CN114432195A - Skin repairing and antibacterial composition, gel, application and preparation method - Google Patents
Skin repairing and antibacterial composition, gel, application and preparation method Download PDFInfo
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- CN114432195A CN114432195A CN202011223845.9A CN202011223845A CN114432195A CN 114432195 A CN114432195 A CN 114432195A CN 202011223845 A CN202011223845 A CN 202011223845A CN 114432195 A CN114432195 A CN 114432195A
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Abstract
The invention relates to the technical field of skin repair and bacteriostasis, in particular to a skin repair bacteriostasis composition and application thereof, a gel prepared from the skin repair bacteriostasis composition and a preparation method thereof. The skin repairing antibacterial composition comprises the following components in parts by weight: 1-3 parts of water-soluble paeonol, 1-3 parts of brown algae extract, 1-3 parts of corydalis yanhusuo root extract, 1-3 parts of cynanchum paniculatum extract, 0.5-1 part of asiaticoside and 1-3 parts of guaiacum extract. The skin repairing antibacterial composition has multiple effects of repairing, moisturizing, diminishing inflammation, inhibiting bacteria and the like, has no irritation to skin, is safe to use, and has the effect of promoting repair of epidermal micro-wounds. The skin repairing antibacterial composition can be made into skin repairing antibacterial products such as washing-free gel, etc.
Description
Technical Field
The invention relates to the technical field of skin repair and bacteriostasis, in particular to a skin repair bacteriostasis composition and application thereof, a gel prepared from the skin repair bacteriostasis composition and a preparation method thereof.
Background
The skin of the hands is exposed for a long time every day and contacts various articles, and is the body part which most contacts bacteria all day long. Bacteria on the skin of the hand can infect lips, eyes, etc. by contact, or enter the body by the route of the hand and mouth, causing physical discomfort or disease. Meanwhile, hands are also very easily infected by bacteria, and skin damage may cause inflammation only locally, but also may cause systemic infection and even endanger life, such as staphylococcus skin scald syndrome. Therefore, daily bacteriostasis and disinfection of hand skin are necessary.
However, in daily life, hands can not be washed anytime and anywhere, so that the washing-free antibacterial product is more practical for daily bacteriostasis.
At present, the hand washing-free hand washing lotion is commonly used for hand bacteriostasis, and the bacteriostasis components of the hand washing lotion mainly comprise medical alcohol, hypochlorous acid, benzalkonium chloride, polyhexamethylene guanidine hydrochloride and the like, wherein the alcohol and the hypochlorous acid have stronger disinfection effect, but have certain irritation to skin and mucous membrane, can cause the skin to be dry to form tiny wounds, and has instability and bad smell. The disinfection effects of benzalkonium chloride and polyhexamethylene guanidine hydrochloride are relatively poor.
Disclosure of Invention
Aiming at the problems that the existing hand bacteriostatic no-wash products have certain irritation or are unstable and have poor disinfection effect, the invention provides a skin repairing bacteriostatic composition, a gel, an application and a preparation method.
In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:
on one hand, the embodiment of the invention provides a skin repairing bacteriostatic composition which comprises the following components in parts by weight: 1-3 parts of water-soluble paeonol, 1-3 parts of brown algae extract, 1-3 parts of corydalis yanhusuo root extract, 1-3 parts of cynanchum paniculatum extract, 0.5-1 part of asiaticoside and 1-3 parts of guaiacum extract.
In the skin repairing antibacterial composition, paeonol has effects of relieving pain and tranquilizing, diminishing inflammation and swelling, and resisting allergy, and has strong antibacterial activity to various human pathogenic bacteria. The water-soluble paeonol can be selected from commercially available products, and is preferably water-soluble clathrate of paeonol prepared from dipotassium glycyrrhizinate and amylopectin as adjuvants.
Brown algae has antioxidant effect, and helps to reduce free radical damage to skin. Fucoidan and fucoidan in brown algae have matrix metalloproteinase inhibitory activity, and can improve and improve skin condition. The brown algae polyphenol can also effectively inhibit the secretion of histamine, and has good anti-inflammatory and anti-allergic effects. The alginate oligosaccharide can increase the softness and smoothness of skin, provide a sebum membrane for the skin, effectively nourish and protect the skin, promote the regeneration of connective tissues and facilitate the natural healing of skin wounds. The fucoidin, brown algae polyphenol and alginate oligosaccharide also have the characteristics of absorbing ultraviolet rays and have the function of resisting photoaging, thereby reducing the degradation of collagen in the dermis and promoting the repair of sunburn. Brown algae also has antibacterial components, which can inhibit bacterial growth.
The corydalis yanhusuo has the effects of activating blood circulation and dissipating blood stasis, and the root extract of the corydalis yanhusuo can relieve and resist allergy and has a conditioning effect on the skin. The paniculate swallowwort root has the effects of clearing away blood poison, clearing heat, cooling blood, dispelling wind and arresting itching, and the extract liquid of the paniculate swallowwort root is beneficial to recovering the health state of skin. The asiaticoside can moisten skin, remove waste cuticle of skin, promote growth of fiber cells, promote collagen synthesis and epithelial cell growth, increase skin elasticity, and has anti-inflammatory effect. The guaiacum extract can promote skin perspiration and toxin expelling, has the effects of astringing skin, moistening and conditioning, relieving itching and swelling, resisting bacteria and inflammation, resisting allergy, promoting tissue regeneration and wound healing and the like, and has high safety.
According to the invention, the components are matched at a specific ratio, so that the obtained composition has multiple effects of repairing, moisturizing, diminishing inflammation, inhibiting bacteria and the like, is nonirritating to skin, is safe to use, and has the effect of promoting repair of epidermal micro-wounds.
Preferably, the skin repairing bacteriostatic composition comprises the following components in parts by weight: 1-1.5 parts of water-soluble paeonol, 1-2 parts of brown algae extract, 1-2 parts of corydalis yanhusuo root extract, 1-2 parts of cynanchum paniculatum extract, 0.5-0.8 part of asiaticoside and 1-2 parts of guaiacum extract.
Preferably, the skin repairing bacteriostatic composition comprises the following components in parts by weight: 1.3 parts of water-soluble paeonol, 2 parts of brown algae extract, 1 part of corydalis yanhusuo root extract, 1 part of cynanchum paniculatum extract, 0.5 part of asiaticoside and 2 parts of guaiacum extract.
Optionally, the preparation method of the brown algae extract comprises the following steps: soaking brown algae in water, performing microwave extraction for 10-15 min under the conditions that the power is 750-850W and the temperature is 40-50 ℃, and filtering to obtain filter residues and a microwave extracting solution; carrying out reflux extraction on the filter residue for 2-3 times by using 25-45% v/v ethanol water solution, each time for 20-30 min, and combining reflux extracting solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the brown algae extracting solution;
the preparation method of the corydalis yanhusuo root extracting solution comprises the following steps: crushing corydalis yanhusuo roots, immersing the crushed corydalis yanhusuo roots into water, performing microwave extraction for 10-15 min under the conditions that the power is 650-750W and the temperature is 30-40 ℃, and filtering to obtain filter residues and a microwave extracting solution; carrying out reflux extraction on the filter residue for 2-3 times by using 35-55% v/v ethanol water solution, each time for 20-30 min, and combining reflux extracting solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the corydalis dentata root extracting solution;
the preparation method of the cynanchum paniculatum extracting solution comprises the following steps: immersing the cynanchum paniculatum in water, performing microwave extraction for 10-15 min under the conditions that the power is 700-800W and the temperature is 30-40 ℃, and filtering to obtain filter residues and a microwave extracting solution; ultrasonically extracting the filter residue with 35-55% v/v ethanol water solution for 2-3 times, wherein each time is 20-30 min, and combining reflux extracting solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the cynanchum paniculatum extracting solution;
the preparation method of the guaiac wood extracting solution comprises the following steps: immersing guaiac wood into 5-15% v/v ethanol water solution, performing microwave extraction for 10-15 min under the conditions of 700-800W of power and 40-60 ℃, and filtering to obtain filter residue and microwave extracting solution; ultrasonically extracting the filter residue with 45-65% v/v ethanol water solution for 2-3 times, each time for 20-30 min, and combining reflux extracting solutions; and combining the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the guaiac wood extracting solution.
The skin repairing and bacteriostasis composition has better repairing, moisturizing, anti-inflammatory and bacteriostasis effects by using the extracting solutions prepared by the preparation method.
Preferably, the skin repairing antibacterial composition further comprises honeysuckle extract, pseudo-ginseng root extract, bletilla striata root extract, horse chestnut extract, forsythia fruit extract and licorice root extract, and the weight parts of the components are as follows: 1-3 parts of honeysuckle extract, 1-3 parts of pseudo-ginseng root extract, 1-3 parts of bletilla striata root extract, 0.5-1 part of horse chestnut extract, 0.5-1 part of forsythia suspensa fruit extract and 1-3 parts of licorice root extract.
The honeysuckle has the effects of clearing heat, expelling toxin, resisting bacteria and diminishing inflammation, and the extract has certain inhibiting and killing effects on common pathogenic bacteria and various viruses of the skin; the Notoginseng radix extract has hemostatic, blood stasis dispelling, repercussive, and analgesic effects; the rhizoma Bletillae root extractive solution has antioxidant, skin whitening and antiinflammatory effects; the horse chestnut extract has the effects of improving skin structure, improving blood circulation, regulating metabolism of epidermis and horny layer, and improving water and oil balance of skin, and is helpful for keeping healthy appearance of skin; fructus forsythiae extractive solution has antioxidant, moisturizing, and anti-inflammatory effects, and can improve dry and rough skin; the Glycyrrhrizae radix extractive solution has antiinflammatory, antibacterial, and antiviral effects. The honeysuckle extract, the pseudo-ginseng root extract, the bletilla striata root extract, the horse chestnut extract, the forsythia fruit extract and the licorice root extract are combined with the water-soluble paeonol, the brown algae extract, the corydalis yanhusuo root extract, the cynanchum paniculatum extract, the asiaticoside and the guaiacum extract according to a specific ratio, so that the obtained skin repair antibacterial composition has stronger antibacterial action and more remarkable repairing and anti-inflammatory effects, and can further promote skin recovery or keep a healthy state.
Preferably, the weight parts of the components are as follows: 1-2 parts of honeysuckle extract, 1-2 parts of pseudo-ginseng root extract, 1-2 parts of bletilla striata root extract, 0.5-0.8 part of horse chestnut extract, 0.5-1 part of forsythia suspensa fruit extract and 1-2 parts of licorice root extract.
Preferably, the weight parts of the components are as follows: 1 part of honeysuckle extract, 1 part of pseudo-ginseng root extract, 1 part of bletilla striata root extract, 0.5 part of horse chestnut extract, 0.5 part of forsythia fruit extract and 1 part of licorice root extract.
Optionally, the preparation method of the honeysuckle extract comprises the following steps: carrying out ultrasonic extraction on honeysuckle at 45-60 ℃ by using 45-65% v/v ethanol water solution, carrying out ultrasonic extraction for 20-30 min at the frequency of 20-35 kHz, and carrying out ultrasonic extraction for 15-20 min at the frequency of 55-70 kHz; filtering after extraction is finished, and concentrating the obtained filtrate to 1.2-1.3 g/ml to obtain the honeysuckle extracting solution;
the preparation method of the pseudo-ginseng root extracting solution comprises the following steps: crushing pseudo-ginseng roots, performing microwave extraction for 10-15 min by using 5-15% v/v ethanol water solution under the conditions that the power is 750-950W and the temperature is 30-40 ℃, and filtering to obtain filter residues and a microwave extracting solution; carrying out reflux extraction on the filter residue for 2-3 times by using 15-25% v/v ethanol water solution, each time for 20-30 min, and combining reflux extracting solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the pseudo-ginseng root extracting solution;
the preparation method of the bletilla striata root extracting solution comprises the following steps: crushing bletilla striata roots, performing microwave extraction for 10-15 min by using 10-20% v/v ethanol water solution under the conditions that the power is 650-850W and the temperature is 30-40 ℃, and filtering to obtain filter residues and a microwave extracting solution; carrying out reflux extraction on the filter residue for 2-3 times by using 35-45% v/v ethanol water solution, each time for 20-30 min, and combining reflux extracting solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the bletilla striata root extracting solution;
the preparation method of the horse chestnut extracting solution comprises the following steps: crushing the branches or trunks of the horse chestnut, and ultrasonically extracting for 2-3 times at the frequency of 30-45 kHz for 20-30 min each time by using 45-65% v/v ethanol water solution at the temperature of 50-70 ℃; filtering after extraction is finished, and concentrating the obtained filtrate to 1.2-1.3 g/ml to obtain the horse chestnut extracting solution;
the preparation method of the forsythia suspense fruit extracting solution comprises the following steps: carrying out ultrasonic extraction on fructus forsythiae by using 15-25% v/v ethanol water solution at 25-35 ℃, extracting for 20-30 min at the frequency of 25-35 kHz, and extracting for 15-25 min at the frequency of 50-70 kHz; filtering after extraction is finished, and concentrating the obtained filtrate to 1.2-1.3 g/ml to obtain the fructus forsythiae fruit extracting solution;
the preparation method of the licorice root extracting solution comprises the following steps: crushing raw liquorice, decocting the crushed raw liquorice for 1-1.5 hours by using water, cooling the decoction to 40-50 ℃, adding cellulase accounting for 0.5-1.5 percent of the mass of the raw liquorice for enzymolysis at 40-50 ℃ for 2-3 hours, heating the mixture to 90-95 ℃, keeping the mixture at the temperature for 5-10 min for enzyme deactivation, and filtering the mixture to obtain enzymolysis liquid and filter residues; ultrasonically extracting the filter residue for 20-30 min by using water at the frequency of 20-30 kHz, and filtering to obtain an ultrasonic extracting solution; and combining the enzymolysis liquid and the ultrasonic extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the licorice root extracting solution.
The honeysuckle extract, the pseudo-ginseng root extract, the bletilla striata root extract, the escargot extract, the forsythia fruit extract and the licorice root extract obtained by the preparation method can further improve the repairing, moisturizing, anti-inflammatory and bacteriostatic effects of the skin repairing and bacteriostatic composition.
In a second aspect, the embodiment of the invention also provides an application of the skin repairing bacteriostatic composition in preparing a skin repairing bacteriostatic product.
The skin repairing antibacterial composition has multiple effects of repairing, moisturizing, diminishing inflammation, inhibiting bacteria and the like, has no irritation to skin, is safe to use, and can be used for preparing skin repairing antibacterial products, such as no-wash gel, shower gel, hand sanitizer, no-wash spray and the like.
In a third aspect, the embodiment of the invention also provides a gel, which comprises the skin repairing antibacterial composition.
Preferably, the gel comprises the following components in parts by weight:
the above preferred raw materials are all compliant components of skin external products, are highly safe, are non-irritating to the skin, and do not cause skin discomfort when used in the above dosage range.
Preferably, the gel comprises the following components in parts by weight:
preferably, the gel comprises the following components in parts by weight:
preferably, the humectant is selected from at least one of propylene glycol, glycerin and butylene glycol;
the preservative is caprylyl hydroximic acid and o-cymene-5-alcohol;
the pH regulator is aminomethyl propanol;
the nutritional supplement is panthenol;
the acrylic acid (ester)/C10-30 alkanol acrylate crosspolymer may be commercially available as carbomer U20 (Luborun USA).
In a fourth aspect, the present invention further provides a method for preparing the above gel, comprising the steps of:
adding acrylic acid (ester)/C10-30 alkanol acrylate cross-linked polymer, poloxamer 407, humectant and PEG-40 hydrogenated castor oil into water, heating to 70-80 ℃, stirring to completely dissolve, adding a pH regulator, stirring uniformly, cooling to 50-55 ℃, adding a nutritional supplement and menthol, stirring and dispersing uniformly for later use;
cooling to 42-45 ℃, adding water-soluble paeonol, brown algae extract, honeysuckle flower extract, pseudo-ginseng root extract, bletilla striata root extract, corydalis yanhusuo root extract, cynanchum paniculatum extract, horse chestnut extract, asiaticoside, fructus forsythiae extract, licorice root extract, guaiacum extract and preservative, uniformly stirring, and cooling to 35-40 ℃ to obtain the gel.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The experimental procedures used in the following examples are all conventional in the art unless otherwise specified.
The paniculate swallowwort root, the honeysuckle, the pseudo-ginseng, the bletilla striata, the forsythia fruit and the raw liquorice used in the following examples all conform to the relevant regulations under each medicinal material item in the text of the Chinese pharmacopoeia (2015 edition).
The raw materials and reagents used in the following examples were obtained commercially, unless otherwise specified, wherein the water-soluble paeonol was obtained from Guangzhou Xiandani chemical science and technology Co., Ltd and was a paeonol water-soluble inclusion compound prepared using dipotassium glycyrrhizinate and pullulan as auxiliary materials.
Examples 1 to 10
The preparation of the skin repairing and bacteriostatic composition comprises the following steps: the following components are included in each example in the following weight portion ratios as shown in table 1:
TABLE 1 examples 1-10 bacteriostatic skin repair compositions
In examples 1 to 3 and 6 to 8, the brown algae extract solution is prepared by the following steps: soaking brown algae in water, performing microwave extraction at 45 deg.C and power of 800W for 12min, and filtering to obtain filter residue and microwave extractive solution; extracting the residue with 35% v/v ethanol water under reflux for 25min for 2 times, and mixing extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.25g/ml to obtain the brown algae extracting solution;
the preparation method of the corydalis yanhusuo root extracting solution comprises the following steps: crushing rhizoma corydalis, soaking in water, performing microwave extraction at power of 700W and temperature of 35 deg.C for 12min, and filtering to obtain filter residue and microwave extract; extracting the filter residue with 45% v/v ethanol water under reflux for 25min for 2 times, and mixing the extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.25g/ml to obtain the corydalis dentata root extracting solution;
the preparation method of the cynanchum paniculatum extract comprises the following steps: soaking radix Cynanchi Paniculati in water, performing microwave extraction at power of 750W and temperature of 35 deg.C for 12min, and filtering to obtain filter residue and microwave extractive solution; ultrasonically extracting the filter residue with 45% v/v ethanol water solution for 2 times, each time for 25min, and mixing the reflux extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.25g/ml to obtain the cynanchum paniculatum extracting solution;
the preparation method of the guaiacum extracting solution comprises the following steps: soaking guaiac wood in 10% v/v ethanol water solution, performing microwave extraction at 50 deg.C and 750W for 12min, and filtering to obtain filter residue and microwave extractive solution; ultrasonically extracting the filter residue with 55% v/v ethanol water solution for 2 times, each time for 25min, and mixing the reflux extractive solutions; and (3) combining the microwave extracting solution and the reflux extracting solution, and concentrating to 1.25g/ml to obtain the guaiac wood extracting solution.
In examples 4 and 9, the brown algae extract is prepared by the following steps: soaking brown algae in water, performing microwave extraction at 50 deg.C and power of 750W for 10min, and filtering to obtain filter residue and microwave extract; reflux-extracting the residue with 25% v/v ethanol water solution for 30min for 2 times, and mixing the extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.3g/ml to obtain the brown algae extracting solution;
the preparation method of the corydalis yanhusuo root extracting solution comprises the following steps: crushing corydalis yanhusuo root, soaking in water, performing microwave extraction at 40 deg.C and 650W for 10min, and filtering to obtain filter residue and microwave extract; extracting the filter residue with 55% v/v ethanol water under reflux for 30min for 2 times, and mixing the extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.3g/ml to obtain the corydalis yanhusuo root extracting solution;
the preparation method of the cynanchum paniculatum extract comprises the following steps: soaking radix Cynanchi Paniculati in water, performing microwave extraction at 40 deg.C and power of 700W for 10min, and filtering to obtain filter residue and microwave extractive solution; ultrasonically extracting the filter residue with 35% v/v ethanol water solution for 3 times, each time for 20min, and mixing the reflux extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2g/ml to obtain the cynanchum paniculatum extracting solution;
the preparation method of the guaiacum extracting solution comprises the following steps: soaking guaiac wood in 5% v/v ethanol water solution, performing microwave extraction at 40 deg.C and power of 800W for 10min, and filtering to obtain filter residue and microwave extractive solution; ultrasonically extracting the filter residue with 65% v/v ethanol water solution for 2 times, each time for 30min, and mixing the reflux extractive solutions; and combining the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2g/ml to obtain the guaiac wood extracting solution.
In examples 5 and 10, the brown algae extract was prepared by the following method: soaking brown algae in water, performing microwave extraction at 850W and 40 deg.C for 15min, and filtering to obtain filter residue and microwave extract; extracting the residue with 45% v/v ethanol water under reflux for 20min for 3 times, and mixing extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2g/ml to obtain the brown algae extracting solution;
the preparation method of the corydalis yanhusuo root extracting solution comprises the following steps: crushing rhizoma corydalis, soaking in water, performing microwave extraction at power of 750W and temperature of 30 deg.C for 15min, and filtering to obtain filter residue and microwave extract; extracting the filter residue with 35% v/v ethanol water under reflux for 20min for 3 times, and mixing the extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2g/ml to obtain the corydalis yanhusuo root extracting solution;
the preparation method of the cynanchum paniculatum extract comprises the following steps: soaking radix Cynanchi Paniculati in water, performing microwave extraction at power of 800W and temperature of 30 deg.C for 15min, and filtering to obtain filter residue and microwave extractive solution; ultrasonically extracting the filter residue with 55% v/v ethanol water solution for 2 times, each time for 30min, and mixing the reflux extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.3g/ml to obtain the cynanchum paniculatum extracting solution;
the preparation method of the guaiac wood extracting solution comprises the following steps: soaking guaiac wood in 15% v/v ethanol water solution, performing microwave extraction at power of 700W and temperature of 60 deg.C for 15min, and filtering to obtain filter residue and microwave extractive solution; ultrasonically extracting the filter residue with 45% v/v ethanol water solution for 3 times, each time for 20min, and mixing the reflux extractive solutions; and (3) combining the microwave extracting solution and the reflux extracting solution, and concentrating to 1.3g/ml to obtain the guaiac wood extracting solution.
In examples 6 to 8, a method for preparing a honeysuckle extract solution comprises: ultrasonic extracting flos Lonicerae with 55% v/v ethanol water solution at 55 deg.C for 25min at 25kHz, and then for 18min at 60 kHz; filtering after extraction is finished, and concentrating the obtained filtrate to 1.25g/ml to obtain the honeysuckle extracting solution;
the preparation method of the pseudo-ginseng root extracting solution comprises the following steps: crushing Notoginseng radix, performing microwave extraction with 10% v/v ethanol water solution at power of 850W and temperature of 35 deg.C for 12min, and filtering to obtain filter residue and microwave extract; reflux-extracting the residue with 20% v/v ethanol water solution for 25min for 2 times, and mixing the extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.25g/ml to obtain the pseudo-ginseng root extracting solution;
the preparation method of the bletilla striata root extracting solution comprises the following steps: crushing rhizoma bletilla, performing microwave extraction with 15% v/v ethanol water solution at power of 750W and temperature of 35 deg.C for 12min, and filtering to obtain filter residue and microwave extract; reflux-extracting the residue with 40% v/v ethanol water solution for 25min for 2 times, and mixing the extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.25g/ml to obtain the bletilla striata root extracting solution;
the preparation method of the horse chestnut extracting solution comprises the following steps: crushing branches or trunks of horse chestnut, and ultrasonically extracting with 55% v/v ethanol water solution at 60 deg.C and 35kHz for 2 times, each time for 25 min; filtering after extraction is finished, and concentrating the obtained filtrate to 1.25g/ml to obtain the horse chestnut extracting solution;
the preparation method of the forsythia suspense fruit extracting solution comprises the following steps: ultrasonically extracting fructus forsythiae with 20% v/v ethanol water solution at 30 deg.C for 25min at 30kHz, and extracting for 20min at 60 kHz; filtering after extraction is finished, and concentrating the obtained filtrate to 1.25g/ml to obtain the forsythia suspense fruit extracting solution;
the preparation method of the licorice root extracting solution comprises the following steps: pulverizing Glycyrrhrizae radix, decocting with water for 1.2h, cooling to 45 deg.C, adding cellulase 1.0% of the Glycyrrhrizae radix, performing enzymolysis at 45 deg.C for 2.5h, heating to 93 deg.C, maintaining for 8min, inactivating enzyme, and filtering to obtain enzymolysis solution and residue; ultrasonically extracting the filter residue for 25min by using water at the frequency of 25kHz, and filtering to obtain an ultrasonic extracting solution; and mixing the enzymolysis liquid and the ultrasonic extraction liquid, and concentrating to 1.25g/ml to obtain the licorice root extraction liquid.
In example 9, the preparation method of the honeysuckle extract solution comprises: ultrasonically extracting flos Lonicerae with 45% v/v ethanol water solution at 60 deg.C, ultrasonically extracting at 20kHz for 30min, and ultrasonically extracting at 70kHz for 15 min; filtering after extraction is finished, and concentrating the obtained filtrate to 1.2g/ml to obtain the honeysuckle extracting solution;
the preparation method of the pseudo-ginseng root extracting solution comprises the following steps: crushing Notoginseng radix, performing microwave extraction with 5% v/v ethanol water solution at power of 950W and temperature of 30 deg.C for 10min, and filtering to obtain filter residue and microwave extract; reflux-extracting the residue with 25% v/v ethanol water solution for 30min for 2 times, and mixing the extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.3g/ml to obtain the seventy root extracting solution;
the preparation method of the bletilla striata root extracting solution comprises the following steps: crushing bletilla striata roots, performing microwave extraction for 10min at the power of 650W and the temperature of 40 ℃ by using 10% v/v ethanol water solution, and filtering to obtain filter residues and microwave extraction liquid; extracting the residue with 45% v/v ethanol water under reflux for 30min for 2 times, and mixing extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.3g/ml to obtain the bletilla striata root extracting solution;
the preparation method of the horse chestnut extracting solution comprises the following steps: crushing branches or trunks of horse chestnut, and ultrasonically extracting with 45% v/v ethanol water solution at 70 deg.C and 30kHz for 2 times, each for 30 min; filtering after extraction is finished, and concentrating the obtained filtrate to 1.3g/ml to obtain the horse chestnut extracting solution;
the preparation method of the forsythia suspense fruit extracting solution comprises the following steps: ultrasonically extracting fructus forsythiae with 15% v/v ethanol water solution at 35 deg.C for 30min at 25kHz, and extracting for 25min at 50 kHz; filtering after extraction is finished, and concentrating the obtained filtrate to 1.2g/ml to obtain the forsythia suspense fruit extracting solution;
the preparation method of the licorice root extracting solution comprises the following steps: pulverizing Glycyrrhrizae radix, decocting with water for 1h, cooling to 40 deg.C, adding cellulase 0.5% of the Glycyrrhrizae radix, performing enzymolysis at 50 deg.C for 2h, heating to 90 deg.C, maintaining for 10min, inactivating enzyme, and filtering to obtain enzymolysis solution and filter residue; extracting the filter residue with water under 20kHz for 30min by ultrasonic extraction, and filtering to obtain ultrasonic extractive solution; and mixing the enzymolysis liquid and the ultrasonic extracting solution, and concentrating to 1.2g/ml to obtain the licorice root extracting solution.
In example 10, the preparation method of the honeysuckle extract solution comprises: carrying out ultrasonic extraction on honeysuckle at 45 ℃ by using 65% v/v ethanol water solution, firstly carrying out ultrasonic extraction for 20min at the frequency of 35kHz, and then carrying out ultrasonic extraction for 20min at the frequency of 55 kHz; filtering after extraction is finished, and concentrating the obtained filtrate to 1.3g/ml to obtain the honeysuckle extracting solution;
the preparation method of the pseudo-ginseng root extracting solution comprises the following steps: crushing Notoginseng radix, performing microwave extraction with 15% v/v ethanol water solution at power of 750W and temperature of 40 deg.C for 15min, and filtering to obtain filter residue and microwave extract; extracting the residue with 15% v/v ethanol water under reflux for 30min for 3 times, and mixing extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2g/ml to obtain the seventy root extracting solution;
the preparation method of the bletilla striata root extracting solution comprises the following steps: crushing rhizoma bletilla, performing microwave extraction with 20% v/v ethanol water solution at 850W and 30 deg.C for 15min, and filtering to obtain filter residue and microwave extract; extracting the residue with 35% v/v ethanol water under reflux for 20min for 3 times, and mixing extractive solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2g/ml to obtain the bletilla striata root extracting solution;
the preparation method of the horse chestnut extracting solution comprises the following steps: crushing branches or trunks of horse chestnut, and ultrasonically extracting with 65% v/v ethanol water solution at 50 deg.C and 45kHz for 3 times, each time for 20 min; filtering after extraction is finished, and concentrating the obtained filtrate to 1.2g/ml to obtain the horse chestnut extracting solution;
the preparation method of the forsythia suspense fruit extracting solution comprises the following steps: ultrasonic extracting fructus forsythiae with 25% v/v ethanol water solution at 35 deg.C for 20min at 35kHz, and extracting for 15min at 70 kHz; filtering after extraction is finished, and concentrating the obtained filtrate to 1.3g/ml to obtain the forsythia suspense fruit extracting solution;
the preparation method of the licorice root extracting solution comprises the following steps: pulverizing Glycyrrhrizae radix, decocting with water for 1.5h, cooling to 50 deg.C, adding cellulase 1.5% of the Glycyrrhrizae radix, performing enzymolysis at 40 deg.C for 3h, heating to 95 deg.C, maintaining for 5min, inactivating enzyme, and filtering to obtain enzymolysis solution and residue; ultrasonically extracting the filter residue for 30min by using water at the frequency of 30kHz, and filtering to obtain an ultrasonic extracting solution; and mixing the enzymolysis liquid and the ultrasonic extraction liquid, and concentrating to 1.3g/ml to obtain the licorice root extraction liquid.
Examples 11 to 20
The invention provides a gel and a preparation method thereof: the following components are included in each example in the following weight portion ratios as shown in table 2:
TABLE 2 examples 11-20 gels
Wherein the brown algae extract, corydalis yanhusuo root extract, paniculate swallowwort root extract, guaiacum extract, honeysuckle extract, notoginseng root extract, bletilla striata root extract, horse chestnut extract, forsythia fruit extract and licorice root extract are prepared by the same method as in example 1.
The preparation method of the gel comprises the following steps:
adding acrylic acid (ester)/C10-30 alkanol acrylate cross-linked polymer, poloxamer 407, humectant and PEG-40 hydrogenated castor oil into water, heating to 80 deg.C, stirring for dissolving completely, adding aminomethyl propanol, stirring, cooling to 50 deg.C, adding panthenol and Mentholum, stirring, and dispersing uniformly;
cooling to 45 ℃, adding water-soluble paeonol, brown algae extract, honeysuckle flower extract, pseudo-ginseng root extract, bletilla striata root extract, corydalis yanhusuo root extract, cynanchum paniculatum extract, horse chestnut extract, asiaticoside, fructus forsythiae extract, licorice root extract, guaiacum extract and preservative, stirring uniformly, and cooling to 40 ℃ to obtain the gel.
Comparative example 1
The comparative example provides a skin repairing bacteriostatic composition, which is prepared by removing brown algae extract on the basis of example 1, and keeping the rest components unchanged, wherein the preparation method of each extract is the same as that of example 1.
Comparative example 2
The comparative example provides a skin repairing and bacteriostatic composition, which is prepared by removing corydalis dentata root extracting solution from the components of the composition of example 1, and keeping the other components unchanged, wherein the preparation method of each extracting solution is the same as that of example 1.
Comparative example 3
The comparative example provides a skin repairing bacteriostatic composition, which is prepared by removing the cynanchum paniculatum extracting solution on the basis of example 1, and keeping the rest components unchanged, wherein the preparation method of each extracting solution is the same as that of example 1.
Comparative example 4
The comparative example provides a skin repairing bacteriostatic composition, which is prepared by removing asiaticoside from the components of example 1, keeping the other components unchanged, and preparing the extracting solutions according to the same method as example 1.
Comparative example 5
The comparative example provides a skin repairing bacteriostatic composition, which is prepared by removing guaiac wood extract from the composition of example 1, and keeping the rest components unchanged, wherein the preparation method of each extract is the same as that of example 1.
Comparative example 6
The comparative example provides a gel and a preparation method thereof, the component proportion of the gel is that the brown algae extracting solution is removed on the basis of the example 1, the content of the water-soluble paeonol, the corydalis yanhusuo root extracting solution, the cynanchum paniculatum extracting solution, the asiaticoside and the guaiacum extracting solution is improved by 34.5 percent, the preparation method of each extracting solution is the same as the example 1, and the preparation method of the gel is the same as the example 11.
Comparative example 7
The comparative example provides a gel and a preparation method thereof, the component proportion of the gel is that on the basis of the example 1, the corydalis yanhusuo root extracting solution is removed, the content of the water-soluble paeonol, the brown algae extracting solution, the cynanchum paniculatum extracting solution, the asiaticoside and the guaiacum extracting solution is improved by 15 percent, the preparation method of each extracting solution is the same as the example 1, and the preparation method of the gel is the same as the example 11.
Comparative example 8
The comparative example provides a gel and a preparation method thereof, wherein the component proportion of the gel is that the cynanchum paniculatum extracting solution is removed on the basis of the gel in the example 1, the content of the water-soluble paeonol, the brown algae extracting solution, the corydalis repens root extracting solution, the asiaticoside and the guaiacum extracting solution is improved by 15 percent, the preparation method of each extracting solution is the same as the example 1, and the preparation method of the gel is the same as the example 11.
Comparative example 9
The comparative example provides a gel and a preparation method thereof, the component proportion of the gel is based on example 1, asiaticoside is removed, the content of water-soluble paeonol, brown algae extract, corydalis yanhusuo root extract, cynanchum paniculatum extract and guaiacum extract is increased by 4%, the preparation method of each extract is the same as example 1, and the preparation method of the gel is the same as example 11.
Comparative example 10
The comparative example provides a gel and a preparation method thereof, wherein the component proportion of the gel is that the guaiacum extracting solution is removed on the basis of the example 1, the content of the water-soluble paeonol, the brown alga extracting solution, the corydalis yanhusuo root extracting solution, the cynanchum paniculatum extracting solution and the asiaticoside is improved by 34.5 percent, the preparation method of each extracting solution is the same as the example 1, and the preparation method of the gel is the same as the example 11.
Test example 1
In the test example, the skin repairing and bacteriostasis compositions prepared in the above examples 1 to 10 and comparative examples 1 to 5 are examined for bacteriostasis effect.
The nutrient agar culture medium adopted in the following experiments comprises the following components in proportion: 10g of peptone, 3g of beef extract, 5g of sodium chloride, 17g of agar and 1000mL of distilled water;
the composition of the Sabouraud's medium is: 10g of peptone, 20g of agar, 40g of glucose and 1L of distilled water.
Test strains: escherichia coli (ATCC25922), Pseudomonas aeruginosa (ATCC 15442).
1. Experimental methods
Slant culture of test bacteria in 5 th generation nutrient agar medium (24h) with 5mL of 0.03mol/L phosphorusWashing thallus Porphyrae with acid salt buffer solution (PBS), diluting with PBS to obtain 5 × 10 concentration4cfu/ml bacterial suspension.
Samples of the skin-repairing antibacterial compositions prepared in examples 1 to 10 and comparative examples 1 to 5 were used as test sample solutions, and 4 tubes of each of the test sample solution (5mL) and the control sample solution (distilled water after sterilization treatment) were taken.
Respectively dripping 100 mu L of the bacterial suspension into each sample solution to be tested and each control sample solution, uniformly mixing, starting timing, acting for 2min, 5min, 10min and 30min, respectively putting the mixed sample solution (0.5mL) into a test tube containing 5mL of PBS by using a sterile pipette, fully and uniformly mixing, respectively sucking 0.5mL, placing the sample solution into two plates, pouring 15mL of nutrient agar medium (escherichia coli) or Sabouraud's agar medium (copper green pseudomonas) cooled to 40-50 ℃, rotating the plates to fully and uniformly stir the plates, culturing the plates for 48h (escherichia coli) or 72h (pseudomonas aeruginosa) at 35 +/-2 ℃ after agar is solidified, and counting viable bacteria colonies.
The experiment was repeated 3 times and the inhibition was calculated as follows:
X＝(A-B)/A×100％；
in the formula: x is the bacteriostasis rate, A is the average colony number of the control sample liquid; b is the average colony number of the sample liquid.
2. Results of the experiment
The average bacteriostasis rate values of the skin repairing bacteriostasis compositions under different action time are recorded, and the results are shown in tables 3 and 4.
TABLE 3 bacteriostatic data on E.coli
TABLE 4 bacteriostatic data on Pseudomonas aeruginosa
As can be seen from the test results in tables 3 and 4, the skin repair antibacterial composition obtained in the embodiments 1 to 10 has a good inhibition effect on Escherichia coli and Pseudomonas aeruginosa, and is superior to the antibacterial effect of the skin repair antibacterial composition obtained in each proportion.
Test example 2
This test example examined the irritation of the gels prepared in examples 11 to 20.
1. Experimental methods
45 new zealand rabbits with intact skins, female rabbits with the weight of 2.0-2.5 kg, are fed by a common-grade rabbit maintaining feed at the temperature of 20-26 ℃ and the relative humidity of 40-70%.
The gels prepared in examples 11 to 20 and comparative examples 6 to 10 were each tested for full skin irritation according to the Disinfection Specification (2002 edition) 2.3.3.3.1 (3 rabbits were used for each sample): about 24 hours before the experiment, removing hairs on two sides of the spinal column of the white rabbit, not damaging the skin, removing hairs in the left side and the right side of the skin by about 3cm multiplied by 3cm respectively, taking 0.5ml of tested gel to be smeared on the skin, then covering the skin with two layers of gauze and a layer of cellophane, fixing the skin by using an adhesive tape, smearing 0.5ml of distilled water on the other side of the skin to be used as a blank control, applying the gel for 4 hours, washing the gel for 1min by using warm water to remove the tested object, observing local reactions of the skin after 1 hour, 24 hours and 48 hours of removing the tested object respectively, and scoring the skin irritation reactions according to a table 5. The results are shown in Table 6.
TABLE 5 skin irritation response Scoring criteria
TABLE 6 test results of once intact skin irritation test of test substances on rabbits
As can be seen from the results in Table 6, the gels obtained in examples 11 to 20 were non-irritating or slightly irritating to the skin of the white rabbits, and the gels obtained in comparative examples 6 to 10 were slightly irritating to the skin of the white rabbits.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. The skin repairing and bacteriostatic composition is characterized by comprising the following components in parts by weight: 1-3 parts of water-soluble paeonol, 1-3 parts of brown algae extract, 1-3 parts of corydalis yanhusuo root extract, 1-3 parts of cynanchum paniculatum extract, 0.5-1 part of asiaticoside and 1-3 parts of guaiacum extract.
2. The skin repair bacteriostatic composition according to claim 1, which comprises the following components in parts by weight: 1-1.5 parts of water-soluble paeonol, 1-2 parts of brown algae extract, 1-2 parts of corydalis yanhusuo extract, 1-2 parts of cynanchum paniculatum extract, 0.5-0.8 part of asiaticoside and 1-2 parts of guaiacum extract.
3. The skin repair bacteriostatic composition according to claim 2, which is characterized by comprising the following components in parts by weight: 1.3 parts of water-soluble paeonol, 2 parts of brown algae extract, 1 part of corydalis yanhusuo root extract, 1 part of cynanchum paniculatum extract, 0.5 part of asiaticoside and 2 parts of guaiacum extract.
4. The skin-repairing bacteriostatic composition according to any one of claims 1 to 3, wherein the preparation method of the brown algae extract comprises the following steps: soaking brown algae in water, performing microwave extraction for 10-15 min under the conditions that the power is 750-850W and the temperature is 40-50 ℃, and filtering to obtain filter residues and a microwave extracting solution; carrying out reflux extraction on the filter residue for 2-3 times by using 25-45% v/v ethanol water solution, each time for 20-30 min, and combining reflux extracting solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the brown algae extracting solution; and/or
The preparation method of the corydalis yanhusuo root extracting solution comprises the following steps: crushing corydalis yanhusuo roots, immersing the crushed corydalis yanhusuo roots into water, performing microwave extraction for 10-15 min under the conditions that the power is 650-750W and the temperature is 30-40 ℃, and filtering to obtain filter residues and a microwave extracting solution; carrying out reflux extraction on the filter residue for 2-3 times by using 35-55% v/v ethanol water solution, each time for 20-30 min, and combining reflux extracting solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the corydalis dentata root extract; and/or
The preparation method of the cynanchum paniculatum extracting solution comprises the following steps: immersing the cynanchum paniculatum in water, performing microwave extraction for 10-15 min under the conditions that the power is 700-800W and the temperature is 30-40 ℃, and filtering to obtain filter residues and a microwave extracting solution; ultrasonically extracting the filter residue for 2-3 times by using 35-55% v/v ethanol water solution, wherein each time is 20-30 min, and combining reflux extracting solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the cynanchum paniculatum extracting solution; and/or
The preparation method of the guaiac wood extracting solution comprises the following steps: immersing guaiac wood into 5-15% v/v ethanol water solution, performing microwave extraction for 10-15 min under the conditions of 700-800W of power and 40-60 ℃, and filtering to obtain filter residue and microwave extracting solution; ultrasonically extracting the filter residue with 45-65% v/v ethanol water solution for 2-3 times, each time for 20-30 min, and combining reflux extracting solutions; and combining the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the guaiac wood extracting solution.
5. The skin repairing bacteriostatic composition according to any one of claims 1 to 3, wherein the skin repairing bacteriostatic composition further comprises a honeysuckle extract, a notoginseng root extract, a bletilla striata root extract, a horse chestnut extract, a forsythia fruit extract and a licorice root extract, and the weight parts of the components are as follows: 1-3 parts of honeysuckle extract, 1-3 parts of pseudo-ginseng root extract, 1-3 parts of bletilla striata root extract, 0.5-1 part of horse chestnut extract, 0.5-1 part of forsythia suspensa fruit extract and 1-3 parts of licorice root extract.
6. The skin repair bacteriostatic composition according to claim 5, wherein the weight parts of the components are as follows: 1-2 parts of honeysuckle extract, 1-2 parts of pseudo-ginseng root extract, 1-2 parts of bletilla striata root extract, 0.5-0.8 part of horse chestnut extract, 0.5-1 part of forsythia suspensa fruit extract and 1-2 parts of licorice root extract.
7. The skin repair bacteriostatic composition according to claim 6, wherein the weight parts of the components are as follows: 1 part of honeysuckle extract, 1 part of pseudo-ginseng root extract, 1 part of bletilla striata root extract, 0.5 part of horse chestnut extract, 0.5 part of forsythia fruit extract and 1 part of licorice root extract.
8. The skin-repairing bacteriostatic composition according to claim 5, wherein the preparation method of the honeysuckle extract comprises the following steps: carrying out ultrasonic extraction on honeysuckle at 45-60 ℃ by using 45-65% v/v ethanol aqueous solution, firstly carrying out ultrasonic extraction for 20-30 min at the frequency of 20-35 kHz, and then carrying out ultrasonic extraction for 15-20 min at the frequency of 55-70 kHz; filtering after extraction is finished, and concentrating the obtained filtrate to 1.2-1.3 g/ml to obtain the honeysuckle extract; and/or
The preparation method of the pseudo-ginseng root extracting solution comprises the following steps: crushing pseudo-ginseng roots, performing microwave extraction for 10-15 min by using a 5-15% v/v ethanol water solution under the conditions that the power is 750-950W and the temperature is 30-40 ℃, and filtering to obtain filter residues and a microwave extracting solution; carrying out reflux extraction on the filter residue for 2-3 times by using 15-25% v/v ethanol water solution, each time for 20-30 min, and combining reflux extracting solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the pseudo-ginseng root extracting solution; and/or
The preparation method of the bletilla striata root extracting solution comprises the following steps: crushing bletilla striata roots, performing microwave extraction for 10-15 min by using 10-20% v/v ethanol water solution under the conditions that the power is 650-850W and the temperature is 30-40 ℃, and filtering to obtain filter residues and a microwave extracting solution; carrying out reflux extraction on the filter residue for 2-3 times by using 35-45% v/v ethanol water solution, each time for 20-30 min, and combining reflux extracting solutions; mixing the microwave extracting solution and the reflux extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the bletilla striata root extracting solution; and/or
The preparation method of the horse chestnut extracting solution comprises the following steps: crushing the branches or trunks of the horse chestnut, and ultrasonically extracting for 2-3 times at the frequency of 30-45 kHz for 20-30 min each time by using 45-65% v/v ethanol water solution at the temperature of 50-70 ℃; filtering after extraction is finished, and concentrating the obtained filtrate to 1.2-1.3 g/ml to obtain the horse chestnut extracting solution; and/or
The preparation method of the forsythia suspense fruit extracting solution comprises the following steps: carrying out ultrasonic extraction on fructus forsythiae by using 15-25% v/v ethanol water solution at 25-35 ℃, extracting for 20-30 min at the frequency of 25-35 kHz, and extracting for 15-25 min at the frequency of 50-70 kHz; filtering after extraction is finished, and concentrating the obtained filtrate to 1.2-1.3 g/ml to obtain the forsythia suspense fruit extracting solution; and/or
The preparation method of the licorice root extracting solution comprises the following steps: crushing raw liquorice, decocting the crushed raw liquorice for 1-1.5 hours by using water, cooling the decoction to 40-50 ℃, adding cellulase accounting for 0.5-1.5 percent of the mass of the raw liquorice for enzymolysis for 2-3 hours at 40-50 ℃, then heating the mixture to 90-95 ℃, keeping the mixture for 5-10 min for enzyme deactivation, and filtering the mixture to obtain enzymolysis liquid and filter residues; ultrasonically extracting the filter residue for 20-30 min by using water at the frequency of 20-30 kHz, and filtering to obtain an ultrasonic extracting solution; and combining the enzymolysis liquid and the ultrasonic extracting solution, and concentrating to 1.2-1.3 g/ml to obtain the licorice root extracting solution.
9. Use of a skin-repairing bacteriostatic composition according to any one of claims 1 to 8 in the preparation of a skin-repairing bacteriostatic product.
10. A gel comprising the skin-repairing bacteriostatic composition of any one of claims 1 to 8.
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