為使貴 審查委員能對本發明之特徵與其特點有更進一步之了解與認同，茲列舉以下較佳之實施例說明如下：請參閱第一圖，為本發明天然色素作為指甲油色料之流程； In order to enable the reviewing committee to have a better understanding and recognition of the features and characteristics of the present invention, the following preferred embodiments are illustrated as follows: Please refer to the first figure, which is a flow of the natural pigment as the nail polish colorant of the present invention;

(一)菌液的培養10 (1) Cultivation of bacterial liquid 10

將紫色桿菌菌株劃線於固體培養基，置於37℃恆溫培養箱中進行培養，並觀察菌體生長情形；取單一菌落接種於含100mL培養液之三角錐形瓶中，置入25℃培養箱，恆溫培養2天，收集足夠菌液量進行紫色桿菌素的萃取。 The purple bacillus strain was streaked into a solid medium, placed in a 37 ° C incubator for culture, and the growth of the cells was observed; a single colony was inoculated into a triangular conical flask containing 100 mL of the culture solution, and placed in a 25 ° C incubator. The culture was carried out at a constant temperature for 2 days, and sufficient bacterial liquid amount was collected for extraction of purple bacillus.

(二)紫色桿菌素的萃取20 (2) Extraction of purple bacillus 20

取一個500mL血清瓶，倒入300mL的紫色桿菌培養液，加入100mL乙酸乙酯至玻璃瓶中，利用攪拌機將兩種液體混勻2天；停止攪拌並靜置2小時，讓溶劑與水分離，利用分液漏斗將上層萃取液與下層 菌液分離，再將上層萃取液中的乙酸乙酯烘乾，加入甲醇進一步的萃取紫色桿菌素，即為紫色桿菌素之粗萃取液。 Take a 500mL serum bottle, pour 300mL of purple bacillus culture solution, add 100mL of ethyl acetate to the glass bottle, mix the two liquids with a blender for 2 days; stop stirring and let stand for 2 hours, let the solvent separate from the water, Use the separatory funnel to separate the upper extract from the lower layer The bacterial liquid is separated, and the ethyl acetate in the upper extract is dried, and the purple extract is further extracted by adding methanol, which is a crude extract of purple bacillus.

(三)紫色桿菌素產量的測定30 (C) Determination of the production of purple bacillus 30

將紫色桿菌素取出加入10mL的甲醇搖晃均勻，取950μL甲醇加入50μL紫色桿菌素做吸光值(577nm)測試，吸取1mL紫色桿菌素至離心機(10,000rpm)離心10分鐘，再將上層萃取液中紫色桿菌素取出做二次離心，接著利用吸光值所測的數據推算出每個離心管中紫色桿菌素的濃度。 The violacein taken Shake added 10mL methanol, methanol was added 950 μ L take 50 μ L violacein do absorbance (577 nm) test, draw 1mL violacein to the centrifuge (10,000 rpm for) rpm for 10 minutes, then The purple bacillus in the upper extract was taken out for secondary centrifugation, and then the concentration of purple bacillus in each centrifuge tube was estimated by using the data measured by the absorbance.

將每管濃度為2μg/mL之萃取液分裝於微量離心管後至烘箱烘乾成為粉末狀，4℃冷藏備用。 After concentration of each tube 2 μ g / mL of the extract was aliquoted into microfuge tubes oven dried to form a powder, 4 ℃ cold standby.

(四)不同精油與紫色桿菌素相溶性之測試40 (4) Testing of the compatibility of different essential oils with purple bacillus 40

將紫色桿菌素從4℃冰箱取出，分別加入200μL的精油(尤加利精油、薰衣草精油、葡萄柚精油、茶樹精油、薄荷精油、天竺葵精油)以振盪儀進行振盪溶解，使紫色桿菌素充分溶解，放置室溫28℃觀察其顏色變化；可觀察到薰衣草精油、薄荷精油及天竺葵精油呈現深藍紫色且體積不變，葡萄柚精油呈現深紫色且體積減少，葡萄柚精油容易因時間增長而增加揮發體積，其餘精油的揮發性則較低，體積不受儲放時間的影響；尤加利精油溶解紫色桿菌素後，顏色快速由紫色轉變成透明淡黃色，茶樹精油溶解紫色桿菌素則呈現金黃色，尤加利及茶樹精油所含的成份能將紫色桿菌素破壞，使得顏色明顯發生改變。 Violacein removed from the refrigerator at 4 ℃, 200 μ L were added essential oils (eucalyptus oil, lavender essential oil, grapefruit oil, tea tree oil, peppermint oil, geranium oil) was dissolved in a shaker oscillate the violacein Fully dissolved, placed at room temperature 28 ° C to observe the color change; can be observed that lavender essential oil, peppermint essential oil and geranium oil are dark blue purple and the volume is unchanged, grapefruit essential oil is dark purple and volume is reduced, grapefruit essential oil is easy to grow due to time Increase the volatile volume, the other essential oils have lower volatility, the volume is not affected by the storage time; after the eucalyptus essential oil dissolves the purple bacillus, the color changes rapidly from purple to transparent pale yellow, and the tea tree essential oil dissolves the purple bacillus. The golden yellow, eucalyptus and tea tree oils contain ingredients that destroy the purple bacillus and cause a noticeable change in color.

(五)指甲油配方之測試50 (5) Testing of nail polish formula 50

請參閱第二圖，將指甲油成分分組為A~E組，A組為乙基纖 維素(Ethyl cellulose)12%、一環氧樹脂10%、一樟腦2%及酒精76%，B組為乙基纖維素15%、一環氧樹脂7%、一樟腦7%及酒精71%，C組為乙基纖維素15%、一環氧樹脂12%、一樟腦7%及酒精66%，D組為乙基纖維素10%、一環氧樹脂7%、一樟腦3%及酒精80%，E組為乙基纖維素10%、一環氧樹脂15%、一樟腦5%及酒精70%，分別配置於15mL離心管中，放置振盪儀振盪至指甲油充分溶解後，取製備好之紫色桿菌素粉末，每管分別加入120μL指甲油，再依序分別加入編號1號甲片為尤加利精油50μL，2號甲片為尤加利精油30μL+酒精20μL，3號甲片為尤加利精油20μL+酒精30μL，4號甲片為薄荷精油50μL，5號甲片為薄荷精油30μL+酒精20μL，6號甲片為薄荷精油20μL+酒精30μL，7號甲片為酒精50μL，8號甲片為甲醇50μL，放置震盪儀震盪，以微量吸量管吸取指甲油塗於透明甲片，放置陰涼處觀察指甲油顏色變化。 Please refer to the second figure, the nail polish components are grouped into groups A~E. Group A is ethyl cellulose (Ethyl cellulose) 12%, epoxy resin 10%, one camphor 2% and alcohol 76%. Group B is Ethylcellulose 15%, epoxide 7%, camphor 7% and alcohol 71%, group C is ethylcellulose 15%, an epoxy resin 12%, a camphor 7% and alcohol 66%, D The group is ethyl cellulose 10%, one epoxy resin 7%, one camphor 3% and alcohol 80%, E group is ethyl cellulose 10%, one epoxy resin 15%, one camphor 5% and alcohol 70% after are arranged in 15mL centrifuge tubes, shaker placed oscillates to fully dissolve the nail polish, a good take prepared powder of violacein, each tube were added 120 μ L Nail, were then added in sequence ID No. 1 a sheet of eucalyptus oil 50 μ L, 2 pieces of eucalyptus oil A 30 μ L + alcohol 20 μ L, 3 A sheet of 20 μ L + eucalyptus oil alcohol 30 μ L, a piece of No. 4 50 peppermint oil μ L, No. 5 A sheet of peppermint essential oil 30 μ L + alcohol 20 μ L, No. 6 A piece of peppermint essential oil 20 μ L + alcohol 30 μ L, No. 7 A sheet for alcohol 50 μ L, 8 A sheet is methanol 50 μ L, instrument placement shock shocks, micro Nail polish applied pipet A transparent sheet, cool place nail polish color change was observed.

1號甲片為指甲油中含有50%尤加利精油，A組甲面呈現深藍紫色有紫色顆粒；D組甲面呈現淡紫色，E組甲面呈現藍紫色表面無顆粒且溶解度佳；B、C組甲面呈現透明藍紫色，表面無光澤且有顆粒；2號甲片為指甲油中含有尤加利精油30%、酒精20%，A組甲面呈現深藍紫色；B、C及E組甲面呈現藍紫色，D組呈現透明淡紫色，C、D及E組甲面呈現無顆粒有光澤；3號甲片為指甲油中含有尤加利精油20%、酒精30%，D組甲面呈現淡紫色且顏色飽和度低，其他組別指甲油呈現深藍紫色，A、B及C組甲面有白色霧狀，E組甲面顏色呈現均勻有光澤；4號甲片為指甲油中含有50%薄荷精油，A、B組顏色為深藍紫色，表面有霧面的紫色顆粒，C、E組顏色呈現透明藍紫色，但E組顏色均勻有光澤，D組顏色呈現透明淡紫色且飽和 度高。 No.1 nail tablet contains 50% eucalyptus essential oil in nail polish, group A has dark blue purple and purple particles in group A; group D presents lavender in group A, and group A has blue-violet surface without particles and good solubility; B Group C has a transparent blue-purple surface with a dull surface and granules; No. 2 nail tablet contains 30% eucalyptus oil and 20% alcohol in nail polish, and group A has a deep blue-violet color; B, C and E Group A showed blue-violet color, group D showed transparent lavender, group C in C, D and E showed no grain and luster; No. 3 piece contained nail oil containing eucalyptus essential oil 20%, alcohol 30%, group D The surface of the nail surface is lavender and the color saturation is low. The nail polishes of other groups are dark blue and purple, the nails of group A, B and C have white mist, the color of nails of group E is even and shiny; the nail of group 4 is nail polish. It contains 50% peppermint essential oil. The color of group A and B is dark blue and purple, and the surface is foggy purple particles. The color of group C and E is transparent blue-violet, but the color of group E is uniform and lustrous, and the color of group D is transparent and lavender. saturation High degree.

5號甲片為指甲油中含有薄荷精油30%、酒精20%，A、B及C組甲面呈現深藍紫色，A、C組甲面有顆粒狀，D組甲片呈現淡紫色且甲面均勻，E組呈現藍紫色甲面均勻有光澤；6號甲片為指甲油中含有薄荷精油20%、酒精30%，A、B、C及E組顏色呈現藍紫色，D組呈現淡紫色，其中D、E組飽和度最佳，甲面顏色均勻有光澤。 No.5 nail piece contains 30% of peppermint essential oil and 20% of alcohol in nail polish. The surface of group A, B and C is dark blue-purple, and the nails of group A and group C are granular, and the group D of group D is lavender and nail surface. Evenly, the E group showed a blue-purple nail surface with uniform luster; the No. 6 nail tablet contained 20% of peppermint essential oil and 30% of alcohol in nail polish, and the color of group A, B, C and E showed blue-violet color, and group D showed lavender. Among them, the D and E groups have the best saturation, and the color of the nail surface is even and shiny.

7號甲片為指甲油中含有50%酒精，甲片顏色以D、E組最好，表面均勻且顏色飽和度高，D組甲面呈現淡紫色，E組甲面呈現透明藍紫色，A、B、C組甲面呈現藍紫色，但表面不均勻且有些許顆粒；8號甲片為指甲油中含有50%甲醇，A組甲片呈現透明藍紫色，D組甲片呈現透明淡紫色，B、C及E組甲片呈現藍紫色，且D、E組表面均勻有光澤。 No.7 nail piece contains 50% alcohol in nail polish. The color of nail piece is best in group D and E. The surface is uniform and the color saturation is high. The surface of group D is lavender, and the surface of group E is transparent blue and purple. Group B and C showed blue-violet, but the surface was uneven and slightly granules; No. 8 nailed with 50% methanol in nail polish, Group A showed transparent blue-violet, and Group A showed transparent lavender The B, C and E groups showed blue-violet color, and the D and E groups were even and shiny.

由上述實驗結果統計為三種表格，以溶解度我們發現D、E組指甲油最好，甲片無顆粒狀且溶解效果最佳；以指甲油成品均勻度來看，E組指甲油最好，顏色呈現均勻有光澤；以指甲油成品色度可發現A組甲片大多以深藍紫色為主；B組甲片大部分以藍紫色為主；C組甲片顏色以藍紫色為主；D組甲片主要以淡紫色為主；E組甲片主要以藍紫色居多；整體而言，指甲油以E組最好，顏色均勻且指甲油易於塗抹於甲片上。 From the above experimental results, there are three kinds of tables. In terms of solubility, we found that the nails of group D and E are the best. The nails of the group A are not granular and the dissolution effect is the best. In terms of the uniformity of finished nail polish, the nail polish of group E is the best. It is uniform and shiny; it can be found that the nails of group A are mostly dark blue and purple; the group A of group B is mainly blue-purple; the color of group A is mainly blue and purple; group D is The tablets are mainly lavender; the E group is mainly blue-purple; overall, the nail polish is best in group E, the color is uniform and the nail polish is easy to apply on the nail.

(六)混合紫色桿菌素與指甲油與精油60 (6) Mixing purple bacillus with nail polish and essential oil 60

最終，將紫色桿菌素搭配低污染的指甲油與薄荷精油製造出本發明之一種天然色素作為指甲油色料；紫色桿菌素是種微生物產生的天然色素，使用上無人工色素的危害，不會使人接觸後造成皮膚過敏，而精油被廣泛運用在各種美容行業中，為一種較天然的溶劑由植物萃取而成， 本發明將兩者結合進行其作為指甲油色料；研究結果發現不同精油溶解紫色桿菌素效果最好的是薄荷精油，顏色屬於藍紫色。 Finally, purple bacillus is combined with low-pollution nail polish and peppermint essential oil to produce a natural pigment of the present invention as a nail polish coloring material; purple bacillus is a natural pigment produced by a microorganism, and no artificial coloring is used, and no It causes skin allergies after contact, and essential oils are widely used in various beauty industries. They are a natural solvent extracted from plants. The invention combines the two as the nail polish coloring material; the research results show that the best effect of dissolving the purple bacillus in different essential oils is the peppermint essential oil, and the color belongs to blue-violet color.

以上所揭，僅為本發明所提供之較佳實施例而已，並非用以限制本創作之實施範圍，凡本技術領域內之相關技藝者根據本創作所為之均等變化，皆應屬本創作所涵蓋之範圍。 The above is only the preferred embodiment provided by the present invention, and is not intended to limit the scope of implementation of the present invention. All of the related art in the art are equivalent to the present invention, and should belong to the present invention. The scope of coverage.