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The invention relates to a sponge spicule, in particular to a preparation method of a high purity sponge spicule.
Sponge animals belong to porous animal doors, commonly known as sponges, which are the simplest multicellular animals. They have no clear organ structure, but the medium-gel layer is well developed, and the cells are pluripotent. Sponge cells exhibit a high degree of independence in sponges compared to other metazoans. Sponge mainly occupies life, and there is a unique ditch system in the body, which is the main channel for life activities such as feeding, breathing, excretion, and reproduction. Because the adult sponge camp is fixed or attached to life, it does not have the ability to exercise.
Sponge animals are widely distributed on the earth and have a huge biomass. It is estimated that there are about 15,000 species of lively species, and about 8,500 sponges have been recorded. Except for 150 kinds of freshwater sponges, the rest are marine species (Hooper, 2002). They survive in large numbers in all seas, including deep seas and polar seas. Sponge coatings on most coastal species grow on the surface of hard materials such as rocks, shells, wood in seawater or coral reefs, which reduces the risk of them being stripped and dismantled by the waves. Upright growing sponges are more common in deep seas where water flow is slow, and erect growth prevents them from being covered by large amounts of sediment in the deep ocean. Most sponge species live in shallower waters, and some species, including most of the six-discharge sponges, inhabit the deep ocean. The uncontaminated coastal and tropical coral reefs are the main living areas of the sponge.
Syringes are secreted by splenocytes in a sponge animal that can enrich and precipitate silicon in a structured manner or calcium. They are a special medium layer of cells that are distributed in the middle layer.
The spicula shape of the sponge is diverse and is an important basis for the classification and identification of sponges. The main component of the bone needle is silica, which accounts for more than 80% of the sponge species, and its main distribution area is shallow sea.
Since the shape of the sponge spicule varies with the type, the particle size is consistent, the number is large, and the composition is stable, and its utilization value is gradually taken seriously. At present, only the extraction method of sponge spicule for scientific research is well known to researchers , but this method exists less processing capacity and high cost. and the purity of the sponge spicule is not high, especially the special protein such as sponge keratin is not completely removed. Therefore, the method is only suitable for the extraction of the sponge spicule for the purpose of morphological classification.
The purpose of the present technology is to provide a method for preparing a high-purity sponge spicule because of the above-mentioned deficiencies in the conventional preparation of a sponge spicule.
This technology including the following step:
1) Washing the sponge and dry it;
2)The dried sponge is crushed, screened, and removing the large particles to get the sponge powder.
3)Adding at least one of acid or hypochlorite to the sponge powder, stirring, heating to 30-110 ° C and holding for 0.5-6 h, stopping the precipitation after heating, collecting the precipitated spicule, and obtaining the crude product of the spicule after washing ;The most important solid impurity in the crude product is sponge keratin;
4)The viscous agent having a mass concentration of 1% to 10% is prepared, and the crude spicule product obtained in the step 3) is mixed with the viscous agent, and a hydrogen peroxide solution having a mass concentration of 1% to 30% is added at the same time. Hydrogen peroxide reacts with sponge keratin in the crude spicule to produce bubbles on the surface of keratin;
5)The viscous agent containing hydrogen peroxide and crude spicule is placed in a vacuum container, and the size of the keratin surface bubble and the escape time are high and low adjustment by adjusting the degree of vacuum. After stewing, the impurities and the spicule are gradually layered. After the pressure is balanced and open the vacuum vessel. the impurities of the upper layer are removed, and the lower layer of the precipitate is washed to obtain a high-purity sponge spicule.
In the step 1), the sponge may be a cultured sponge, or a natural sponge having a mineral spicule obtained by means of natural sea area collection, or a sponge remaining from the active material; which washed can tear the sponge into 0.5 ~ 5.0 cm3 size, rinse with water to remove sediment, impurities, and symbiotic organisms; the drying can be removed by centrifugation or extrusion to remove excess water, and then dried or dried.
In step 2), the sieving may pass through a screen of 15 to 60 mesh.
In step 3), the acid may be selected from one of hydrochloric acid, nitric acid, sulfuric acid, phosphoric acid, etc.; the precipitation may be carried out by natural precipitation for 0.5 to 6 hours or centrifugation, and the centrifugal force of the centrifugation is <1000 g; Rinse with water for 1 to 4 times.
In step 4), the viscous agent may be selected from one of glycerin, cellulose, polyacrylic acid and the like.
The invention adopts a method of co-heat treatment of the sample with acid or hypochlorite, suspension of keratin with hydrogen peroxide bubbles, and the like, and significantly improves the acquisition rate and purity of the sponge spicule.
Specific mode of implementation
The remaining sponge residue was extracted from the sponge active product, and dried in a fume hood to remove the organic solvent. The powdery sponge residue was obtained by filtration through a 40-mesh, 80-mesh two-stage sieve. Hydrochloric acid was added to start the digestion according to the ratio of the sponge residue to the mass ratio of hydrochloric acid of 1:2, and the digestion vessel was sealed while heating in a boiling water bath. After 2 hours, the digested spicule was rinsed twice with water. Remove the spicule crude product. In the crude product, the content of impurities accounts for about 12% of the dry weight of the product. A 2% cellulose gel solution was prepared, 100 g of the crude spicule product was mixed with 50 mL of hydrogen peroxide, and 1 L of the newly prepared gel solution was quickly added, after stirred and dispersed, and then placed in a vacuum vessel and evacuated. After keeping the 10min in a vacuum state, the vacuum was quickly released, and the impurities floating on the surface were quickly taken out. The purity of the sponge spicule precipitated at the bottom of the gel solution can reach 95% to 98%.
Sample two: spicule precipitated at the bottom of the gel solution can reach 95% to 98%.
Use freshly obtained sponge as a material to tear, clean, and remove impurities and other creatures. The washed sponge is dried and smashed. A powdered sponge residue was obtained through a 20-mesh and 60-mesh two-stage sieve. According to the mass volume ratio of sponge powder to sodium hypochlorite at 1:4, sodium hypochlorite solution was added, heated to 60 ℃ to 70℃ in the ventilation system, and stirred continuously during digestion. After 4 h, the heating was stopped, and it was naturally cooled and precipitated. The supernatant was poured out and the precipitate was washed 2 to 3 times. The purity of the sponge spicule obtained at this time is about 82%. Prepare of 6% alginic acid gel solution 2L, stir 500g of crude sponge spicule and 100mL of hydrogen peroxide, quickly put into the gel solution, After stir and disperse, it was placed in a vacuum vessel and evacuated. Keep the vacuum for 20 minutes, the vacuum state was quickly released, and the impurities floating on the surface were taken out. The purity of the spicule can reach 98% by microscopic examination.
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