanol = 5:4:2 by volume was used as the developing solvent. The plate was developed, taken out, dried, and examined under a 365nm ultraviolet lamp. A fluorescent spot was detected in the chromatogram of the medicinal material of Zhushagen, but no fluorescent spot of the same color was detected at the corresponding position of the spot in the chromatogram of the medicinal material of Xiaoluosan. Whether the preparation contained Zhushagen was determined by whether the spot existed at that position in the test sample of the preparation.

[0048] Example 7 Thin layer identification method confirmation test 1. Inspection method; Take 0.4 g of powdered Radix Angelicae Dahuricae and Radix Cinnabarifolia, add 40 mL of methanol, and sonicate for 30 minutes. Cool, filter, and evaporate the filtrate to dryness. Dissolve the residue in 2 mL of methanol to prepare the test solution. According to the thin-layer chromatography method (General Method 0502), spot 5–8 μl of each test solution on a single silica gel G thin-layer plate using chloroform-ethyl acetate (20:1) as the developing solvent. Develop the plate, remove the plate, air dry, and examine the plate under 365 nm UV light to observe if there are any differences in the spots between the two test samples.

[0049] 2. Comparison and inspection of 10 batches of Cinnabar Root and 10 batches of Osmanthus fragrans medicinal materials; According to the above test method, 10 batches of cinnabar root and 10 batches of small umbrella medicinal materials were tested respectively, and the thin layer chromatography was compared. The results showed that in the chromatograms of the 10 batches of cinnabar root medicinal materials, the corresponding position of the fluorescent spot was displayed, and in the chromatograms of the 10 batches of small umbrella medicinal materials, no fluorescent spot of the same color was detected, confirming the repeatability and feasibility of the method. The test results are shown in Figure 1 、 2 .

[0050] Figure 1 The following are the thin layer chromatography chromatograms of three batches of medicinal materials, namely, Zhushagen and Xiaoluosan; the order of spotting is: from left to right, 5 μl of Xiaoluosan batch 20240101 medicinal materials, 5 μl of Xiaoluosan batch 20240201 medicinal materials, 5 μl of Xiaoluosan batch 20240301 medicinal materials, 5 μl of Zhushagen batch 20230101 medicinal materials, 5 μl of Zhushagen batch 20230201 medicinal materials, and 5 μl of Zhushagen batch 20230301 medicinal materials; Figure 2 Thin layer chromatography spectra of 7 batches of medicinal materials Zhushagen and Xiaoluosan; the order of spotting: from left to right is 5μl of Xiaoluosan 20240501 batch of medicinal materials, 5μl of Xiaoluosan 20240502 batch of medicinal materials, 5μl of Xiaoluosan 20240601 batch of medicinal materials, 5μl of Xiaoluosan 20240602 batch of medicinal materials, 5μl of Xiaoluosan 20240603 batch of medicinal materials, 5μl of Xiaoluosan 20240701 batch of medicinal materials, 5μl of Xiaoluosan 20240702 batch of medicinal materials, 5μl of Zhushagen 20230401 batch of medicinal materials , 5 μl of medicinal materials from batch 20230501 of Cinnabar root, 5 μl of medicinal materials from batch 20230601 of Cinnabar root, 5 μl of medicinal materials from batch 20230701 of Cinnabar root, 5 μl of medicinal materials from batch 20230702 of Cinnabar root, 5 μl of medicinal materials from batch 20230801 of Cinnabar root, 5 μl of medicinal materials from batch 20230802 of Cinnabar root; 3. Specificity inspection According to the inspection method in step 1 above (spread distance 13cm), the finished product of detumescent and analgesic tincture (made from cinnabar root medicinal material) and the negative sample of detumescent and analgesic tincture lacking small umbrella were inspected and tested. The preparation method of the test sample is as follows: measure 25mL of the finished product of detumescent and analgesic tincture, add 50mL of methanol, ultrasonicate for 30 minutes, let cool, filter, evaporate the filtrate to dryness, add 3mL of methanol to the residue to dissolve it, centrifuge, and take the supernatant as the test sample solution; the negative sample was prepared in the same way as the test sample. The inspection results showed that in the chromatogram of the finished product of detumescent and analgesic tincture (made from cinnabar root medicinal material), the same color fluorescent spot appeared at the corresponding position of the chromatogram of cinnabar root medicinal material. In the chromatogram of the negative sample of detumescent and analgesic tincture lacking small umbrella, the same color fluorescent spot was not detected at the corresponding position of the chromatogram of cinnabar root medicinal material, confirming that the method has good specificity. The results showed that Figure 3 ; Figure 3 Thin layer chromatography chromatograms of the finished product of detumescence and analgesia tincture (made from Cinnabar root) and the negative sample lacking Xiaoluosan; order of spotting: from left to right: 5μl of Xiaoluosan batch 20240101 medicinal materials, 5μl of Cinnabar root batch 20230101 medicinal materials, 5μl of the finished product of detumescence and analgesia tincture (made from Cinnabar root), 5μl of the negative sample lacking Xiaoluosan, 5μl of the negative sample lacking Xiaoluosan, 5μl of the negative sample lacking Xiaoluosan, 5μl of the negative sample lacking Xiaoluosan, and 5μl of the finished product of detumescence and analgesia tincture (made from Cinnabar root).

[0051] 4. Conclusion The above experimental studies show that the thin layer chromatography method of the present invention can better distinguish the medicinal materials of Xiaoluosan and Zhushagen by testing them separately. There is a certain difference in the quality of the two medicinal materials. The thin layer method can also better distinguish the type of Xiaoluosan medicinal materials used in the finished product of swelling and analgesic tincture.

[0052] Example 8 Confirmation test of high performance liquid chromatography fingerprint identification method 1. Experimental materials and instruments 1.1 Reagents Methanol (analytical grade) was purchased from Chengdu Kelong Chemical Co., Ltd.; n-butanol (analytical grade) was purchased from Chengdu Kelong Chemical Co., Ltd.; methanol (chromatographic grade) was purchased from Xilong Science Co., Ltd.; fresh ultrapure water was prepared in the laboratory.

[0053] 1.2 Sample Samples: Anti-swelling and analgesic tincture (made from the medicinal material of Rhizoma Cibotii), Anti-swelling and analgesic tincture (made from the medicinal material of Cinnabar Root) 1.3 Instruments and Equipment KQ-500DA CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); LC-40D high-performance liquid chromatograph (Shimadzu Enterprise Management (China) Co., Ltd.); HH digital display constant temperature water bath (Changzhou Ronghua Instrument Manufacturing Co., Ltd.).

[0054] 2. Experimental methods 2.1 Sample preparation and pretreatment Preparation of test solution: 25 mL of the preparation made from Cinnabar Root and Rhizoma Cinnabarinii were measured and evaporated to dryness, the residue was dissolved in 40 mL of n-butanol, transferred to a 50 mL volumetric flask, and ultrasonically treated for 40 minutes. After cooling, it was diluted to the scale and filtered. 25 mL of the filtrate was accurately measured and evaporated to dryness. The residue was dissolved in methanol and diluted to a 10 mL volumetric flask. It was filtered through a 0.45 μ microporous membrane and analyzed.

[0055] 2.2 Chromatographic conditions Chromatographic column: SHIMADZU Shim-pack GIST C18 (4.6×25 mm, 5 μm); column temperature: 45°C; flow rate: 1 mL / min; injection volume: 10 μL; detection wavelength: 275 nm; mobile phase: water (phase A)-methanol (phase B) binary gradient elution system, gradient elution program: 0-5 min, 8%-15% B; 5-15 min, 15%-30% B; 15-25 min, 30%-60% B; 25-35 min, 60% B; 35-35.1 min, 60%-8%; 35.1-45 min, 8%.

[0056] 3. Results and Discussion Comparative analysis of fingerprints of preparations of Radix Angelicae Dahuricae and Radix Cinnabarinii The fingerprints of the anti-swelling and analgesic tincture (made from the medicinal material of Rhizoma Cibotii) and the anti-swelling and analgesic tincture (made from the medicinal material of Cinnabar Root) collected by the above-mentioned liquid chromatography method are as follows: Figure 4 、 5 The fingerprint comparison of the two preparations is shown in Figure 6 As shown, Figure 6The upper middle one is the medicinal preparation of Luosan, and the lower one is the medicinal preparation of Zhushagen.

[0057] The above fingerprint comparison results show that the fingerprints of the two preparations of Xiaoluosan and Zhushagen are quite different. Although the main peak (RT=13.105±0.1min) of the two preparations at a wavelength of 275nm is the same component, the other key components detected are different to varying degrees. For example, in the fingerprint of the swelling and analgesic tincture (made from Xiaoluosan medicinal material), the peaks of RT=16.004min, RT=17.021min, RT=17.648min, RT=20.203min, and RT=21.324min were not detected in the fingerprint of the swelling and analgesic tincture (made from Zhushagen medicinal material).

[0058] 4. Summary The above experimental studies show that the high performance liquid chromatography method of the present invention can better distinguish the preparations made from the medicinal materials of Cinnabar Root and Small Parasol by testing the fingerprints of the preparations made from the medicinal materials of Small Parasol and Cinnabar Root respectively. There are significant differences in the fingerprints of the two preparations, which proves that the quality of the preparations made from the two medicinal materials is quite different.

Claims

1. A method for detecting and distinguishing Root of Cinnabarinum and Rhizoma Cibotii in a pharmaceutical preparation, characterized by: High performance liquid chromatography was used to construct the fingerprint for detection, and the chromatographic conditions were as follows: Chromatographic column: C18 column; Detection wavelength: 275nm; Mobile phase: Mobile phase A is water, mobile phase B is methanol, gradient elution is used, the mobile phase ratios are all volume percentages, and the elution procedure is as follows: 0-5 min, mobile phase A 92-85%, mobile phase B 8-15%; 5-15 min, mobile phase A 85%→70%, mobile phase B 15%→30%; 15-25 min, mobile phase A 70%→40%, mobile phase B 30%→60%; 25-35 min, mobile phase A 40%, mobile phase B 60%; 35.1-35 min, mobile phase A 40%→92%, mobile phase B 60%→8%; 35.1-45min, mobile phase A is 92%, mobile phase B is 8%.

2. The method for detecting and distinguishing between Radix Cinnabarinii and Radix Rhizoma Cinnabarinii in the pharmaceutical preparation according to claim 1, characterized in that: The specifications of the C18 chromatographic column are: 4.6×25 mm, 5 μm; the chromatographic conditions also include: column temperature: 40-50°C; flow rate: 0.8-1.3 mL / min.

3. The method for detecting and distinguishing between Radix Cinnabarinii and Radix Rhizoma Cinnabarinii in the pharmaceutical preparation according to claim 1, characterized in that: The C18 chromatographic column is: SHIMADZU Shim-pack GIST C18 chromatographic column.

4. The method for detecting and distinguishing between Radix Cinnabarinii and Radix Rhizoma Cinnabarinii in the pharmaceutical preparation according to claim 1, characterized in that: The method also includes a thin layer chromatography identification method for detection, using the medicinal materials of Xiao Luosan and Zhu Shagen as references, including the following steps: The test samples made from the medicinal materials of Xiaoluosan, Zhushagen, and preparations were respectively spotted on the same silica gel G thin layer plate, and chloroform-ethyl acetate-methanol was used as the developing solvent in a volume ratio of 4-6:3-5:1-3. The plate was developed, taken out, dried, and examined under a 365nm ultraviolet lamp. A fluorescent spot was detected in the chromatogram of the medicinal material of Zhushagen, and no fluorescent spot of the same color was detected at the corresponding position of the spot in the chromatogram of the medicinal material of Xiaoluosan. Based on whether the preparation test sample has the spot at that position, it can be distinguished whether the preparation contains Zhushagen.

5. The method for detecting and distinguishing Root of Cinnabarinum and Rhizoma Cibotii in the pharmaceutical preparation according to claim 1 or 4, characterized in that: The pharmaceutical preparation is a liquid preparation.

6. The method for detecting and distinguishing Root of Cinnabarinum and Rhizoma Cibotii in a pharmaceutical preparation according to claim 1, characterized in that: In the high performance liquid chromatography method for constructing the fingerprint, the method for preparing the test solution comprises the following steps: Preparations made from the medicinal materials of Cinnabar Root and Rhizoma Cibotii were taken respectively; If it is a solid preparation, grind the solid preparation containing 0.212-0.318g of cinnabar root or Radix Aconiti Lateralis Preparata and place it in a 50mL volumetric flask. Add 35-42mL of n-butanol and sonicate for 30-50 minutes. After cooling, dilute to the mark with n-butanol, shake, filter, accurately measure 25mL of the filtrate, evaporate to dryness, dissolve the residue with methanol and dilute to a 10mL volumetric flask. Filter through a 0.45μm microporous filter membrane to obtain the product. If it is a liquid preparation, 20-30 mL of the liquid preparation of the cinnabar root or the small umbrella medicinal material is measured, evaporated to dryness, the residue is dissolved with 30-41 mL of n-butanol, transferred to a 50 mL volumetric flask, ultrasonically treated for 30-50 minutes, cooled, diluted to the scale with n-butanol, shaken, filtered, accurately measured 25 mL of the filtrate, evaporated to dryness, the residue is dissolved with methanol and diluted to a 10 mL volumetric flask, filtered with a 0.45 μm microporous filter membrane, and the product is obtained.

7. The method for detecting and distinguishing between Radix Cinnabarinii and Radix Rhizoma Cinnabarinii in a pharmaceutical preparation according to claim 5, characterized in that: In the thin layer chromatography identification method, the test solution preparation method is as follows: The preparation of the test solution of the medicinal material of Radix Cinnabarinii and the test solution of the medicinal material of Radix Cinnabarinii comprises the following steps: Take 0.2-0.6g of powder of Radix Angelicae Dahuricae and Radix Cinnabarifolia respectively, add 30-50mL of methanol respectively, and ultrasonically treat for 20-40 minutes. Let cool, filter, evaporate the filtrate to dryness, and dissolve the residue in 1-3mL of methanol respectively. The method for preparing the test solution of the preparation comprises the following steps: Take 20-30 mL of the liquid preparation made from the medicinal materials of Cinnabar Root and Small Parashorea chinensis, add 40-60 mL of methanol, ultrasonicate for 20-40 minutes, cool, filter, evaporate the filtrate to dryness, add 2-4 mL of methanol to dissolve the residue, centrifuge, and take the supernatant as the test solution.

8. The method for detecting and distinguishing Radix Cinnabarinii and Radix Rhizoma Cinnabarinii in the pharmaceutical preparation according to claim 1 or 4, characterized in that: The pharmaceutical preparation is a swelling-reducing and analgesic tincture prepared using cinnabar root or Rhizoma Cinnabarinii as one of the raw materials; Prepared from the following raw materials: 40-100 parts of costus root, 40-100 parts of siler, 40-100 parts of schizonepeta, 40-100 parts of asarum, 40-100 parts of acanthopanax bark, 40-100 parts of cassia twig, 40-100 parts of cyperus rotundus, 40-100 parts of ligusticum chuanxiong, 40-100 parts of cyperus rotundus, 70-120 parts of angelica dahurica, 40-100 parts of zedoaria, 70-120 parts of red eucommia bark, 130-180 parts of datura, 130-180 parts of cinnabar root, 130-180 parts of zymophora japonica, 120-160 parts of yellow twig, 130-180 parts of gardenia, 70-120 parts of trigonosciadium japonicum, 30-60 parts of agarwood, 60-100 parts of camphor, and 60-100 parts of menthol; Or prepared from the following raw materials: 40-100 parts of costus root, 40-100 parts of siler, 40-100 parts of schizonepeta, 40-100 parts of asarum, 40-100 parts of acanthopanax bark, 40-100 parts of cassia twig, 40-100 parts of cyperus rotundus, 40-100 parts of ligusticum chuanxiong, 40-100 parts of cyperus rotundus, 70-120 parts of angelica dahurica, 40-100 parts of zedoaria, 70-120 parts of red eucommia, 130-180 parts of radix scutellariae, 130-180 parts of radix scutellariae, 130-180 parts of zizyphus chinensis, 120-160 parts of truncatum, 130-180 parts of gardenia, 70-120 parts of trigonosciadium japonicum, 30-60 parts of aquilariae, 60-100 parts of camphor, and 60-100 parts of menthol.

9. The method for detecting and distinguishing Root of Cinnabarinum and Rhizoma Cibotii in a pharmaceutical preparation according to claim 8, characterized in that: The following characteristic peaks were detected in the fingerprint of the swelling and analgesic tincture prepared with Xiaoluosan as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; auxiliary characteristic peaks: RT = 16.004 min, RT = 17.021 min, RT = 17.648 min, RT = 20.203 min, RT = 21.324 min; The following characteristic peaks were detected in the fingerprint of the swelling-reducing and analgesic tincture prepared with cinnabar root as one of the raw materials: Main peak: RT = 13.105 ± 0.1 min; there is no characteristic peak at the position of the above-mentioned auxiliary characteristic peaks.

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