CN111854339B - Freeze-drying method of collagen-containing skin-whitening, anti-wrinkle and skin-care freeze-dried powder - Google Patents
Freeze-drying method of collagen-containing skin-whitening, anti-wrinkle and skin-care freeze-dried powder Download PDFInfo
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- CN111854339B CN111854339B CN202010631341.4A CN202010631341A CN111854339B CN 111854339 B CN111854339 B CN 111854339B CN 202010631341 A CN202010631341 A CN 202010631341A CN 111854339 B CN111854339 B CN 111854339B
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F26—DRYING
- F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
- F26B5/00—Drying solid materials or objects by processes not involving the application of heat
- F26B5/04—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum
- F26B5/06—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum the process involving freezing
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- 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
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
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Abstract
The invention discloses a freeze-drying method of collagen-containing skin-whitening, anti-wrinkle and skin-care freeze-dried powder. According to the invention, the water content of the final product is controlled to be 5-8% by regulating and controlling the heating speed, temperature, vacuum degree and holding time of the pre-freezing, primary sublimation and analytic drying stages in the freeze drying process, and the obtained freeze-dried powder has the advantages of stable properties, attractive appearance, high collagen activity and the like.
Description
Technical Field
The invention belongs to the technical field of cosmetics, and relates to a freeze-drying method of collagen-containing skin-whitening, anti-wrinkle and skin-care freeze-dried powder.
Background
The vacuum freeze drying technology is to sublimate water from ice directly at relatively low temperature (-10 deg.c to-50 deg.c) to dry without surface tension and deformation, and to dewater material. Because the vacuum freeze-drying technology is carried out in a low-temperature and low-pressure environment, most biological reactions are stopped, liquid water does not exist in the treatment process, and the water is directly sublimated in a solid state, so that the original structure and shape of the material are protected to the greatest extent, a high-quality dried product with both appearance and internal quality is finally obtained, and the activity of active substances (such as collagen) in the freeze-dried product can be fully reserved due to the biological reaction stop. The vacuum freeze drying technology is applied to the cosmetic industry to obtain high-quality skin care products.
Because the collagen has bioactivity, the bioactivity needs to be maintained to the maximum extent, and the freeze-dried product has compact internal crystallization, beautiful appearance and no obvious phenomena of collapse, atrophy, bubble formation and the like, the freeze-drying process has extremely strict requirements. In the existing freeze-drying process of products containing collagen, the drying and sublimation temperature is controlled to be higher (higher than 40 ℃), collagen is easy to inactivate, meanwhile, the water control is not strict, and the conditions of product collapse, atrophy or large crystal particles and loose product appearance often occur, so that the products are not beautiful, and the bioactivity does not reach the standard. Therefore, the development of a freeze-drying process is urgently needed, and the quality requirements of the collagen-containing skin-whitening, anti-wrinkle and skin-care freeze-dried powder can be met.
Disclosure of Invention
The invention aims to provide a freeze-drying method of collagen-containing skin-whitening anti-wrinkle skin-care freeze-dried powder. According to the method, the pre-freezing temperature, the sublimation temperature and the vacuum degree during drying are regulated, the moisture of the freeze-dried powder is controlled to be 5-8%, the obtained product is stable in property and attractive in appearance, and the activity of the collagen is kept at a high level.
The technical scheme for realizing the purpose of the invention is as follows:
the freeze-drying method of the collagen-containing skin-whitening, anti-wrinkle and skin-care freeze-dried powder comprises the following steps:
placing the whitening anti-wrinkle skin care stock solution containing the collagen into a freeze-drying device, and carrying out the following procedures:
step 1, pre-freezing:
the temperature of the heat conducting oil reaches-5 to-10 ℃ in 2 to 10 minutes at the first stage, and is kept for 0.5 to 1 hour;
the temperature of the heat conducting oil in the second stage reaches-35 to-45 ℃ in 20 to 40 minutes, and is kept for 2 to 3 hours;
step 2, primary sublimation:
the temperature of the heat conducting oil in the first stage reaches-30 to-20 ℃ in 0.5 to 1 hour, the vacuum degree is kept at the limit vacuum, and the vacuum degree is kept for 5 to 10 hours;
the temperature of the second stage heat conducting oil reaches-20 to-10 ℃ within 0.5 to 1 hour, the vacuum degree is 20 to 30 Pa, and the second stage heat conducting oil is kept for 5 to 10 hours;
in the third stage, the temperature of the heat conducting oil reaches-10 ℃ to 0 ℃ in 0.5 to 1 hour, the vacuum degree is 30 Pa to 40 Pa, and the heat conducting oil is kept for 5 hours to 10 hours;
the temperature of the heat conducting oil at the fourth stage reaches 0-5 ℃ in 0.5-1 hour, the vacuum degree is 40-50 Pa, and the heat conducting oil is kept for 2-3 hours;
and 3, resolving and drying:
the temperature of the heat conducting oil in the first stage reaches 5-20 ℃ in 0.5-1 hour, the vacuum degree is 40-50 Pa, and the heat conducting oil is kept for 3-4 hours;
the temperature of the second stage heat conducting oil reaches 20-35 ℃ in 0.5-1 hour, the vacuum degree is 30-40 Pa, and the second stage heat conducting oil is kept for 2-3 hours;
step 4, pressure rise test: setting pressure rise to be 3 Pa/min, wherein the pressure rise is qualified, and the freeze-drying is finished, wherein the water content of the freeze-dried powder is 5-8%.
The whitening anti-wrinkle skin care stock solution containing collagen comprises the following components in percentage by mass:
| collagen protein | 0.5％ |
| Hyaluronic acid sodium salt | 2.0％ |
| Mannitol | 0.3％ |
| Pullulan polysaccharide | 0.3％ |
| Trehalose | 0.6％ |
| Hydroxypropyl cyclodextrins | 0.5％ |
| Nicotinamide | 0.15％ |
| Water (W) | Balance of |
The inventor finds that in the pre-freezing process, if the vacuum pumping treatment is directly carried out, gas dissolved in water escapes due to reduction of external pressure to form bubbles, so that cavities appear in the interior and on the surface of the raw material, and the appearance of the product is affected. Therefore, the invention keeps normal pressure operation during pre-freezing, and performs stage pre-freezing at different temperatures and time, and the finally obtained product has compact internal crystal and beautiful appearance.
Compared with the prior art, the invention has the following remarkable effects:
vacuum drying is generally divided into 2 stages of sublimation drying (one time sublimation) and resolution drying. Sublimation drying is mainly directed at free water in the material; the desorption drying is mainly to remove the adsorbed water strongly bonded to the solid. The vacuum degree and temperature during the drying process directly influence the drying process and the final moisture of the product, and the moisture in turn influences the biological activity of the collagen. The invention selects reasonable pre-freezing and sublimation temperatures and vacuum degrees, optimizes the freeze-drying curve, improves the product properties, enhances the product stability, keeps the biological activity of the collagen at a higher level, and ensures that the quality standard of the freeze-dried product meets various indexes specified by the national drug standard.
Drawings
FIG. 1 is a diagram of a lyophilized powder prepared in example 1;
FIG. 2 is a diagram of a lyophilized powder prepared in comparative example 1;
fig. 3 is a diagram of a lyophilized powder entity prepared in example 2 and comparative example 2.
Detailed Description
The invention is further illustrated by the following examples and figures.
In the following examples and comparative examples, the adopted collagen-containing whitening anti-wrinkle skin care stock solution consists of the following components in percentage by mass:
| collagen protein | 0.5％ |
| Hyaluronic acid sodium salt | 2.0％ |
| Mannitol | 0.3％ |
| Pullulan polysaccharide | 0.3％ |
| Trehalose | 0.6％ |
| Hydroxypropyl cyclodextrins | 0.5％ |
| Nicotinamide | 0.15％ |
| Water (W) | Balance of |
Example 1
(1) Pre-freezing:
the temperature of the heat conducting oil reaches-10 ℃ in 10 minutes at the first stage, and the heat conducting oil is kept for 1 hour;
the temperature of the heat conducting oil in the second stage reaches minus 45 ℃ in 40 minutes, and the heat conducting oil is kept for 3 hours;
(2) and carrying out primary sublimation:
the temperature of the heat conducting oil reaches-30 ℃ in 1 hour at the first stage, the vacuum degree is limited to be vacuum, and the heat conducting oil is kept for 10 hours;
the temperature of the heat conducting oil in the second stage reaches-20 ℃ in 1 hour, the vacuum degree is 30 Pa, and the heat conducting oil is kept for 10 hours;
in the third stage, the temperature of the heat-conducting oil reaches-5 ℃ in 1 hour, the vacuum degree is 40 Pa, and the heat-conducting oil is kept for 10 hours;
the temperature of the heat conducting oil at the fourth stage reaches 5 ℃ in 1 hour, the vacuum degree is 50 Pa, and the heat conducting oil is kept for 3 hours;
(3) and (3) analysis and drying:
the temperature of the heat conducting oil reaches 20 ℃ in 1 hour, the vacuum degree is 50 Pa, and the heat conducting oil is kept for 4 hours;
the temperature of the second stage heat-conducting oil reaches 35 ℃ in 1 hour, the vacuum degree is 40 Pa, and the second stage heat-conducting oil is kept for 3 hours;
(4) and pressure rise test: the pressure is increased by 3 Pa/min, the pressure is increased to be qualified, the freeze-drying is finished, the water content of the freeze-dried powder is 5-7%, the internal crystallization of the product is compact, the surface is smooth, and the phenomena of obvious collapse, atrophy, bubble formation and the like are avoided, as shown in figure 1.
Comparative example 1
(1) Pre-freezing:
the temperature of the heat conducting oil reaches-8 ℃ in 10 minutes at the first stage, and the heat conducting oil is kept for 1 hour;
the temperature of the heat conducting oil in the second stage reaches minus 40 ℃ in 40 minutes, and the heat conducting oil is kept for 3 hours;
(2) and carrying out primary sublimation:
the temperature of the heat conducting oil reaches-25 ℃ in 2 hours at the first stage, the vacuum degree is limited to be vacuum, and the heat conducting oil is kept for 10 hours;
the temperature of the heat conducting oil in the second stage reaches-18 ℃ in 2 hours, the vacuum degree is 30 Pa, and the heat conducting oil is kept for 10 hours;
in the third stage, the temperature of the heat conducting oil reaches-10 ℃ in 2 hours, the vacuum degree is 40 Pa, and the heat conducting oil is kept for 10 hours;
the temperature of the heat conducting oil at the fourth stage reaches 0 ℃ in 2 hours, the vacuum degree is 50 Pa, and the heat conducting oil is kept for 3 hours;
(3) and (3) analysis and drying:
the temperature of the heat-conducting oil reaches 15 ℃ in 2 hours, the vacuum degree is 50 Pa, and the heat-conducting oil is kept for 4 hours;
the temperature of the second stage heat-conducting oil reaches 30 ℃ in 2 hours, the vacuum degree is 40 Pa, and the second stage heat-conducting oil is kept for 3 hours;
(4) and pressure rise test: the pressure is increased by 3 Pa/min, the pressure is increased to be qualified, the freeze-drying is finished, the water content of the freeze-dried powder is 8.5-10%, the product has loose internal crystals and is easy to fall off, the appearance is general, and partial samples have collapse or atrophy phenomena, as shown in figure 2.
Example 2
(1) Pre-freezing:
the temperature of the heat conducting oil reaches-10 ℃ in 5 minutes at the first stage, and the heat conducting oil is kept for 1 hour;
the temperature of the heat conducting oil in the second stage reaches-45 ℃ in 25 minutes, and is kept for 3 hours;
(2) and carrying out primary sublimation:
the temperature of the heat conducting oil reaches-30 ℃ in 0.5 hour at the first stage, the vacuum degree is limited to be vacuum, and the heat conducting oil is kept for 10 hours;
the temperature of the second stage heat-conducting oil reaches-20 ℃ in 0.5 hour, the vacuum degree is 30 Pa, and the second stage heat-conducting oil is kept for 10 hours;
the temperature of the heat-conducting oil in the third stage reaches-5 ℃ in 0.5 hour, the vacuum degree is 40 Pa, and the heat-conducting oil is kept for 10 hours;
the temperature of the heat conducting oil in the fourth stage reaches 5 ℃ in 0.5 hour, the vacuum degree is 50 Pa, and the heat conducting oil is kept for 3 hours;
(3) and (3) analysis and drying:
the temperature of the heat-conducting oil in the first stage reaches 20 ℃ in 0.5 hour, the vacuum degree is 50 Pa, and the heat-conducting oil is kept for 4 hours;
the temperature of the second stage heat-conducting oil reaches 35 ℃ in 0.5 hour, the vacuum degree is 40 Pa, and the second stage heat-conducting oil is kept for 3 hours;
(4) and pressure rise test: the pressure is increased by 3Pa/1 min, the pressure is increased to be qualified, the freeze-drying is finished, the water content of the freeze-dried powder is 6-8%, the internal crystallization of the product is compact, the appearance is attractive, and the phenomena of obvious collapse, atrophy, bubble formation and the like are avoided.
Comparative example 2
(1) Pre-freezing:
the temperature of the heat conducting oil reaches-10 ℃ in 2 minutes at the first stage, and the heat conducting oil is kept for 1 hour;
the temperature of the heat conducting oil in the second stage reaches-45 ℃ in 20 minutes, and is kept for 3 hours;
(2) and carrying out primary sublimation:
the temperature of the heat conducting oil reaches-30 ℃ in 20 minutes at the first stage, the vacuum degree is limited to be vacuum, and the heat conducting oil is kept for 10 hours;
the temperature of the second stage heat-conducting oil reaches-20 ℃ in 20 minutes, the vacuum degree is 30 Pa, and the second stage heat-conducting oil is kept for 10 hours;
in the third stage, the temperature of the heat-conducting oil reaches-0 ℃ in 20 minutes, the vacuum degree is 40 Pa, and the heat-conducting oil is kept for 10 hours;
the temperature of the heat conducting oil at the fourth stage reaches 5 ℃ in 20 minutes, the vacuum degree is 50 Pa, and the heat conducting oil is kept for 3 hours;
(3) and (3) analysis and drying:
the temperature of the heat conducting oil in the first stage reaches 20 ℃ in 20 minutes, the vacuum degree is 50 Pa, and the heat conducting oil is kept for 4 hours;
the temperature of the second stage heat-conducting oil reaches 35 ℃ in 20 minutes, the vacuum degree is 40 Pa, and the second stage heat-conducting oil is kept for 3 hours;
(4) and pressure rise test: setting pressure to rise for 3 Pa/min, ensuring that the pressure rises to be qualified, finishing freeze-drying, ensuring that the water content of freeze-dried powder is 2.5-4.5%, ensuring that the internal crystallization of the product is compact and the appearance is common, and ensuring that part of samples have bubbles, as shown in figure 3, the left sample in figure 3 is a product of a comparative example 2, and the right sample is a product of an example 2.
Selecting the freeze-dried powder prepared by a control group (stock solution before freeze-drying), examples 1-2 and comparative examples 1-2 to perform a cell proliferation experiment, and investigating the biological activity of collagen in the freeze-dried powder, wherein the specific experimental steps are as follows:
1. collecting human epidermal cells with good growth condition, digesting to obtain single cell suspension, counting, and adjusting cell density to 104and/mL, uniformly inoculating the cells into a 96-well cell culture plate, adding 200 mu L of cell suspension into each well, adding samples (blank group, control group (stock solution before freeze drying), freeze-dried powder of example 1, freeze-dried powder of comparative example 1, freeze-dried powder of example 2 and freeze-dried powder of comparative example 2) diluted by 10 times in equal amount after the cells are attached to the wall, setting 5 multiple wells for each sample, placing the sample at 37 ℃ and 5% CO2Incubation was continued in the incubator for 24, 48, 72, 96, 120, 144, 168 h.
2. To each time node, 20 μ L of MTT solution (formulated with PBS at pH 7.4) was added to the corresponding well and incubation continued for 4h, terminating the culture.
3. The plate was quickly inverted, the supernatant discarded, 150. mu.L of DMSO was added to each well, and the mixture was shaken for 10min to completely dissolve the MTT bluish-purple crystals.
4. The absorbance (OD) of each well was measured at 490nm using an ELISA, and the absorbance of wells containing no inoculated cells and medium alone was set to zero as a blank, and the average of 6 wells was taken.
5. The cell growth curve was plotted with the culture time as abscissa and the absorbance as ordinate, and the cell proliferation rate of each sample was calculated, and the results are shown in table 1.
TABLE 1 cell proliferation Rate of samples
| Sample (I) | Blank group | Control group | Example 1 | Comparative example 1 | Example 2 | Comparative example 2 |
| Cell proliferation rate | 100％ | 137.21％ | 133.33％ | 113.78％ | 134.28％ | 109.43％ |
The results of cell proliferation experiments show that the results of the embodiment 1 and the embodiment 2 are ideal, and the biological activity of the collagen can be maintained to a greater extent, which indicates that the water content of the product can be controlled to be 5-8% by adopting the freeze-drying process disclosed by the invention, so that the appearance of the product can be ensured to be attractive, and the biological activity of the collagen can be ensured.
Claims (2)
1. The freeze-drying method of the collagen-containing skin-whitening, anti-wrinkle and skin-care freeze-dried powder is characterized by comprising the following steps:
placing the whitening anti-wrinkle skin care stock solution containing the collagen into a freeze-drying device, and carrying out the following procedures:
(1) pre-freezing:
the temperature of the heat conducting oil reaches-10 ℃ in 10 minutes at the first stage, and the heat conducting oil is kept for 1 hour;
the temperature of the heat conducting oil in the second stage reaches minus 45 ℃ in 40 minutes, and the heat conducting oil is kept for 3 hours;
(2) and carrying out primary sublimation:
the temperature of the heat conducting oil reaches-30 ℃ in 1 hour at the first stage, the vacuum degree is limited to be vacuum, and the heat conducting oil is kept for 10 hours;
the temperature of the heat conducting oil in the second stage reaches-20 ℃ in 1 hour, the vacuum degree is 30 Pa, and the heat conducting oil is kept for 10 hours;
in the third stage, the temperature of the heat-conducting oil reaches-5 ℃ in 1 hour, the vacuum degree is 40 Pa, and the heat-conducting oil is kept for 10 hours;
the temperature of the heat conducting oil at the fourth stage reaches 5 ℃ in 1 hour, the vacuum degree is 50 Pa, and the heat conducting oil is kept for 3 hours;
(3) and (3) analysis and drying:
the temperature of the heat conducting oil reaches 20 ℃ in 1 hour, the vacuum degree is 50 Pa, and the heat conducting oil is kept for 4 hours;
the temperature of the second stage heat-conducting oil reaches 35 ℃ in 1 hour, the vacuum degree is 40 Pa, and the second stage heat-conducting oil is kept for 3 hours;
(4) and pressure rise test: setting pressure rise to be 3 Pa/min, wherein the pressure rise is qualified, and the water content of the freeze-dried powder is 5-7 percent after freeze-drying is finished;
the whitening anti-wrinkle skin care stock solution containing collagen consists of the following components in percentage by mass:
0.5 percent of collagen protein,
2.0 percent of sodium hyaluronate,
0.3 percent of mannitol,
0.3 percent of pullulan polysaccharide,
0.6 percent of trehalose,
0.5 percent of hydroxypropyl cyclodextrin,
0.15 percent of nicotinamide,
the balance of water.
2. The freeze-drying method of the collagen-containing skin-whitening, anti-wrinkle and skin-care freeze-dried powder is characterized by comprising the following steps:
placing the whitening anti-wrinkle skin care stock solution containing the collagen into a freeze-drying device, and carrying out the following procedures:
(1) pre-freezing:
the temperature of the heat conducting oil reaches-10 ℃ in 5 minutes at the first stage, and the heat conducting oil is kept for 1 hour;
the temperature of the heat conducting oil in the second stage reaches-45 ℃ in 25 minutes, and is kept for 3 hours;
(2) and carrying out primary sublimation:
the temperature of the heat conducting oil reaches-30 ℃ in 0.5 hour at the first stage, the vacuum degree is limited to be vacuum, and the heat conducting oil is kept for 10 hours;
the temperature of the second stage heat-conducting oil reaches-20 ℃ in 0.5 hour, the vacuum degree is 30 Pa, and the second stage heat-conducting oil is kept for 10 hours;
the temperature of the heat-conducting oil in the third stage reaches-5 ℃ in 0.5 hour, the vacuum degree is 40 Pa, and the heat-conducting oil is kept for 10 hours;
the temperature of the heat conducting oil in the fourth stage reaches 5 ℃ in 0.5 hour, the vacuum degree is 50 Pa, and the heat conducting oil is kept for 3 hours;
(3) and (3) analysis and drying:
the temperature of the heat-conducting oil in the first stage reaches 20 ℃ in 0.5 hour, the vacuum degree is 50 Pa, and the heat-conducting oil is kept for 4 hours;
the temperature of the second stage heat-conducting oil reaches 35 ℃ in 0.5 hour, the vacuum degree is 40 Pa, and the second stage heat-conducting oil is kept for 3 hours;
(4) and pressure rise test: setting pressure rise to be 3Pa/1 min, wherein the pressure rise is qualified, and the water content of the freeze-dried powder is 6-8 percent after freeze-drying is finished;
the whitening anti-wrinkle skin care stock solution containing collagen consists of the following components in percentage by mass:
0.5 percent of collagen protein,
2.0 percent of sodium hyaluronate,
0.3 percent of mannitol,
0.3 percent of pullulan polysaccharide,
0.6 percent of trehalose,
0.5 percent of hydroxypropyl cyclodextrin,
0.15 percent of nicotinamide,
the balance of water.
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| CN109959227A (en) * | 2017-12-14 | 2019-07-02 | 青岛华迈士药业有限公司 | A kind of method of urokinase injection lyophilized preparation freeze-drying curve optimization |
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