CN1436524A - Application of bamboo leaf general flavone as antisenility skin care factor in cosmetics - Google Patents

Abstract

The present invention aims at providing one kind of natural plant-originated antisenility skin care factor for cosmetics preparation. Bamboo leaf general flavone has the beautifying effect of resisting free radical, resisting oxidation, resisting radiation, resisting bacteria, bacteriostasis, promoting proliferation of skin cells, preventing skin from oxidation damage, reducing melanin synthesis and delaying skin senility; and it has special scent,no irritation to skin and no allergic reaction. It may be applied in various cosmetics, such as nourishing agent, sun block skin care agent, anti-wrinkle whitening agent, face cleaning cream, shampoo, bathing cream, etc.

Description

Folium Bambusae total flavones is as the application of the defying age skin protection factor in cosmetics

Technical field

The invention belongs to the daily cosmetics field.The present invention relates to a kind of Folium Bambusae total flavones as the application of the defying age skin protection factor in cosmetics, purpose is the defying age skin protection factor that a kind of natural plants source is provided for cosmetic industry.Specifically have free radical resisting, antioxidation, radioprotective, antibiotic, antibacterial and whiten, the Folium Bambusae total flavones of crease-resistant, spot-removal function is as the application of the defying age skin protection factor in daily cosmetics.

Background technology

Cosmetics are popular very strong commodity, and its consumption follows social current closely and moves, and have distinct characteristics of the times.Along with going deep into of medical research, consumers in general hold more careful attitude to the cosmetics that use the synthetics manufacturing, and in recent years, Cosmetic Manufacture begins naturalization of raw material, form world's upsurge of exploitation natural resources.The lifting of the quality of the life that is accompanied by economic development and comes; make how the slow down aging process becomes the most popular topic; everybody wishes to have health, beautiful, young skin, and what need is the skin care item that can not only protect skin and help delaying decrepitude of skin.Thereby current skin care item market has occurred more and more for alleviating the cosmetics that aging sign is developed.

Skin is to safeguard human life and healthy pith, directly contacts with extraneous, and be the first road barrier of body, very easily be subjected to Effect of Environmental and cause damaging.The aging action that brings except the age, stress, pollution, sunlight and ultraviolet irradiation all are to cause that the major reason of wrinkle or gauffer appears in skin.

Skin aging mainly shows two aspects: the 1. skin formation wrinkle that follows the string gradually; 2. skin appearance produces various mottles such as chloasma, senile plaque gradually.According to the free-radical theory of aging, in the process of skin aging, the damage of radical pair skin is a key factor.Reactive oxygen free radical and unsaturated fatty acid effect excessive in the body generate malonaldehyde materials such as (MDA), and effect such as the protein on MDA and the cell membrane generates brown pigment, and being deposited in just becomes various mottles on the skin.Free radical also can make skin interior collagen fiber, elastic fiber interlinkage and degeneration, become fragile, follows the string, and when the water deficient in the skin, makes the elastic fiber fracture easily and forms wrinkle.The profound reason of skin aging comprises that also the renewal metabolism of epidermal keratinocytes slows down, and dermal fibroblast multiple fission speed reduces.The old people compares with adolescence, the epiderm skin attenuation, and skin keratin forms cell and fibroblast obviously reduces, and therefore promotes that the propagation of old and feeble Skin Cell is one of pursuing one's goal of many skin care item.If chemicals can reduce the generation of skin free radical; improve the activity of SOD; again Skin Cell had certain proliferation function; skin histology is upgraded accelerate, collagen fiber generate and increase, the moisture of protection skin; thereby slow down skin aging [Huang Hansheng effectively; the anti-aging skin care item, daily chemical industry collected translation, 1994 (2): 35-38].

The skin care item industry is consistent to be thought, fruitful ground nursing skin must be noted that following 2 points: 1. should bring into use the moistening skin care item in one's early years; 2. must defend skin to be subjected to ultraviolet injury.Although obtained certain progress alleviating the technical elements that the skin microgroove manifests, the best way remains appearance that delays wrinkle and the appearance that as far as possible reduces wrinkle.The Mausner of Chanel company thinks, the aging of social population and the deterioration of environment will cause the aging of people's skin and be on the rise, and that people will more and more need will be long-acting, modulability and preventative skin care item efficiently.Elizabeth Arden company thinks, the trend of skin care item industry is that the direction to Cosmeceuticals (meaning is functional nutrient type cosmetics) develops.

The residing special industry background of the characteristics of the times of aged tendency of population and cosmetics provides wide space for the application of plant extract with physiologically actives such as free radical resisting, antioxidation, radioprotective, antibiotic, antiinflammatories.The chain reaction that brings of bovine spongiform encephalopathy in recent years in addition, the animal derived skin protection factor very easily causes the psychological fear of people, and is therefore growing to the demand of phytochemistry element.The vegetalitas cosmetic cosmetics of having released both at home and abroad have kind more than 50 by the floristics branch that adds, that concrete effect can reduce is wrinkle resistant, whiten, speckle dispelling, antipruritic, preserve moisture, aspect such as fat-reducing.The extract of these natural plants has characteristics such as side effect is little, curative effect is good, and the cosmetics of making had both kept the characteristic of cosmetics, had good curing, health-care effect again.

Flavone compound (Flavonoids is hereinafter to be referred as flavonoid) is distributed widely in plant kingdom, up to the present, has found kind more than 2000, and most forms with glycosides exist, and minority exists with free state, and its physiological action and biological activity are diversified.Conjugacy owing to its structure all shows strong absorption to ultraviolet and visible light, and stablizes at visible and ultra-violet (UV) band inner height.That the flavonoid of natural origin also has is antibiotic, anti-photosensitive, antioxidation, detoxify and effect such as brighten, can remove the free radical in the skin, avoid the radical pair cells injury, promote the metabolism of skin, improve the elasticity of skin, the generation of delaying skin wrinkle, the calmness of minimizing pigment, moist skin, reach significant whitening effect [Wang Jianguo, Liu Haifeng, Wang Jianxin, the research that sunscreen Parsol1789 photolysis suppresses, fragrance flavor and cosmetic, 2002 (1): 17-20].For example, extensively be present in Quercetin in vegetable and the fruit and have free radical resisting, antioxidation, antibiotic, antiviral and antiinflammatory, antianaphylactic biological activity and pharmacological action.Luteolin has very strong inhibitory action to H.Sui s virus, and concentration can suppress the growth of staphylococcus and bacillus subtilis at 1: 35000 o'clock, bacterium such as mucositis, white beads, distortion are also had inhibitory action; Antiinflammatory usefulness is arranged, can treat dermatitis with the phospholipid compatibility; Penetration on skin is strong, but hyaluronidase inhibitor is used the physiologically active that can prolong this material together with hyaluronic acid; Can suppress and eliminate skin splash (as senile plaque).Can make the fingernail glazed layer keep smooth with a small amount of luteolin (0.01%) in the nial polish.Rutin has antioxygen, antiinflammatory, anti-Stomatovirus effect.Rutin cooperates with 50: 1 with limonene has good inhibition effect to active oxygen, can powerful absorb ultraviolet, can be used for sun-proof, brighten, hair care category cosmetics [Wang Jianxin, natural activity cosmetics, China Light Industry Press, 1997:214].In the whitening type cosmetics, add 0.1～0.2% soybean isoflavone and can prevent skin aging, melanism and prevention brown patch; Adding 2% can absorb ultraviolet and keep hair gloss in suppurative mastitis.Radix Consolidae in the anthocyan have very strong antioxygen and antiinflammatory action, can suppress scalp or skin and excessively cause the empty courses of infection that causes of skin because of sebum secretion, and consumption is 1% in the astringent; The asterin that obtains from China aster adds in astringent can provide denseness and soft and smooth sense preferably, and deodorant, astringent effect [Wang Qiuan, the physiological function of natural flavone compounds and application thereof, fragrance flavor and cosmetic, 1999 (1): 28-33] are arranged.[recognize entirely by Huang Xingyu, fourth because of various active that it had and nontoxic, harmless safety thereof have wide practical use in medicine, food and cosmetics for flavonoid, Quercetin and derivant thereof effect and the mechanism in cosmetics, fragrance flavor and cosmetic, 1998 (3): 17-18].

The present plant flavone class extract of industrialization development is as Semen Ginkgo extrac, tea polyphenols, Semen Vitis viniferae extract, Cortex Pini Massonianae extract, licoflavone, soybean isoflavone, Radix Puerariae isoflavone etc., many cosmetic industries that have been applied to.For example, over nearly 30 years, Semen Ginkgo has been carried out research extensively and profoundly both at home and abroad, many patentability medicines, cosmetics and health product [Liu Yan, Chen Jinghua, the applied research of Semen Ginkgo in anti-aging cosmetics, daily chemical industry have been developed, 2002,32 (4): 79-81].Ginkgetin is strong reactive oxygen free radical scavenger, can protect Skin Cell not to be subjected to snperoxiaized the influence, thereby prolongs the life-span of Skin Cell, strengthens its antidotal ability; Bilobalide also can quicken metabolism, improves blood circulation, strengthens cell viability; In addition, Semen Ginkgo extrac also has broad-spectrum bactericidal action, common pathogenic bacterium of infecting skin such as staphylococcus aureus, bacillus pyocyaneus and acrothesium floccosum are all had the obvious suppression effect, and this antibacterial action can show just when extremely low concentration.The feature of this high-efficiency antimicrobial as the functional raw material of cosmetics, is advantageous conditions [Japan: the spy opens flat 5-70333,1993-03-23 for little removing thousand spring etc., skin preparations for extenal use] very for it.

In China, add the tea extract history of existing more than ten years in toothpaste and the shampoo.Now, Mitsui agricultural company has released the multiple product that contains tea polyphenols (tea catechin class), is used for foods and cosmetics [the former man of virtue and ability of levying etc., food industry, 1995,38 (2): 71].Flavonoid in the tea mainly is flavonol and flavonol, and its major function is antioxidation, anticancer, antimicrobial and deodorization.Catechin belongs to flavanol compound, accounts for 20～30% of green tea dry weight, and main flavonol comprises Quercetin, kaempferol and myricetin in the tea, accounts for 2～3% of tea dissolubility extract.Antibiotic and the deodorization of catechin has delayed rot (the anti-dental caries) of tooth, and improved the breath freshening degree, therefore, can in toothpaste, collutory, chewing gum and flavorants, add, also can be applicable in other article of everyday use, as all containing tea extract in many shampoos, moist cream, perfume and the sunscreen, it is believed that they can play sedation to skin, and prevent that as antioxidant skin is subjected to the attack [Jia Xudong of free radical, teas flavonoid function and application thereof, foreign medical science hygiology fascicle, 2001,28 (6): 369-371].

The main component of Radix Puerariae comprises osajin, triterpenoid saponin and alkaloid etc., and wherein of greatest concern is isoflavones components such as puerarin, daidzein.Isoflavone like substance in the Radix Puerariae is the catalytic activity of restraint of tyrosinase significantly, interrupts the melanocyte oxidizing process, suppresses the generation and the formation of melanocyte, thereby prevents chloasma, the plain deposition of day sunburn isochrome.So it is another skin depigmentation component that comes from green plants that Radix Puerariae is described as by international cosmetics circle, and be used for cosmetics by the Japan of cosmetics lead in the development of science and technology, Japanese Kao company is applied to fair complexion cream [Tang Chunhong, Chen Qi with Radix Puerariae isoflavone as active substance, the research and development present situation of domestic and international Pueraria lobota alimentary health-care function, Food Additives Used in China, 2002, (6): 56～58].

Flavonoids of Glycyrrhiza from the oil-soluble extract that licorice is obtained through refining has the acne of skin and to suppress active.The exploitation of its water soluble preparation has also obtained success, and the preventive and therapeutic effect of acne, acne has all been obtained proof on human body: the acne (antagonism androgen receptor) that acne that testosterone-the 5a reductase causes and androgen are caused all has interception; The microbial acne of acne is also had preventive and therapeutic effect, is that the acne of reason also has interception [Yamamoto is advanced, the new raw material information of cosmetics---the water soluble preparation of flavonoids of Glycyrrhiza, 2001, p12-13] to lipase and phosphorus lipase.

Other flavonoid such as hesperidin can prevent that ultraviolet from causing the Skin Cell lipid peroxidation and the erythema and the skin carcinoma that cause, this means that it can be used as the desirable feedstock of sun care preparations [Ou Shiyi, the pharmacological action of hesperidin, Chinese crude drug, 2002,25 (7): 531-533].Contain flavonoid glycoside, Coumarins, organic acid etc. in the Sarcandra glabra effective constituent, Jiangxi Herba Pileae Scriptae group company tries out [Hu Guoshun in products such as shampoo, cleansing milk, bath gel, sunscreen cream and toothpaste with its extract, the performance of sarcandra glabra fluid extract and the application in cosmetics, daily chemical industry, 2000 (4): 61-64].

China have the title of " bamboo kingdom ", has very abundant bamboo resource and bamboo culture of long standing and well established.Have within the border bamboo class more than 40 belong to 400 surplus kind, about 4,000,000 ha of bamboo grove area.According to incompletely statistics, China has that population more than 100,000,000 is all or part of to obtain living expenses from the processing of bamboo grove and bamboo grove product.Bamboo plant not only has higher economic value as the important component part of the forest reserves, and has ecology and social benefit widely.Bamboo is subject to people's attention day by day with characteristics such as its unique biological, ecology and multipurposes, is just bringing into play more and more important effect in the sustainable development of China strategy.

China is being in the leading level in the world aspect the research and development of bamboo effective ingredient.Folium Bambosae extract is a kind of plant flavonoids preparation that Zhang Ying equals the exploitation nineties in 20th century, and its patent of invention " a kind of health-care beer (ZL 98 1 04563.4) that adds bamboo leaf flavone " and " extracting the production method (ZL 98 1 04564.2) of flavonoid compound extract or powder from Folium Bambusae " were respectively at the mandate with calendar year 2001 acquisition State Patent Office in 2000.A large amount of studies show that, Folium Bambosae flavone has biological effectiveness such as good free radical resisting, antioxidation, defying age, antibiotic, antiviral and protection cardiovascular and cerebrovascular vessel, the old degenerative disease of control.And with the preparation quality and the pure and fresh fragrant and sweet bamboo local flavor of its abundant raw material sources, clear and definite functional factor, compellent safety, efficient stable, [Zhang Ying in recent years shows up prominently in functional food and Medicines and Health Product field, natural function additive---Folium Bambosae extract, meticulous and specialty chemicals, 2002,10 (7): 20～22].

The functional factor of Folium Bambosae extract mainly is a C-glucosides flavone, and four kinds of main Folium Bambusae carbon glycosides flavone are respectively orientin (Orientin), Lutonaretin (Homoorientin), apigenin-8-C-glucoside (Vitexin) and Saponaretin (Isovitexin).Carbon glycosides flavone is compared with oxygen glycosides flavone, has the outstanding advantage of following several respects: (1) Stability Analysis of Structures is difficult for being degraded; (2) can go deep into lesions position, directly bring into play curative effect; (3) hydrophilic strengthens, and helps food, medicine, cosmetic development.International academic community is paid close attention to carbon glycosides flavone since the nineties, this field belongs to up-to-date research forward position.

Japan starts from the seventies in 20th century to the research and development of bamboo effective ingredient, [claims bamboo grass (English) Bamboo grass, i.e. Sasaalbomarginta Makino ﹠amp but a large amount of work mainly concentrates on the class draft type bamboo with Japan's resource characteristic; Shibata etc.] on, relate to the Phyllostachys kind on a small quantity.NishinaAtsuyoshi etc. 1991 once detected antibacterial activity composition 2 from the peel of stem of Phyllostachys pubescens Mazei ex H.de Lebaie, 6-methoxyl group 1,4-benzoquinone [Jof Agric ﹠amp; Food Chem, 1991,39 (2): 266-269]; Applied for patent " the antibiotic paper that contains Folium Bambosae extract " [JP Patent 9961697] in 1997; Applied for again " containing V in 1998 _B1Food antibacterial with bamboo extract " patent [JP Patent 99269020].Watanabe Mayumi etc. applied for the patent [JP Patent 98310992] of " the antibiotic paper that contains bamboo grass extract " in 1997.YamanakaSatoshi etc. (1998) have reported the antibacterial activity and the application [Gekkan FudoKemikaru, 1998,14 (9): 57-60] thereof of bamboo dry distillation liquid.Nishina Atsuro etc. declared the patent [JP Patent 97278662] of " anti-allergic agent that contains bamboo extract " in 1996.Kiyooka Takatoshi (1995) has declared the patent [JP Patent 9794290] of " preparing deodorant with organic solvent extractionprocess from the bamboo skin ".Sato T. etc. reported that a kind of Folium Bambosae extract solution was used for the treatment of gingiva [NipponShishubyo Gakkai Kaishi, 1986,28 (2): 752-757] in 1986.Particularly what deserves to be mentioned is, the Japan Patent of KenjiMatsui etc. (nineteen ninety) application, point out to use one or more organic solvents, the effective ingredient that extracts from the bamboo of Phyllostachys such as mottled bamboo, Phyllostachys pubescens Mazei ex H.de Lebaie, golden Phyllostachys pubescens Mazei ex H.de Lebaie can suppress multiple dandruff and generate bacterium, prevent and treat skin aging, and can suppress lipid peroxidation and stable prod [JP Patent 03251518].But in this patent documentation, raw material sources only refer to bamboo (bamboo), do not refer to Folium Bambusae or full bamboo, and do not illustrate which kind of chemical compound effective ingredient is.Up to now, not spell out the bamboo effective ingredient relevant with such use be flavone compound to the document of Ri Ben patent and open report.

Open on the basis that English and partner thereof formerly study, recently again the Folium Bambusae total flavones function relevant with skin physiology carried out systematic study, mainly reduce the following aspects.

The antioxidation of 1 Folium Bambusae total flavones, anti-aging effects

Zhang Ying etc. had once studied henon bamboo [Phyllostachys nigra var.hnonis (Bean) Stepf exRendle] the antidotal biological effectiveness of leaf extract.The result shows that high dose group can significantly strengthen resistivity (normal pressure hypoxia endurance test, p＜0.01) and the anti-fatigue ability (swimming test, p＜0.01) of mice to nonspecific stimulation; Learning capacity to normal mouse has certain facilitation (electric maze method, p＜0.05); Activity to intravital superoxide dismutase of Aged Mice (SOD) and glutathione peroxidase (GSH-px) has remarkable inducing action; Suppressed significantly Aged Mice plasma lipid peroxide (LPO) generation, reduced the content of liver lipofuscin (LF); By measuring the variation of hydroxyproline content in mouse skin and the tail tendon; just demonstrate positive-effect during low dosage to collagen protein; showing has antioxidation, suppresses lipid peroxidation product, removes activity [Zhang Ying, the Tang Lili of lipofuscin and protection collagen protein; henon bamboo leaf extract experimental study about anti-senility; bamboo research transactions; 1997,16 (4): 62-67].

The free radical resisting of 2 Folium Bambusae total flavones, radiation resistance

Calendar year 2001, Master degree candidate---the Xu Bing of the biophysics of Zhejiang University specialty, by laboratory facilities such as faint chemiluminescence analysis and fluorophotometric analysiss, comparative study tea polyphenols, Semen Ginkgo extrac and Folium Bambusae total flavones three kind of plant flavonoid remove O ₂ ^-Active and right with OH ⁶⁰Co-r irradiation and CuSO ₄-Phen-Vc-H ₂O ₂The protective effect of DNA damage due to the OH in the-DNA system.

2.1 raw material

2.1.1 sample source

Tea polyphenols: content 99%, lot number 991227, Zhejiang University's agricultural provides with the Yang Xianqiang professor of biotechnology institute; Semen Ginkgo extrac: total flavones glucosides content 〉=24%, lot number 20000041, herbal pharmaceutical factory in Ningbo City's produces; Folium Bambusae total flavones: total flavones glucosides content 〉=24%, lot number 980625, Zhejiang University's biosystem engineering and professor Zhang Ying of Food Science institute provide.With tri-distilled water said sample is mixed with the stock solution of 1mg/mL respectively, stand-by.

2.1.2 medicine and reagent

Calf thymus DNA, hypoxanthine, xanthine oxidase, luminol, Phen (Phen), fluorescent probe (EB) are U.S. Sigma company product; The medicine of potassium nitrate, copper sulfate, preparation PBS and CBS and reagent etc. are analytical pure; The zymosan suspension is made by oneself from active dry yeast; Vc and hydrogen peroxide solution are now with the current.

2.1.3 instrument

RF-5000 type spectrofluorophotometer, day island proper Tianjin company produces; BPCL-II superweak luminescence instrument, Chinese Academy of Sciences's biophysics manufacturing; ⁶⁰The Co-r irradiation bomb is examined agricultural institute irradiation center by Zhejiang University and is provided.

2.2 experimental technique

2.1.1 remove reactive oxygen free radical (O ₂ ^-And OH) ability is measured and is adopted Vc-Cu respectively ²⁺-H ₂O ₂-zymosan system produces OH, hypoxanthine-xanthine oxidase-luminol system produces O ₂ ^-, suppressing with the sample of debita spissitudo scope, the superweak luminescence instrument is measured luminous value blank and the test system, calculates suppression ratio, and further calculates various sample to OH and O by equation of linear regression ₂ ^-503nhibiting concentration (IC ₅₀).

2.1.2 the protective effect to DNA damage is measured

2.1.2.1 EB-DNA system fluorescence intensity detection method

In 1mL fluorescence cup, add 25 μ g NDA and 2 μ g EB, add certain density sample solution again, insufficient section is supplied with the potassium nitrate solution of 10mmol/L, leaves standstill the mensuration of carrying out fluorescence intensity behind the 10min.Test condition is: slit Ex=5nm, Em=5nm, excitation wavelength 520nm, medium scanning speed, sweep limits is 550～720nm, measures near the fluorescence intensity (580.8nm) of emission maximum under the different condition, and calculates the suppression ratio I of sample to fluorescence intensity.

2.1.2.2 superweak luminescence detection method

With 0.1mol/L CBS (pH=10.5) preparation CuSO ₄-Phen-DNA solution 10mL (adds 10 ^-2The Phen 490 μ l, 10 of mol/L ^-2The CuSO of mol/L ₄70 μ l, 4 * 10 ^-4The DNA 35 μ l of g/mL), get above-mentioned solution 858 μ l, add 10 ^-2Mol/L Vc solution 42 μ l obtain the mixed liquor that volume is 900 μ l.After in mixed liquor, adding the sample of variable concentrations, supply with CBS that to make cumulative volume be 1000 μ l, insert in the light-emitting appearance measuring chamber.Last H with 200 μ l3% ₂O ₂Start reaction, write down chemiluminescent kinetic curve.

2.1.3 the irradiation damage of anti-DNA experiment

The dna solution (containing DNA100 μ g) that adds 0.4mL in 5mL finger-type plastic centrifuge tube adds the various sample of 0,20,40,80,160 μ g/mL, with the distilled water polishing to 2mL.The variable concentrations of each sample is all established three parallel samples, and above-mentioned serial vial is carried out the gamma-radiation irradiation. ⁶⁰The source strength of CO is about 144000Ci (April 12 calendar year 2001).Five dosage are established in experiment altogether, are respectively 0,0.8,2,8,20Gy, but identical close rate is all adopted in all processing, is 0.5Gy/min.After finishing, irradiation measures by the method among the 2.1.1 rapidly in the 2.5h.

2.3 result

2.3.1 remove O ₂ ^-Comparison with the OH ability

The performance that table 1 three kind of plant flavone extract samples are removed reactive oxygen free radical compares

The extract kind	Active constituent content (%)	Remove the IC of OH 50????(mg/mL)	Remove O 2 -IC 50????(μg/mL)
				Tea polyphenols	????99％	??????1.04	???????3.08
Semen Ginkgo extrac	????≥24％	??????1.43	???????3.43
				Folium Bambusae total flavones	????≥24％	??????1.55	???????3.59

Relatively three kinds of samples are removed O ₂ ^-With the ability of OH, Folium Bambusae total flavones and Semen Ginkgo extrac are very approaching, and tea polyphenols slightly is better than them, infer by relevant with its high-purity (table 1).

2.3.2 influence to EB-DNA system fluorescent characteristic

Three kind of plant flavone extracts are significant concentration dependence to the inhibition of EB-DNA system fluorescence intensity, from IC ₅₀Value compares, and Folium Bambusae total flavones and Semen Ginkgo extrac are very approaching, all are better than tea polyphenols (table 2).Three kinds of natural plants flavone extracts of table 2 sample is to the suppression ratio of EB-DNA system fluorescence intensity

The extract kind	Sample solution concentration (mg/mL)					????IC 50???(mg/mL)
							??0.25	???0.50	???0.75	???1.00	???1.25
	Tea polyphenols	??14.5％	???40.1％	???51.2％	???68.0％							???90.1％	?????0.71
Semen Ginkgo extrac						??37.8％	???51.7％	???61.7％	???69.5％	???82.1％	?????0.50
	Folium Bambusae total flavones	??36.1％	???51.0％	???59.8％	???64.2％							???81.4％	?????0.55

Flavone compound may with the EB competition binding site on dna molecular, the EB molecule that is embedded on the DNA base pair is pulled down, like this, the concentration of sample is high more, and is just few more in conjunction with the EB molecule on the DNA, the fluorescence intensity of EB-DNA is just low more.

2.3.3 protective effect to the DNA radiation damage

The gamma-irradiation of table 3 various dose is to the influence of EB-DNA fluorescent characteristic

Dosage (Gy)	??????0	???????0.8	????2	????????8	???????20
						Fluorescence intensity (λ 2＝580.8)	??180.25±8.2	???168.82±7.1	?151.55±7.5	??132.65±1.8	???122.43±3.2
Suppression ratio (%)	??????0	????????6.3	?????15.9	?????26.4	????????32.1

Table 3 shows that irradiation can produce direct effect and indirect effect to dna solution.Direct effect can cause that the dna molecular hydrogen bond destroys, Double helix is untied, base comes off, single double-strand break etc.; Indirect effect mainly is because free radical, the especially OH that is produced during irradiation attacks the DNA chain, causes the damage of dna molecular.The result causes EB and DNA binding site to reduce, and fluorescence intensity level also just reduces.

Table 4, table 5 and table 6 have shown the influence to the EB-DNA fluorescence intensity under various dose of variety classes, variable concentrations sample respectively.Therefore flavonoid can effectively be removed free radical, and the radical damage of the dna molecular that irradiation is caused has significant protective effect.Tea polyphenols, Semen Ginkgo extrac, Folium Bambusae total flavones are in the concentration range of 20～160 μ g/mL, under different dosage, all can make the fluorescence intensity of EB-DNA remain on higher level, illustrate that they have the effect of protection DNA irradiation damage, and the display density dependence.Look down upon greatly from the protective effect of three kinds of extracts, tea polyphenols slightly is better than Semen Ginkgo extrac and Folium Bambusae total flavones, and the latter two are close.

The influence of the tea polyphenols of table 4 variable concentrations EB-DNA fluorescence intensity during to various dose irradiation

Irradiation dose (Gy)	Sample solution concentration (μ g/mL)
						????0	????20	????40	?????80	?????160
	????0.8	??168.8±7.1	?173.1±7.7	??174.8±3.2	??175.9±8.9						??176.1±6.3
????2						??151.6±7.5	?164.6±8.2	??170.2±7.0	??172.1±4.5	???173.5±9.0
	????8	??132.7±1.8	?153.1±3.6	??159.7±6.2	??167.4±9.3						???170.1±6.7
????20						??122.4±3.2	?140.9±4.5	??144.8±5.4	??147.5±6.0	???151.1±5.8

The influence of the Semen Ginkgo extrac of table 5 variable concentrations EB-DNA fluorescence intensity during to various dose irradiation

Irradiation dose (Gy)	Sample solution concentration (μ g/mL)
						????0	????20	????40	????80	????160
	????0.8	??168.8±7.1	??170.3±2.6	???172.6±9.9	??173.7±7.5						??175.2±7.1
????2						??151.6±7.5	??160.4±8.8	???165.2±6.5	??168.7±3.9	??170.5±9.4
	????8	??132.7±1.8	??148.6±7.6	???154.3±3.4	??160.8±7.2						??162.6±5.8
????20						??122.4±3.2	??135.5±5.9	???138.2±8.1	??143.9±3.4	??146.6±4.6

The influence of the Folium Bambusae total flavones of table 6 variable concentrations EB-DNA fluorescence intensity during to various dose irradiation

Irradiation dose (Gy)	Sample solution concentration (μ g/mL)
						????0	????20	????40	????80	????160
	???0.8	??168.8±7.1	???169.9±2.6	??172.0±7.9	??173.3±2.8						???174.5±7.3
????2						??151.6±7.5	???157.9±8.7	??161.3±6.6	??166.2±9.1	???168.5±8.4
	????8	??132.7±1.8	???150.7±6.4	??152.5±5.7	??157.3±7.4						???160.2±6.5
????20						???122.4±3.2	???130.2±9.7	??134.6±2.1	??140.3±4.8	???142.5±2.9

2.3.4 to CuSO ₄-Phen-Vc-H ₂O ₂-DNA chemiluminescence effect of kinetics

CuSO ₄-Phen-Vc-H ₂O ₂The chemical luminophor of-DNA ties up in the alkaline medium, Phen (Phen) under the catalysis of metal ion, can and H ₂O ₂Effect produces chemiluminescence, and its maximum emission wavelength is in 445～450nm scope.According to pertinent literature, O ₂Be to cause that Phen generates the principal element of chemiluminescent substance, is in preceding peak position on the luminescence kinetics curve.Simultaneously, the OH that produces in the system causes the DNA damage fracture, produces one and postpones the luminous slow chemiluminescence in Phen itself, and maximum emission wavelength is between 380～420nm, this is luminous to be the characteristic reaction of free guanine, is in the peak position, back on the luminescence kinetics curve.

This luminescence system is under the situation that tea polyphenols, Semen Ginkgo extrac, three kinds of samples of Folium Bambusae total flavones exist, and leading peak, back peak-to-peak value descend quite obvious, and its 503nhibiting concentration is about 0.2 μ g/mL.The decline that causes luminescence peak be since antioxidant for clearing start the OH etc. of chain reaction, reduced the DNA damage that .OH etc. causes, think that flavonoid has played the effect of prevention type antioxidant.Find out also that simultaneously the time of occurrence of preceding peak-to-peak value does not almost change, then peak-to-peak value lags behind, and with the increase of antioxidant concentration, shift time prolongs behind the peak value.Think that causing the time-delay of postpeak luminescence peak is because sample has been removed the intermediate state free radical of free chain reaction, chain reaction is interrupted, play the effect of protection DNA radical damage, illustrate that flavonoid has been the antioxidant of chain scission.Proved that flavonoidss such as tea polyphenols, Semen Ginkgo extrac, Folium Bambusae total flavones are simultaneously as one-level antioxidant and secondary antioxidants and the [Xu Bing that works; several Natural antioxidants are to protective effect and the molecular radiobiology Study on Mechanism thereof of DNA; Zhejiang University's master thesis, 2001].

Antibacterial, the antiinflammatory action of 3 Folium Bambusae total flavones

3.1 experiment raw material

3.1.1 bacterium source

Enterococcus faecalis (E.faecalis), micrococcus scarlatinae (S.pyogenes), staphylococcus epidermidis (S.epidermidis), proteus vulgaris (P.vulgaris), Klebsiella Pneumoniae (K.pneumoniae) are tested in preceding 3 months separating obtained from the clinical urogenital tract specimen of Zhejiang Provincial People's Hospital.Standard strain staphylococcus aureus S.aureus (ATCC25923), escherichia coli E.coli (ATCC25922) are provided by Clinical Laboratory center, Zhejiang Province.

3.1.2 laboratory animal

ICR mice (cleaning level), body weight 18～21g, male are provided by Zhejiang Province's Experimental Animal Center.

3.1.3 sample

Folium Bambusae total flavones: total flavones glucosides content 〉=50%, be cryodesiccated sepia crystalline powder, put storage in the exsiccator, face the time spent and prepare, the ultrasound wave hydrotropy with distilled water.

3.1.4 medicine and reagent

2% Oleum Tiglii: it is formulated with 2% Oleum Tiglii, 20% dehydrated alcohol, 5% distilled water and 73% ether to face the time spent; Indometacin: crude drug, be so kind as to give by Ningbo Medicine Mfg. Factory, it is formulated with the 1%CMC-Na suspendible to face the time spent; Dimethylbenzene: Hangzhou chemical reagent factory; M-H meat soup, M-H agar are available from factory of the military region, Zhejiang Province logistics check institute; 0.22 the antibacterial filter membrane of μ m is a U.S. Milipore company product; Newborn calf serum: super no mycoplasma, available from Sijiqing Bioengineering Material Inst., Hangzhou City.

3.1.5 instrument and equipment

Shell/JB 3200 type CO ₂Cultivate phase, U.S. Sheldon company product; DG-3022A type enzyme-linked immunosorbent assay instrument, East China Electronics Co., Ltd pipe factory produces; The biological inverted microscope of COIC type, the optical instrument factory, Chongqing makes; SC-1 type horizontal laminar flow clean work station, Suzhou clean technology institute is made; The CQ250 ultrasonic cleaner, 726 ultrasonic instrument factories of middle ship head office No. 7 Institute produce.

3.2 method and result

3.2.1 bacteriostasis

Carry out with reference to " Department of Medical Administration of Ministry of Health of the People's Republic of China whole nation Clinical Laboratory rule of operation " second edition related content.

Micrococcus scarlatinae is undertaken by the pharmaceutical liquid dilution method: add the calf serum of 10% amount deactivation in sterilization M-H meat soup, add after sample makes 25mg/mL concentration again, the aseptic filtration of big syringe dress antibacterial filter membrane is handled through sterilizing.Its bacteria-free filtrate dilutes every pipe 1mL by 2 times of successives with sterilization M-H blood serum medium liquid.Each pipe add cultivated through 4～6 hours young age streptococcus pyogenes liquid 0.05mL (total bacteria containing amount is 10 ⁴C.f.u) put 35 ℃ of calorstats and cultivate 20～24h after, with its minimum medicament contg pipe that bacterial growth do not occur as its MIC value.

Other antibacterials carry out with medicine two-fold dilution method: after M-H agar, the autoclaving of each bottle packing dissolved, be cooled to about 55 ℃, add not commensurability sample, make it to become respectively 25.0,12.5,6.25,3.125,1.56,0.78 and 0.39mg/mL concentration, the M-H agar of establishing a not dosing is as adding positive growth control ware behind the bacterium.In each ware, drip 6 kinds (seeing the 3.1.1 bacterium source) then through (always bacteria containing amount is 10 than the turbid bacterium liquid 0.05mL that decide bacteria concentration ⁴C.f.u), paint little garden speckle (be 4～6h children age bacterium), cover ware after doing with inoculating loop, put 35 ℃ of constant temperature culture 16～20h after, with its minimum sample solution concentration pipe that bacterial growth do not occur as its MIC value.

The bacteriostasis of table 7 Folium Bambusae total flavones

Test strain	Minimal inhibitory concentration (MIC value)
		Enterococcus faecalis E.faecalis	??????3.125mg/ml
Micrococcus scarlatinae S.pyogenes	??????3.125mg/ml
		Staphylococcus epidermidis S.epidermidis	??????0.78mg/ml
Proteus vulgaris P.vulgaris	??????1.56mg/ml
		Klebsiella Pneumoniae K.pneumoniae	??????25.0mg/ml
Staphylococcus aureus S.aureus	??????1.56mg/ml
		Escherichia coli E.coli	??????25.0mg/ml

The result shows that Folium Bambusae total flavones all has in various degree inhibitory action (table 7) to above-mentioned 7 kinds of antibacterials.

3.2.2 antiinflammatory action

3.2.2.1 mouse knoting oil is caused the influence of auricle edema

Mice is divided into 5 groups at random by body weight, 12 every group, is provided with as follows: 1. matched group: equal-volume normal saline, 0.8mL/20g; 2. indometacin group: indometacin 10mg/kg; 3. the BLTF-01 of high dose group: 10mg/mL, dosage is 400mg/kg; 4. middle dosage group: the BLTF-01 of 5mg/mL, dosage 200mg/kg; 5. the BLTF-01 of low dose group: 2.5mg/mL, dosage 100mg/kg.Animal p.o. every day administration 1 time, continuous 7d (indometacin only administration 1 time), 1h after the last administration, mouse right ear is coated with 2% Oleum Tiglii 0.02mL/, causes inflammation, puts to death behind the 4h, cuts left and right sides ear along auricle, sweep away left and right sides auricle with the 9mm trepan, weigh, make the swelling rate with the difference note of left and right sides auricle weight.

Experimental result shows, mouse stomach gives the Folium Bambusae total flavones of 200mg/kg and 400mg/kg dosage after continuous 7 days, auricle edema due to the Oleum Tiglii is had significantly (* p＜0.05) and the extremely remarkable inhibitory action of (p＜0.01), and be tangible dose-dependence (table 8).Table 8 Folium Bambusae total flavones is to the influence of auricle edema due to the mouse knoting oil

Group	Dosage (mg/kg body weight)	Number of animals (only)	Auricle edema rate (mg)
				The blank group	Normal saline	?????13	????19.8±2.9
The indometacin matched group	????10	?????12	????14.8±5.1**
				High dose group	????400	?????12	????15.8±2.7**
Middle dosage group	????200	?????12	????16.9±2.8*
				Low dose group	????100	?????12	????18.2±3.9

Annotate: compare * p＜0.05 with matched group; * p＜0.01.

3.2.2.2 influence to mice caused by dimethylbenzene xylene auricle edema

Mice is divided into 5 groups at random by body weight, 10 every group, same 3.2.2.1 is set.Animal p.o. every day administration 1 time, continuous 7d (indometacin only administration 1 time), 30min after the last administration, mouse right ear is coated with dimethylbenzene 0.05mL/, causes inflammation, put to death behind the 15min, cut left and right sides ear, sweep away left and right sides auricle with the 9mm trepan along auricle, weigh, make the swelling rate with the difference note of left and right sides auricle weight.And with matched group relatively, judge curative effect.

Table 9 Folium Bambusae total flavones is to the influence of auricle edema due to the mice dimethylbenzene

Group	Dosage (mg/kg body weight)	Number of animals (only)	Auricle edema rate (mg)
				The blank group	Normal saline	????10	??????11.4±4.9
The indometacin matched group	????10	????10	??????6.4±2.3**
				High dose group	????400	????10	??????7.8±2.2*
Middle dosage group	????200	????10	??????11.1±3.4
				Low dose group	????100	????10	??????11.1±3.0

Annotate: compare * p＜0.05 with matched group; * p＜0.01.

Experimental result shows that mouse stomach gives the Folium Bambusae total flavones of 400mg/kg dosage after continuous 7 days, and auricle edema has significantly the inhibitory action (table 9) of (* p＜0.05) due to the xylol.

The skin physiology activity of 4 Folium Bambusae total flavones

4.1 experiment material and method

4.1.1 sample

Folium Bambusae total flavones: pale brown toner end, general flavone content are 28.7%.Sample dissolves with DMSO, and through 0.22 μ m filter membrane sucking filtration degerming, for being tried concentration, final concentration is 0.5%, 0.05% and 0.005% ,-4 ℃ of preservations with serum-free DMEM dilution.With serum-free DMEM in contrast.

4.1.2 reagent, vessel, instrument

DMEM culture medium, keratinocyte (K-SFM) culture medium (Gibco BRL company), insulin (U.S. Sigma company), trypsin U.S. Difco company) calf serum (NBS, Hangzhou Ilex purpurea Hassk.[I.chinensis Sims company), tetramethyl azo azoles (MTT, U.S. Fluka company), dimethyl sulfoxine (DMSO, Shanghai biochemical research institute); Change cortisone, penicillin, streptomycin, Ficoll, 96 orifice plates and 24 orifice plates, reach 35mm culture dish and 25cm2 culture bottle (U.S. Corning company).Microplate reader (BIO-TEK company), pH meter, super-clean bench, incubator, 722 spectrophotometers.

4.1.3 cell culture

4.1.3.1 skin keratin forms cell former be commissioned to train foster

Under aseptic condition, peel off SD rat (3 age in days) skin of back, place 4 ℃ of refrigerator 30～60min adherent skin graft, vertically add 0.25% pancreatin lentamente, in 4 ℃ of refrigerator overnight (10～11h), use the PBS rinsing then, and with in the DMEM culture fluid that contains 10%NBS and pancreatin.Separate epidermis and corium, brush is got epidermis base layer keratinocyte, centrifugal removal fibroblast.With the K-SFM culture medium cell density is adjusted to 1 * 10 ⁶/ mL is inoculated in 96 orifice plates and 24 orifice plates, changes first not good liquor behind the 24h, changes liquid every 1d later on.Treat that cell grows to subconfluence, add each concentration sample behind the hungry 24h of processing.

4.1.3.2 it is foster that skin flbroblast former is commissioned to train

Isolating corium is continued digestion 90min, with among the DMEM that contains 10%NBS and pancreatin, shred in DMEM and blow and beat, 100 mesh filter screens filter, and wash cell 2 times, adjust cell density to 2 * 10 with the DMEM culture fluid that contains 10%NBS ⁶/ mL is inoculated in 96 orifice plates.Change first not good liquor behind the 24h, later per 2～3d changes liquid.When cell grows to 80% fusion, add each concentration behind the hungry 24h of processing and tried thing.

4.1.3.3 the B-16 melanoma cells is cultivated

Adjust cell number to 1 * 10 with the DMEM culture medium that contains 15%NBS ⁴Individual/ml, be inoculated in culture bottle and 35mm culture dish.

4.1.4 cell proliferation vitality test

96 orifice plates every hole inoculation 2 * 10 ⁵Individual cell changes liquid and adds each concentration and tried thing after cultivation reaches subconfluence, each concentration is 4 samples.Each hole of 24h adds 10 μ l MTT after the dosing, continue to cultivate 4h, abandons supernatant, add 100 μ l lysates after, on microplate reader in 570mm, the detection of 630nm place.

4.1.5 lipid peroxidation product (MDA) and superoxide dismutase (SOD) are measured

MDA measures and adopts TBA colorimetry, SOD to measure employing nitrite reducing process, and test kit builds up biotech firm by Nanjing and provides.

4.1.6 melanocyte tyrosinase activity and synthetic melanocyte are measured

4.1.6.1 tyrosinase activity is measured

With B16 cell density furnishing 1.0 * 10 ⁴Individual cell, each 35mm culture dish inoculation 1.0mL cell suspension is cultivated 24h..Add each concentration during the cell place multiplication phase and tried thing, harvesting during 24h with PBS flushing twice, adds 2.0Ml PBS (including 0.1%Triton-X), behind 37 ℃, 30min, takes out cell debris liquid 0.1mL, carries out protein determination.In raffinate, add the PBS 2.0mL that contains 0.1%L-DOPA, hatch 1h for 37 ℃, the colorimetric determination of 400nm place.

Measure 4.1.6.2 melanocyte is synthetic

Cell density is transferred title 1.0 * 10 ⁴Individual cell is inoculated in and cultivates 24h in the culture bottle, adds to be tried thing, and 48h changes L liquid, is subjected to test solution effect 72h harvesting.Cell is washed with PBS, use trypsinization.Collect cytolysis in 2mL PBS, take out 0.2mL and measure protein content.In residue 0.9mL Cell sap, add the NaOH of 1.8mL, 4mol/L, 100 ℃ are heated 20min down, in the colorimetric determination of 400nm place.

4.1.7 safety evaluatio test

Carry out according to safety evaluatio program and method in " the cosmetics health standard " of health ministry promulgation in 1999.Being tried substrate concentration is 10%.Skin irritation test: get and tried thing 0.2ml and be applied on the skin.Once a day, each 1h, continuous 14 days; Eye irritant test: will be tried thing 0.1ml and splash in the conjunctival sac, in contamination back 1,24,48,72h and animal eyes being checked in the 4th day, the 7th day.

4.2 result

4.2.1 proliferation function to Skin Cell

Table 10 shows that Folium Bambusae total flavones has the effect that promotes skin cell proliferation in 0.005%～0.05% dosage range, wherein the proliferation function that skin keratin is formed cell when 0.005% concentration significantly (p＜0.05), during in 0.05% concentration extremely significantly (p＜0.01); The multiplication capacity of skin flbroblast compared with matched group also has significant difference (0.005% o'clock, p＜0.05).

Table 10 Folium Bambusae total flavones is to the influence of two kinds of skin cell proliferation abilities

Sample solution concentration (%)	Keratinocyte	Fibroblast
			Blank	????0.293±0.054	????0.337±0.075
????0.0005	????0.449±0.136	????0.436±0.074
			????0.005	????0.536±0.133*	????0.507±0.059*
????0.05	????0.628±0.092**	?????0.398±0.066

Annotate: * p＜0.05, * * p＜0.01, compared with the control.

4.2.2 to Skin Cell MDA growing amount and SOD is active influences table 11 as seen, Folium Bambusae total flavones has the lipid peroxidation product of attenuating MDA and generates, improves the active effect of antioxidase SOD in 0.0005%～0.005% dosage range, wherein 0.005% dosage group has significance (p＜0.05) compared with the control.

4.2.3 skin whitening efficacy

B16 melanocyte tumor cell tryrosinase inhibition test result shows that Folium Bambusae total flavones does not have obvious influence to this enzymatic activity; And learn to have a significant inhibitory effect (seeing Table 12) to melanocyte is synthetic during 0.005%～0.05% concentration by melanocyte synthetic test result.

Table 11 Folium Bambusae total flavones forms cell MDA to skin keratin and generates and the active influence of SOD

Sample solution concentration (%)	?????MDA(nmol/mL)	????SOD(U/mL)
			Blank	?????1.66±0.22	????2.94±0.085
????0.0005	?????1.40±0.53	????3.00±0.17
			????0.005	?????1.13±0.13*	????3.29±0.16*
????0.05	?????2.26±0.81	????2.49±0.16

Annotate: * p＜0.05, compared with the control.Table 12 Folium Bambusae total flavones is to B16 melanocyte tumor cell tyrosinase activity and the synthetic influence of melanocyte

Sample solution concentration (%)	Tyrosinase activity (OD/mg albumen)	Melanocyte synthesizes (OD/mg albumen)
			Blank	????5.81±0.31	????1.72±0.38
????0.0005	????6.50±0.47	????1.12±0.23
			????0.005	????6.83±1.34	????1.05±0.13*
????0.05	????6.29±0.54	????0.78±0.07**
			????0.5	????5.28±2.40	????2.00±0.56

Annotate: * p＜0.05, * * p＜0.01, compared with the control.

4.2.4 cutaneous safety evaluation test

The skin irritation test and the eye irritant test result of Folium Bambusae total flavones are all negative, show no skin and eye irritation.

4.3 conclusion

Skin irritation test and eye irritant test be understand exotic with situation that skin or mucosa contacts under, whether to skin generation stimulation detrimental effect or cause mucosa generation inflammatory lesion, be the essential items for inspection of cosmetics and raw materials for production safety evaluatio thereof.Test result shows that the skin of Folium Bambusae total flavones and eye conjunctiva irritant reaction are all negative, show under the external preparation for skin condition, can not produce stimulation to human body skin and mucosa.Test shows, Folium Bambusae total flavones has good biological action to Skin Cell, can promote skin keratin to form cell and fibroblasts proliferation ability, the while SOD activity improving, reduce oxidative damage, the active substance that the prompting Folium Bambusae total flavones can be used as delaying decrepitude of skin is applied in the cosmetics.The skin whitening efficacy test finds that Folium Bambusae total flavones can reduce the content of the synthetic melanocyte of melanocyte, and prompting may have the white-skinned face function to skin.

Summary of the invention

The purpose of this invention is to provide a kind of Folium Bambusae total flavones as the application of the defying age skin protection factor in cosmetics.

Folium Bambusae total flavones is applied in the cosmetics as the defying age skin protection factor.

Advantage of the present invention is: provide a kind of source wide, safe and efficient, the economic and practical defying age skin protection factor---Folium Bambusae total flavones, its characteristic relevant with the skin physiology function has been carried out systematic study, show that it has free radical resisting, antioxidation, radioprotective, antibiotic, antibacterial, promote skin cell proliferation, prevent the oxidative damage of skin, it is synthetic to reduce melanocyte, beauty functions such as delaying decrepitude of skin, and can give product with the bamboo special delicate fragrance, such as the nutrition moisturizing agent, the sun-proof skin care agent, the wrinkle resistance, whitening agent, facial milk cleanser, shampoo, there is wide application prospect in daily cosmetics fields such as bath gel.

Description of drawings

Fig. 1 is the infrared spectrum (through pressing potassium bromide troche) of Folium Bambusae total flavones;

Fig. 2 is the uv atlas (being dissolved in specpure methanol) of Folium Bambusae total flavones.

Specific implementation method

The Folium Bambusae total flavones of indication of the present invention is the flavones preparation of the different accuracy that obtains from the leaf of grass family (Graminae), Bambusoideae (Bambusoideae), Phyllostachys (Phyllostachys Sieb.et Zucc) kind, existing relating in the binomial patent of invention (patent No. is respectively ZL 98 1 04564.2 and ZL 98 1 04563.4) of its production technology before Zhang Ying.It is to be noted, the Folium Bambusae total flavones of this patent indication both can be the product that adopts above-mentioned patent technology to obtain, and also can be further to use new and high technology and the refining Folium Bambosae flavone goods that obtain of combined method thereof such as absorption～desorbing, membrance separation, supercritical fluid extraction on this basis.

The outward appearance of Folium Bambusae total flavones is yellow or pale brown toner end (also can extractum form exist), total flavones glucosides content (in butt) can change between 10～90%, and wherein the content of orientin, Lutonaretin, apigenin-8-C-glucoside and four kinds of main carbon glycosides flavone of Saponaretin accounts for more than 50% of total flavones glucosides content.Infrared spectrogram behind pressing potassium bromide troche shows, Folium Bambusae total flavones 3400,2925,1610,1118,1077cm ^-1Deng near have characteristic to absorb (seeing accompanying drawing I); After it is dissolved in spectroscopic pure methanol, in the wave-length coverage of 200～600nm, scan, spectrogram is presented at the last one absworption peak between 240～280nm, once strong absworption peak between 340～380nm meets the typical characteristic (seeing accompanying drawing II) of flavone compound.

The chemical reagent of Folium Bambusae total flavones is differentiated: the 0.5g that materialses is dissolved in the ethanol of 100mL95%, 1. gets above-mentioned solution 1 mL, adds 1%FeCl ₃2～3 of-alcoholic solution should show navy blue or blue purple.2. get above-mentioned solution 1mL, add 1%AlCl ₃2～3 of-alcoholic solution should be foresythia.3. get BLTF0.5g, add the 10mL ether, ultrasound wave auxiliary extraction 30s filters.Get filtrate 1mL, put volatilize ether in 70～90 ℃ the water-bath after, add 2% meta-dinitro-benzent solution (with the preparation of 95% ethanol) and each 1mL of KOH aqueous solution of 2.5mol/L successively, blush should appear immediately, put into above-mentioned hot bath, become royal purple rapidly.

The related current research of the present invention shows that Folium Bambusae total flavones is non-stimulated to skin and mucosa, anaphylactic reaction; Have free radical resisting, antioxidation and the radioprotective activity that can compare favourably with tea polyphenols and Semen Ginkgo extrac; In 0.005%～0.05% dosage range, can promote the propagation of Skin Cell significantly, and can suppress the synthetic of melanocyte significantly; In 0.0005%～0.005% dosage range, can significantly lower MDA and generate, and improve the activity of SOD, possess as the plain abundant and necessary condition of a kind of safe, efficient, economic phytochemistry as the defying age skin protection factor.

In view of free radical resisting that Folium Bambusae total flavones had, antioxidation, defying age, radioprotective, antibiotic, antibacterial, antiinflammatory, whiten, effects such as multi-biological such as crease-resistant, meet security requirement simultaneously as cosmetics additive, can be used as the functional component of defying age skin protection cosmetics, be applied in such as in the multiple daily cosmetics such as nutrition moisturizing agent, sun-proof skin care agent, wrinkle resistance, whitening agent, facial milk cleanser, shampoo, bath gel.

The functional component in Folium Bambusae total flavones, the skin protection factor of the present invention can suitably cooperate, make the product of various different purposes with the component in being generally used for cosmetics.These other component can comprise various vitamin and derivant thereof, hyaluronic acid, surfactant, wetting agent, chelating agen, pH regulator agent, excipient, other UV absorbent, antiseptic, dyestuff, spice etc.

Cosmetics of the present invention can be made various forms, as oily cosmetics, emulsion cosmetic, water quality cosmetics.

Beauty and skin care effects such as cosmetics Pear Power of the present invention, crease-resistant, sun-proof, antiinflammatory, speckle dispelling and the bean of dispelling.

The present invention will be illustrated by following non-limiting examples.Unit in the following formulation examples all be weight percentage (w/%).

Embodiment 1 nutrition face beautifying cream (frost)

Cera Flava 13～17

Ceresin(e) 13～17

Vaseline 15～20

Lanoline 3～7

Olive oil 8～12

Tween-20 2～6

Spice 0.1～1

Folium Bambusae total flavones 0.005～0.05

Water adds to 100.

This product to skin have antibiotic, improve looks, improve blood circulation, enhance metabolism, strengthen skin vitality, the moistening epidermis, suppress different shape sebum secretion, and activating cell, anti-ly organize effect such as poison.

The embodiment 2 crease-resistant honey that brighten

L-ascorbic acid and derivant 1～3 thereof

Olive oil 10～20

Isopropyl myristate 3～7

NPE 0.1～1

Glycerol 3～7

Hyaluronic acid 1～3

Folium Bambusae total flavones 0.005～0.05

Ethanol 5～10

Water adds 100.

Effects such as that this product has is crease-resistant, whiten, preserve moisture, speckle dispelling.

Embodiment 3 sunscreen cream

16～octadecanol 6～10

White oil 26 ^#3～6

Isopropyl palmitate 3～6

Dimethicone 1～2

Monoglyceride 2～3

Wool grease 0.1～1

Polyethylene Glycol-6,000 0.1～1

Glycerol 2～6

Folium Bambusae total flavones 0.005～0.05

SPP-200 (phosphide quaternary ammonium compound cationic emulsifier) 2～3

Essence, antiseptic 0.1～1

Water adds to 100.That this product has is sun-proof, defying age and skin-care effect.

Embodiment 4 cleansing milk

Vitamin E and derivant 0.05～0.5 thereof

Glyceryl tristearate 6～10

Spermol 1～3

Liquid paraffin 8～16

Folium Bambusae total flavones 0.01～0.05

Triethanolamine 1～3

Propylene glycol 4～8

Water adds to 100.That this product has is antibacterial, suppress effects such as different shape sebum secretion, antiinflammatory, the bean of dispelling.

Embodiment 5 bath gel

Both sexes Laurel acidic group hydroxypropyl sulfonate 20～30

Cocinic acid amido propyl hydroxypropyl sulfonation betanin 7～13

Lauryl ether sulfate (30%) 8～12

Lauroyl amido diethanolamine 0.5～1.5

Folium Bambusae total flavones 0.01～0.5

Water adds to 100.

This product has the harmful bacterium that suppresses skin surface, the activation Skin Cell, and the effect of conditioning skin and prevention skin carcinoma, and make the health after the bath have a kind of pleasant delicate fragrance for a long time.Embodiment 6 shampoos

Sodium lauryl sulphate 40～60

Hydroxypropyl cellulose 0.5～1.5

Oil base diglycollic amide 3～7

Cationic animal proteinum 3～7

Polyethylene Glycol wool grease 1～3

Methyl hydroxybenzoate 0.1～0.3

Quaternary ammonium compound-15 0.05～0.15

Poly-aminopropyl biguanide xylene chloride phenol 0.05～0.15

Soybean phospholipid 0.1～1

Folium Bambusae total flavones 0.01～0.5

Water adds to 100.

This product also can prevent keratic loss in the hair washing process except that can making shinny, soft, the easy comb of hair, prevent the withered and yellow bifurcated of hair, the control scalp itch, thus play the effect of protecting hair, and make the hair after washing have a kind of simple and elegant fragrance for a long time.

Obviously, according to above-mentioned example, the present invention can have many improvement and change.Therefore, should be understood that in the scope of appended claim that except that described herein, the present invention also can implement with additive method.

Claims (8)

1. a Folium Bambusae total flavones is characterized in that as the application of the defying age skin protection factor in cosmetics Folium Bambusae total flavones is applied in the cosmetics as the defying age skin protection factor.

2. Folium Bambusae total flavones according to claim 1 is characterized in that as the application of the defying age skin protection factor in cosmetics said cosmetics are nutrition face beautifying cream (frost), crease-resistant honey, sunscreen cream, cleansing milk, bath gel, the shampoo of brightening.

3. Folium Bambusae total flavones according to claim 2 is characterized in that as the application of the defying age skin protection factor in cosmetics the weight percent of said nutrition face beautifying cream (frost) consists of:

Cera Flava 13～17

Ceresin(e) 13～17

Vaseline 15～20

Lanoline 3～7

Olive oil 8～12

Tween-20 2～6

Spice 0.1～1

Folium Bambusae total flavones 0.005～0.05

Water adds to 100.

4. Folium Bambusae total flavones according to claim 2 is characterized in that as the application of the defying age skin protection factor in cosmetics the said crease-resistant weight percent that brightens honey consists of:

L-ascorbic acid and derivant 1～3 thereof

Olive oil 10～20

Isopropyl myristate 3～7

NPE 0.1～1

Glycerol 3～7

Hyaluronic acid 1～3

Folium Bambusae total flavones 0.005～0.05

Ethanol 5～10

Water adds 100.

5. Folium Bambusae total flavones according to claim 2 is characterized in that as the application of the defying age skin protection factor in cosmetics the weight percent of said sunscreen cream consists of:

16～octadecanol 6～10

White oil 26 ^#3～6

Isopropyl palmitate 3～6

Dimethicone 1～2

Monoglyceride 2～3

Wool grease 0.1～1

Polyethylene Glycol-6,000 0.1～1

Glycerol 2～6

Folium Bambusae total flavones 0.005～0.05

SPP-200 (phosphide quaternary ammonium compound cationic emulsifier) 2～3

Essence antiseptic 0.1～1

Water adds to 100.

6. Folium Bambusae total flavones according to claim 2 is characterized in that as the application of the defying age skin protection factor in cosmetics the weight percent of said cleansing milk consists of:

Vitamin E and derivant 0.05～0.5 thereof

Glyceryl tristearate 6～10

Spermol 1～3

Liquid paraffin 8～16

Folium Bambusae total flavones 0.01～0.05

Triethanolamine 1～3

Propylene glycol 4～8

Water adds to 100.

7. Folium Bambusae total flavones according to claim 2 is characterized in that as the application of the defying age skin protection factor in cosmetics the weight percent of said bath gel consists of:

Both sexes Laurel acidic group hydroxypropyl sulfonate 20～30

Cocinic acid amido propyl hydroxypropyl sulfonation betanin 7～13

Lauryl ether sulfate (30%) 8～12

Lauroyl amido diethanolamine 0.5～1.5

Folium Bambusae total flavones 0.01～0.5

Water adds to 100.

8. Folium Bambusae total flavones according to claim 2 is characterized in that as the application of the defying age skin protection factor in cosmetics the weight percent of said shampoo consists of:

Poly-aminopropyl biguanides xylene chloride phenol 0.05～0.15 soybean lecithin 0.1～1 bamboo leave flavone 0.01～0.5 water of lauryl sodium sulfate 40～60 hydroxypropyl celluloses 0.5～1.5 oil base diglycollic amide 3～7 cationic animal protein 3～7 polyethylene glycol wool greases 1～3 methyl hydroxybenzoate 0.1～0.3 quaternary ammonium compounds-15 0.05～0.15 adds to 100.

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