Meridian conditioning health-care paste and purification method thereof

Abstract

The invention discloses a meridian conditioning health-care paste and a purification method thereof, and belongs to the technical field of traditional Chinese medicines. Comprising the following components: frankincense, angelica sinensis, myrrh, borneol, radix angelicae, rheum officinale, artificial musk, golden cypress, radix saposhnikoviae, safflower carthamus, resina draconis, liquorice, asarum, coptis chinensis, notopterygium root, radix angelicae pubescentis, radix paeoniae rubra, cinnamon, wormwood essential oil, garter snake, black ant, earthworm, ground beetle, corydalis tuber, flos caryophylli, pseudo-ginseng, polygonum cuspidatum, liquidambar formosana hance, peach kernel, garden balsam stem, vaseline, glycerol, hirudin, centipede, dung beetle, nidus vespae and ligusticum wallichii. The formula provided by the invention is prepared from data mining of medication rules of a plurality of external pain-relieving traditional Chinese medicine prescriptions in combination with a traditional Chinese medicine syndrome differentiation and treatment theory. According to the purification method, modern ultrasonic low-temperature extraction is adopted to fully extract effective components, and the problem that high-temperature denatured effective components of animal medicinal materials are difficult to extract is solved.

Description

Technical Field

[0001] The invention relates to a meridian regulating health-care paste and a purification method thereof, belonging to the technical field of traditional Chinese medicines. Background Art

[0002] Existing meridian conditioning health creams generally use frying to extract active ingredients, but frying has the problems of high extraction temperature, thermally unstable ingredients are easily destroyed by high temperature, active ingredient proteins or polypeptides in animal drugs are easily denatured, and effective ingredients are difficult to extract. Therefore, existing meridian conditioning health creams generally have low effective ingredient content, slow onset, large side effects, and are easy to delay the disease. Therefore, there is an urgent need for a meridian conditioning health cream with high effective ingredient content and fast onset and a purification method thereof. Summary of the invention

[0003] The purpose of the present invention is to provide a meridian regulating health cream in view of the problems existing in the prior art. The formula of the health cream is derived from data mining of the medication rules of various external analgesic Chinese medicine prescriptions and is formulated in combination with the theory of syndrome differentiation and treatment of traditional Chinese medicine.

[0004] Another object of the present invention is to provide a method for purifying a meridian regulating health cream, which uses modern ultrasonic low-temperature extraction to fully extract effective ingredients, thereby solving the problem of difficulty in extracting effective ingredients from animal medicinal materials that are denatured at high temperatures.

[0005] Another object of the present invention is to provide an application of a meridian regulating health-care cream.

[0006] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0007] A meridian regulating health cream, based on dry medicinal materials, includes the following ingredients: frankincense, angelica, myrrh, borneol, angelica dahurica, rhubarb, artificial musk, phellodendron, siler, safflower, dragon's blood, licorice, asarum, coptis, notopterygium, angelica pubescens, red peony root, cinnamon, wormwood essential oil, black snake, black ant, earthworm, earthworm, Corydalis yanhusuo, clove, Panax notoginseng, Polygonum cuspidatum, Smilax glabra, peach kernel, Radix scutellariae, vaseline, glycerin, leech, centipede, dung beetle, beehive, and Ligusticum chuanxiong.

[0008] Preferably, a meridian conditioning health cream, based on dry medicinal materials, includes the following ingredients in parts by weight: 5-10 grams of frankincense, 10-15 grams of angelica, 5-10 grams of myrrh, 1-3 grams of borneol, 5-10 grams of angelica, 5-10 grams of rhubarb, 0.5-1 gram of artificial musk, 5-10 grams of phellodendron, 5-10 grams of siler, 5-10 grams of safflower, 5-10 grams of dragon's blood, 5-10 grams of liquorice, 5-10 grams of asarum, 5-10 grams of coptis, 10-15 grams of notopterygium, 10-15 grams of angelica root, 10-15 grams of red peony root, 10-10 grams of safflower root, 5-10 grams of dracaena, 5-10 grams of licorice, 5-10 grams of asarum, 5-10 grams of coptis root, 10-15 grams of notopterygium, 10-15 grams of angelica root, 10-10 grams of red peony root, 10-10 grams of safflower root, 5-10 grams of dracaena, 5-10 grams of safflower root, 5-10 grams of dracaena, 5-10 grams of dracaena root, 5-10 grams of safflower root, 5-10 grams of dracaena ... 0-15 grams, 3-6 grams of cinnamon, 1-3 grams of mugwort essential oil, 1-5 grams of black snake, 1-5 grams of black ant, 1-5 grams of earthworm, 1-5 grams of earthworm, 5-10 grams of Corydalis, 5-10 grams of clove, 5-10 grams of Panax notoginseng, 5-10 grams of Polygonum cuspidatum, 5-10 grams of Rhizoma Smilacis Glabrae, 5-10 grams of peach kernel, 5-10 grams of Herba Polygoni Multiflori, 100-200 grams of Vaseline, 20-100 grams of glycerin, 1-3 grams of hirudin, 1-5 grams of centipede, 1-5 grams of dung beetle, 1-5 grams of beehive, and 1-5 grams of Chuanxiong.

[0009] A method for purifying a meridian regulating health cream comprises the following steps:

[0010] Step 1, crushing the drugs except borneol, artificial musk, wormwood essential oil, vaseline, glycerin and hirudin, and sieving to obtain mixed medicinal material powder;

[0011] Step 2: subject the mixed medicinal powder to ultrasonic low-temperature extraction, immerse the mixed medicinal powder in deionized water, the amount of deionized water added is 10 to 30 times the weight of the mixed medicinal powder, the ultrasonic power is 100 to 400 W, the reaction temperature is 55 to 60 ° C, the pH is 6 to 8, the amount of cellulose immobilized enzyme added is equivalent to 10 to 12 wt% of the weight of the mixed medicinal powder, and the ultrasonic reaction time is 1 to 2 hours; after the ultrasonic reaction is completed, boil in water for 2 to 3 minutes, filter, take the filtrate, and concentrate it at low temperature to a thick paste, collect the paste, and the density of the thick paste is 1.3 to 1.4 g / cm 3 , obtaining an extract;

[0012] Step 3: Evenly mix the extract with borneol, hirudin, artificial musk, wormwood essential oil, vaseline and glycerin, and pour the mixture into a special container that has been cleaned and disinfected in advance.

[0013] Preferably, the method for preparing the cellulose immobilized enzyme comprises the following steps:

[0014] S01, 4-5 g of microcrystalline cellulose is dissolved in 10-15 mL of DMSO, and then 40-50 mL of acetone is added dropwise while stirring to obtain a microcrystalline cellulose solution, and the microcrystalline cellulose solution is added dropwise into 100-150 mL of 0-4°C cold water while stirring to obtain microcrystalline cellulose particles, which are washed with water and dried for later use;

[0015] S02, taking microcrystalline cellulose particles, placing them in 70-80 mL of a 0.3-0.4 mol / L NaOH solution, and keeping them warm at 20-30° C. for at least 30 min to obtain hydroxyl-activated microcrystalline cellulose particles;

[0016] S03, taking half of the hydroxy-activated microcrystalline cellulose particles, adding them to deionized water with an alkaline pH value, the amount of deionized water being equivalent to 10 to 15 times the mass of half of the hydroxy-activated microcrystalline cellulose particles, stirring in a boiling water bath until dissolved, to obtain a hydroxy-activated microcrystalline cellulose solution;

[0017] S04, adding amino acids to the hydroxy-activated microcrystalline cellulose solution, wherein the amount of the amino acids added is 1 / 25 to 1 / 20 of the hydroxy-activated microcrystalline cellulose solution, reacting under magnetic stirring at 70 to 80° C. for at least 1 hour, freeze-drying and then crushing to obtain amino acid-modified microcrystalline cellulose;

[0018] S05, dissolving β-glucosidase and alkaline protease in a mass ratio of 1:1 in a phosphate buffer with a concentration of 0.2-0.5 mol / L and a pH of 6-8 to form an enzyme solution with a mass concentration of 3-5 mg / mL, adding hydroxy-activated microcrystalline cellulose particles and amino acid-modified microcrystalline cellulose into a centrifuge tube, and then adding the enzyme solution into the centrifuge tube, wherein the amount of the enzyme solution added is equivalent to 1 / 4-1 / 3 of the total weight of the hydroxy-activated microcrystalline cellulose particles and the amino acid-modified microcrystalline cellulose; placing the centrifuge tube in an oscillator and oscillating at room temperature at 100-150 rpm for at least 30 minutes, taking it out, centrifuging it, and obtaining a lower precipitate;

[0019] S06, the lower precipitate is dissolved in a phosphate buffer with a concentration of 0.2-0.5 mol / L and a pH of 6-8, and the amount of phosphate buffer added is 20-30 times the weight of the lower precipitate; then a cross-linking agent is added, and the concentration of the cross-linking agent is 0.5-4wt%; after mixing evenly, it is placed in a refrigerator at 0-4°C for cross-linking reaction overnight, and the next day, it is taken out, centrifuged, and the lower layer is taken out and rinsed with distilled water at room temperature, and dried at room temperature to obtain cellulose immobilized enzyme, which is stored at 0-4°C for future use.

[0020] Preferably, the cross-linking agent comprises glutaraldehyde.

[0021] Preferably, the amino acid comprises L-serine, glycine or alanine.

[0022] Preferably, the alkaline protease comprises Alcalase 2.4L.

[0023] Preferably, the temperature of low temperature concentration is 50-60°C.

[0024] The invention discloses an application of a meridian regulating health-care cream in preparing medicines for resisting joint pain, muscle pain, chronic pain and dysmenorrhea.

[0025] Joint pain includes rheumatoid arthritis and osteoarthritis; muscle pain includes muscle pain caused by sprains, strains, and excessive fatigue; chronic pain includes chronic cervical spondylosis and lumbar spondylosis.

[0026] In the formula of the present invention, wormwood essential oil has the effects of warming the meridians and dispelling cold, and relieving pain; frankincense has the effects of promoting blood circulation and removing blood stasis, and relieving pain; angelica has the effects of nourishing blood, promoting blood circulation, and relieving pain; myrrh has the effects of promoting blood circulation and removing blood stasis, relieving pain, and reducing swelling; borneol has the effects of clearing away heat and detoxifying, and relieving pain; angelica has the effects of promoting blood circulation, relieving pain, and unblocking meridians; rhubarb has the effects of purging fire and detoxifying, promoting blood circulation and removing blood stasis; artificial musk has the effects of relaxing muscles and tendons, relieving pain, and reducing swelling; phellodendron has the effects of clearing heat and detoxifying, and relieving pain. Dampness-drying and analgesic effects; Saposhnikovia divaricata: has the effects of dispelling wind and cold and relieving pain; Safflower: has the effects of promoting blood circulation and removing blood stasis and relieving pain; Xuejie: ​​has the effects of cooling blood and relieving pain; Licorice: has the effects of clearing away heat and detoxifying and relieving pain; Asarum: has the effects of removing rheumatism and relieving pain; Coptis chinensis: has the effects of clearing away heat and detoxifying and relieving pain; Notopterygium wilfordii: has the effects of removing rheumatism and relieving pain; Duhuo: has the effects of removing rheumatism and relieving pain; Red Peony Root: has the effects of promoting blood circulation and relieving pain; Cinnamon Bark: has the effects of warming the meridians and dispelling cold and relieving pain.

[0027] The meridian conditioning health cream of the present invention is suitable for the following groups of people:

[0028] Arthritis patients: joint pain caused by rheumatoid arthritis, osteoarthritis, etc.

[0029] Patients with muscle pain: muscle pain caused by sprains, strains, overwork, etc.

[0030] Patients with chronic pain: such as chronic pain caused by chronic cervical spondylosis, lumbar spondylosis, etc.

[0031] Patients with dysmenorrhea: Topical application can relieve dysmenorrhea symptoms.

[0032] Indications of the meridian regulating health cream of the present invention:

[0033] Cervical spondylosis, frozen shoulder, lumbar disc herniation, stomach cold, rheumatism, rheumatoid arthritis, leg cramps, synovitis, arthritis, trauma, knee pain, meniscus injury, dysmenorrhea, tennis elbow, tenosynovitis, uterine cold, cold hands and feet, etc.

[0034] The use method of the meridian conditioning health care cream of the present invention:

[0035] Apply an appropriate amount of meridian conditioning health cream evenly to the area where pain relief is needed, and then gently massage to help the drug ingredients penetrate and absorb. The number of times per day can be adjusted appropriately according to the degree of pain and personal constitution.

[0036] Notes on the meridian conditioning health cream of the present invention:

[0037] Perform a skin test before use to ensure it does not cause an allergic reaction.

[0038] If allergic symptoms such as redness, swelling, itching, etc. occur at the application site, please stop using it immediately and consult a professional doctor.

[0039] Do not use the meridian conditioning health cream on open wounds or mucous membranes.

[0040] Pregnant women, breastfeeding women and people with allergies should use it under the guidance of a doctor.

[0041] Keep the meridian regulating health cream out of the reach of children to avoid accidental ingestion.

[0042] The meridian regulating health-care cream of the present invention is developed by combining a pain-relieving formula that has been passed down for more than 2,000 years with new technology, and is made of a variety of animal medicines + plant medicines, etc., and takes effect in 5 minutes.

[0043] The present invention has the following beneficial effects:

[0044] The meridian regulating health cream of the present invention has a formula derived from data mining of the medication rules of various external analgesic Chinese medicine prescriptions and is formulated in combination with the theory of syndrome differentiation and treatment of traditional Chinese medicine. It is made of various animal medicines + plant medicines, etc., and takes effect in 5 minutes.

[0045] The invention discloses a purification method for meridian regulating health cream, which uses modern ultrasonic low-temperature extraction to fully extract effective ingredients, thus solving the problem that effective ingredients of animal medicinal materials denatured at high temperature are difficult to extract. In the purification method, cellulose-immobilized enzyme enzymatic ultrasonic hydrolysis is used, which not only significantly increases the content of polypeptides and flavonoids in the cream, improves the efficacy, and relieves pain in 5 minutes, but also has a detoxification effect. Under the ultrasonic low-temperature extraction conditions of the invention, each drug synergistically reduces toxicity and increases efficacy. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 It is a finished paste diagram of the present invention;

[0047] Figure 2 This is a diagram of the medicinal materials of Comparative Example 1;

[0048] Figure 3 This is a finished paste image of Comparative Example 1;

[0049] Figure 4 is the standard curve of rutin reference;

[0050] Figure 5 is the standard curve of the standard protein. DETAILED DESCRIPTION

[0051] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments. The following embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0052] Example 1

[0053] A meridian regulating health cream, based on dry medicinal materials, includes the following ingredients: frankincense, angelica, myrrh, borneol, angelica dahurica, rhubarb, artificial musk, phellodendron, siler, safflower, dragon's blood, licorice, asarum, coptis, notopterygium, angelica pubescens, red peony root, cinnamon, wormwood essential oil, black snake, black ant, earthworm, earthworm, Corydalis yanhusuo, clove, Panax notoginseng, Polygonum cuspidatum, Smilax glabra, peach kernel, Radix scutellariae, vaseline, glycerin, leech, centipede, dung beetle, beehive, and Ligusticum chuanxiong.

[0054] Preferably, a meridian regulating health cream, based on dry medicinal materials, includes the following ingredients in parts by weight: 7 grams of frankincense, 12 grams of angelica, 8 grams of myrrh, 2 grams of borneol, 6 grams of angelica dahurica, 8 grams of rhubarb, 0.7 grams of artificial musk, 6 grams of phellodendron, 7 grams of siler, 7 grams of safflower, 7 grams of dragon's blood, 6 grams of licorice, 6 grams of asarum, 6 grams of coptis, 12 grams of notopterygium, 12 grams of angelica pubescens, 12 grams of red peony root, 4 grams of cinnamon, 2 grams of wormwood essential oil, 3 grams of black snake, 3 grams of black ant, 3 grams of earthworm, 3 grams of earthworm, 7 grams of Corydalis yanhusuo, 7 grams of clove, 8 grams of Panax notoginseng, 8 grams of Polygonum cuspidatum, 8 grams of Scutellaria baicalensis, 8 grams of peach kernel, 8 grams of Herba speranskiae radix, 150 grams of vaseline, 50 grams of glycerin, 2 grams of leech, 3 grams of centipede, 3 grams of dung beetle, 3 grams of beehive, and 3 grams of Chuanxiong.

[0055] A method for purifying a meridian regulating health cream comprises the following steps:

[0056] Step 1, crushing the drugs except borneol, artificial musk, wormwood essential oil, vaseline, glycerin and hirudin, and sieving to obtain mixed medicinal material powder;

[0057] Step 2: subject the mixed medicinal powder to ultrasonic low-temperature extraction, immerse the mixed medicinal powder in 20 times the amount of deionized water, the ultrasonic power is 300W, the reaction temperature is 58°C, the pH is 7, the amount of cellulose immobilized enzyme added is 10wt% of the weight of the mixed medicinal powder, and the ultrasonic reaction time is 1.5h; after the ultrasonic reaction is completed, boil in water for 2min, filter, take the filtrate, and concentrate at 55°C to a thick paste, collect the paste, and the density of the thick paste is 1.3g / cm 3 , obtaining an extract;

[0058] Step 3: Evenly mix the extract with borneol, hirudin, artificial musk, wormwood essential oil, vaseline and glycerin, and pour the mixture into a special container that has been cleaned and disinfected in advance.

[0059] The preparation method of cellulose immobilized enzyme comprises the following steps:

[0060] S01, 4 g of microcrystalline cellulose was dissolved in 12 mL of DMSO, and then 45 mL of acetone was added dropwise while stirring to obtain a microcrystalline cellulose solution, and the microcrystalline cellulose solution was added dropwise into 120 mL of 2°C cold water while stirring to obtain microcrystalline cellulose particles, which were washed with water and dried for later use;

[0061] S02, taking microcrystalline cellulose particles, placing them in 75 mL of a 0.3 mol / L NaOH solution, and keeping them at 25° C. for 30 min to obtain hydroxyl-activated microcrystalline cellulose particles;

[0062] S03, taking half of the hydroxyl-activated microcrystalline cellulose particles, adding them to 12 times the amount of deionized water with an alkaline pH value, and stirring them in a boiling water bath until dissolved to obtain a hydroxyl-activated microcrystalline cellulose solution;

[0063] S04, adding L-serine to the hydroxy-activated microcrystalline cellulose solution, wherein the amount of L-serine added is 1 / 25 of that of the hydroxy-activated microcrystalline cellulose solution, reacting under magnetic stirring at 75° C. for 1 hour, freeze-drying and then pulverizing to obtain amino acid-modified microcrystalline cellulose;

[0064] S05, dissolving β-glucosidase and alkaline protease Alcalase 2.4L in a mass ratio of 1:1 in a phosphate buffer solution with a concentration of 0.3mol / L and a pH of 6-8 to form an enzyme solution with a mass concentration of 4mg / mL, adding hydroxy-activated microcrystalline cellulose particles and amino acid-modified microcrystalline cellulose into a centrifuge tube, and then adding the enzyme solution into the centrifuge tube, wherein the amount of the enzyme solution added is equivalent to 1 / 4 of the total weight of the hydroxy-activated microcrystalline cellulose particles and the amino acid-modified microcrystalline cellulose; placing the centrifuge tube in an oscillator at room temperature and 120rpm for 30min, taking it out, centrifuging it, and obtaining a lower precipitate;

[0065] S06, the lower precipitate is dissolved in 25 times the amount of phosphate buffer with a concentration of 0.4 mol / L and a pH of 6 to 8; then the cross-linking agent glutaraldehyde is added, and the concentration of the cross-linking agent is 2wt%; after mixing evenly, it is placed in a refrigerator at 2°C for cross-linking reaction overnight, and the next day, it is taken out, centrifuged, and the lower layer is taken out and rinsed with distilled water at room temperature, and dried at room temperature to obtain cellulose immobilized enzyme, which is stored at 2°C for future use.

[0066] The invention discloses an application of a meridian regulating health-care cream in preparing medicines for resisting joint pain, muscle pain, chronic pain and dysmenorrhea.

[0067] Joint pain includes rheumatoid arthritis and osteoarthritis; muscle pain includes muscle pain caused by sprains, strains, and excessive fatigue; chronic pain includes chronic cervical spondylosis and lumbar spondylosis.

[0068] like Figure 1 Shown is a picture of the meridian regulating health cream obtained in this embodiment.

[0069] Example 2

[0070] A meridian regulating health cream, based on dry medicinal materials, includes the following ingredients: frankincense, angelica, myrrh, borneol, angelica dahurica, rhubarb, artificial musk, phellodendron, siler, safflower, dragon's blood, licorice, asarum, coptis, notopterygium, angelica pubescens, red peony root, cinnamon, wormwood essential oil, black snake, black ant, earthworm, earthworm, Corydalis yanhusuo, clove, Panax notoginseng, Polygonum cuspidatum, Smilax glabra, peach kernel, Radix scutellariae, vaseline, glycerin, leech, centipede, dung beetle, beehive, and Ligusticum chuanxiong.

[0071] Preferably, a meridian regulating health cream, based on dry medicinal materials, includes the following ingredients in parts by weight: 5 grams of frankincense, 10 grams of angelica, 5 grams of myrrh, 1 gram of borneol, 5 grams of angelica dahurica, 5 grams of rhubarb, 0.5 grams of artificial musk, 5 grams of phellodendron, 5 grams of saposhnikovia divaricata, 5 grams of safflower, 5 grams of dragon's blood, 5 grams of licorice, 5 grams of asarum, 5 grams of coptis chinensis, 10 grams of notopterygium, 10 grams of angelica pubescens, 10 grams of red peony root, 3 grams of cinnamon, 1 gram of wormwood essential oil, 1 gram of black snake, 1 gram of black ant, 1 gram of earthworm, 1 gram of earthworm, 5 grams of Corydalis yanhusuo, 5 grams of cloves, 5 grams of Panax notoginseng, 5 grams of Polygonum cuspidatum, 5 grams of Scutellaria baicalensis, 5 grams of peach kernel, 5 grams of Herba speranskiae radix, 100 grams of vaseline, 20 grams of glycerin, 1 gram of hirudin, 1 gram of centipede, 1 gram of dung beetle, 1 gram of beehive, and 1 gram of Chuanxiong.

[0072] A method for purifying a meridian regulating health cream comprises the following steps:

[0073] Step 1, crushing the drugs except borneol, artificial musk, wormwood essential oil, vaseline, glycerin and hirudin, and sieving to obtain mixed medicinal material powder;

[0074] Step 2: subject the mixed medicinal powder to ultrasonic low-temperature extraction, immerse the mixed medicinal powder in 10 times the amount of deionized water, the ultrasonic power is 100 W, the reaction temperature is 55 ° C, the pH is 6, the amount of cellulose immobilized enzyme added is 11wt% of the weight of the mixed medicinal powder, and the ultrasonic reaction time is 1h; after the ultrasonic reaction is completed, boil in water bath for 3min, filter, take the filtrate, and concentrate at 50 ° C to a thick paste, collect the paste, and the density of the thick paste is 1.4g / cm 3 , obtaining an extract;

[0075] Step 3: Evenly mix the extract with borneol, hirudin, artificial musk, wormwood essential oil, vaseline and glycerin, and pour the mixture into a special container that has been cleaned and disinfected in advance.

[0076] The preparation method of cellulose immobilized enzyme comprises the following steps:

[0077] S01, 5 g of microcrystalline cellulose was dissolved in 10 mL of DMSO, and then 40 mL of acetone was added dropwise while stirring to obtain a microcrystalline cellulose solution, and the microcrystalline cellulose solution was added dropwise into 100 mL of 0°C cold water while stirring to obtain microcrystalline cellulose particles, which were washed with water and dried for later use;

[0078] S02, taking microcrystalline cellulose particles, placing them in 70 mL of a 0.4 mol / L NaOH solution, and keeping them at 20° C. for 50 min to obtain hydroxyl-activated microcrystalline cellulose particles;

[0079] S03, taking half of the hydroxyl-activated microcrystalline cellulose particles, adding them to 10 times the amount of deionized water with an alkaline pH value, and stirring them in a boiling water bath until dissolved to obtain a hydroxyl-activated microcrystalline cellulose solution;

[0080] S04, adding glycine to the hydroxy-activated microcrystalline cellulose solution, wherein the amount of glycine added is 1 / 20 of that of the hydroxy-activated microcrystalline cellulose solution, reacting under magnetic stirring at 70° C. for 2 h, freeze-drying and then crushing to obtain amino acid-modified microcrystalline cellulose;

[0081] S05, dissolving β-glucosidase and alkaline protease Alcalase 2.4L in a mass ratio of 1:1 in a phosphate buffer solution with a concentration of 0.2mol / L and a pH of 6-8 to form an enzyme solution with a mass concentration of 3mg / mL, adding hydroxy-activated microcrystalline cellulose particles and amino acid-modified microcrystalline cellulose into a centrifuge tube, and then adding the enzyme solution into the centrifuge tube, wherein the amount of the enzyme solution added is equivalent to 1 / 3 of the total weight of the hydroxy-activated microcrystalline cellulose particles and the amino acid-modified microcrystalline cellulose; placing the centrifuge tube in an oscillator at room temperature and 100rpm for 40min, taking it out, centrifuging it, and obtaining a lower layer of precipitate;

[0082] S06, the lower precipitate is dissolved in 20 times the amount of phosphate buffer with a concentration of 0.2 mol / L and a pH of 6 to 8; then the cross-linking agent glutaraldehyde is added, and the concentration of the cross-linking agent is 0.5wt%; after mixing evenly, it is placed in a refrigerator at 0°C for cross-linking reaction overnight, and the next day, it is taken out, centrifuged, and the lower layer is taken out and rinsed with distilled water at room temperature, and dried at room temperature to obtain cellulose immobilized enzyme, which is stored at 0°C for future use.

[0083] The invention discloses an application of a meridian regulating health-care cream in preparing medicines for resisting joint pain, muscle pain, chronic pain and dysmenorrhea.

[0084] Joint pain includes rheumatoid arthritis and osteoarthritis; muscle pain includes muscle pain caused by sprains, strains, and excessive fatigue; chronic pain includes chronic cervical spondylosis and lumbar spondylosis.

[0085] Example 3

[0086] A meridian regulating health cream, based on dry medicinal materials, includes the following ingredients: frankincense, angelica, myrrh, borneol, angelica dahurica, rhubarb, artificial musk, phellodendron, siler, safflower, dragon's blood, licorice, asarum, coptis, notopterygium, angelica pubescens, red peony root, cinnamon, wormwood essential oil, black snake, black ant, earthworm, earthworm, Corydalis yanhusuo, clove, Panax notoginseng, Polygonum cuspidatum, Smilax glabra, peach kernel, Radix scutellariae, vaseline, glycerin, leech, centipede, dung beetle, beehive, and Ligusticum chuanxiong.

[0087] Preferably, a meridian conditioning health cream, based on dry medicinal materials, includes the following ingredients in parts by weight: 10 grams of frankincense, 15 grams of angelica, 10 grams of myrrh, 3 grams of borneol, 10 grams of angelica dahurica, 10 grams of rhubarb, 1 gram of artificial musk, 10 grams of phellodendron, 10 grams of siler, 10 grams of safflower, 10 grams of dragon's blood, 10 grams of licorice, 10 grams of asarum, 10 grams of coptis, 15 grams of notopterygium, 15 grams of angelica dahurica grams, red peony root 15 grams, cinnamon 6 grams, mugwort essential oil 3 grams, black snake 5 grams, black ant 5 grams, earthworm 5 grams, earthworm 5 grams, Corydalis 10 grams, clove 10 grams, Panax notoginseng 10 grams, Polygonum cuspidatum 10 grams, Rhizoma Smilacis Glabrae 10 grams, peach kernel 10 grams, Herba Polygoni Multiflori 10 grams, Vaseline 200 grams, glycerin 100 grams, hirudin 3 grams, centipede 5 grams, dung beetle 5 grams, beehive 5 grams, and Ligusticum chuanxiong 5 grams.

[0088] A method for purifying a meridian regulating health cream comprises the following steps:

[0089] Step 1, crushing the drugs except borneol, artificial musk, wormwood essential oil, vaseline, glycerin and hirudin, and sieving to obtain mixed medicinal material powder;

[0090] Step 2: subject the mixed medicinal powder to ultrasonic low-temperature extraction, immerse the mixed medicinal powder in 30 times the amount of deionized water, the ultrasonic power is 400W, the reaction temperature is 60°C, the pH is 8, the amount of cellulose immobilized enzyme added is 12wt% of the weight of the mixed medicinal powder, and the ultrasonic reaction time is 2h; after the ultrasonic reaction, boil in water for 3min, filter, take the filtrate, and concentrate at 50°C to a thick paste, collect the paste, and the density of the thick paste is 1.4g / cm 3 , obtaining an extract;

[0091] Step 3: Evenly mix the extract with borneol, hirudin, artificial musk, wormwood essential oil, vaseline and glycerin, and pour the mixture into a special container that has been cleaned and disinfected in advance.

[0092] The preparation method of cellulose immobilized enzyme comprises the following steps:

[0093] S01, 5 g of microcrystalline cellulose was dissolved in 15 mL of DMSO, and then 50 mL of acetone was added dropwise while stirring to obtain a microcrystalline cellulose solution, and the microcrystalline cellulose solution was added dropwise into 150 mL of 4°C cold water while stirring to obtain microcrystalline cellulose particles, which were washed with water and dried for later use;

[0094] S02, taking microcrystalline cellulose particles, placing them in 80 mL of a 0.4 mol / L NaOH solution, and keeping them at 30° C. for 40 min to obtain hydroxyl-activated microcrystalline cellulose particles;

[0095] S03, taking half of the hydroxyl-activated microcrystalline cellulose particles, adding them to 15 times the amount of deionized water with an alkaline pH value, and stirring them in a boiling water bath until dissolved to obtain a hydroxyl-activated microcrystalline cellulose solution;

[0096] S04, adding alanine to the hydroxy-activated microcrystalline cellulose solution, wherein the amount of alanine added is 1 / 20 of that of the hydroxy-activated microcrystalline cellulose solution, reacting under magnetic stirring at 80° C. for 1.5 h, freeze-drying and then crushing to obtain amino acid-modified microcrystalline cellulose;

[0097] S05, dissolving β-glucosidase and alkaline protease Alcalase 2.4 L in a mass ratio of 1:1 in a phosphate buffer solution with a concentration of 0.5 mol / L and a pH of 6-8 to form an enzyme solution with a mass concentration of 5 mg / mL, adding hydroxy-activated microcrystalline cellulose particles and amino acid-modified microcrystalline cellulose into a centrifuge tube, and then adding the enzyme solution into the centrifuge tube, wherein the amount of the enzyme solution added is equivalent to 1 / 3 of the total weight of the hydroxy-activated microcrystalline cellulose particles and the amino acid-modified microcrystalline cellulose; placing the centrifuge tube in an oscillator and oscillating it at room temperature at 150 rpm for 50 min, taking it out, centrifuging it, and obtaining a lower layer of precipitate;

[0098] S06, the lower precipitate is dissolved in 30 times the amount of phosphate buffer with a concentration of 0.5 mol / L and a pH of 6 to 8; then a cross-linking agent glutaraldehyde is added, and the concentration of the cross-linking agent is 4 wt%; after mixing evenly, it is placed in a refrigerator at 4°C for cross-linking reaction overnight. The next day, it is taken out, centrifuged, and the lower layer is taken out and rinsed with distilled water at room temperature, and dried at room temperature to obtain cellulose immobilized enzyme, which is stored at 4°C for future use.

[0099] The invention discloses an application of a meridian regulating health-care cream in preparing medicines for resisting joint pain, muscle pain, chronic pain and dysmenorrhea.

[0100] Joint pain includes rheumatoid arthritis and osteoarthritis; muscle pain includes muscle pain caused by sprains, strains, and excessive fatigue; chronic pain includes chronic cervical spondylosis and lumbar spondylosis.

[0101] Comparative Example 1

[0102] The difference between this comparative example and Example 1 is only that:

[0103] A purification method for meridian regulating health cream:

[0104] Step 1: Grind the above-mentioned Chinese medicines except mugwort essential oil, vaseline and glycerin into fine powder and set aside.

[0105] Step 2: Heat sesame oil to 60°C, add the Chinese medicine powder into the sesame oil, and simmer over low heat until the drug components are fully dissolved and extracted, the time is 2 hours. The amount of sesame oil added is enough to cover the Chinese medicine powder.

[0106] Step three: filter the medicinal oil after stewing and remove the residue.

[0107] Step 4: Add mugwort essential oil to the filtered medicinal oil and mix thoroughly.

[0108] Step 5: Place the mixed medicinal oil and mugwort essential oil into a suitable container and seal it for storage.

[0109] like Figure 2 As shown in FIG. 1 , it is a diagram of the medicinal materials of this comparative example. In this comparative example, sesame oil is used as a base oil to enhance the penetration of the drug. Figure 3 Shown is the finished paste obtained in this comparative example.

[0110] Comparative Example 2

[0111] The difference between this comparative example and Example 1 is only that:

[0112] During ultrasonic low-temperature extraction, the added enzyme is not a cellulose-immobilized enzyme, but a 1:1 mass ratio of β-glucosidase and alkaline protease without cellulose immobilization. The added amount of β-glucosidase and alkaline protease is equivalent to 10wt% of the weight of the mixed medicinal material powder.

[0113] The meridian regulating health cream of this embodiment and the comparative example was tested and analyzed to detect the contents of polypeptides and flavonoids therein. The test results are shown in Table 2 below.

[0114] 1. Determination of total flavonoids content

[0115] 1. Experimental Materials

[0116] 1.1 Reagents

[0117] Rutin Reference

[0118] 70% ethanol

[0119] 5%NaNO2

[0120] 10%Al(NO3)3

[0121] 4% NaOH

[0122] 1.2 Instruments

[0123] Microplate reader (1510; Thermo Fisher Scientific Inc.), centrifuge (TD5M; Shanghai Huxiangyi Centrifuge Instrument Co., Ltd.), CNC ultrasonic cleaner (KH-800DE; Kunshan Hechuang Ultrasonic Instrument Co., Ltd.)

[0124] 2. Methods

[0125] 2.1 Preparation of test solution

[0126] Take 2g of the test ointment (i.e., Examples 1 to 3 and Comparative Examples 1 to 2) respectively, add 20mL of 70% ethanol, perform ultrasonic extraction for 30min at room temperature and 100W, centrifuge at 3000r / min for 10min, take the supernatant, filter, place the filtrate in a 50mL volumetric flask, and add water to make up to volume to obtain the test ointment flavonoid extract.

[0127] 2.2 Preparation of reference solution

[0128] Accurately weigh 10 mg of rutin reference substance dried to constant weight at 100°C, dissolve it in 70% ethanol, place it in a 50 mL volumetric flask, and add water to make up to volume to obtain a 0.2 mg / mL rutin reference solution.

[0129] 2.3 Solution color development

[0130] Accurately measure 1 mL of the reference solution and the test solution, place them in a 10 mL volumetric flask, first add 0.4 mL of 5% NaNO2 solution, shake well, let stand for 6 min, then add 0.4 mL of 10% Al(NO3)3 solution, shake well, let stand for 6 min, finally add 4.0 mL of 4% NaOH solution, add water to the scale line of the volumetric flask, shake well and let stand for 15 min. Use pure water plus corresponding reagents as the blank control group and wait for detection.

[0131] 2.4 Determination of measurement wavelength

[0132] According to the color development method of "2.3", the reference solution, test solution and blank control solution (pure water) were developed respectively, and then scanned at a wavelength of 200-800nm ​​in a 1cm cuvette. It was found that both the reference solution and the sample solution showed a maximum absorption peak at 508nm, so 508nm was selected as the measurement wavelength.

[0133] 2.5 Drawing of standard curve

[0134] Accurately pipette 0, 1.0, 1.5, 2.0, 2.5, and 3.0 mL of the reference solution into a 10 mL volumetric flask, respectively, and use pure water as a blank control. After color development according to the method in "2.3", measure its absorbance at a wavelength of 508 nm, and draw a standard curve with the concentration of rutin reference as the horizontal axis and the absorbance as the vertical axis. Perform linear regression, and the standard curve is as follows: Figure 4 shown.

[0135] 2.6 Determination of total flavonoid content

[0136] Accurately measure 1.0 mL of each test solution and place it in a 10 mL volumetric flask. Add 1.0 mL of ultrapure water to the blank group. After color development according to the method in "2.3", use the blank group as a control to quickly measure the average absorbance of the test solution at 508 nm. Calculate the corresponding flavonoid concentration C from the linear correlation equation, and then convert it into the mass of total flavonoids in the extract to calculate the total flavonoid content (X, mg / g). The formula is as follows:

[0137] X=C×250

[0138] Wherein, C is calculated based on the standard curve fitting equation and the unit is mg / mL.

[0139] 3. Results

[0140] See Table 2 below.

[0141] 2. BCA kit to measure peptide content

[0142] 1. Preparation of BCA working solution: According to the number of samples, prepare an appropriate amount of BCA working solution by adding 50 volumes of BCA Reagent A to 1 volume of BCA Reagent B (50:1) and mix thoroughly.

[0143] Specifically, prepare: 10mL+200uL.

[0144] 2. Draw the standard curve: Take an ELISA plate and add reagents according to the data in Table 1:

[0145] Table 1 Data table of standard proteins

[0146] Hole No. 0 1 2 3 4 5 6 7 Standard protein (μL) 0 1 2 4 8 12 16 20 Deionized water (μL) 20 19 18 16 12 8 4 0 BCA working solution (μL) 200 200 200 200 200 200 200 200 Corresponding protein content (μg) 0 1 2 4 8 12 16 20

[0147] The standard curve was added according to Table 1, standard protein concentration: 1 mg / mL

[0148] 3. Sample preparation: Take 2 g of the test ointment, add 50 mL of ultrapure water, extract by ultrasonic at room temperature and 100 W for 30 min, and centrifuge at 3000 r / min for 10 min.

[0149] Sample addition: Add 20 μL of ointment sample supernatant and 200 μL of BCA working solution respectively. The sample addition and standard curve addition are carried out simultaneously.

[0150] 4. Mix by oscillation and place at 37℃ for 60 min.

[0151] 5. Use an enzyme-labeled instrument to measure the absorbance at A562nm, and use the absorbance without BSA as the blank control (the first marker).

[0152] 6. Draw a standard curve with protein concentration as the horizontal axis and absorbance as the vertical axis. The standard curve is as follows: Figure 5 shown.

[0153] 7. According to the measured absorbance value, the polypeptide content of the sample is calculated on the standard curve. The polypeptide content is shown in Table 2 below.

[0154] Table 2 Data table of peptide and flavonoid content

[0155] Peptide content (%) Flavonoid content (mg / g) Example 1 1.422 8.272 Example 2 1.587 9.134 Example 3 1.492 9.058 Comparative Example 1 0.051 3.112 Comparative Example 2 0.068 4.257

[0156] The meridian regulating health-care cream of this embodiment is safe and non-irritating to the skin.

[0157] It should be understood that in order to streamline the present disclosure and aid in understanding one or more of the various inventive aspects, in the above description of exemplary embodiments of the present invention, various features of the present invention are sometimes grouped together into a single embodiment, figure, or description thereof. However, this disclosed method should not be interpreted as reflecting the intention that the claimed invention requires more features than those expressly recited in each claim. Rather, as reflected in the claims, inventive aspects lie in less than all of the features of the previously disclosed embodiments. Therefore, the claims that follow the detailed description are hereby expressly incorporated into the detailed description, with each claim itself serving as a separate embodiment of the present invention.

[0158] Although the present invention has been described according to a limited number of embodiments, it will be apparent to those skilled in the art, with the benefit of the above description, that other embodiments may be envisioned within the scope of the invention thus described. In addition, it should be noted that the language used in this specification is selected primarily for readability and teaching purposes, rather than for explaining or defining the subject matter of the present invention. Therefore, many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the appended claims. The disclosure of the present invention is illustrative, not restrictive, with respect to the scope of the present invention, which is defined by the appended claims.

[0159] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A meridian regulating health cream, characterized in that: Based on dry medicinal materials, the following ingredients are included: frankincense, angelica, myrrh, borneol, angelica dahurica, rhubarb, artificial musk, phellodendron, siler, safflower, dragon's blood, licorice, asarum, coptis, notopterygium, angelica dahurica, red peony root, cinnamon, mugwort essential oil, black snake, black ant, earthworm, earthworm, Corydalis yanhusuo, clove, Panax notoginseng, knotweed, rhubarb, peach kernel, radix polygoni aviculare, vaseline, glycerin, leech, centipede, dung beetle, beehive, and Chuanxiong.

2. The meridian regulating health cream according to claim 1, characterized in that: Based on dry medicinal materials, the following ingredients are included in weight: 5-10 grams of frankincense, 10-15 grams of angelica, 5-10 grams of myrrh, 1-3 grams of borneol, 5-10 grams of angelica dahurica, 5-10 grams of rhubarb, 0.5-1 grams of artificial musk, 5-10 grams of phellodendron, 5-10 grams of saposhnikovia, 5-10 grams of safflower, 5-10 grams of dragon's blood, 5-10 grams of licorice, 5-10 grams of asarum, 5-10 grams of coptis, 10-15 grams of notopterygium, 10-15 grams of angelica dahurica, 10-15 grams of red peony root, 5-10 grams of meat 3-6 grams of cinnamon, 1-3 grams of mugwort essential oil, 1-5 grams of black snake, 1-5 grams of black ant, 1-5 grams of earthworm, 1-5 grams of earthworm, 5-10 grams of Corydalis, 5-10 grams of clove, 5-10 grams of Panax notoginseng, 5-10 grams of Polygonum cuspidatum, 5-10 grams of Rhizoma Smilacis Glabrae, 5-10 grams of peach kernel, 5-10 grams of Herba Polygoni Multiflori, 100-200 grams of Vaseline, 20-100 grams of glycerin, 1-3 grams of hirudin, 1-5 grams of centipede, 1-5 grams of dung beetle, 1-5 grams of beehive, and 1-5 grams of Ligusticum chuanxiong.

3. A purification method for a meridian conditioning health cream according to claim 1 or 2, characterized in that: The following steps are involved: Step 1, crushing the drugs except borneol, artificial musk, wormwood essential oil, vaseline, glycerin and hirudin, and sieving to obtain mixed medicinal material powder; Step 2: subject the mixed medicinal powder to ultrasonic low-temperature extraction, immerse the mixed medicinal powder in deionized water, the amount of deionized water added is 10 to 30 times the weight of the mixed medicinal powder, the ultrasonic power is 100 to 400 W, the reaction temperature is 55 to 60 ° C, the pH is 6 to 8, the amount of cellulose immobilized enzyme added is equivalent to 10 to 12 wt% of the weight of the mixed medicinal powder, and the ultrasonic reaction time is 1 to 2 hours; after the ultrasonic reaction is completed, boil in water for 2 to 3 minutes, filter, take the filtrate, and concentrate it at low temperature to a thick paste, collect the paste, and the density of the thick paste is 1.3 to 1.4 g / cm 3 , obtaining an extract; Step 3: Evenly mix the extract with borneol, hirudin, artificial musk, wormwood essential oil, vaseline and glycerin, and pour the mixture into a special container that has been cleaned and disinfected in advance.

4. The purification method according to claim 3, characterized in that The preparation method of cellulose immobilized enzyme comprises the following steps: S01, 4-5 g of microcrystalline cellulose is dissolved in 10-15 mL of DMSO, and then 40-50 mL of acetone is added dropwise while stirring to obtain a microcrystalline cellulose solution, and the microcrystalline cellulose solution is added dropwise into 100-150 mL of 0-4°C cold water while stirring to obtain microcrystalline cellulose particles, which are washed with water and dried for later use; S02, taking microcrystalline cellulose particles, placing them in 70-80 mL of a 0.3-0.4 mol / L NaOH solution, and keeping them warm at 20-30° C. for at least 30 min to obtain hydroxyl-activated microcrystalline cellulose particles; S03, taking half of the hydroxy-activated microcrystalline cellulose particles, adding them to deionized water with an alkaline pH value, the amount of deionized water being equivalent to 10 to 15 times the mass of half of the hydroxy-activated microcrystalline cellulose particles, stirring in a boiling water bath until dissolved, to obtain a hydroxy-activated microcrystalline cellulose solution; S04, adding amino acids to the hydroxy-activated microcrystalline cellulose solution, wherein the amount of the amino acids added is 1 / 25 to 1 / 20 of the hydroxy-activated microcrystalline cellulose solution, reacting under magnetic stirring at 70 to 80° C. for at least 1 hour, freeze-drying, and then pulverizing to obtain amino acid-modified microcrystalline cellulose; S05, dissolving β-glucosidase and alkaline protease in a mass ratio of 1:1 in a phosphate buffer with a concentration of 0.2-0.5 mol / L and a pH of 6-8 to form an enzyme solution with a mass concentration of 3-5 mg / mL, adding hydroxy-activated microcrystalline cellulose particles and amino acid-modified microcrystalline cellulose into a centrifuge tube, and then adding the enzyme solution into the centrifuge tube, wherein the amount of the enzyme solution added is equivalent to 1 / 4-1 / 3 of the total weight of the hydroxy-activated microcrystalline cellulose particles and the amino acid-modified microcrystalline cellulose; placing the centrifuge tube in an oscillator and oscillating at room temperature at 100-150 rpm for at least 30 minutes, taking it out, centrifuging it, and obtaining a lower precipitate; S06, the lower precipitate is dissolved in a phosphate buffer with a concentration of 0.2-0.5 mol / L and a pH of 6-8, and the amount of phosphate buffer added is 20-30 times the weight of the lower precipitate; then a cross-linking agent is added, and the concentration of the cross-linking agent is 0.5-4wt%; after mixing evenly, it is placed in a refrigerator at 0-4°C for cross-linking reaction overnight, and the next day, it is taken out, centrifuged, and the lower layer is taken out and rinsed with distilled water at room temperature, and dried at room temperature to obtain cellulose immobilized enzyme, which is stored at 0-4°C for future use.

5. The purification method according to claim 4, characterized in that Cross-linking agents include glutaraldehyde.

6. The purification method according to claim 4, characterized in that The amino acids include L-serine, glycine or alanine.

7. The purification method according to claim 4, characterized in that Alkaline proteases include Alcalase 2.4L.

8. The purification method according to claim 3, characterized in that The temperature of low temperature concentration is 50-60°C.

9. Use of the meridian regulating health cream according to claim 1 or 2 in the preparation of drugs for resisting joint pain, muscle pain, chronic pain and dysmenorrhea.

10. The use according to claim 9, characterized in that: Joint pain includes rheumatoid arthritis and osteoarthritis; muscle pain includes muscle pain caused by sprains, strains, and excessive fatigue; chronic pain includes chronic cervical spondylosis and lumbar spondylosis.

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