all technical and scientific terms used in the invention , unless otherwise specified , have the common meanings that are understood by person skilled in the art . the foregoing detailed description of the invention and the specific examples are provided herein for the purpose of illustration only , and the invention is not limited to the preferred embodiments shown . it should be understood that any changes or modifications within the spirit of the invention shall be included in the scope of present invention . take 80 g of the dried powder of sea grape and 1 , 600 g of distilled water , the sea grape used is caulerpa lentillifera , blend thoroughly ( the temperature is 85 ° c ., the time is 90 minutes ), followed by filtration using 5 - μm and 1 - μm filter membranes to give the fluid of sea grape extract ( i ); the obtained fluid is then subjected to the second filtration ; the first membrane is the ew membrane made by ge and microfiltration eliminates large particles ; the second membrane is the pw membrane made by ge and ultrafiltration eliminates tiny particles ; the third membrane is the dk membrane made by ge and nanofiltration eliminates salts ; the filtrate is then freeze - dried after filtration to give the sea grape extract ( i ). the abovementioned caulerpa lentillifera may be replaced by other species of sea grapes such as caulerpa taxifolia . release of the allergic mediator , β - hexosaminidase , of the cells treated with only sea grape extract ( i ) or refined sea grape extract ( i ) in the absence of the stimulant ( a23187 (−)) ( fig1 ). comparisons of the sea grape extract ( i ) alone group , refined sea grape ( i ) alone group and the control group . use of the sea grape extract ( i ) alone and the refined sea grape ( i ) alone did not increase the release of β - hexosaminidase , suggesting the sea grape extract ( i ) and refined sea grape ( i ) do not trigger allergy . calcium ionophore a23187 stimulates degranulation of cells to release the allergic mediator , β - hexosaminidase , the concentration of β - hexosaminidase is determined by enzyme - linked immunosorbent assay ( elisa ). from the ctr results of fig1 , calcium ionophore a23187 induces release of β - hexosaminidase . according to the results of fig2 , the sea grape extract ( i ) can inhibit release of β - hexosaminidase induced by calcium ionophore a23187 ( a23187 (+) group ), 2 mg / ml of the sea grape extract ( i ) showed 8 % reduction of the released β - hexosaminidase and 4 mg / ml of the sea grape extract ( i ) showed 25 % reduction of the released β - hexosaminidase ; the β - hexosaminidase inhibition effect of refined sea grape extract ( i ) is more significant , the refined sea grape extract ( i ) at 1 mg / ml inhibited 45 % of β - hexosaminidase release , 4 mg / ml of the refined sea grape extract ( i ) inhibited 82 % of β - hexosaminidase release without affecting cell viability ( fig1 ). specific amount of distilled water is boiled to 50 ˜ 60 ° c . before addition of surfactant , mixed thoroughly . the emollient is added , mix thoroughly . stop heating . cool to 30 ° c . before addition of the refined sea grape extract ( i ), orange citrus sinensis , sodium anisate , mixed thoroughly to give the shampoo mousse containing the refined sea grape extract ( i ). the formula of the shampoo mousse is shown in table 1 . the ama laboratories inc . certified by the food and drug administration ( fda ) of the u . s . was retained to conduct the clinical trial to assess skin irritation and sensitivity of the shampoo mousse ( the formula is shown in table 1 ) containing the refined sea grape extract ( i ), the number of subjects enrolled is 52 , the number of subjects who completed the trial is 50 , the subjects were aged 20 - 68 years , and the gender as well as race are shown in table 2 . the procedure of the clinical trial : the shampoo mousse of table 1 was diluted 10 fold in water .). 0 . 2 ml of the diluted shampoo mousse sample onto the semi - occlusive , hypoallergenic patch . the patch is then applied directly to the skin of the infrascapulart regions of the back , to the right or left of the midline and the subject is dismissed with instructions not to wet or expose the test area to direct sunlight . after 24 hours the patch is removed by the subjects at home . the procedure is repeated until a series of nine consecutive 24 hour exposures have been made for every monday , wednesday , and friday for three consecutive weeks . in the event of an adverse reaction , the area of erythema and edema is measured . as shown in table 3 , a total of 50 subjects had completed the study . all subjects had no allergic reaction to the shampoo mousse containing refined sea grape extract ( i ), indicating the shampoo mousse containing refined sea grape extract ( i ) causes no skin irritation and no allergy . take 1 . 5 kg of fresh sea grape and 1 . 5 kg of distilled water , the sea grape used is caulerpa lentillifera , blend and mix thoroughly ( temperature is 85 ° c ./ time is 30 minutes ), followed by filtration using 5 - μm and 1 - μm filter membranes to give the fluid of sea grape extract ( ii ); the sea grape extract ( ii ) was then subjected to further filtration ; the first membrane is the ew membrane made by ge and microfiltration eliminates large particles ; the second membrane is the pw membrane made by ge and ultrafiltration eliminates tiny particles ; the third membrane is the dk membrane made by ge and nanofiltration eliminates salts ; the filtrate is then freeze - dried to give the refined sea grape extract ( ii ). the abovementioned caulerpa lentillifera may be replaced by other species of sea grapes , such as caulerpa taxifolia . based on the results of fig3 , the refined sea grape extract ( ii ) can inhibit the release of β - hexosaminidase induced by calcium ionophore a23187 ( a23187 (+) group ), 1 mg / ml of the refined sea grape extract ( ii ) can inhibit 40 % of β - hexosaminidase release , 2 mg / ml of the refined sea grape extract ( ii ) can inhibit 60 % of β - hexosaminidase release , and 4 mg / ml of the refined sea grape extract ( ii ) can inhibit 80 % of β - hexosaminidase release . tale 1 . 5 kg of fresh sea grape and 1 . 5 kg of distilled water , the sea grape used is caulerpa lentillifera , blend thoroughly ( the temperature is around 25 ° c ./ time is around 60 min ), followed by filtration by using 5 - μm and 1 - μm filter membrane to give the sea grape extract ( iii ), the sea grape extract ( iii ) is then subjected to filtration ; the first membrane is the ew membrane made by ge and microfiltration eliminates large particles ; the second membrane is the pw membrane made by ge and ultrafiltration eliminates tiny particles ; the third membrane is the dk membrane made by ge and nanofiltration eliminates salts ; the filtrate was then freeze - dried to give the refined sea grape extract ( iii ). the abovementioned caulerpa lentillifera may be replaced by other species of sea grapes , such as caulerpa taxifolia . based on the results of fig3 , the refined sea grape extract ( iii ) can inhibit the release of β - hexosaminidase induced by calcium ionophore a23187 ( a23187 (+) group ), 1 mg / ml of the refined sea grape extract ( iii ) can inhibit 33 % of β - hexosaminidase release , 2 mg / ml of the refined sea grape extract ( ii ) can inhibit 56 % of β - hexosaminidase release , 4 mg / ml of the refined sea grape extract ( ii ) can inhibit 75 % of β - hexosaminidase release .
present invention relates to an allergy - inhibiting sea grape extract , its preparation method and thereof . the preparation method includes following steps : the sea grape is blended in distilled water in the weight ratio of 1 : 1 ˜ 20 , mix thoroughly , stir at 20 - 100 ° c . for 30 ˜ 120 minutes . the fluid obtained after the first filtration is the sea grape extract . the sea grape extract can be subjected to the second filtration by using ew , pw and dk membranes and freeze - dried to give a refined sea grape extract . the invention exhibits superior allergy - inhibiting effect ; moreover , the prepared extract does not cause allergy and can be used in cosmetics and food .