CN102973483B - Bee venom composition with wrinkle removing effect - Google Patents

Bee venom composition with wrinkle removing effect Download PDF

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Publication number
CN102973483B
CN102973483B CN201210559366.3A CN201210559366A CN102973483B CN 102973483 B CN102973483 B CN 102973483B CN 201210559366 A CN201210559366 A CN 201210559366A CN 102973483 B CN102973483 B CN 102973483B
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China
Prior art keywords
apis
venenum apis
hyaluronic acid
skin
wrinkle
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CN102973483A (en
Inventor
陈华
孙恢礼
张偲
潘剑宇
蔡冰娜
孙小钰
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South China Sea Institute of Oceanology of CAS
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South China Sea Institute of Oceanology of CAS
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Priority to CN201210559366.3A priority Critical patent/CN102973483B/en
Priority to PCT/CN2013/070958 priority patent/WO2014094379A1/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/65—Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73—Polysaccharides
    • A61K8/735—Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations

Abstract

The invention discloses a bee venom composition with a wrinkle removing effect. The composition comprises wrinkle removing cosmetic matrix components and bee venom. The bee venom is applied to wrinkle removing cosmetics for the first time, and can act on the epidermis, the dermis and the muscle tissues at the same time to realize deep wrinkle removal. By using good synergistic effect of the bee venom, hyaluronic acid and marine animal collagen peptide, the effects of moisturizing, improving dryness and laxity of skin, improving the elasticity of the skin, deeply removing wrinkles and the like are remarkably enhanced. The bee venom composition with the wrinkle removing effect is definite in effect component, stable in quality and diverse in variety, and can meet the requirements of different genders, ages and skin types.

Description

A kind of Venenum apis compositions with wrinkle removal effect
Technical field:
The invention belongs to cosmetic field, be specifically related to a kind of Venenum apis compositions with wrinkle removal effect.
Background technology:
Aging is the irresistible natural process of life, be subject to the impact of the many factors such as heredity, environment, social mentality, wrinkle is the outward manifestation of body aging on skin, repeatedly shrink with cutis laxa, ultraviolet radiation, gravity, facial expression muscle and the factor such as the tension force of muscle own relevant.Generally speaking, there is initial wrinkle in forehead and lower eyelid from 25 years old, within 30 years old, outwards occurs fish tail shape fine wrinkle from paropia, before 35 years old ear, occurs longitudinal wrinkles, and before 40 years old ear, wrinkle increases, and throat occurs vertical wrinkle downwards from jaw.Within 45 years old, lower eyelid wrinkle is more obvious, and red lip starts attenuation.50 years old facial wrinkles are deepened, and the back of the hand wrinkle increases, and before the bridge of the nose, ear, wrinkle also appears in jaw portion, throat is vertical gauffer obviously, as chicken neck shape.Within 55 years old, start to occur that senile plaque, 60 years old deutostoma week be shown in radial wrinkle, within 75 years old, red lip becomes very thin, and skin of dorsum of hand is as creping tissue sample.Therefore,, from 25 years old, wrinkle is almost on very close terms with people.
Wrinkle is not only declaring publicly and is being difficult to obliterated years vestige, and has a strong impact on even people social and psychological of skin beauty, easily causes anxiety, depression, feels oneself inferior and the negative emotions such as panic, thereby affect quality of life, is personage's the formidable enemy of liking to be beautiful.For reducing flesh age, dispel wrinkle, remaining youthful forever, use that to have the cosmetics of remarkable wrinkle removal effect essential.But Wrinkle removing cosmetic is a feast for the eyes on the market at present, advertisement is also very tempting, and effect is barely satisfactory.And how containing forbidden amphetamines such as heavy metal, hormones, there is certain potential safety hazard in the cosmetics of some wrinkle removal effect highly significants.And comparatively safe product also can only act on skin surface, can not go deep into skin corium even muscular tissue play a role, wrinkle removal effect not obvious.This is like repairing the roads, and only allows evenness of road surface, and roadbed is but also unguyed.Therefore, act on epidermis, corium and muscular tissue deep layer Wrinkle removing cosmetic if develop simultaneously, bring glad tidings to the personage that likes to be beautiful.
Summary of the invention:
The object of the invention is to have potential potential safety hazard, only can act on skin surface for existing Wrinkle removing cosmetic, can not go deep into the skin corium problems such as even muscular tissue plays a role, wrinkle removal effect is not remarkable, a kind of safer, deep layer wrinkle removal that can simultaneously act on epidermis, corium and muscular tissue is provided, and wrinkle removal effect has the Venenum apis compositions of wrinkle removal effect significantly.
The Venenum apis compositions with wrinkle removal effect of the present invention, comprises Wrinkle removing cosmetic substrate composition, it is characterized in that, also contains Venenum apis.
Preferably, described Venenum apis is prepared by the following method, gets thick Venenum apis water dissolution, and then is undertaken, after ultrafiltration, filtrate lyophilization being made to Venenum apis by the ultrafilter membrane of molecular cut off 10000Da.
This Venenum apis purification process is being simplified Venenum apis purifying process, provides cost savings simultaneously, can also, to having melittin, the effective enrichment of apamin isoreactivity material and concentrated of remarkable wrinkle removal effect, remove hyaluronidase, phospholipase A 2deng the material that easily causes skin allergy, inflammation and pain, ensure safety, without skin irritation and irritated in, wrinkle removal effect is significantly strengthened.This purification process has solved the not yet effectively problem such as enrichment of active substance in Venenum apis in prior art with wrinkle removal effect, has filled up the blank that Venenum apis is applied in Wrinkle removing cosmetic, and has given full play to its wrinkle removal effect.
Preferably, described Venenum apis, its content accounts for 0.00001%～2% of Venenum apis compositions gross mass.
Described Venenum apis is preferably prepared by the following method: get thick Venenum apis and add 10-30 times of volume water and fully dissolve, the centrifugal 10-30min of 5000-10000g, supernatant is undertaken, after ultrafiltration, filtrate lyophilization being made to Venenum apis by the ultrafilter membrane of molecular cut off 10000Da.
For strengthening moisturizing, increasing skin elasticity, deep layer wrinkle-dispelling function, the Venenum apis compositions with wrinkle removal effect of the present invention preferably also contains hyaluronic acid, and its content accounts for the 0.01%-2% of Venenum apis compositions gross mass.Further preferably also contain marine animal collagen peptide, its content accounts for the 0.01%-5% of Venenum apis compositions gross mass.
Described marine animal collagen peptide refers to one or more the combination in cod collagen peptide, sharkskin collagen peptide, holothurian collagen peptide, collagen from squid skin peptide.It is commercially available, also can be according to existing method preparation.
Hyaluronic acid of the present invention is commercially available.
Described thick Venenum apis can be the thick Venenum apis of wasp and/or the thick Venenum apis of Apis, can own use collector of bee venom collection, also can be from commercial.
The Venenum apis compositions with wrinkle removal effect of the present invention is prepared by the following method, by Venenum apis taking powdery form or be mixed with in advance mass fraction as 10%-50% aqueous solution, with other composition homogenizing or mix, can, according to prepared Venenum apis compositions kind according to conventional technique preparation, can be made into the Wrinkle removing cosmetics such as cream frost, emulsion, water preparation, gel, gel, aerosol or facial film.
The Venenum apis composition stable with wrinkle removal effect of the present invention is good, coating evenly, spreadability is good.
The Venenum apis compositions that the present invention has wrinkle removal effect can be used as ordinary cosmetics or has the function cosmetics of wrinkle removal effect.If for dispelling wrinkle of skin, can dispel face, near the eyes, the wrinkle of neck neck, trunk and extremity.
Form reason by analyzing skin physiological structure and wrinkle, the inventor finds that skin mesostroma forms hydration grid structure and determining configuration and the physical property of skin, i.e. the vision appearance such as skin-tightening degree, elasticity, water humidity.If substrate the Nomenclature Composition and Structure of Complexes is in health status, cutaneous manifestations for filling, compact, water profit, high resilience.Therefore, the main path of the crease-resistant skin that compacts maintains and protects dermal matrix the Nomenclature Composition and Structure of Complexes and subcutaneous muscular tissue exactly.
Solution of the present invention just based on inventor to the various influence factors of wrinkle and form mechanism, and understanding to each functional activity material beauty and skin care effect, in conjunction with modern cosmetics and pharmacological research achievement, from numerous active substances, optimize and there is Venenum apis, hyaluronic acid and the marine animal collagen peptide of significantly dispelling wrinkle effect, and carry out scientific composition and collaborative composite, optimize formula and the technique of the cosmetic composition with remarkable wrinkle removal effect.
The present invention has the Venenum apis in the Venenum apis compositions of wrinkle removal effect, and it contains abundant melittin, apamin, catecholamine, free amino acid and glycerol isoreactivity composition.Its molecular weight is little, and immunogenicity is low, is difficult for producing anaphylaxis.
Melittin is neurotropic, can be optionally have an effect with motor nerve ending presynaptic membrane around, suppress the release of presynaptic membrane to neurotransmitter acetylcholine, make excited conduction block in nerve synapse, thereby cause arranged property paralysis of flaccid muscles, and suppress peripheral nerve conduction of impulse.Crows-feet, volume stricture of vagina, glabella stricture of vagina etc. are many to be formed because facial expression muscle shrinks tractive skin for a long time, after melittin is had an effect with the nervus facialis of domination facial expression muscle, and facial expression muscle flaccid paralysis, hypomyotonia, wrinkle is obviously desalinated or is disappeared.Melittin also has the effects such as obvious antioxidation, anti-ultraviolet radiation, can eliminate scytitis, suppress swelling, delaying skin aging.
Apamin can increase capillary permeability, improves microcirculation, promotes epidermis cell metabolism, accelerates the old and new's Skin Cell and substitutes, and makes turgor, high resilience.It can also nutrition repair muscular dystrophy cell, and Skin Cell is rejuvenated again.Catecholamine has antiinflammation to the damaged skin causing because of illumination, acne/acne etc., is beneficial to wound healing, reduces cicatrix.
Serine, glycine, threonine and alanine in free amino acid is Moisture factor natural in horny layer, and methionine and cysteine are keratin important composition compositions, and glycine, proline and alanine are collagen protein important composition compositions.Arginine energy vasodilator, promotes skin blood to flow and metabolism.Glutamatergic promotes cell proliferation and differentiation, promotes collagen protein to generate and wound healing.Ornithine can improve blood flow, promotes cell proliferation and skin regeneration.Methionine and histidine have antioxidant activity, and glutamic acid and cysteine not only have antioxidant activity, also have restraint of tyrosinase activity, can protect skin to avoid ultraviolet injury, and check melanin generates.Glycerol can absorb airborne moisture, prevents moisture of skin evaporation, thereby reaches moistening effect, auxiliary delay skin aging.
The hyaluronic acid that the present invention has in the Venenum apis compositions of wrinkle removal effect claims again Hyaluronic Acid, is commonly called as hyaluronic acid on Cosmetic Market, is one of main matrix composition of people's epidermis and corium.It is the best material of moisture retention that occurring in nature is found at present, is called as optimal nature moisturizing factor, can improve the wrinkle generating because of dry.Exogenous interpolation hyaluronic acid can supplement skin endogenous hyaluronic acid, and the micromolecule amount hyaluronic acid skin corium that can penetrate to the skin, maintains moisture of skin, promotes epidermal cell proliferation, differentiation and wound healing.It can also form thin film at skin surface, makes skin tender, smooth, increases elasticity, slow down aging.Hyaluronic acid has high wellability and moisture retention to keratodermatitis, can also promote Venenum apis, marine animal collagen peptide isoreactivity composition to be more easily absorbed by the skin, thereby strengthens wrinkle removal effect.
The present invention has marine animal collagen peptide and the keratodermatitis structural similarity in the Venenum apis compositions of wrinkle removal effect, and therefore it and skin have the good compatibility, affinity and permeability.It can infiltrate skin epidermis, and the effect of performance nature moisturizing factor, can also be filled between dermal matrix, forms one deck rete as thin as a wafer at skin surface, to increase skin tension, unfolds tiny wrinkle.Owing to containing the abundant hydrophilic group such as hydroxyl, amino, marine animal collagen peptide also has good moisture retention, can promote skin flbroblast collagen and hyaluronic generation, enhance metabolism, repairing skin tissue, thereby effectively prevent and reduce wrinkle of skin or relax, make skin soft and moist smooth, recover elasticity.
It is distinguished that the Venenum apis compositions that the present invention has wrinkle removal effect is dispelled wrinkle effect, wherein Venenum apis and hyaluronic acid, marine animal collagen peptide have good synergistic function, can make moisturizing, improve xerosis cutis and lax, increase skin elasticity, deep layer wrinkle removal texts significantly strengthens.It is clear and definite that the present invention has the Venenum apis compositions functional component of wrinkle removal effect, and steady quality is numerous in variety, can meet different sexes, age, skin quality crowd's demand.
Generally speaking, because of facial expression muscle, to shrink the wrinkle causing particularly effective to dispelling for the Venenum apis compositions with wrinkle removal effect that only contains Venenum apis in the present invention, as wrinkles on one's forehead, bridge of the nose stricture of vagina, glabella river word stricture of vagina, crows-feet, buccal wrinkle etc.; It is particularly effective to dispelling the wrinkle causing because of hydropenia or habitual expression that the present invention contains Venenum apis and the hyaluronic Venenum apis compositions with wrinkle removal effect, as dry stricture of vagina, microgroove, eye pouch stricture of vagina etc.; It is particularly effective that the Venenum apis compositions with wrinkle removal effect that the present invention contains Venenum apis, hyaluronic acid and marine animal collagen peptide reduces to the subcutaneous fat of expressing one's feelings, often making up because of Exposure to Sunlight, hydropenia, habituation or even excessively fat-reducing causing the wrinkle causing.
It should be noted that, because wrinkle is one of the most ticklish skin problem, the Venenum apis compositions with wrinkle removal effect equally also can be used as ordinary cosmetics and uses, and generates, and dispel existing dry stricture of vagina, microgroove to delay wrinkle.That is to say, the Venenum apis compositions that the present invention has wrinkle removal effect both can be used as ordinary cosmetics, can be used as again having the function cosmetics of wrinkle removal effect.
Compared with prior art, the present invention has following beneficial effect:
(1), for the special physiology of skin structure and the sensitivity to cosmetics composition used thereof, the Venenum apis of the Venenum apis compositions with wrinkle removal effect in the present invention is preferred pure natural is primary raw material.Although it is very rare to add at present the cosmetics of Venenum apis, Venenum apis is included already in " international cosmetic raw material standard Chinese catalogue (2010 editions) ", and it is safe, feasible that Venenum apis is applied to cosmetics by this explanation.In addition, have in the Venenum apis compositions of wrinkle removal effect in the present invention, the Venenum apis using is through purification process, the material that easily causes skin allergy, inflammation and pain in thick Venenum apis is effectively removed, and by highly enriched the composition such as melittin, apamin, thereby give play to more significant wrinkle removal effect.Meanwhile, the Venenum apis in the present invention can be absorbed by the skin rapidly, does not cause skin allergy, also can not bring any burden to skin.
(2) the Venenum apis compositions that the present invention has wrinkle removal effect from replenishing water and preserving moisture, promote microcirculation, protection skin to avoid ultraviolet injury, remove free radical, promote the many-sides such as collagen protein and elastic fiber synthesize, diminish inflammation to play a role; can also have an effect with the nervus facialis of domination facial expression muscle; make facial expression muscle flaccid paralysis, hypomyotonia, thus wrinkle removal wrinkle.The Venenum apis compositions that the present invention has wrinkle removal effect can act on epidermis, corium and muscular tissue simultaneously, heavy agonistic muscle skin elastic structure, alleviate or eliminate established wrinkle or pleat seal, dynamic property and Combination wrinkle are played to the wrinkle removal effect suitable with facelift, and effect being lasting.
(3) the present invention has the Venenum apis compositions of wrinkle removal effect; it is numerous in variety; be applicable to the crowd of different sexes, age and skin quality, be particularly useful for the personage that likes to be beautiful who enjoys wrinkle to perplex because of the subcutaneous fat minimizing that Exposure to Sunlight, skin hydropenia, habituation are expressed one's feelings, often made up or even excessively fat-reducing causes for a long time.The Venenum apis compositions that the present invention has wrinkle removal effect can be used as dispelling ordinary cosmetics or the function cosmetics of face and Neck wrinkles, also can be used as dispelling ordinary cosmetics or the function cosmetics of other body part wrinkle.
(4) the present invention has utilized the stabilisation synergistic function between Venenum apis and hyaluronic acid, marine animal collagen peptide, and its effect of dispelling wrinkle of skin is significantly strengthened.
Detailed description of the invention
The invention will be further described below, but embodiments of the present invention are not limited to following embodiment introduction, all variations being equal to of doing according to method of the present invention or the flexible category that all should be considered as the present invention's protection.
Raw materials used below, except special indicating, be commercially available.
Embodiment 1
Adopt collector of bee venom to gather the thick Venenum apis of wasp, add 25 volume water and fully dissolve, the centrifugal 15min of 8000g, supernatant is by the ultrafilter membrane ultrafiltration of molecular cut off 10000Da, and filtrate lyophilization makes Venenum apis.
The Venenum apis compositions with wrinkle removal effect of the present embodiment, by gross mass percent 100%, is made up of following component:
A phase: carbomer 0.2%, glycerol 4%, propylene glycol 3%, deionized water surplus.
B phase: ceteareth-2 1.5%, ceteareth-21 2%, Jojoba oil 3%, cetearyl alcohol 2%, glyceryl monostearate 1%, caprylic/capric triglyceride 4%, ethylhexyl palmitate 3%, polydimethylsiloxane 2%, isopropyl myristate 3%, ethyl hydroxybenzoate 0.1%.
The form that C phase: Venenum apis 0.1%(is mixed with mass fraction 50% Venenum apis solution in advance with deionized water is added).
D phase: azone/Cremophor RH40 1%, triethanolamine 0.2%, phenoxyethanol/methyl hydroxybenzoate/butyl hydroxybenzoate/ethyl hydroxybenzoate/propyl hydroxybenzoate/oxybenzene isobutyl ester 0.1%.
Technical process: heat respectively A phase and B to 80 DEG C, under stirring, B is added to A mutually in.Stirring is cooled to 45 DEG C, C is added in the mixture of A, B phase, is uniformly mixed after 10min, adds D phase, mixes homogenizing and make the wrinkle eliminating ointment frost of the Venenum apis compositions with wrinkle removal effect of the present embodiment-contain Venenum apis.
The cream frost of present embodiment preparation is glossy, exquisite quality, be evenly distributed, profit do not separate mutually.Be coated on the face, easily absorb, after continuing to use one month, wrinkles on one's forehead, bridge of the nose stricture of vagina and buccal wrinkle are most of to disappear or shoals.
Embodiment 2
Adopt collector of bee venom to gather the thick Venenum apis of Apis, add 10 times of volume water and fully dissolve, the centrifugal 30min of 5000g, supernatant is undertaken, after ultrafiltration, filtrate lyophilization being made to Venenum apis by the ultrafilter membrane of molecular cut off 10000Da.
The Venenum apis compositions with wrinkle removal effect of the present embodiment, by gross mass percent 100%, is made up of following component:
A phase: carbomer 0.1%, glycerol 5%, propylene glycol 3%, deionized water surplus.
B phase: ceteareth-2 1.2%, ceteareth-21 1.5%, Parleam 3%, caprylic/capric triglyceride 3%, ethylhexyl palmitate 3%, polydimethylsiloxane 2%, ethyl hydroxybenzoate 0.1%.
C phase: Venenum apis 2%(adds with the form of powder).
D phase: azone/Cremophor RH40 1%, triethanolamine 0.1%, phenoxyethanol/methyl hydroxybenzoate/butyl hydroxybenzoate/ethyl hydroxybenzoate/propyl hydroxybenzoate/oxybenzene isobutyl ester 0.1%.
Technical process: heat respectively A phase and B to 80 DEG C, under stirring, slowly B is added to A mutually in.Stirring is cooled to 55 DEG C, C is added in the mixture of A, B phase, is uniformly mixed after 10min, adds D phase, mixes homogenizing and make the wrinkle removal emulsion of the Venenum apis compositions with wrinkle removal effect of the present embodiment-contain Venenum apis.
The emulsion feel exquisiteness of present embodiment preparation, coating are evenly, profit do not separate, easily absorbs.Be coated in neck neck place, after continuing to use one month, Neck wrinkles obviously shoals.
Embodiment 3
Buy the thick Venenum apis of wasp from Wuhan Yi Min bee product company limited, add 30 times of volume water and fully dissolve, the centrifugal 10min of 10000g, supernatant is undertaken, after ultrafiltration, filtrate lyophilization being made to Venenum apis by the ultrafilter membrane of molecular cut off 10000Da.
The Venenum apis compositions with wrinkle removal effect of the present embodiment, by gross mass percent 100%, is made up of following component:
A phase: HANSHENGJIAO 0.4%, acrylate/C 10-30alkyl acrylate cross-linked polymer 0.6%, EDTA-2Na0.1%, glycerol 5%, propylene glycol 5%, butanediol 5%, deionized water surplus.
B phase: Venenum apis 0.00001%(with distilled water be mixed with in advance mass fraction 10% Venenum apis solution form add), deionized water 20%.
C phase: Cremophor RH40 ester 1%, triethanolamine 0.2%, essence 0.1%, phenoxyethanol/methyl hydroxybenzoate/butyl hydroxybenzoate/ethyl hydroxybenzoate/propyl hydroxybenzoate/oxybenzene isobutyl ester 0.1%.
Technical process: mix A phase component and be heated to 80 DEG C stirring, while being cooled to 40 DEG C, add and be pre-mixed uniform B phase, stir 10min to transparent, add the wrinkle-removing gel of the Venenum apis compositions with wrinkle removal effect that makes the present embodiment after C phase stirring and evenly mixing-contain Venenum apis.
The gel of present embodiment preparation is transparent, tissue is fine and smooth, without caking, under-10 DEG C and 40 DEG C of conditions, without paste separation.Be coated near the eyes on skin, very easily absorb, continue to use one month, microgroove disappears, and crows-feet obviously shoals.
Embodiment 4
The thick Venenum apis of Apis of buying from Shen Feng Science and Technology Development Co., Ltd. of Fujian Province, adds 15 times of volume water and fully dissolves, the centrifugal 20min of 6000g, and supernatant is undertaken, after ultrafiltration, filtrate lyophilization being made to Venenum apis by the ultrafilter membrane of molecular cut off 10000Da.
The Venenum apis compositions with wrinkle removal effect of the present embodiment, by gross mass percent 100%, is made up of following component:
A phase: HANSHENGJIAO 0.2%, acrylate/C 10-30alkyl acrylate cross-linked polymer 0.6%, glycerol 5%, propylene glycol 3%, deionized water surplus.
B phase: Venenum apis 0.01%(with deionized water be mixed with in advance mass fraction 50% Venenum apis solution form add), from Liuzhou Liuhua Group Dali Hyaluronic Acid Co., Ltd. buy hyaluronic acid 2%, deionized water 30%.
C phase: azone/Cremophor RH40 1%, triethanolamine 0.3%, phenoxyethanol/methyl hydroxybenzoate/butyl hydroxybenzoate/ethyl hydroxybenzoate/propyl hydroxybenzoate/oxybenzene isobutyl ester 0.1%.
Technical process: mix A phase component and be heated to 80 DEG C stirring, while being cooled to 45 DEG C, add and be pre-mixed uniform B phase, stir homogenizing 3min, under room temperature, be sequentially added into the mutually each component of C, stir and make the wrinkle removal gel of the Venenum apis compositions with wrinkle removal effect of the present embodiment-contain Venenum apis.
The gel of preparing in present embodiment, quality is slim and graceful, and sensation is nice and cool.Be coated near the eyes with ear before on skin, absorb rapidly, continue to use one month, near the eyes with ear before dry stricture of vagina and the little microgroove at skin place disappear, eye pouch stricture of vagina obviously shoals.
Embodiment 5
The method of preparing Venenum apis according to embodiment 1 and 2 makes respectively wasp aptoxin and Apis Venenum apis.
The Venenum apis compositions with wrinkle removal effect of the present embodiment, by gross mass percent 100%, is made up of following component:
A phase: glycerol 5%, propylene glycol 3%, butanediol 2%, deionized water surplus.
B phase: wasp aptoxin 1%, Apis Venenum apis 1%(Venenum apis are all added with the form of powder), hyaluronic acid 0.5%, the deionized water 38% bought from Hai Hua bio tech ltd, Jiangsu.
C phase: azone/Cremophor RH40 1%, triethanolamine 0.2%, phenoxyethanol/methyl hydroxybenzoate/butyl hydroxybenzoate/ethyl hydroxybenzoate/propyl hydroxybenzoate/oxybenzene isobutyl ester 0.1%.
Technical process: 35 DEG C, first mix A phase, then by the B group mixing in advance add A mutually in, after being uniformly mixed, add C phase component, mix the wrinkle removal water preparation that makes the Venenum apis compositions with wrinkle removal effect of the present embodiment-contain Venenum apis.
The water preparation of present embodiment preparation is without precipitation, high, the easy absorption of transparency, non-stimulated.Spread upon face, lip or neck neck, continue to use one month, lip and neck neck place skin texture shoal, and the dry stricture of vagina of face disappears, and expression stricture of vagina shoals.
Embodiment 6
Prepare the method for Venenum apis according to embodiment 2, make Apis Venenum apis.
The Venenum apis compositions with wrinkle removal effect of the present embodiment, by gross mass percent 100%, is made up of following component:
Venenum apis 0.00001%(adds taking the form of powder), hyaluronic acid 0.01%, concentration ethanol 9%, perfume 1%, antiseptic 0.1%, the deionized water surplus as 75%.30 DEG C, stir, make the wrinkle removal aerosol of the Venenum apis compositions with wrinkle removal effect of the present embodiment-contain Venenum apis.
The aerosol good stability of present embodiment preparation, the globule exquisiteness of spraying without precipitation, formation.Be sprayed on trunk and extremity, continue to use one month, skin becomes water profit, and dry stricture of vagina and microgroove disappear.
Embodiment 7
The method of preparing Venenum apis according to embodiment 3, makes wasp aptoxin.
The Venenum apis compositions with wrinkle removal effect of the present embodiment, by gross mass percent 100%, is made up of following component:
A phase: carbomer 0.1%, glycerol 5%, propylene glycol 3%, deionized water surplus.
B phase: ceteareth-2 1.2%, ceteareth-21 1.5%, Parleam 3%, caprylic/capric triglyceride 3%, ethylhexyl palmitate 3%, polydimethylsiloxane 2%, ethyl hydroxybenzoate 0.1%.
C phase: the cod collagen peptide 0.5% of wasp aptoxin 0.2%, hyaluronic acid 1%, the micro-Eco Science Technology of the Cacumen et folium clerodendri mandarinori (Clerodendron mandarinorum Diels) difficult to understand company limited purchase from Qingdao.
D phase: azone/Cremophor RH40 1%, triethanolamine 0.1%, phenoxyethanol/methyl hydroxybenzoate/butyl hydroxybenzoate/ethyl hydroxybenzoate/propyl hydroxybenzoate/oxybenzene isobutyl ester 0.1%.
Technical process: heat respectively A phase and B to 80 DEG C, under stirring, slowly B is added to A mutually in.Stirring is cooled to 45 DEG C, C is added in the mixture of A, B phase, is uniformly mixed after 10min, adds D phase, mixes homogenizing and make the wrinkle removal emulsion of the Venenum apis compositions with wrinkle removal effect of the present embodiment-contain Venenum apis.
The emulsion feel exquisiteness of present embodiment preparation, coating are evenly, profit do not separate.Be coated in the back of the hand place, very easily absorb, after continuing to use one month, it is smooth, flexible that skin of dorsum of hand becomes, and wrinkle obviously shoals.
Embodiment 8
The method of preparing Venenum apis according to embodiment 1 and 2 makes respectively wasp aptoxin and Apis Venenum apis.
The Venenum apis compositions with wrinkle removal effect of the present embodiment, by gross mass percent 100%, is made up of following component:
A phase: HANSHENGJIAO 0.4%, acrylate/C 10-30alkyl acrylate cross-linked polymer 0.6%, EDTA-2Na0.1%, glycerol 5%, propylene glycol 5%, butanediol 5%, deionized water surplus.
B phase: sharkskin collagen peptide 0.01%, the deionized water 26% of wasp aptoxin 1%, Apis Venenum apis 1%, hyaluronic acid 0.01%, the following bio tech ltd purchase from Qingdao.
C phase: Cremophor RH40 ester 1%, triethanolamine 0.2%, essence 0.1%, phenoxyethanol/methyl hydroxybenzoate/butyl hydroxybenzoate/ethyl hydroxybenzoate/propyl hydroxybenzoate/oxybenzene isobutyl ester 0.1%.
Technical process: mix A phase component and be heated to 80 DEG C stirring, while being cooled to 45 DEG C, add and be pre-mixed uniform B phase, stir 10min to transparent, add the wrinkle-removing gel of the Venenum apis compositions with wrinkle removal effect that makes the present embodiment after C phase stirring and evenly mixing-contain Venenum apis.
The gel of present embodiment preparation is transparent, tissue is fine and smooth, without caking, under-10 DEG C and 40 DEG C of conditions, without paste separation.Be coated near the eyes and neck neck place, very easily absorb, continue to use one month, eye pouch stricture of vagina disappears, and crows-feet and Neck wrinkles obviously shoal.
Embodiment 9
Prepare the method for Venenum apis according to embodiment 4, make Apis Venenum apis.
The Venenum apis compositions with wrinkle removal effect of the present embodiment, by gross mass percent 100%, is made up of following component:
A phase: HANSHENGJIAO 0.2%, acrylate/C 10-30alkyl acrylate cross-linked polymer 0.6%, glycerol 5%, propylene glycol 3%, deionized water surplus.
B phase: Apis Venenum apis 0.00001%, hyaluronic acid 2%, collagen from squid skin peptide 1.5%, holothurian collagen peptide 1.5%, deionized water 34.8%.
C phase: azone/Cremophor RH40 1%, triethanolamine 0.3%, phenoxyethanol/methyl hydroxybenzoate/butyl hydroxybenzoate/ethyl hydroxybenzoate/propyl hydroxybenzoate/oxybenzene isobutyl ester 0.1%.
Technical process: mix A phase component and be heated to 80 DEG C stirring, while being cooled to 45 DEG C, add and be pre-mixed uniform B phase, stir homogenizing 3min, under room temperature, be sequentially added into the mutually each component of C, stir and make the wrinkle removal gel of the Venenum apis compositions with wrinkle removal effect of the present embodiment-contain Venenum apis.
The gel of preparing in present embodiment, quality is slim and graceful, and sensation is nice and cool.Be coated in abdominal part, can absorb rapidly, continue to use one month, because the abdominal part that excessively fat-reducing or liposuction the cause skirt that wrinkles obviously shoals, skin become compact flexible.
Embodiment 10
The method of preparing Venenum apis according to embodiment 1, makes wasp aptoxin.
The Venenum apis compositions with wrinkle removal effect of the present embodiment, by gross mass percent 100%, is made up of following component:
Venenum apis 1%, hyaluronic acid 2%, cod collagen peptide 5%, nanometer Margarita powder 10%, Semen Armeniacae Amarum powder 11%, Mel 5%, antiseptic 1%, deionized water surplus.Under room temperature, stir, make the wrinkle removal facial film of the Venenum apis compositions with wrinkle removal effect of the present embodiment-contain Venenum apis.
The facial film of present embodiment preparation is direct-edible, mouthfeel is good, also can directly spread on face, supplements nutrient and moisture to skin.Continue to use one month, it is smooth, soft and moist, flexible that skin becomes, and dry stricture of vagina, microgroove and expression stricture of vagina disappear, and degree of depth wrinkle obviously shoals.
Embodiment 11
Following various samples are carried out to skin allergy, zest test, and to removing free radical ability and improving wrinkle of skin effect (after using continuously 8 weeks) and evaluate: the thick Venenum apis solution of mass fraction 0.5% wasp, the thick Venenum apis solution of mass fraction 0.5% Apis, mass fraction 0.5% wasp aptoxin solution (are prepared the method for Venenum apis according to embodiment 1, make wasp aptoxin), mass fraction 0.5% Apis Venenum apis solution (prepares the method for Venenum apis according to embodiment 2, make Apis Venenum apis), solvent is deionized water.
Skin allergy, zest test and improve wrinkle of skin effect assessment experimental subject: 20 of healthy volunteers, each 10 of man, female, age 38-41 year, without serious systemic disease, without immunodeficiency or autoimmune disease, without activeness anaphylactic disease, to skin-care cosmetics without severe allergy history, in nearly 1 month, do not used hormone medicine or immunosuppressant, do not participated in other clinical trials, do not smear the external preparation such as any cosmetics.Be divided at random 4 groups, every group of 5 people.
Use wrinkle of skin tester SV600 to carry out the evaluation of wrinkle removal effect, first copy the wrinkle of skin model at measured position with silicon mould, computer carries out grade separation and statistical disposition according to the wrinkle of skin degree of depth.Skin mean roughness R3(before calculation sample uses, after using is: 5 equal length, the measured R1 value of different segment are got to arithmetic mean number, and wherein R1 is illustrated in the distance between the interior peak value of certain length and minimum, and μ m).Result uses Δ R3(: the difference of the skin mean roughness before using sample and after use sample) expression, numerical value is higher, represents that wrinkle improves effect better.
Table 1 each laboratory sample skin hypersensitivity test and antioxidation, wrinkle removal effect are evaluated
Due to hyaluronidase, phospholipase A in thick Venenum apis solution (i.e. the thick Venenum apis solution of 0.5% wasp, the thick Venenum apis solution of 0.5% Apis) 2deng the material not yet in effect the removing that easily cause skin allergy, inflammation or sensation of pricking, so thick Venenum apis solution has compared with obvious stimulation skin, easily cause the allergic symptoms such as redness, pruritus.And Venenum apis after purification, due to anaphylactogen and cause that the material of scytitis and pain is effectively dispelled, so the Venenum apis solution (i.e. 0.5% wasp aptoxin solution, 0.5% Apis Venenum apis solution) after purification is minimum to skin irritation, does not also cause skin allergy, can relievedly use.
Simultaneously, because the wrinkle removal such as melittin, apamin composition has obtained effectively concentrated and enrichment, purified Venenum apis solution (i.e. 0.5% wasp aptoxin solution, 0.5% Apis Venenum apis solution) is removed hydroxy radical and DPPH free radical ability, and improve the ability of wrinkle of skin, be all significantly better than thick Venenum apis solution (i.e. the thick Venenum apis solution of 0.5% wasp, the thick Venenum apis solution of 0.5% Apis).
In addition, compared with deriving from the Venenum apis solution (i.e. the thick Venenum apis of 0.5% Apis, 0.5% Apis Venenum apis solution) of Apis, Venenum apis solution (i.e. the thick Venenum apis solution of 0.5% wasp, the 0.5% wasp aptoxin solution) antioxidation that derives from wasp is more remarkable with the ability of dispelling wrinkle.Therefore, wasp aptoxin wrinkle removal effect outline is better than Apis Venenum apis.
Embodiment 12
Following various samples are improved to the effects such as wrinkle of skin and evaluate (using continuously 8 weeks).
The percent of the present embodiment refers to mass percent.
The method that the wasp aptoxin of the present embodiment is prepared Venenum apis according to embodiment 1 makes, and the method that Apis Venenum apis is prepared Venenum apis according to embodiment 2 makes.
Cosmetic base: HANSHENGJIAO 0.2%, acrylate/C 10-30alkyl acrylate cross-linked polymer 0.6%, glycerol 5%, propylene glycol 3%, azone/Cremophor RH40 1%, triethanolamine 0.3%, phenoxyethanol/methyl hydroxybenzoate/butyl hydroxybenzoate/ethyl hydroxybenzoate/propyl hydroxybenzoate/oxybenzene isobutyl ester 0.1%, deionized water surplus.
Contain thick Venenum apis cosmetic composition (be sample A, thick Venenum apis content is mass fraction 2%):
A1: the thick Venenum apis 2% of wasp, cosmetic base 98%;
A2: the thick Venenum apis 2% of Apis, cosmetic base 98%;
A3: the thick Venenum apis 0.5% of wasp, the thick Venenum apis 1.5% of Apis, cosmetic base 98%.
Contain Venenum apis cosmetic composition (be sample B, Venenum apis content is mass fraction 2%):
B1: wasp aptoxin 2%, cosmetic base 98%;
B2: Apis Venenum apis 2%, cosmetic base 98%;
B3: wasp aptoxin 0.5%, Apis Venenum apis 1.5%, cosmetic base 98%.
Contain hyaluronic acid cosmetic composition (being sample C): hyaluronic acid 2%, cosmetic base 98%.
Containing Venenum apis and hyaluronic acid cosmetic composition (be sample D, Venenum apis, hyaluronic total content are mass fraction 2%, the difference of D1-D4 be Venenum apis kind and Venenum apis different from the ratio of hyaluronic acid contents):
D1: wasp aptoxin 0.00001%, hyaluronic acid 1.99999%, cosmetic base 98%;
D2: Apis Venenum apis 0.1%, hyaluronic acid 1.9%, cosmetic base 98%;
D3: wasp aptoxin 0.5%, hyaluronic acid 1.5%, cosmetic base 98%;
D4: wasp aptoxin 0.99%, Apis Venenum apis 1%, hyaluronic acid 0.01%, cosmetic base 98%.
(be sample E containing hyaluronic acid and marine animal collagen peptide cosmetic composition, the total content of hyaluronic acid and marine animal collagen peptide is mass fraction 2%, the difference of E1-E4 be the kind of marine animal collagen peptide and marine animal collagen peptide different from the ratio of hyaluronic acid contents):
E1: hyaluronic acid 0.01%, cod collagen peptide 1.99%, cosmetic base 98%;
E2: hyaluronic acid 0.5%, sharkskin collagen peptide 1%, holothurian collagen peptide 0.5%, cosmetic base 98%;
E3: hyaluronic acid 1%, cod collagen peptide 0.5%, sharkskin collagen peptide 0.5%, cosmetic base 98%;
E4: hyaluronic acid 1.99%, collagen from squid skin peptide 0.01%, cosmetic base 98%;
Contain Venenum apis, hyaluronic acid and marine animal collagen peptide cosmetic composition (be sample F, the total content of Venenum apis, hyaluronic acid and marine animal collagen peptide is mass fraction 2%):
F1: wasp aptoxin 0.00001%, hyaluronic acid 0.99999%, cod collagen peptide 1%, cosmetic base 98%;
F2: Apis Venenum apis 0.1%, hyaluronic acid 1.4%, sharkskin collagen peptide 0.5%, cosmetic base 98%;
F3: wasp aptoxin 0.4%, Apis Venenum apis 0.4%, hyaluronic acid 0.01%, cod collagen peptide 1.19%, cosmetic base 98%;
F4: wasp aptoxin 1%, hyaluronic acid 0.99%, collagen from squid skin peptide 0.01%, cosmetic base 98%;
F5: Apis Venenum apis 0.5%, hyaluronic acid 0.5%, cod collagen peptide 0.5%, holothurian collagen peptide 0.5%, cosmetic base 98%;
F6: wasp aptoxin 0.5%, Apis Venenum apis 1%, hyaluronic acid 0.2%, sharkskin collagen peptide 0.3%, cosmetic base 98%.
Choose have dry stricture of vagina, 66 of the healthy volunteers of the wrinkle puzzlement such as microgroove, crows-feet, expression stricture of vagina, cervical region stricture of vagina, wherein 24 of male, 42 of women, age 38-43 year, average 40.7 years old, without serious systemic disease, immunodeficiency or autoimmune disease, to cosmetics without allergy, facial NAI, do not use hormone medicine or immunosuppressant, do not participate in other clinical trials, do not smear external preparation.Be divided at random 22 groups, be cosmetic base group, contain thick Venenum apis cosmetic composition group (sample A1-A3), contain Venenum apis cosmetic composition group (sample B1-B3), contain hyaluronic acid cosmetic composition group (sample C), contain Venenum apis and hyaluronic acid cosmetic composition group (sample D1-D4), contain hyaluronic acid and marine animal collagen peptide cosmetic composition group (sample E1-E4), contain Venenum apis, hyaluronic acid and marine animal collagen peptide cosmetic composition group (sample F 1-F6), 3 every group.
Each group volunteer is with after warm water cleaning skin, at face, near the eyes, cervical region, the back of the hand (same position) evenly smear corresponding cosmetics, every day each 1 time sooner or later, after using 8 weeks continuously, use moisture of skin measuring instrument Corneometer CM825 to measure the water content (%) in skin of dorsum of hand horny layer, use epidermis moisture loss analyzer TM300 to measure skin of dorsum of hand water loss amount [TEML, g/ (hm 2)].Use wrinkle of skin tester SV600 to test wrinkle of skin near the eyes and (first copy the wrinkle of skin model at measured position with silicon mould, computer carries out grade separation and statistical disposition according to the wrinkle of skin degree of depth), calculation sample use before, use after skin mean roughness R3(: 5 equal length, the measured R1 value of different segment are got to arithmetic mean number, wherein R1 is illustrated in the distance between the interior peak value of certain length and minimum, and μ m).Result uses Δ R3(: the difference of the skin mean roughness before using sample and after use sample) expression, numerical value is higher, represents that wrinkle improves effect better.Use skin elasticity tester MPA580 to test facial skin elasticity (the constant 450mpa of being of negative pressure in test process), result cancels the recovery value of negative pressure skin after 0.1 second with springback capacity Ur() with the ratio value representation of the maximum tension amount Uf of skin, numerical value more approaches 1, illustrates that skin elasticity is better.
Measurement result is in table 2, wherein sample A(is containing thick Venenum apis cosmetic composition) meansigma methods organized for sample A1-A3, sample B(is containing Venenum apis cosmetic composition) meansigma methods organized for sample B1-B3, sample D(is containing Venenum apis and hyaluronic acid cosmetic composition) meansigma methods organized for sample D1-D4, sample E(is containing hyaluronic acid and marine animal collagen peptide cosmetic composition) be the meansigma methods of sample E1-E4 group, sample F (containing Venenum apis, hyaluronic acid and marine animal collagen peptide cosmetic composition) is the meansigma methods that sample F 1-F6 organizes.
Compared with " cosmetic base " group, using continuously other cosmetic composition after 8 weeks, skin beauty water is right obvious increase, and epidermis water loss amount significantly reduces, and it is obvious that elasticity and wrinkle improve degree.
Compared with " containing thick Venenum apis cosmetic composition-A ", using continuously " containing Venenum apis cosmetic composition-B " after 8 weeks, it is more obvious that wrinkle of skin improves effect.Compare with " containing hyaluronic acid cosmetic composition-C " and " containing Venenum apis cosmetic composition-B ", using continuously " containing Venenum apis and hyaluronic acid cosmetic composition-D " after 8 weeks, skin beauty water is right, and to improve effect more obvious with wrinkle.Compare with " containing Venenum apis cosmetic composition-B " with " containing hyaluronic acid and marine animal collagen peptide cosmetic composition-E ", using continuously " containing Venenum apis, hyaluronic acid and marine animal collagen peptide cosmetic composition-F " after 8 weeks, it is more obvious that keratodermatitis water content, elasticity and wrinkle improve effect, crows-feet and expression stricture of vagina shoal and even disappear, and it is smoother and flexible that skin becomes.
The each cosmetic sample of table 2 improves the evaluation of skin effect
As can be seen here, the Venenum apis in the present invention improve skin beauty water right, reduce water loss amount, improve skin elasticity and wrinkle texts is significantly better than thick Venenum apis.In addition, the Venenum apis in the present invention and hyaluronic acid, and Venenum apis and hyaluronic acid, marine animal collagen peptide have significant synergistic function, can make that skin beauty water is right, elasticity and wrinkle etc. improve effect and greatly improve.
In sum, it is distinguished that the Venenum apis compositions with wrinkle removal effect in the present invention is dispelled wrinkle effect, and Venenum apis and hyaluronic acid and marine animal collagen peptide have good collaborative complex role, can make to improve xerosis cutis, lax, wrinkle texts significantly strengthens.
Above-described embodiment is preferably embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and principle, substitutes, combination, simplify; all should be considered as equivalent substitute mode, be included in protection scope of the present invention.

Claims (1)

1. a Venenum apis compositions with wrinkle removal effect, is characterized in that, concrete formula is:
(1), by gross mass mark 100%, formed by wasp aptoxin 0.00001%, hyaluronic acid 1.99999% and cosmetic base 98%;
Or (2) are by gross mass mark 100%, are made up of Apis Venenum apis 0.1%, hyaluronic acid 1.9% and cosmetic base 98%;
Or (3) are by gross mass mark 100%, are made up of wasp aptoxin 0.5%, hyaluronic acid 1.5% and cosmetic base 98%;
Or (4) are by gross mass mark 100%, are made up of wasp aptoxin 0.99%, Apis Venenum apis 1%, hyaluronic acid 0.01% and cosmetic base 98%;
Or (5) are by gross mass mark 100%, are made up of wasp aptoxin 0.00001%, hyaluronic acid 0.99999%, cod collagen peptide 1% and cosmetic base 98%;
Or (6) are by gross mass mark 100%, are made up of Apis Venenum apis 0.1%, hyaluronic acid 1.4%, sharkskin collagen peptide 0.5% and cosmetic base 98%;
Or (7) are by gross mass mark 100%, are made up of wasp aptoxin 0.4%, Apis Venenum apis 0.4%, hyaluronic acid 0.01%, cod collagen peptide 1.19% and cosmetic base 98%;
Or (8) are by gross mass mark 100%, are made up of wasp aptoxin 1%, hyaluronic acid 0.99%, collagen from squid skin peptide 0.01% and cosmetic base 98%;
Or (9) are by gross mass mark 100%, are made up of Apis Venenum apis 0.5%, hyaluronic acid 0.5%, cod collagen peptide 0.5%, holothurian collagen peptide 0.5% and cosmetic base 98%;
Or (10) are by gross mass mark 100%, are made up of wasp aptoxin 0.5%, Apis Venenum apis 1%, hyaluronic acid 0.2%, sharkskin collagen peptide 0.3% and cosmetic base 98%;
Described cosmetic base, by gross mass mark 100%, comprises HANSHENGJIAO 0.2%, acrylate/C 10-30alkyl acrylate cross-linked polymer 0.6%, glycerol 5%, propylene glycol 3%, azone/Cremophor RH40 1%, triethanolamine 0.3%, phenoxyethanol/methyl hydroxybenzoate/butyl hydroxybenzoate/ethyl hydroxybenzoate/propyl hydroxybenzoate/oxybenzene isobutyl ester 0.1%, deionized water surplus;
Described wasp aptoxin is by thick wasp Venenum apis, adds 25 volume water and fully dissolves, and the centrifugal 15min of 8000g, supernatant is by the ultrafilter membrane ultrafiltration of molecular cut off 10000Da, and filtrate lyophilization makes wasp aptoxin;
Described Apis Venenum apis is by thick Apis Venenum apis, adds 10 times of volume water and fully dissolves, and the centrifugal 30min of 5000g, supernatant is undertaken, after ultrafiltration, filtrate lyophilization being made to Apis Venenum apis by the ultrafilter membrane of molecular cut off 10000Da.
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