Composition for external skin application containing pine knot extract and method for preparation of the same
A technology of composition and extract, applied in skin care preparations, medical preparations containing active ingredients, drug combinations, etc., can solve problems such as adverse skin reactions and achieve the effect of preventing skin aging
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Embodiment 1
[0030] The turpentine is washed with pure water, dried and finely ground into a powder. Turpentine powder (100 g) was mixed with 50% aqueous ethanol (1 L). The mixture was extracted by boiling for 12 hours in an extractor equipped with a cooling condenser. The extract was filtered through a 300-mesh filter cloth. The filtrate was kept at 4-15°C for 7 days and filtered through Whatman No. 2 filter paper. The filtered extract was placed in a 3 L separating funnel and after addition of ethyl acetate (1 L) was mixed well by shaking. The mixture was left at normal temperature. After complete separation into two layers, the upper layer (ethyl acetate layer) was collected. Two more extractions were performed from the lower layer (aqueous layer) using a separating funnel. The collected upper layers were combined and concentrated at 50° C. under reduced pressure using a distillation apparatus equipped with a cooling condenser. After drying, 22.1 g (dry weight) of extract were obt...
experiment example 1
[0037] [Experimental example 1] Skin cell proliferation effect of pine knot extract
[0038] The skin cell proliferation effect of pine knot extract was tested. First, normal human fibroblasts were seeded on a 96-well microplate at 1×10 per well 4 cells. Seeded cells were cultured in Dulbecco's modified Eagle's medium (DMEM) for 24 hours. After replacing the medium with serum-free DMEM containing 500 μg / mL of the pine knot extract in Example 1 or the extracts in Comparative Examples 1-3, respectively, the cells were further cultured for 24 hours. After adding 5 mg / mL 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium (MTT) (10 μL per well), the medium was removed after 4 hours. After adding dimethyl sulfoxide (100 μL per well), the absorbance was measured at 570 nm with a microplate reader.
[0039] Cell proliferation was calculated by Equation 1 and the results are given in Table 1.
[0040] [equation 1]
[0041] Cell proliferation (%)=[(absorbance after treatment with...
experiment example 2
[0047] [Experimental Example 2] Stimulation of collagen biosynthesis
[0048] The collagen biosynthesis effect of the pine knot extract was tested as follows. Normal human fibroblasts were seeded on a 96-well microplate, 1×10 per well 4 cells. Seeded cells were cultured in DMEM for 24 hours. After replacing the medium with serum-free DMEM containing 500 μg / mL of the pine knot extract in Example 1 or the extracts in Comparative Examples 1-3, respectively, the cells were further cultured for 48 hours. 24 hours before the final incubation, 50 μg / mL ascorbic acid was added to promote collagen synthesis. Then, after washing each well, the cells were further cultured for 24 hours followed by replacement of the medium with serum-free DMEM. The supernatant of each well was collected, and the amount of collagen synthesis was measured using a collagenolytic IC-peptide (PICP) kit (Takara, Kyoto, Japan). The results are given in Table 2.
[0049] Table 2
[0050] [Table 2]
[0051...
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