Preparation method and application of zinc-rich hot spring algae extract

Abstract

The invention relates to a preparation method and application of a zinc-rich hot spring algae extract, and belongs to the technical field of microalgae. Zinc-containing habitat water is used as a culture medium water source to carry out zinc-rich culture on the hot spring algae; subsequently, efficiently extracting trace elements such as zinc, strontium and the like in the microalgae through a carbonization combined bionic extraction technology; no organic solvent is used in the whole extraction process, so that the environmental burden is small. The obtained zinc-rich hot spring algae extract can be applied to skin care products for repairing, resisting aging and controlling oil.

Description

Technical Field

[0001] The present invention belongs to the technical field of microalgae, and relates to a preparation method and application of a zinc-rich hot spring algae extract. Background Art

[0002] In recent years, the application of natural skin care ingredients has attracted more and more attention from consumers. Hot spring algae refer to those living in high-temperature environments rich in minerals such as hot springs, thermal springs, and geothermal springs. These algae include Scenedesmus, Volvox, Chlorella, Chlamydomonas, Chroococcus, Aphanocapsa, Dunaliella, Lyngbya, Nostoc, etc. Ordinary algae are often rich in polysaccharides and proteins, and have good effects of moisturizing and nourishing the skin. Hot spring algae are rich in minerals, trace elements, and some antioxidant components, etc. These components have good stability at high temperatures and have effects such as promoting the repair of the skin barrier, anti-inflammatory, oil control, antioxidant, and anti-aging. Therefore, hot spring algae extracts have good application prospects in the field of skin care.

[0003] Most of the existing technologies are to extract active substances such as polysaccharides, proteins, and polypeptides from algae and apply them to skin care. For example, CN116751313B introduces a method for extracting SDEC-18 polysaccharide from Chlorella, and CN118146294A discloses a method for preparing spirulina polypeptide using compound protease. Some technologies disclose the enrichment of trace elements in algae. For example, CN119081869A realizes the selenium-rich culture of Chlamydomonas through high-density fermentation, but there are few technologies for enriching and culturing and extracting zinc elements in algae. As an essential trace element for the human body, zinc element has effects such as anti-inflammatory, antioxidant, promoting the repair of the skin barrier, and regulating sebum secretion, and can be widely applied to skin care.

[0004] The existing technologies do not fully utilize algae to enrich zinc elements. When enriching other elements (taking selenium enrichment as an example), in order to avoid the inhibition of selenium elements on the growth of algae, it is usually necessary to gradually add selenium elements to domesticate the algae, and this process is time-consuming. When extracting selenium elements from algae, no targeted extraction technology is adopted, and the selenium content of the algae extract obtained by the general extraction process is low. Summary of the Invention

[0005] The present invention relates to a preparation method and application of a zinc-rich hot spring algae extract, belonging to the technical field of microalgae. The present invention uses zinc-containing habitat water as the culture medium water source to carry out zinc-rich culture on hot spring algae; subsequently, through carbonization combined with biomimetic extraction technology, trace elements such as zinc and strontium in microalgae are efficiently extracted; and no organic solvents are used in the whole extraction process, which has a small environmental burden. The obtained zinc-rich hot spring algae extract can be applied to skin care products for repair, anti-aging, and oil control.

[0006] The object of the present invention can be achieved by the following technical solutions:

[0007] A method for preparing a zinc-rich hot spring alga extract, comprising the following steps:

[0008] (1) Inoculate hot spring algae into a BG11 medium with habitat water as the water source, adjust the temperature to 22-25 °C, the light intensity to 1500-2000 lux, and the pH to 7-7.5, and perform aeration culture. End the enrichment culture when the dry weight of the algal cells reaches 40 g / L;

[0009] (2) Add zinc sulfate to the above medium so that the added zinc content is 2-5 mg / L, adjust the light intensity to 2500-3000 lux, continue the culture and sample every day, and stop the culture until the dry weight of the algal cells no longer increases to obtain zinc-rich hot spring algae;

[0010] (3) Collect the zinc-rich hot spring algae and dry them, add them to a muffle furnace, carbonize them at a temperature of 700-800 °C for a holding time of 2-4 h, and take out the sample after cooling;

[0011] (4) Add the carbonized hot spring algae sample to the feed liquid, adjust the pH to 5.0-6.5, stir at 20-30 °C for 4-6 h, centrifuge to obtain the supernatant, and concentrate it under reduced pressure through a microfiltration membrane until it is the same weight as the carbonized hot spring algae sample, which is the zinc-rich hot spring algae extract.

[0012] As a preferred technical solution of the present invention, the hot spring algae in step (1) is one of Chlamydomonas and Chlorella.

[0013] As a preferred technical solution of the present invention, the habitat water in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, <0.05 mg / L of strontium, 0.011 mg / L of iron, 0.0012 mg / L of selenium, and the balance is water and unavoidable impurity elements.

[0014] As a preferred technical solution of the present invention, the mass ratio of the carbonized hot spring algae sample to the feed liquid in step (4) is 1 g:20-25 mL.

[0015] As a preferred technical solution of the present invention, the feed liquid in step (4) is a biomimetic mineralization agent solution, which is composed of amino acids, nicotinamide adenine dinucleotide and deionized water; the amino acid is one of glycine, glutamic acid and methionine; the amino acid content is 0.2-0.5 mol / L, and the nicotinamide adenine dinucleotide content is 0.004-0.01 mol / L.

[0016] Furthermore, the above-mentioned zinc-rich hot spring alga extract is applied to skin care products for repair, anti-aging and oil control.

[0017] The beneficial effects of the present invention:

[0018] (1) The process of the present invention is simple, easy to operate and highly efficient. By using the hot spring habitat water as the water source and adding it to the zinc-rich medium, since the habitat water itself contains zinc, the hot spring algae can better adapt to the zinc-rich environment and rapidly enrich zinc elements during the growth process.

[0019] (2) During the process of extracting the active substances from the hot spring algae of the present invention, first, the carbonization reaction is used to fully destroy the cell structure of the algae, and then a mineralization inducer is used to gently extract active ingredients such as zinc elements by simulating the biological environment, obtaining a zinc-rich hot spring algae extract.

[0020] (3) Throughout the process of the present invention, no organic solvents are used, which places a small burden on the environment. Moreover, the obtained zinc-rich hot spring algae extract can be applied to skin care products, and its by-product, the hot spring algae biochar, can be applied to the environmental and energy fields after washing and drying, realizing the diversified utilization of microalgae products. Specific Embodiments

[0021] To further elaborate on the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the following, in combination with embodiments, details the specific embodiments, structures, features and their effects according to the present invention as follows.

[0022] In the following examples and comparative examples, the BG11 medium was purchased from Qingdao Haibo Biotechnology Co., Ltd.; Chlamydomonas and Chlorella were purchased from Shanghai Guangyu Biotechnology Co., Ltd.

[0023] Example 1

[0024] A preparation method of a zinc-rich hot spring algae extract, comprising the following steps:

[0025] (1) Inoculate the hot spring algae into the BG11 medium with the habitat water as the water source, adjust the temperature to 22 °C, the light intensity to 1500 lux, and the pH to 7.0, and conduct aeration culture. Stop the enrichment culture when the dry weight of the algal cells reaches 40 g / L.

[0026] (2) Add zinc sulfate to the above medium so that the added zinc content is 5 mg / L, adjust the light intensity to 2500 lux, continue the culture and take samples every day. Stop the culture until the dry weight of the algal cells no longer increases to obtain zinc-rich hot spring algae.

[0027] (3) Collect the zinc-rich hot spring algae and dry them, add them to a muffle furnace, carbonize them at a temperature of 800 °C for a holding time of 2 h, and take out the sample after cooling.

[0028] (4) Add the carbonized hot spring algae sample to the feed liquid, adjust the pH to 6.0, stir at 25 °C for 5 h, centrifuge to obtain the supernatant, and concentrate it under reduced pressure through a microfiltration membrane until it has the same weight as the carbonized hot spring algae sample, which is the zinc-rich hot spring algae extract.

[0029] The thermophilic alga described in step (1) is Chlamydomonas.

[0030] The habitat water described in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, <0.05 mg / L of strontium, 0.011 mg / L of iron, 0.0012 mg / L of selenium, and the balance is water and inevitable impurity elements.

[0031] The ratio of the carbonized thermophilic alga sample to the liquid material in step (4) is 1 g:20 mL.

[0032] The liquid material described in step (4) is a biomimetic mineralization agent solution, which is composed of amino acid, nicotinamide adenine dinucleotide and deionized water; the amino acid is glycine; the content of the amino acid is 0.2 mol / L, and the content of nicotinamide adenine dinucleotide is 0.006 mol / L.

[0033] The application of the zinc-rich thermophilic alga extract is in skin care products for repair, anti-aging and oil control.

[0034] Example 2

[0035] A preparation method of a zinc-rich thermophilic alga extract, comprising the following steps:

[0036] (1) Inoculate the thermophilic alga into the BG11 medium using the habitat water as the water source, adjust the temperature to 25 °C, the light intensity to 2000 lux, and the pH to 7.5, and perform aeration culture. Stop the enrichment culture when the dry weight of the algal cells reaches 40 g / L.

[0037] (2) Add zinc sulfate to the above medium so that the added zinc content is 3 mg / L, adjust the light intensity to 2700 lux, continue the culture and take samples every day. Stop the culture until the dry weight of the algal cells no longer increases to obtain the zinc-rich thermophilic alga.

[0038] (3) Collect the zinc-rich thermophilic alga and dry it, add it to a muffle furnace, carbonize it at a temperature of 750 °C for a holding time of 3 h, and take out the sample after cooling.

[0039] (4) Add the carbonized thermophilic alga sample to the liquid material, adjust the pH to 6.5, stir at 25 °C for 5 h, centrifuge to obtain the supernatant, and concentrate it under reduced pressure through a microfiltration membrane to the same weight as the carbonized thermophilic alga sample, which is the zinc-rich thermophilic alga extract.

[0040] The thermophilic alga described in step (1) is Chlamydomonas.

[0041] The habitat water described in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, <0.05 mg / L of strontium, 0.011 mg / L of iron, 0.0012 mg / L of selenium, and the balance is water and inevitable impurity elements.

[0042] The mass ratio of the carbonized hot spring algae sample to the liquid material in step (4) is 1 g: 20 mL.

[0043] The liquid material in step (4) is a biomimetic mineralization agent solution, which is composed of amino acid, nicotinamide adenine dinucleotide and deionized water; the amino acid is glycine; the content of the amino acid is 0.5 mol / L, and the content of nicotinamide adenine dinucleotide is 0.01 mol / L.

[0044] The application of the zinc-rich hot spring algae extract is in skin care products for repair, anti-aging and oil control.

[0045] Example 3

[0046] A preparation method of a zinc-rich hot spring algae extract includes the following steps:

[0047] (1) Inoculate hot spring algae into BG11 medium with habitat water as the water source, adjust the temperature to 23 °C, the light intensity to 1700 lux, and the pH to 7.3, and conduct aeration culture. When the dry weight of the algal cells reaches 40 g / L, terminate the enrichment culture;

[0048] (2) Add zinc sulfate to the above medium so that the added zinc content is 2 mg / L, adjust the light intensity to 3000 lux, continue the culture and sample every day. Stop the culture until the dry weight of the algal cells no longer increases to obtain zinc-rich hot spring algae;

[0049] (3) Collect the zinc-rich hot spring algae and dry them, add them to a muffle furnace, carbonize them at a temperature of 700 °C for a holding time of 4 h, and take out the sample after cooling;

[0050] (4) Add the carbonized hot spring algae sample to the liquid material, adjust the pH to 5.5, stir at 30 °C for 4 h, centrifuge to obtain the supernatant, and concentrate it under reduced pressure through a microfiltration membrane until it has the same weight as the carbonized hot spring algae sample, which is the zinc-rich hot spring algae extract.

[0051] The hot spring algae in step (1) is Chlorella vulgaris.

[0052] The habitat water in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, <0.05 mg / L of strontium, 0.011 mg / L of iron, 0.0012 mg / L of selenium, and the balance is water and inevitable impurity elements.

[0053] The mass ratio of the carbonized hot spring algae sample to the liquid material in step (4) is 1: 20 mL.

[0054] The feed liquid described in step (4) is a biomimetic mineralizing agent solution, which is composed of amino acid, nicotinamide adenine dinucleotide and deionized water; the amino acid is glutamic acid; the content of the amino acid is 0.4 mol / L, and the content of nicotinamide adenine dinucleotide is 0.004 mol / L.

[0055] The application of the zinc-rich hot spring algae extract is in skin care products for repair, anti-aging and oil control.

[0056] Comparative Example 1

[0057] A preparation method of a zinc-rich hot spring algae extract includes the following steps:

[0058] (1) Inoculate hot spring algae into BG11 medium with deionized water as the water source, adjust the temperature to 22 °C, the light intensity to 1500 lux, and the pH to 7.0, and conduct aeration culture. End the enrichment culture when the dry weight of algal cells reaches 40 g / L;

[0059] (2) Add zinc sulfate to the above medium so that the added zinc content is 5 mg / L, adjust the light intensity to 2500 lux, continue to culture and take samples every day. Stop culturing until the dry weight of algal cells no longer increases to obtain zinc-rich hot spring algae;

[0060] (3) Collect the zinc-rich hot spring algae and dry them, add them to a muffle furnace, carbonize them at a temperature of 800 °C for a holding time of 2 h, and take out the sample after cooling;

[0061] (4) Add the carbonized hot spring algae sample to the feed liquid, adjust the pH to 6.0, stir at 25 °C for 5 h, centrifuge to obtain the supernatant, and concentrate it under reduced pressure through a microfiltration membrane to the same weight as the carbonized hot spring algae sample, which is the zinc-rich hot spring algae extract.

[0062] The hot spring algae described in step (1) is Chlamydomonas.

[0063] The ratio of the carbonized hot spring algae sample to the feed liquid described in step (4) is 1 g:20 mL.

[0064] The feed liquid described in step (4) is a biomimetic mineralizing agent solution, which is composed of amino acid, nicotinamide adenine dinucleotide and deionized water; the amino acid is glycine; the content of the amino acid is 0.2 mol / L, and the content of nicotinamide adenine dinucleotide is 0.006 mol / L.

[0065] The application of the zinc-rich hot spring algae extract is in skin care products for repair, anti-aging and oil control.

[0066] Comparative Example 2

[0067] A preparation method of a zinc-rich hot spring algae extract includes the following steps:

[0068] (1) Inoculate the hot spring algae into the BG11 medium with habitat water as the water source, adjust the temperature to 22 °C, the light intensity to 1500 lux, and the pH to 7.0, and conduct aerated culture. End the enrichment culture when the dry weight of the algal cells reaches 40 g / L;

[0069] (2) Add zinc sulfate to the above medium so that the added zinc content is 7 mg / L, adjust the light intensity to 2500 lux, continue the culture and take samples every day. Stop the culture when the dry weight of the algal cells no longer increases to obtain zinc-rich hot spring algae;

[0070] (3) Collect the zinc-rich hot spring algae and dry them, add them to a muffle furnace, carbonize them at a temperature of 800 °C for a holding time of 2 h, and take out the sample after cooling;

[0071] (4) Add the carbonized hot spring algae sample to the feed liquid, adjust the pH to 6.0, stir at 25 °C for 5 h, centrifuge to obtain the supernatant, filter it through a microfiltration membrane and concentrate it under reduced pressure to the same weight as the carbonized hot spring algae sample, which is the zinc-rich hot spring algae extract.

[0072] The hot spring algae described in step (1) are Chlamydomonas.

[0073] The habitat water described in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, <0.05 mg / L of strontium, 0.011 mg / L of iron, 0.0012 mg / L of selenium, and the balance is water and unavoidable impurity elements.

[0074] The ratio of the carbonized hot spring algae sample to the feed liquid described in step (4) is 1 g:20 mL.

[0075] The feed liquid described in step (4) is a biomimetic mineralization agent solution, which is composed of amino acids, nicotinamide adenine dinucleotide, and deionized water; the amino acid is glycine; the content of the amino acid is 0.2 mol / L, and the content of nicotinamide adenine dinucleotide is 0.006 mol / L.

[0076] The application of the zinc-rich hot spring algae extract is in skin care products for repair, anti-aging, and oil control.

[0077] Comparative Example 3

[0078] A preparation method of a zinc-rich hot spring algae extract, comprising the following steps:

[0079] (1) Inoculate the hot spring algae into the BG11 medium with habitat water as the water source, adjust the temperature to 22 °C, the light intensity to 1500 lux, and the pH to 7.0, and conduct aerated culture. End the enrichment culture when the dry weight of the algal cells reaches 40 g / L;

[0080] (2) Add zinc sulfate to the above medium so that the added zinc content is 5 mg / L. Adjust the light intensity to 2500 lux and continue the cultivation. Take samples every day until the dry weight of the algal cells no longer increases, then stop the cultivation to obtain zinc-rich Thermus thermophilus algae;

[0081] (3) Collect the zinc-rich Thermus thermophilus algae and dry them. Add them to a muffle furnace and carbonize them at a temperature of 600 °C for 2 hours of heat preservation. Take out the sample after cooling;

[0082] (4) Add the carbonized Thermus thermophilus algae sample to the feed liquid, adjust the pH to 6.0, stir at 25 °C for 5 hours, centrifuge to obtain the supernatant, and concentrate it under reduced pressure through a microfiltration membrane until it has the same weight as the carbonized Thermus thermophilus algae sample, which is the zinc-rich Thermus thermophilus algae extract.

[0083] The Thermus thermophilus algae described in step (1) is Chlamydomonas reinhardtii.

[0084] The habitat water described in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, <0.05 mg / L of strontium, 0.011 mg / L of iron, 0.0012 mg / L of selenium, and the balance is water and inevitable impurity elements.

[0085] The ratio of the carbonized Thermus thermophilus algae sample to the feed liquid described in step (4) is 1 g:20 mL.

[0086] The feed liquid described in step (4) is a biomimetic mineralization agent solution, which is composed of amino acids, nicotinamide adenine dinucleotide, and deionized water; the amino acid is glycine; the amino acid content is 0.2 mol / L, and the nicotinamide adenine dinucleotide content is 0.006 mol / L.

[0087] The application of the zinc-rich Thermus thermophilus algae extract is in skin care products for repair, anti-aging, and oil control.

[0088] Comparative Example 4

[0089] A preparation method of a zinc-rich Thermus thermophilus algae extract, comprising the following steps:

[0090] (1) Inoculate Thermus thermophilus algae into a BG11 medium using habitat water as the water source, adjust the temperature to 22 °C, the light intensity to 1500 lux, and the pH to 7.0, and conduct aeration cultivation. Stop the enrichment cultivation when the dry weight of the algal cells reaches 40 g / L;

[0091] (2) Add zinc sulfate to the above medium so that the added zinc content is 5 mg / L. Adjust the light intensity to 2500 lux and continue the cultivation. Take samples every day until the dry weight of the algal cells no longer increases, then stop the cultivation to obtain zinc-rich Thermus thermophilus algae;

[0092] (3) Collect the zinc-rich hot spring algae and dry them. Add them to a muffle furnace and carbonize them at a temperature of 800 °C for a holding time of 2 h. After cooling, take out the sample;

[0093] (4) Add the carbonized hot spring algae sample to the feed liquid, adjust the pH to 6.0, stir at 25 °C for 5 h, centrifuge to obtain the supernatant, and concentrate it under reduced pressure through a microfiltration membrane until it has the same weight as the carbonized hot spring algae sample, which is the zinc-rich hot spring algae extract.

[0094] The hot spring algae described in step (1) is Chlamydomonas.

[0095] The habitat water described in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, <0.05 mg / L of strontium, 0.011 mg / L of iron, and 0.0012 mg / L of selenium, with the balance being water and unavoidable impurity elements.

[0096] The ratio of the carbonized hot spring algae sample to the feed liquid described in step (4) is 1 g:20 mL.

[0097] The feed liquid described in step (4) is a biomimetic mineralization agent solution, which is composed of amino acids, nicotinamide adenine dinucleotide, and deionized water; the amino acid is glycine; the amino acid content is 0.1 mol / L, and the nicotinamide adenine dinucleotide content is 0.006 mol / L.

[0098] The application of the zinc-rich hot spring algae extract is in skin care products for repair, anti-aging, and oil control.

[0099] Control Example 5

[0100] A preparation method of a zinc-rich hot spring algae extract, comprising the following steps:

[0101] (1) Inoculate the hot spring algae into a BG11 medium using the habitat water as the water source, adjust the temperature to 22 °C, the light intensity to 1500 lux, and the pH to 7.0, and perform aeration culture. End the enrichment culture when the dry weight of the algal cells reaches 40 g / L;

[0102] (2) Add zinc sulfate to the above medium so that the added zinc content is 5 mg / L, adjust the light intensity to 2500 lux, continue the culture and take samples every day, and stop the culture until the dry weight of the algal cells no longer increases to obtain the zinc-rich hot spring algae;

[0103] (3) Collect the zinc-rich hot spring algae and dry them. Add them to a muffle furnace and carbonize them at a temperature of 800 °C for a holding time of 2 h. After cooling, take out the sample;

[0104] (4) Add the carbonized hot spring algae sample to the feed solution, adjust the pH to 6.0, stir at 25 °C for 5 h, centrifuge to obtain the supernatant, and concentrate it under reduced pressure through a microfiltration membrane until it has the same weight as the carbonized hot spring algae sample, which is the zinc-rich hot spring algae extract.

[0105] The hot spring algae described in step (1) is Chlamydomonas.

[0106] The habitat water described in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, <0.05 mg / L of strontium, 0.011 mg / L of iron, 0.0012 mg / L of selenium, and the balance is water and inevitable impurity elements.

[0107] The ratio of the carbonized hot spring algae sample to the feed solution in step (4) is 1 g:20 mL.

[0108] The feed solution described in step (4) is a biomimetic mineralization agent solution, which is composed of amino acids, nicotinamide adenine dinucleotide, and deionized water; the amino acid is glycine; the amino acid content is 0.2 mol / L, and the nicotinamide adenine dinucleotide content is 0.002 mol / L.

[0109] The application of the zinc-rich hot spring algae extract is in skin care products for repair, anti-aging, and oil control.

[0110] Comparative Example 6

[0111] A preparation method of a zinc-rich hot spring algae extract, comprising the following steps:

[0112] (1) Inoculate the hot spring algae into the BG11 medium with habitat water as the water source, adjust the temperature to 22 °C, the light intensity to 1500 lux, and the pH to 7.0, and perform aeration culture. When the dry weight of the algal cells reaches 40 g / L, the enrichment culture ends;

[0113] (2) Add zinc sulfate to the above medium so that the added zinc content is 5 mg / L, adjust the light intensity to 2500 lux, continue to culture and sample every day, and stop culturing until the dry weight of the algal cells no longer increases to obtain zinc-rich hot spring algae;

[0114] (3) Collect the zinc-rich hot spring algae and dry them, add them to the feed solution, adjust the pH to 6.0, stir at 25 °C for 5 h, centrifuge to obtain the supernatant, and concentrate it under reduced pressure through a microfiltration membrane until it has the same weight as the carbonized hot spring algae sample, which is the zinc-rich hot spring algae extract.

[0115] The hot spring algae described in step (1) is Chlamydomonas.

[0116] The habitat water described in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, < 0.05 mg / L of strontium, 0.011 mg / L of iron, 0.0012 mg / L of selenium, and the balance is water and inevitable impurity elements.

[0117] The ratio of the carbonized hot spring algae sample to the liquid material described in step (4) is 1 g:20 mL.

[0118] The liquid material described in step (4) is a biomimetic mineralization agent solution, which is composed of amino acid, nicotinamide adenine dinucleotide and deionized water; the amino acid is glycine; the content of the amino acid is 0.2 mol / L, and the content of nicotinamide adenine dinucleotide is 0.006 mol / L.

[0119] The application of the zinc-rich hot spring algae extract is in skin care products for repair, anti-aging and oil control.

[0120] Comparative Example 7

[0121] A preparation method of a zinc-rich hot spring algae extract, comprising the following steps:

[0122] (1) Inoculate the hot spring algae into the BG11 medium with the habitat water as the water source, adjust the temperature to 24 °C, the light intensity to 1700 lux, and the pH to 7.2, and conduct aeration culture. When the dry weight of the algal cells reaches 40 g / L, the enrichment culture ends;

[0123] (2) Add zinc sulfate to the above medium so that the added zinc content is 3 mg / L, adjust the light intensity to 2800 lux, continue to culture and take samples every day. When the dry weight of the algal cells no longer increases, stop the culture to obtain zinc-rich hot spring algae;

[0124] (3) Collect the zinc-rich hot spring algae and dry them, add them to a muffle furnace, and carbonize them at a temperature of 750 °C for a holding time of 3 h. After cooling, take out the sample;

[0125] (4) Add the carbonized hot spring algae sample to the liquid material, adjust the pH to 6.0, stir at 25 °C for 5 h, centrifuge to obtain the supernatant, and concentrate it under reduced pressure through a microfiltration membrane to the same weight as the carbonized hot spring algae sample, which is the zinc-rich hot spring algae extract.

[0126] The hot spring algae described in step (1) is Chlamydomonas.

[0127] The habitat water described in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, < 0.05 mg / L of strontium, 0.011 mg / L of iron, 0.0012 mg / L of selenium, and the balance is water and inevitable impurity elements.

[0128] The mass ratio of the carbonized hot spring algae sample to the liquid material described in step (4) is 1:22.

[0129] The feed liquid described in step (4) is a biomimetic mineralizing agent solution, which is composed of amino acids and deionized water; the amino acid is glycine; the content of the amino acid is 0.307 mol / L.

[0130] The application of the zinc-rich hot spring algae extract is in skin care products for repair, anti-aging, and oil control.

[0131] Comparative Example 8

[0132] A preparation method of a zinc-rich hot spring algae extract includes the following steps:

[0133] (1) Inoculate hot spring algae into BG11 medium with habitat water as the water source, adjust the temperature to 24 °C, the light intensity to 1700 lux, and the pH to 7.2, and conduct aeration culture. End the enrichment culture when the dry weight of the algal cells reaches 40 g / L.

[0134] (2) Add zinc sulfate to the above medium so that the added zinc content is 3 mg / L, adjust the light intensity to 2800 lux, continue the culture and take samples every day. Stop the culture until the dry weight of the algal cells no longer increases to obtain zinc-rich hot spring algae.

[0135] (3) Collect the zinc-rich hot spring algae and dry them, add them to a muffle furnace, carbonize them at a temperature of 750 °C for a holding time of 3 h, and take out the sample after cooling.

[0136] (4) Add the carbonized hot spring algae sample to the feed liquid, adjust the pH to 6.0, stir at 25 °C for 5 h, centrifuge to obtain the supernatant, filter it through a microfiltration membrane, and concentrate it under reduced pressure to the same weight as the carbonized hot spring algae sample, which is the zinc-rich hot spring algae extract.

[0137] The hot spring algae described in step (1) is Chlamydomonas.

[0138] The habitat water described in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, <0.05 mg / L of strontium, 0.011 mg / L of iron, 0.0012 mg / L of selenium, and the balance is water and unavoidable impurity elements.

[0139] The mass ratio of the carbonized hot spring algae sample to the feed liquid described in step (4) is 1:22.

[0140] The feed liquid described in step (4) is a biomimetic mineralizing agent solution, which is composed of nicotinamide adenine dinucleotide and deionized water; the content of nicotinamide adenine dinucleotide is 0.307 mol / L.

[0141] The application of the zinc-rich hot spring algae extract is in skin care products for repair, anti-aging, and oil control.

[0142] Performance test:

[0143] Zinc content test: Referring to the method of GB 5009.14-2017, the zinc content in the hot spring algae extract in Examples 1-3 and Comparative Examples 1-8 was detected.

[0144] Repair performance test: Prepare HaCaT cells in the logarithmic growth phase and inoculate them into the culture plate at a cell density of 6×10 5 cells. Incubate in a constant temperature incubator at 37°C and 5% (v / v) CO2 for 24 h. Use a pipette tip to scratch the cells, wash the cells 3 times with PBS, and then add 2 mL of a 0.5 wt% sample deionized aqueous solution and continue to culture for 24 h. Use an inverted microscope to take pictures of the migrated cells in each group, and use image pro plus software to calculate the average value of the scratch area. The cell migration rate formula for each group is as follows:

[0145] Cell migration rate = (Initial scratch area - Final scratch area) / Initial scratch area × 100%.

[0146] Anti-aging performance test: Test through an in vitro experiment to promote the production of type I collagen. Implant 2 ml of human skin fibroblasts cultured to the logarithmic phase into a culture plate with a cover glass at a concentration of 3×10 5 cells / ml. The cover glass was treated with polylysine before culture. After culturing with DMEM culture medium containing 10% FBS for 24 h, add the sample solutions of Examples 1-3 and Comparative Examples 1-8 at a mass percentage concentration of 0.1%, and add an equal volume of serum to the control group. After culturing for 7 d, detect the production of type I collagen and calculate the growth rate.

[0147] Oil control effect test: Select 55 volunteers with oily skin and randomly divide them into 11 groups. Use the 0.5 wt% sample aqueous solutions prepared in Examples 1-3 and Comparative Examples 1-8 to replace the lotion they usually use, and use it twice a day, morning and evening, for 28 consecutive days. Then use a Sebumeter SM 815 sebum tester to measure the change in the average facial sebum content of the volunteers.

[0148]

[0149]

[0150] It can be seen from the test results that in the present invention, zinc-rich hot spring algae extract is obtained by using zinc-containing habitat water as the culture medium water source for zinc-rich culture of hot spring algae and carbonization combined with bionic extraction. The cell migration promotion rates of the hot spring algae extracts obtained in Examples 1-3 are significantly higher than those treated in Comparative Examples 1-8, indicating that the hot spring algae extract of the present invention has excellent repair efficacy; the promotion effect on collagen expression is much higher than that in Comparative Examples 1-8, indicating that the hot spring algae extract prepared by the method of the present invention has better anti-aging efficacy; and it can effectively reduce the sebum content of oily skin.

[0151] The above are only the preferred embodiments of the present invention, and do not impose any form of limitation on the present invention. Although the present invention has been disclosed above with the preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some changes or modifications to equivalent embodiments by using the technical content disclosed above within the scope of the technical solution of the present invention. However, as long as it does not depart from the content of the technical solution of the present invention, any indirect modification, equivalent change and modification made to the above embodiments based on the technical essence of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A preparation method of zinc-rich hot spring alga extract, characterized in that: It includes the following steps: (1) Inoculate thermophilic algae into BG11 medium with habitat water as the water source, adjust the temperature to 22 - 25 °C, light intensity to 1500 - 2000 lux, pH to 7 - 7.5, and conduct aerated cultivation. End the enrichment cultivation when the dry weight of algal cells reaches 40 g / L; (2) Add zinc sulfate to the above medium so that the added zinc content is 2 - 5 mg / L, adjust the light intensity to 2500 - 3000 lux, continue the cultivation and sample every day. Stop the cultivation when the dry weight of algal cells no longer increases to obtain zinc-rich thermophilic algae; (3) Collect the zinc-rich thermophilic algae and dry them, then put them into a muffle furnace, carbonize at a temperature of 700 - 800 °C for a heat preservation time of 2 - 4 h, and take out the sample after cooling; (4) Add the carbonized thermophilic algae sample to the feed liquid, adjust the pH to 5.0 - 6.5, stir at 20 - 30 °C for 4 - 6 h, centrifuge to obtain the supernatant, filter it through a microfiltration membrane and then concentrate it under reduced pressure to the same weight as the carbonized thermophilic algae sample, which is the zinc-rich thermophilic algae extract.

2. The preparation method of a zinc-rich hot spring alga extract according to claim 1, characterized in that: The thermophilic algae described in step (1) is one of Chlamydomonas and Chlorella.

3. The preparation method of a zinc-rich hot spring alga extract according to claim 1, characterized in that: The habitat water described in step (1) contains 0.008 mg / L of zinc, 0.487 mg / L of silicon, < 0.05 mg / L of strontium, 0.011 mg / L of iron, 0.0012 mg / L of selenium, and the balance is water and inevitable impurity elements.

4. The preparation method of a zinc-rich hot spring alga extract according to claim 1, characterized in that: The mass ratio of the carbonized thermophilic algae sample to the feed liquid described in step (4) is 1 g: 20 - 25 mL.

5. The preparation method of a zinc-rich hot spring algal extract according to claim 1, characterized in that: The feed liquid described in step (4) is a biomimetic mineralization agent solution, which is composed of amino acid, nicotinamide adenine dinucleotide and deionized water; the amino acid is one of glycine, glutamic acid and methionine; the content of the amino acid is 0.2 - 0.5 mol / L, and the content of nicotinamide adenine dinucleotide is 0.004 - 0.01 mol / L.

6. Use of a zinc-rich hot spring alga extract prepared by the preparation method according to any one of claims 1-5, characterized in that: It is applied to skin care products for repair, anti-aging and oil control.

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