CN106974876A - A kind of method that eye care solution is prepared by horseshoe crab cell pyrolysis liquid - Google Patents
A kind of method that eye care solution is prepared by horseshoe crab cell pyrolysis liquid Download PDFInfo
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- CN106974876A CN106974876A CN201710147889.XA CN201710147889A CN106974876A CN 106974876 A CN106974876 A CN 106974876A CN 201710147889 A CN201710147889 A CN 201710147889A CN 106974876 A CN106974876 A CN 106974876A
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- horseshoe crab
- cell pyrolysis
- care solution
- eye care
- pyrolysis liquid
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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
- A61K8/96—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
- 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
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
-
- 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
-
- 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
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/005—Preparations for sensitive skin
-
- 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 present invention relates to a kind of method that eye care solution is prepared by horseshoe crab cell pyrolysis liquid, including：The first step, the preparation of horseshoe crab cell pyrolysis liquid；Second step, the preparation of T-1 freeze-dried powder；3rd step, 1 is pressed by above-mentioned T-1 freeze-dried powder:1‑1:300 ratio is added in purifying water-soluble liquid, obtains T-1 eye care solution, and further 0.1 10 parts of Sodium Hyaluronate of addition, 0.1 5 parts of the borneol in 100 parts of T-1 eye care solution.The horseshoe crab cell pyrolysis liquid (T-1) of the present invention is natural materials, it is green, natural, with the antibacterial effect of cleaning, product is in addition to containing T-1 made from this method, also containing nutritional ingredients such as several amino acids, nourishing, wrinkle resistant, reparation skin injury can be played a part of, simultaneously, Sodium Hyaluronate is efficient water lock NMF, and group is repaired containing skin, assigns product water and moistens soft skin sense, the borneol contained has effect of clearing liver and improving vision, this method is simple and easy to apply, and composition cooperative effect is good, with great market value.
Description
Technical field
It is specifically a kind of that eye care solution is prepared by horseshoe crab cell pyrolysis liquid the present invention relates to eye care solution technical field
Method.
Background technology
Eyes and skin around eyes are very tender, and the skin around eyes is especially soft and fine slim, and has many pleats
Wrinkle, therefore skin moisture evaporation speed near the eyes, while skin sweat gland and sebaceous glands are less near the eyes, are particularly easy to dry water shortage.
And modern people because make up, swimming, for a long time in face of a variety of external circumstances such as computer and mobile phone, surrounding environment haze be serious
, there is eyes drying, eyes are itched, eye part skin is itched, eye in influence so that eyes and skin around eyes are greatly affected
A variety of situations such as dry skin, livid ring around eye, pouch, eye edema, eye wrinkle-removing, and the production such as eye ointment, eye cream on the market at present
Product moisturizing, cleaning, it is antibacterial go to itch in terms of do not dabble also, this cannot also meet the use needs of current people.
Horseshoe crab, is subordinate to Arthropoda Arthropoda, is a kind of Large Arthropodss lived in ocean, early ancient raw
Ordovician period in generation occurs, so far the existing history of more than 400,000,000 years, is that the animal kingdom has one of unique evolutionary degree " living fossil ".
East horseshoe crab (being called Tachypleus tridentatus) Tachypleus tridentatus wherein in the limulus of east be distributed in China coast Zhejiang,
Fujian, Taiwan, Guangxi and Guangdong.The blood of horseshoe crab can extract TAL, later again find horseshoe crab element to fungi, influenza virus,
Oral cavity herpes virus and Immune Deficiency Syndrome viral (HIV) are resistant.So horseshoe crab be can Devoting Major Efforts To Developing preciousness sea
Foreign medical animal resource.
Contain the compositions such as abundant T-1, SOD, several amino acids, a variety of peptides in horseshoe crab cell pyrolysis liquid.Horseshoe crab element is 1988
A kind of 17 amino acid that year Nakamura T. etc. is obtained in Chinese limulus blood cell chip with acid extraction method and containing 2 disulfide bond
Beta sheet structure antibacterial peptide.Horseshoe crab element antibacterial peptide be primarily present in the little particle in limulus blood cell, by and bacterial lipolysaccharide
Compound is formed, at low concentrations with regard to the growth of Gram-negative and positive bacteria can be suppressed.Horseshoe crab element is suitable under low pH, high temperature
It is stable, it is a kind of antibacterial substance for having very much an application potential.Itself it is exactly in addition amino acid, with reparation, health-care efficacy.
SOD (superoxide dismutase) is a kind of active material for coming from life entity, can eliminate organism and be produced in metabolic processes
Raw harmful substance.Constantly supplementing human body SOD has the special-effect of anti-aging.
The content of the invention
It is an object of the invention to provide the new application of horseshoe crab cell pyrolysis liquid, horseshoe crab cell pyrolysis liquid is applied to eye-care
Field, is specifically to provide a kind of method that eye care solution is prepared by horseshoe crab cell pyrolysis liquid.
The present invention realizes that the technical scheme that its technical purpose is used is as follows：
A kind of method that eye care solution is prepared by horseshoe crab cell pyrolysis liquid, including step are as follows：
The first step, the preparation of horseshoe crab cell pyrolysis liquid
(1) a certain amount of limulus blood cell is taken by 1:1-1:15, which add lysate, carries out freezing-thawing and cracking；
(2) after cracking, centrifuged horseshoe crab cell pyrolysis liquid；
Second step, the preparation of T-1 freeze-dried powder
(1) 100 parts of horseshoe crab cell pyrolysis liquids are taken uniformly to be mixed with 0.1-3 parts of excipient；
(2) and then by above-mentioned mixed solution is canned freezed into bottle, obtain T-1 freeze-dried powder finished product；
3rd step, the preparation of eye care solution
Above-mentioned T-1 freeze-dried powder is pressed 1:1-1:300 ratio is added in purifying water-soluble liquid, obtains T-1 eye
Conditioning liquid.
Moreover, the optimal proportion scope of T-1 freeze-dried powder and purified water is in the T-1 eye care solution：1:10-
1:200。
Moreover, the centrifuge RPMs of step (2) are 3000-5000 revs/min in the first step.
Moreover, the excipient in the step of second step (1) is the lactose for removing thermal source.
Moreover, the concrete operations freezed in the step of second step (2) are：Under conditions of vacuum is 10Pa, -50
DEG C pre-freeze 1 hour, then temperature programming, product reaches 24 DEG C, opens permeation valve, starts redrying, and then product at the uniform velocity rises
Temperature reaches 31 DEG C, cracks vacuum, product export.
Moreover, further adding Sodium Hyaluronate 0.1- in the T-1 eye care solution of 100 parts of the 3rd step
10 parts and 0.1-5 parts of borneol.
Advantages of the present invention and effect are：
1st, horseshoe crab cell pyrolysis liquid (T-1) of the invention is natural materials, green, natural；With the antibacterial work of cleaning
With.
2nd, product of the present invention is in addition to containing T-1, also containing nutritional ingredients such as several amino acids, can play nourishing, prevent
Wrinkle, the effect for repairing skin injury.
3rd, the Sodium Hyaluronate added in the lysate of the present invention is efficient water lock NMF, and base is repaired containing skin
Group, assigns product water and moistens soft skin sense；The borneol contained has effect of clearing liver and improving vision.
4th, the present invention is directly formed using the cracking of limulus blood cell lysate, and method is simple and easy to apply, and composition cooperative effect is good, tool
There are cleaning, nourishing, wrinkle resistant, eye health protection effect, applied to eye-care field, with great market value.
Brief description of the drawings
Fig. 1 is 1:1 ratio pure water dilution T-1 freeze-dried powder reads ball in staphylococcus aureus, Escherichia coli, white
2 minutes and the bacteriostasis rate test photo figure of 20 minutes bacteriostatic tests on bacterium；
Fig. 2 is 1:5 ratio pure water dilution T-1 freeze-dried powder reads ball in staphylococcus aureus, Escherichia coli, white
2 minutes and the bacteriostasis rate test photo figure of 20 minutes bacteriostatic tests on bacterium；
Fig. 3 is 1:10 ratio pure water dilution T-1 freeze-dried powder reads ball in staphylococcus aureus, Escherichia coli, white
2 minutes and the bacteriostasis rate test photo figure of 20 minutes bacteriostatic tests on bacterium；
Fig. 4 is 1:15 ratio pure water dilution T-1 freeze-dried powder reads ball in staphylococcus aureus, Escherichia coli, white
2 minutes and the bacteriostasis rate test photo figure of 20 minutes bacteriostatic tests on bacterium；
Fig. 5 is that 2 minute and 20 minute of the high purity water in staphylococcus aureus, Escherichia coli, Candida albicans are antibacterial
The bacteriostasis rate test photo figure of experiment.
Fig. 6 is 1:1 to 1:Staphylococcus aureus of the sample of 300 dissolving ratios at 2 minutes, Escherichia coli, white
Read the curvilinear motion comparison diagram of coccus bacteriostasis rate.
Embodiment
With reference to specific embodiment, the present invention is further described, and its specific embodiment is construed as only lifting
Example explanation, is not limited, it is impossible to limit protection scope of the present invention with following illustrations.
A kind of method that eye care solution is prepared by horseshoe crab cell pyrolysis liquid, it is as follows that the method comprising the steps of：
The first step, the preparation of horseshoe crab cell pyrolysis liquid
(1) a certain amount of limulus blood cell is taken by 1:1-1:15, which add lysate, carries out freezing-thawing and cracking；
(2) after cracking, centrifuged horseshoe crab cell pyrolysis liquid；
Wherein, centrifuge RPMs are 3000-5000 revs/min；
Second step, the preparation of T-1 freeze-dried powder
(1) 100 parts of horseshoe crab cell pyrolysis liquids are taken uniformly to be mixed with 0.1-3 parts of excipient；
(2) then it is canned freezed into bottle, obtain freeze-dried powder finished product；
Wherein, the excipient is removes the lactose of thermal source, and the lyophilisation condition is -50 DEG C of pre-freezes 1 hour, vacuum
10Pa, then temperature programming, product reaches 24 DEG C, opens permeation valve, starts redrying, and products temperature reaches 31 DEG C, can be with
Crack vacuum, product export.
3rd step, the preparation of eye care solution
Above-mentioned T-1 freeze-dried powder is pressed 1:1-1:300 ratio is added in purifying water-soluble liquid, obtains T-1 eye
Conditioning liquid.
In the specific implementation of the present invention, hyaluronic acid is further added in 100 parts of T-1 eye care solutions
0.1-10 parts of sodium, 0.1-5 parts of borneol so that the T-1 eye care solution has efficient water lock moisture-keeping functions, and clear liver and improve vision work(
Effect.
Bacteriostatic test is carried out to above-mentioned T-1 freeze-dried powder
The first kind is tested, by above-mentioned T-1 freeze-dried powder respectively to Escherichia coli, staphylococcus aureus, Candida albicans
The bacteriostatic test of progress 2 minutes and 20 minutes, experimental procedure is as follows：
1) 4 inclined-planes, 2 inclined-planes of sabouraud culture medium are made of nutrient agar first.Each bacterium two manages-and nutrient agar is used for
Staphylococcus aureus and Escherichia coli, sabouraud culture medium are used for Candida albicans.
2) phosphate buffer (PBS), PH=7.2,0.03mol/L buffer solution are prepared：The water of disodium hydrogen phosphate 12,
7.13g；Potassium dihydrogen phosphate, 1.36g；Plus 1L distilled water, 121 degree sterilizing 15min it is standby.
3) bacterium is taken from the staphylococcus aureus of line, Escherichia coli, Candida albicans flat board, by its stroke on inclined-plane
On, inclined-plane is placed in 37 degree of biochemical cultivation cases and cultivates 24h.
4) bacterium washed in two times in a test tube with 2mlPBS, two test tubes have used 4mlPBS all to wash out it, prepare
Bacteria suspension is standby.
5) sample-horseshoe crab cell pyrolysis liquid (T-1) dry powder of checking will be thought, with water for injection 1:1、1:5、1:10、1:
15 dilutions, are then dispensed into two test tubes, 5 kinds of sample liquids are constituted together with comparison high purity water group.
6) bacteria suspension 100ul drops are taken in 5ml sample liquids, start timing, each timing of effect 2min, 20min is once.2min、
Respectively take 0.5ml sample liquids to put into 5ml PBS test tubes during 20min, be sufficiently mixed to obtain A.Then gradient is done in dilution, is 2-4
Individual gradient.Therefrom take 1ml sample liquids to be added in 9ml PBS test tubes, be sufficiently mixed to obtain B, take 1ml sample liquids to be added to 9ml from B
In PBS test tubes, the like.
7) and then the above-mentioned gradient done is inhaled into 0.5ml to be put into flat board, adds the nutrient agar culture of 15ml 40-50 degree
Liquid or Sharpe nutrient solution, after being well mixed, put it into 37 degree of 48h and do count plate.72h is waited if fungi.
8) the bacterium bacteriostasis rate of 2 minutes is computed more than 85%, and the bacteriostasis rate of 20 minutes is more than 77%；2 minutes and
The fungi bacteriostasis rate of 20 minutes is more than 60%.
Equations of The Second Kind experiment is that the healthy population chosen between 30 20-55 Sui carries out experiment on probation, records their impression,
Observation uses rear effect.
Embodiment 1
Limulus blood cell and decomposition agent press 1:10 cracking freeze thawing obtain limulus blood cell lysate, the freeze-dried powder of the lysate with it is molten
Solve liquid 1:1 is mixed to get test specimen, and the test specimen is carried out into Escherichia coli, staphylococcus aureus, Candida albicans
Bacteriostatic test, result of the test is shown in Table 1, and photo is shown in Fig. 1,5.
Embodiment 2
Experimental procedure is consistent with embodiment 1, except that freeze-dried powder and lysate are according to 1:5 ratios are mixed to get experiment
Sample.Result of the test is shown in Table 1, and photo is shown in Fig. 2,5.
Embodiment 3
Experimental procedure is consistent with embodiment 1, and different freeze-dried powders and lysate are according to 1:10 ratios are mixed to get experiment
Sample.Result of the test is shown in Table 1, and photo is shown in Fig. 3,5.
Embodiment 4
Experimental procedure is consistent with embodiment 1, and different freeze-dried powders and lysate are according to 1:15 ratios are mixed to get experiment
Sample.
Result of the test is shown in Table 1, and photo is shown in Fig. 4,5.
Table 1 is purified water, purified water by 1:1、1:5、1:10、1:15 dilution T-1 freeze-dried powders are to Escherichia coli, golden yellow
Color staphylococcus, the bacteriostasis rate of Candida albicans.
Table 1
Embodiment 5
Experimental procedure is consistent with embodiment 1, except that the ratio for choosing freeze-dried powder and lysate is 1:1、1:5、1:
10、1:15、1:20、1:50、1:100、1:200、1:300 staphylococcus aureuses at 2 minutes, Escherichia coli, white are read
The contrast of coccus bacteriostasis rate.Result of the test is shown in Fig. 6.
Embodiment 6
A kind of T-1 eye care solution, specifically includes the following component constituted by mass fraction：
Freeze-dried powder：5 parts, 95 parts of purified water.
It is test crowd to choose the healthy population (male or female) between 20-55 Sui, first cleans both hands and face, eye skin
Skin, pours into clean cup 5ml by eye care solution, then cup is buckled on eyes, then faces upward head and continues in 20 seconds by tight cup
Body, does not allow liquid outflow.Discovery eye part skin moistening is smooth after each use, and eyes are dry and astringent to be alleviated, continuous using after 30 days, sends out
Existing eye part skin is substantially more smooth flexible, and small microgroove disappears, and eyes are bright, moistening.Compared with using common eye cream, skin
Wettability increase, elasticity increase, eyes are more comfortable.
Embodiment 7
A kind of T-1 eye care solution, specifically includes the following component constituted by mass fraction：
Freeze-dried powder：5 parts, 5 parts of purified water.
It is test crowd to choose the healthy population (male or female) between 20-55 Sui, first cleans both hands and face, eye skin
Skin, pours into clean cup 5ml by eye care solution, then cup is buckled on eyes, then faces upward head and continues in 20 seconds by tight cup
Body, does not allow liquid outflow.Found after use, compared with Example 6, freeze-dried powder concentration is increased to 1:1, find eye after each use
Portion's skin wet is smooth, and eyes are dry and astringent to be alleviated, but because freeze-dried powder concentration is big, has 2 people to feel that eyes have excitement in 30 people, but
Still in tolerance interval.Continuous using after 30 days, discovery eye part skin is substantially more smooth flexible, and small microgroove disappearance, eyes are bright
Bright, moistening.Compared with using common eye cream, wettability increase, the elasticity increase of skin, eyes are more comfortable.
Embodiment 8
A kind of T-1 eye care solution, specifically includes the following component constituted by mass fraction：
Freeze-dried powder：5 parts, 85 parts of purified water, 10 parts of Sodium Hyaluronate.
It is test crowd to choose the healthy population (male or female) between 20-55 Sui, is increased after Sodium Hyaluronate, is found every
The time increase by 20% of secondary moisturizing, the lubricious sense of skin further increase.Make the sense of touch of people more preferable.
Embodiment 9
A kind of T-1 eye care solution, specifically includes the following component constituted by mass fraction：
Freeze-dried powder：5 parts, 85 parts of purified water, 10 parts of Sodium Hyaluronate, 5 parts of borneol.
It is test crowd to choose the healthy population (male or female) between 20-55 Sui, is increased after borneol, it is found that eyes have cool
Cool sensation, more comfortable, the speed increase by 35% that eyes are dry and astringent, pain, pruritis mitigate, makes eyes more comfortable, bright.
Synthesis human feeling's effect of 5 fungistatic effect and embodiment 6- embodiments 9, determines the horseshoe crab element in conjunction with the embodiments
The optimal proportion scope of T-1 freeze-dried powder and purified water is in peptide eye care solution：1:10-1:200.
Claims (6)
1. a kind of method that eye care solution is prepared by horseshoe crab cell pyrolysis liquid, it is characterised in that as follows including step：
The first step, the preparation of horseshoe crab cell pyrolysis liquid
(1) a certain amount of limulus blood cell is taken by 1:1-1:15, which add lysate, carries out freezing-thawing and cracking；
(2) after cracking, centrifuged horseshoe crab cell pyrolysis liquid；
Second step, the preparation of T-1 freeze-dried powder
(1) 100 parts of horseshoe crab cell pyrolysis liquids are taken uniformly to be mixed with 0.1-3 parts of excipient；
(2) and then by above-mentioned mixed solution is canned freezed into bottle, obtain T-1 freeze-dried powder finished product；
3rd step, the preparation of eye care solution
Above-mentioned T-1 freeze-dried powder is pressed 1:1-1:300 ratio is added in purifying water-soluble liquid, obtains T-1 eye-care
Liquid.
2. the method according to claim 1 that eye care solution is prepared by horseshoe crab cell pyrolysis liquid, it is characterised in that：The horseshoe crab
The optimal proportion scope of T-1 freeze-dried powder and purified water is in plain peptide eye care solution：1:10-1:200.
3. the method according to claim 1 that eye care solution is prepared by horseshoe crab cell pyrolysis liquid, it is characterised in that：Described
The centrifuge RPMs of step (2) are 3000-5000 revs/min in one step.
4. the method according to claim 1 that eye care solution is prepared by horseshoe crab cell pyrolysis liquid, it is characterised in that：Described
Excipient in the step of two steps (1) is the lactose for removing thermal source.
5. the method according to claim 1 that eye care solution is prepared by horseshoe crab cell pyrolysis liquid, it is characterised in that：Described
Lyophilized concrete operations are in the step of two steps (2)：Under conditions of vacuum is 10Pa, -50 DEG C of pre-freezes 1 hour, Ran Houcheng
Sequence is heated up, and product reaches 24 DEG C, opens permeation valve, starts redrying, and then product is at the uniform velocity warming up to up to 31 DEG C, is cracked true
Sky, product export.
6. the method according to claim 1 that eye care solution is prepared by horseshoe crab cell pyrolysis liquid, it is characterised in that：100
0.1-5 parts of 0.1-10 parts of Sodium Hyaluronate and borneol are further added in the T-1 eye care solution of 3rd step of part.
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Application publication date: 20170725 |