A low temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate and a cosmetic gel

Abstract

The application discloses a low-temperature polyol-polyose protective acetyl hexapeptide-8 concentrate and a cosmetic gel, and sequentially limits starting materials, separation or conversion steps, process endpoints, stabilization conditions and batch release parameters to prepare an acetyl hexapeptide-8 protective concentrate, wherein the core quality range comprises 0.05%-0.15% acetyl hexapeptide-8 content, not more than 2.0% related peptide impurities and not more than 0.8% methionine oxidation related peaks, and the active substance is further prepared into a gel according to specific component proportions and adding sequences. The application limits the sources of raw materials, the sequence of steps and the process endpoints, so that the acetyl hexapeptide-8 content, the related peptide impurities and the methionine oxidation related peaks form a reviewable quality window, which is beneficial to improving batch consistency. By connecting the standardized active substance with specific product formulations, adding sequences, temperature and pH boundaries, the risk of crystallization, aggregation, oxidation, odor or stratification in the gel is reduced.

Description

Technical Field

[0001] This invention relates to the fields of cosmetic raw material preparation, quality standardization, and cosmetic formulation technology, specifically a low-temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate and a cosmetic gel. Background Technology

[0002] When acetyl hexapeptide-8 protected concentrate is used in beauty care products, its raw material identity, key impurities, molecular or particle state, and ionic or oxidative load can all affect formulation compatibility. Delivery based solely on raw material name or a single total content cannot simultaneously reflect batch-to-batch variations in acetyl hexapeptide-8 content, related peptide impurities, methionine oxidation-related peaks, and other indicators.

[0003] The operating conditions and process endpoints of nanofiltration washing jointly affect the content of acetyl hexapeptide-8, related peptide impurities, and methionine oxidation-related peaks. If only a single step is retained or only a single content indicator is delivered, it is difficult to determine whether the above quality boundaries are simultaneously met, and it is also not conducive to reproducing the formulation and application of subsequent gels.

[0004] Therefore, it is necessary to combine the identity of starting materials, the sequence of key processes, the quality release range, and the boundaries of the finished product formulation into a reproducible overall solution, and to define the technical characteristics described in the title using detectable parameters such as acetyl hexapeptide-8 content, related peptide impurities, and methionine oxidation-related peaks. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a low-temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate and cosmetic gel, which reduces batch fluctuations caused by key impurities or unstable physical states, and enables the obtained raw materials to enter the gel preparation process within a uniform release range.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a low-temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate, wherein the acetyl hexapeptide-8 content is 0.05% to 0.15%, the related peptide impurities are not higher than 2.0%, the methionine oxidation-related peak is not higher than 0.8%, the pH is 5.3 to 5.7, the viscosity is 800 to 2500 mPa·s, and the total bacterial count / total mold and yeast count is not higher than 100 / 50 CFU / g.

[0007] A cosmetic care composition comprising, by weight of total composition, 4.8% to 7.2% of acetyl hexapeptide-8 protective concentrate and a cosmetically acceptable carrier.

[0008] Preferably, the composition comprises, by total mass, 4.8%–7.2% acetyl hexapeptide-8 protected concentrate, 3.2%–4.8% glycerol, 4%–6% 1,3-propanediol, 1.2%–1.8% betaine, 0.20%–0.30% polyacrylate crosspolymer-6, 0.08%–0.12% stearin gum, 0.64%–0.96% phenoxyethanol / ethylhexylglycerin complex (9:1 by mass), 0.12%–0.18% citric acid / sodium citrate buffer (1:2 by mass), and the balance being purified water.

[0009] Preferably, the composition is a gel; by total mass, it comprises 6.00% acetyl hexapeptide-8 protected concentrate, 4.00% glycerol, 5.00% 1,3-propanediol, 1.50% betaine, 0.25% polyacrylate crosspolymer-6, 0.10% stearin gum, 0.80% phenoxyethanol / ethylhexylglycerin complex (mass ratio 9:1), 0.15% citric acid / sodium citrate buffer (mass ratio 1:2), and 82.20% purified water, with the total of the above components being 100%.

[0010] A method for preparing the beauty care composition includes the following: The purified water was mixed evenly with glycerol, 1,3-propanediol and betaine; polyacrylate cross-linked polymer-6 and hard dextran gum were pre-dispersed and added and hydrated for 20-40 min; acetyl hexapeptide-8 protective concentrate, phenoxyethanol / ethylhexylglycerin complex and citric acid / sodium citrate buffer were added at a temperature not exceeding 35°C to adjust the pH to 5.3-5.7; the mixture was degassed for 5-10 min under a vacuum of -0.08 to -0.06 MPa and then filled.

[0011] A method for preparing the low-temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate includes the following steps: (1) Dissolve acetyl hexapeptide-8 in citrate buffer water with pH 5.2 to 5.8 at 18 to 25°C to make the peptide concentration 0.8% to 1.5%; (2) Use a nanofiltration membrane with a molecular weight cutoff of 300-500 Da to collect the retentate and wash it with a low shear method for 2-3 membrane volumes to control free salt and short-chain peptide impurities; (3) Add trehalose, mannitol and 1,3-propanediol to the nanofiltration retentate, with the mass ratio of the three to acetyl hexapeptide-8 being 8-15:3-8:20-35:1; (4) After prehydrating pullulan and hydroxypropyl cellulose, add peptide solution at a temperature not higher than 28°C; based on the total mass of the obtained protective concentrate, the amount of pullulan is 0.8% to 1.2% and the amount of hydroxypropyl cellulose is 0.2% to 0.4% to form a protective system; (5) Degas under a vacuum of -0.08 to -0.06 MPa and adjust the pH to 5.3 to 5.7; (6) Based on the measured content of acetyl hexapeptide-8, a standardized concentrate with a peptide content of 0.05% to 0.15% was prepared; The resulting protected concentrate was tested and found to simultaneously meet the requirements that peptide impurities are not higher than 2.0% and methionine oxidation-related peaks are not higher than 0.8%.

[0012] Preferably, the starting material is acetyl hexapeptide-8 (Ac-Glu-Glu-Met-Gln-Arg-Arg-NH2) with an HPLC purity of not less than 98.0%.

[0013] Preferably, the obtained acetyl hexapeptide-8 protected concentrate simultaneously meets the following quality parameters: acetyl hexapeptide-8 content is 0.05% to 0.15%, related peptide impurities are not higher than 2.0%, methionine oxidation-related peak is not higher than 0.8%, pH is 5.3 to 5.7, viscosity is 800 to 2500 mPa·s, and total bacterial count / total mold and yeast count is not higher than 100 / 50 CFU / g.

[0014] Compared with the prior art, the technical solution of this application has the following beneficial effects: 1. By limiting the source of raw materials, the order of steps, and the process endpoint, this invention enables the content of acetyl hexapeptide-8, related peptide impurities, and methionine oxidation-related peaks to form a verifiable quality window, which is beneficial to improving batch consistency.

[0015] 2. By linking standardized active ingredients with specific finished product formulations, addition sequences, temperature, and pH boundaries, this invention helps reduce the risks of crystallization, aggregation, oxidation, off-odors, or stratification in gels.

[0016] 3. The process steps, process endpoints and quality indicators of the present invention correspond to each other, which is beneficial to production process control, batch verification and subsequent formula application. Attached Figure Description

[0017] Figure 1 This is a flowchart of the preparation method of the low-temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate of the present invention; Figure 2 This is a flowchart of the preparation method of the beauty care composition of the present invention; Figure 3 This is a graph showing the test results of the low-temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate of the present invention. Detailed Implementation

[0018] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] Please see Figure 1-3 In this embodiment, a low-temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate and a cosmetic gel are prepared using the following process chain: The starting material was acetyl hexapeptide-8 (Ac-Glu-Glu-Met-Gln-Arg-Arg-NH2) with an HPLC purity of not less than 98.0%.

[0020] The preparation steps are as follows: (1) Dissolve acetyl hexapeptide-8 in citrate buffer water with pH 5.2 to 5.8 at 18 to 25°C to make the peptide concentration 0.8% to 1.5%.

[0021] (2) Use a nanofiltration membrane with a molecular weight cutoff of 300-500 Da to collect the retentate and wash it with a low shear for 2-3 membrane volumes to control free salt and short-chain peptide impurities.

[0022] (3) Add trehalose, mannitol and 1,3-propanediol to the nanofiltration retentate, with the mass ratio of the three to acetyl hexapeptide-8 being 8-15:3-8:20-35:1.

[0023] (4) After prehydrating pullulan and hydroxypropyl cellulose, add peptide solution at a temperature not higher than 28°C; based on the total mass of the obtained protective concentrate, the amount of pullulan is 0.8% to 1.2% and the amount of hydroxypropyl cellulose is 0.2% to 0.4% to form a protective system.

[0024] (5) Degas under a vacuum of -0.08 to -0.06 MPa and adjust the pH to 5.3 to 5.7.

[0025] (6) Based on the measured content of acetyl hexapeptide-8, a standardized concentrate with a peptide content of 0.05% to 0.15% was prepared.

[0026] The resulting acetyl hexapeptide-8 protected concentrate should simultaneously meet the quality ranges listed in Table 1.

[0027] Table 1 Standardized Active Ingredient Quality Release Range The starting material is acetyl hexapeptide-8 with an HPLC purity of not less than 98.0%. The raw material batches should have traceable sources, inspection records, and storage conditions; purified water and all preservatives should meet cosmetic production standards.

[0028] The term "acetyl hexapeptide-8 protected concentrate" as used herein refers to an aqueous concentrate processed according to the process described in this application and simultaneously meeting all release requirements in Table 1. Unless otherwise stated, percentage content is a mass percentage relative to the total mass of the concentrate; membrane molecular weight cutoff is the nominal value specified by the membrane manufacturer for the specified aqueous system.

[0029] When low-temperature preparation and nanofiltration washing are involved, the temperature, time, stirring speed, membrane material, effective area, transmembrane pressure difference, circulation flow rate and constant volume washing volume should be recorded in the batch record.

[0030] II. Detection Methods like Figure 3 As shown, two parallel samples were prepared for each batch of key content and key impurities. Acetyl hexapeptide-8 and related peptides were detected using C18 reversed-phase UPLC at 214 nm; the content of acetyl hexapeptide-8 was determined using the external standard method. Related peptide impurities were analyzed using the peak area normalization method, calculated as "the sum of the peak areas of all peptide-related impurities excluding the acetyl hexapeptide-8 main peak / the sum of the peak areas of all peptide-related peaks × 100%"; the methionine oxidation-related peak was reported separately as a designated peak among the related peptide impurities, calculated as "the area of ​​this designated peak / the sum of the peak areas of all peptide-related peaks × 100%", and cross-confirmed by LC-MS retention time and mass-to-charge ratio. Viscosity was measured at 25°C using a rotational viscometer, and rotor, rotation speed, and shear time were recorded; microbiological items were analyzed using the plate count method. The system suitability and spiked recovery rate of batch B17-01 met the preset standards.

[0031] Example 1: Determining the preparation of batch-protected acetyl hexapeptide-8 concentrate The batch number for this embodiment is B17-01. The starting material, acetyl hexapeptide-8, was 11.20 g, with an HPLC purity of 98.9%, related peptide impurities of 1.21%, and a methionine oxidation-related peak of 0.34%. The target batch size of the resulting concentrate was 9.80 kg.

[0032] 1. Add 11.20 g of acetyl hexapeptide-8 to 922.1 g of 20 mmol / L citrate buffer water with pH 5.50, and stir at 21.5℃ and 300 rpm for 30 min to make the peptide mass concentration 1.20%.

[0033] 2. A nanofiltration membrane with a molecular weight cutoff of 400 Da was used to collect the retentate. The membrane area was 0.10 m², the feed temperature was 22±2℃, the transmembrane pressure difference was 0.90 MPa, and the circulation flow rate was 1.2 L / min. Three membrane volumes were washed with purified water at low shear constant volume.

[0034] 3. Add 128.8 g of trehalose, 61.6 g of mannitol and 308.0 g of 1,3-propanediol to the nanofiltration retentate in sequence, and stir at 22±2℃ and 400 rpm for 30 min; the mass ratio of the three to acetyl hexapeptide-8 is 11.5∶5.5∶27.5∶1.

[0035] 4. 98.0 g of pullulan and 29.4 g of hydroxypropyl cellulose were pre-hydrated in 1.20 kg of purified water for 30 min. The nanofiltration retentate was added at 27 °C and 300 rpm, and the mixture was stirred for another 40 min to form a homogeneous protective system. Based on a target batch size of 9.80 kg, the pullulan and hydroxypropyl cellulose were 1.00% and 0.30%, respectively.

[0036] 5. Make up the volume to 9.80 kg with purified water, degas at 23°C and a vacuum of -0.075 MPa for 8 min, and adjust the pH to 5.49 with citrate buffer.

[0037] 6. After degassing, stir at 200 rpm for 10 min, and take three samples from the top, middle and bottom to confirm the homogeneity; the relative standard deviation of the acetyl hexapeptide-8 content at the three points is 1.1%, and batch B17-01 acetyl hexapeptide-8 protected concentrate is obtained.

[0038] The finished product weight of batch B17-01 was 9.80 kg. Release was carried out according to Table 1 and the testing methods in this section, and the results are shown in Table 3. This batch was used in Example 2 after all items were within the range of Table 1.

[0039] Example 2: Beauty Care Composition Batch B17-F01 gel was prepared with a total mass of 10.00 kg. The actual amount of each component added was the mass percentage listed in Table 2 multiplied by 100 g.

[0040] Table 2 Composition of Beauty Care Compositions The preparation method is as follows: purified water, glycerol, 1,3-propanediol and betaine were mixed at 25℃; polyacrylate cross-linked polymer-6 and hard dextran gum were pre-dispersed and added and hydrated for 30 min; batch B17-01 concentrate, preservative compound and buffer were added at 30℃ to make the pH 5.53; degassing was carried out under vacuum of -0.075 MPa for 8 min and then filled to obtain batch B17-F01.

[0041] Comparative Example 1: Key Step Omission Solution The same peptide solution and polyol ratio as in Example 1 were used, but the trehalose and mannitol protective components were omitted. Except for the differences, the starting material batches, feed amounts, equipment, sampling times, and testing methods remained consistent.

[0042] Verification results Table 3. Verification results of the examples and comparative examples The above results are used to evaluate process repeatability, contribution of key steps, and physical stability of the formulation; the technical effects of this invention are evaluated based on improvements in physicochemical quality and stability, and do not involve disease treatment effects.

[0043] It should be noted that, in this document, relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0044] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A low-temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate, characterized in that, The content of acetyl hexapeptide-8 is 0.05% to 0.15%, the content of related peptide impurities is not higher than 2.0%, the methionine oxidation-related peak is not higher than 0.8%, the pH is 5.3 to 5.7, the viscosity is 800 to 2500 mPa·s, and the total colony count / total mold and yeast count is not higher than 100 / 50 CFU / g.

2. A beauty care composition, characterized in that, The composition comprises, by weight, 4.8% to 7.2% of the acetyl hexapeptide-8 protective concentrate of claim 1 and a cosmetically acceptable carrier.

3. The beauty care composition according to claim 2, characterized in that, The composition comprises, by total mass, 4.8%–7.2% of the acetyl hexapeptide-8 protected concentrate of claim 1, 3.2%–4.8% of glycerol, 4%–6% of 1,3-propanediol, 1.2%–1.8% of betaine, 0.20%–0.30% of polyacrylate crosspolymer-6, 0.08%–0.12% of stearin gum, 0.64%–0.96% of phenoxyethanol / ethylhexylglycerin complex (9:1 by mass), 0.12%–0.18% of citric acid / sodium citrate buffer (1:2 by mass), and the balance being purified water.

4. The beauty care composition according to claim 2, characterized in that, The composition is a gel.

5. The beauty care composition according to claim 2, characterized in that, The composition is a gel; by total mass, it comprises 6.00% acetyl hexapeptide-8 protected concentrate as described in claim 1, 4.00% glycerol, 5.00% 1,3-propanediol, 1.50% betaine, 0.25% polyacrylate crosspolymer-6, 0.10% stearin gum, 0.80% phenoxyethanol / ethylhexylglycerin complex (mass ratio 9:1), 0.15% citric acid / sodium citrate buffer (mass ratio 1:2), and 82.20% purified water, with the total of the above components being 100%.

6. The beauty care composition according to claim 2, characterized in that, The preparation methods include the following: Purified water, glycerol, 1,3-propanediol, and betaine were mixed evenly. Polyacrylate cross-linked polymer-6 and hard dextran gum were pre-dispersed and added to the mixture, followed by hydration for 20–40 min. The acetyl hexapeptide-8 protective concentrate, phenoxyethanol / ethylhexylglycerin complex, and citric acid / sodium citrate buffer were added at a temperature not exceeding 35°C to adjust the pH to 5.3–5.

7. The mixture was degassed for 5–10 min under a vacuum of -0.08 to -0.06 MPa and then filled into containers.

7. A method for preparing the low-temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate of claim 1, characterized in that, Includes the following steps: (1) Dissolve acetyl hexapeptide-8 in citrate buffer water with pH 5.2 to 5.8 at 18 to 25°C to make the peptide concentration 0.8% to 1.5%; (2) Use a nanofiltration membrane with a molecular weight cutoff of 300-500 Da to collect the retentate and wash it with a low shear method for 2-3 membrane volumes to control free salt and short-chain peptide impurities; (3) Add trehalose, mannitol and 1,3-propanediol to the nanofiltration retentate, with the mass ratio of the three to acetyl hexapeptide-8 being 8-15:3-8:20-35:1; (4) After prehydrating pullulan and hydroxypropyl cellulose, add peptide solution at a temperature not higher than 28°C; based on the total mass of the obtained protective concentrate, the amount of pullulan is 0.8% to 1.2% and the amount of hydroxypropyl cellulose is 0.2% to 0.4% to form a protective system; (5) Degas under a vacuum of -0.08 to -0.06 MPa and adjust the pH to 5.3 to 5.7; (6) Based on the measured content of acetyl hexapeptide-8, a standardized concentrate with a peptide content of 0.05% to 0.15% was prepared; The resulting protected concentrate was tested and found to simultaneously meet the requirements that peptide impurities are not higher than 2.0% and methionine oxidation-related peaks are not higher than 0.8%.

8. The method for preparing a low-temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate according to claim 7, characterized in that, The starting material is acetyl hexapeptide-8 (Ac-Glu-Glu-Met-Gln-Arg-Arg-NH2) with an HPLC purity of not less than 98.0%.

9. A method for preparing a low-temperature polyol-polysaccharide protected acetyl hexapeptide-8 concentrate according to claim 7 or 8, characterized in that, The obtained acetyl hexapeptide-8 protected concentrate simultaneously meets the following quality parameters: acetyl hexapeptide-8 content is 0.05% to 0.15%, related peptide impurities are not higher than 2.0%, methionine oxidation-related peak is not higher than 0.8%, pH is 5.3 to 5.7, viscosity is 800 to 2500 mPa·s, and total bacterial count / total mold and yeast count is not higher than 100 / 50 CFU / g.

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