重要平地造林樹種之保健活性成分分析(2011-2015) Analysis of the health active ingredient of important tree species in plains afforestation.

2011年
本研究第一年選取茄苳(Bischofia jabanica)及樟樹(Cinnamomum campho)兩物種，作為活性成分分析之材料，試材於2-3月間採集自北部地區，分別進行5%乙醇、95%乙醇、甲醇及熱水萃取。茄苳及樟樹總體收率以甲醇萃取物之收率最佳，茄苳之5%乙醇抽出物具有最佳的抗氧化活性，茄苳葉部不同萃取方法之抽出物物，以5%EtOH萃取方式具有良好的清除能力，樟樹不同部位之萃取物，葉部熱水萃取的粗萃物最具有良好的清除能力。茄苳B u t a n o l可溶部之D P P H 自由基半數抑制濃度（IC50）最佳，為IC50＝8.78±2.07μg/mL、粗抽出物IC50＝17.06±1.00 μg/mL、EA可溶部IC5 0＝19.17±2.15 μg/mL、 水可溶部IC5 0＞100 μg/mL；樟樹葉部Butanol可溶部最佳IC5 0＝18.60±2.95 μg/mL、其次EA可溶部IC5 0＝20.35±2.56
μg/mL、粗抽出物IC50＝22.96±2.56 μg/mL、水可溶部IC50＞100 μg/mL。抑制酪氨酸酶活性試驗中，茄苳5% EtOH、葉部之熱水抽出物具有較佳活性，樟樹兩種品系，本樟及芳樟熱水萃取物皆具有極佳的抑制活性；從可溶部的角度，茄苳抽出物之水可溶部及樟樹抽出物之正丁醇可溶部具有相對較佳活性。海蝦毒性試驗，本樟熱水抽出物及芳樟乙醇抽出物在濃度500 ppm和100 ppm並不具毒性。茄苳5%乙醇抽出物，具有較高的抗發炎效果，且對於細胞存活率並無影響；茄苳5%乙醇抽出物之乙酸乙酯可溶部及芳樟葉子正丁醇可溶部，有相對較佳之抗發炎活性及較高細胞存活率。綜合以上結果茄苳乙醇抽出物具有較佳的抗氧化、抗發炎及抑制酪氨酸酶活性，具有更進一步研究之潛力。

2011
This first year of research, selected Bishop Wood (Bischofia javanica) and camphor tree (Cinnamomum camphora) , two species as material for the active constituent analysis.Gathering material from February to March in the north areas, carries on 5% ethyl alcohol, 95% ethyl alcohol, the
methyl alcohol and the hot water extracts separately.The B. javanica and C. camphora overall receiving rate were best by the methyl alcohol
extracts, the B. javanica 5% ethyl alcohol extracts have the best antioxidant activities.B. jabanica leaf extracts with different extraction methods, 5% EtOH extraction method has a good free radicals remove ability. Extracts of different parts of camphor, hot water crude extracts of leaf had the most good scavenging abilities.Butanol soluble fraction of B. javanica, scavenged 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals effectively with IC50 (half inhibitory concentration) best for IC50 = 8.78 ± 2.07μg/mL, crude extracts IC50 = 17.06 ± 1.00 μg /mL, EA soluble Department IC50 = 19.17 ± 2.15 μg / mL, water-soluble fraction IC50＞ 100 μg / mL.Butanol soluble fraction of camphor leaves best IC50 = 18.60 ± 2.95 μg / mL, followed by the EA soluble fraction IC50 = 20.35 ± 2.56 μg / mL, crude extracts IC50 = 22.96 ± 2.56 μg /mL, the fraction of water soluble IC50＞ 100 μg / mL.Tyrosinase Inhibition activity test, B. javanica 5% EtOH, the hot water leaf extracts had a better activities of two strains of camphor tree, both camphor and linalool type of hot water extracts had excellent inhibitory activity; as to fraction activities , B. javanica water-soluble extracts and camphor n-butanol soluble extracts had a relatively better activity. Shrimp toxicity test, the camphor type of hot water extracts and linalool type of ethanol extracts at a concentration of 500 ppm and 100 ppm were not toxic.B. javanica 5% ethanol extracts, with high anti-inflammatory effects, were no effect on cell survival.B. javanica 5% ethanol extracts of acetate soluble fraction and linalool type leaves of the f n-butanol soluble fraction , there were relatively better of the anti-inflammatory activity and a higher survival rate. Based on the above results, B.
javanica ethanol extracts had better anti-oxidation, anti-inflammatory and inhibit tyrosinase activity, with further research potential.

2012年
本研究針對桃花心木及苦楝進行研究，試材分別來自台灣中部、南部、北部、及中國大陸，分別從細胞內及細胞外抗氧化、抗木材腐朽菌、抗發炎、抗登格勒病媒蚊幼蟲及細胞內外之美白活性進行篩選。抗氧化部分不管是細胞外或細胞內，大葉桃花心木皆有最佳的抗氧化能力，作為民俗藥用植物的大葉桃花心木種子在Ethyl acetate可溶部IC50=8.63 μg/mL，細胞內抗氧化在濃度50 ppm、100 ppm在細胞誘導傷害實驗中具有保護效果；抗菌性活性中，苦楝各部位對於L. S.木材腐朽菌具有100%之抑制效果，黃葉萃取物對於G. T.腐朽菌可達到80 %之抑制效果；抗病媒蚊幼蟲方面，川楝皮及苦楝枝條抽出物有較佳之抑制效果；抗發炎方面，利用RAW264.7巨噬細胞的抑制NO生成效果來篩選大葉桃花心木、小葉桃花心木、苦楝果實、川楝子及川楝皮粗萃物及不同可溶部的抗發炎功效，在濃度50及100 ppm，大葉桃花心木種子EtOH之粗萃物比正對照組Curcumin有更明顯的抑制由LPS 誘發細胞生成的NO，可有效修復發炎反應效果，但對細胞具有微量的毒性；美白活性方面分別從抑制細胞內、外酪氨酸活性方面及抑制皮膚黑色素瘤細胞存活率進行評估，大葉桃花心木果核及種子MeOH萃取之不同可溶部皆具有抑制細胞外酪胺酸酶之活性，果核以Butanol可溶部(濃度12.5-50 ppm分別為9.12 %、8.03 %及0.73 %)最佳，果核在濃度50 ppm Butanol可溶部(39.10 %)及在濃度12.5 ppm、25 ppm、50 ppm，果核Water可溶部(50.13 %、31.42 %、25.24 %)及種子的Ethyl acetate可溶部及Butanol可溶部，則具有抑制細胞內酪胺酸活性，苦楝主要活性成分印楝素對於細胞外具有活性，並且對於皮膚細胞毒性較低，值得進一步評估。
2012
This study for mahogany and Chinaberry test as a materials were from central China. Intracellular and extracellular antioxidant , anti- wood
decay fungi , anti-inflammatory , anti- dengue mosquito larvae and intracellular and extracellular whitening activities were screening.
Antioxidant part whether extracellular or intracellular the Swietenia macrophylla had best antioxidant capacity.As a folk medicinal plants of
big-leaf mahogany seed, Ethyl acetate soluble fraction IC50 = 8.63 μg /mL, and intracellular antioxidant protective effect at a concentration
were of 50 ppm , 100 ppm of cell injury.100 % of the inhibitory effect of various parts of the LS wood decay fungus, and yellow leaves extract for GT decay fungi can reach 80 % of the inhibitory effect of antibacterial properties.The anti mosquito larvae , TSN skins and chinaberry branches extract showed better inhibition effect.Anti-inflammatory assay used RAW264.7 macrophages NO production inhibitory effect to screen large leaf mahogany , lobular mahogany , neem fruit , Toosendan TSN skin crude extract and soluble anti-inflammatory efficacy.Concentrations of 50 and 100 ppm, big-leaf mahogany seed EtOH crude extract more obvious than the positive control group Curcumin inhibited LPS -induced cell -generated NO fixes the effects of inflammation , but has traces of toxicity to cells. Whitening active from the inhibition of intracellular and extracellular tyrosine activity and suppression of cutaneous melanoma cell survival evaluate different soluble fraction of the MeOH extract of big-leaf mahogany or nut and seed are inhibit extracellular tyrosinase enzyme of activities. Stone Butanol soluble fraction ( 12.5-50 ppm concentration were 9.12% , 8.03% and 0.73% ) were best in a concentration of 50 ppm Butanol soluble fraction ( 39.10% ) and the concentration of 12.5 ppm , 25 ppm , 50 ppm, stone Water soluble fraction ( 50.13% , 31.42% , 25.24% ) and seed Ethyl acetate soluble fraction Butanol soluble fraction , with inhibition of intracellular tyrosine.The Chinaberry main active ingredient, azadirachtin had extracellular active and less toxic skin
cells , worthy of further evaluation.

2013
本研究利用白千層(Melaleuca leucadendra)與櫸木(Zelkova serrata)MeOH及EtOH溶劑以不同濃度萃取及熱水萃取之粗萃取物，粗萃物利用液相-液相分配分為Hexane、Ethyl acetate、Butanol及Water等4個可溶部。至於生物活性評估則包括DPPH自由基清除能力、海蝦致死試驗、抗發炎活性試驗、細胞內美白試驗、抗腐朽菌試驗、抗病媒蚊幼蟲試驗。結果發現白千層枝、葉EtOH萃取之Butanol可溶部具有少量的DPPH自由基清除能力，中部櫸木葉60 %、95 % EtOH萃取粗萃物，抑制率分別為IC50= 33.21 ± 0.84 μg/mL、IC50= 75.82 ± 1.44 μg/mL，清除DPPH自由基之抑制率較好，其次是中櫸木葉95 % EtOH萃取粗萃物，北、中、南枝、葉EtOH萃取粗萃物之不同可溶部，枝EtOH萃取之不同可溶部皆以Ethyl acetate可溶部，葉EtOH萃取之不同可溶部Butanol可溶部有較好的清除DPPH自由基之效果。由海蝦致死試驗結果發現，白千層精油具有顯著的毒殺作用，然而葉EtOH萃取粗萃物在24、48、72小時對海蝦並無太大毒性，櫸木粗萃物海蝦毒性測試隨時間和粗萃物及不同可溶部濃度的增加，海蝦的死亡率增高。白千層精油和澳洲茶樹精油相比較，尤其以濃度50 ppm白千層精油較佳，皆能有效抑制LPS誘發RAW 264.7小鼠巨噬細胞所生成之一氧化
氮(NO)，顯示其具良好之抗發炎活性。抑制細胞內酪胺酸酶活性結果顯示，皆有50 %-80 %之抑制效果，細胞內黑色素含量結果顯示，濃度愈高，抑制黑色素效果愈佳，與對照組Kojin acid比較，皆具有55%之抑制黑色素合成效果。抗白線斑蚊幼蟲試驗結果顯示，精油可完全抑制幼蟲活性，抗腐朽菌試驗結果顯示，白千層、櫸木粗萃物及不同可溶部及精油，由低濃度到高濃度，皆對Trametes versicolo白腐菌、Gloeophyllum trabeum、Laetiporus sulphureus褐腐菌及Phellinus noxius褐根腐菌皆具有不錯的抗菌活性，綜合上述試驗結果顯示，白千層葉EtOH萃取粗萃物對細胞不具有毒性，不具抗發炎活性，但與白千層精油相比，精油皆具有不錯的抑制細胞內酪氨酸酶活性、抑制細胞黑色素含量，抗腐朽、抗病媒蚊及抗發炎活性。
2013
In this study, Melaleuca (Melaleuca leucadendra) and Beech (Zelkova serrata) MeOH and EtOH crude extracts of different concentrations of
solvent extraction and hot water extraction , crude extract by liquid - liquid distribution into Hexane, Ethyl acetate, 4 soluble fraction Butanol and Water , etc. As for biological activity assessment includes DPPH radical scavenging ability , shrimp lethality test, the antiinflammatory activity of the test , the test cell whitening , anti- rot test , anti- mosquito larvae test . Butanol soluble fraction was found Melaleuca branches, leaves with a small amount of EtOH extract of DPPH radical scavenging ability , Central Beech Leaves 60%, 95% EtOH extract crude extract , the inhibition rates were IC50 = 33.21 ± 0.84 μg / mL , IC50 = 75.82 ± 1.44 μg / mL, clear inhibition of DPPH radical better rate , followed by the different soluble in beech leaves 95% EtOH extract crude extract , northern, central, Nanzhi leaf extract crude extract of EtOH Ministry of different soluble portion of the branch 's begin with Ethyl acetate EtOH extract the soluble portion , different soluble portion Butanol extraction of the soluble portion of the leaf EtOH have better scavenging DPPH radicals . The shrimp lethality test results showed that Melaleuca oil has significant toxic effects , but leaves EtOH extract crude extract at 48 and 72 hours is not much for shrimp toxicity , beech crude extract with shrimp toxicity test increase the time and
crude extract and different concentrations of the soluble portion ,shrimp mortality increased. Melaleuca oil and tea tree oil compared to Australia , particularly in the concentration of 50 ppm oil Melaleuca better , able to inhibit LPS -induced RAW 264.7 murine macrophage nitric oxide generated by one (NO), which has a good display the antiinflammatory activity. Inhibition of tyrosinase activity results in the cells showed , there are 50% -80 % of the inhibitory effect on melanin content in cells showed that the higher the concentration , the better the effect of melanin , Kojin acid comparison with the control group ,both with 55% the inhibition of melanin synthesis results. Anti- Aedes albopictus larvae test results show that essential oils can be completely inhibited larvae activity , antifungal test results show that Melaleuca ,beech and crude extracts and essential oils of different soluble portion , from low concentration to high concentration , both for Trametes versicolo white rot fungi , Gloeophyllum trabeum, Laetiporus sulphureus brown rot fungi and brown root rot fungus Phellinus noxius both have good antibacterial activity, integrated above test results show that the melaleuca leaf EtOH extract crude extract is not toxic to cells , does not have anti-inflammatory activity , but compared with Melaleuca oil , essential oils are having a good inhibition of intracellular tyrosinase activity , inhibiting cell melanin content , anti- rot, anti- mosquito
and anti -inflammatory activity.

2014
本研究以經濟造林樹種光蠟樹( F r a x i n u s f o r m o s a n a)及印度紫檀(Pterocarpus indicus)為標的。利用MeOH、EtOH萃取得到粗萃取物及不同可溶部(液相-液相層析)，進行DPPH自由基清除能力、抗發炎活性、細胞內外美白活性試驗、毒性試驗及氣相層析儀活性成分試驗等生物活性評估，目的開發出具有價值之商品。由DPPH自由基清除能力結果發現，北區光蠟樹葉部位EtOH萃取之粗萃物抑制率較好，有56.74 ± 1.73 %的抑制率，印度紫檀枝部位MeOH萃取之粗萃物抑制率較高，有33.86 ± 1.42 %的抑制率。抗發炎活性方面發現，巨噬細胞經由LPS處理24小時後，光蠟樹北中南區及印度紫檀粗萃物及不同可溶部在濃度100、50、25 ppm皆具有細胞增生之功能，無毒殺效果，只有中部及北部光蠟樹枝EtOH萃取之不同可溶部，Hex、EA層具有些許的毒殺，光蠟樹種子部位EtOH萃取之粗萃物及不同可溶部，具有良好細胞修復發炎之反應，其餘在高濃度有微量的修復發炎反應。抑制細胞外酪胺酸酶活性結果顯示，北區光蠟樹葉部位MeOH及EtOH萃取之粗萃物，抑制酪胺酸酶活性較佳。黑色素細胞瘤細胞存活性篩選方面，光蠟樹及印度紫檀粗萃物沒有明顯完全抑制黑色素細胞瘤之生長，不同可溶部在高濃度對黑色素細胞存活有些許的毒殺約50-70 %，低濃度毒性不強約70-95 %。細胞內直接及間接抑制酪胺酸酶方面，光蠟樹及印度紫檀之不同可溶部與對照組市售麴酸相比較，皆有抑制細胞內酪胺酸酶的活性，抑制活性大小依序為Water可溶部＞ But可溶部＞EA可溶部＞Hex可溶部，只有印度紫檀葉部位不同可溶部Hex及EA可溶部不具直接抑制酪胺酸酶活性。黑色素合成抑制方面，葉部之不同可溶部皆具有抑制黑色素合成之效果，GC-MS成分分析中有苯乙醇成份存在，綜合結果，光蠟樹葉部位皆可應用於美白產品的應用開發。本研究本年度發表研討會輪文”Whitening and anti-inflammatory activity of mahogany”一篇，並成功技轉「高效防蚊配方」。

2014
In this study, we selected the economic planting tree species,Fraxinus formosana and Indian rosewood (Pterocarpus indicus) for the subject .Using MeOH, EtOH extract to obtain a crude extract and different soluble portion ( liquid - liquid chromatography ) , conducted DPPH radical scavenging , anti-inflammatory activity, both inside and outside the cell whitening activity test , toxicity test and a gas chromatograph activity assessment tests and other bioactive components , the purpose of the development of a valuable commodity .By DPPH radical scavenging ability was found that parts of Fraxinus formosana EtOH leaf crude extracts inhibition rate is better, there are 56.74 ± 1.73% inhibition rate , higher crude extract Indian rosewood sticks parts of MeOH extraction rate of inhibition , there are 33.86 ± 1.42% inhibition rate . Anti-inflammatory activity found that macrophages by LPS treatment for 24 hours, the northern and central region of Fraxinus formosana tree and Indian rosewood crude extracts and different concentrations of the soluble portion 100,50,25 ppm in both cell proliferation function , nontoxic effect , only the central and northern branches of different Fraxinus formosana EtOH extraction of the soluble portion , Hex, EA layer having a little poison , Fraxinus formosana seed extraction of crude extracts parts of EtOH and different soluble portion thereof , has a good inflammatory response of cells to repair , remaining at a high concentration of a trace repair inflammation reaction. Inhibit tyrosinase activity outside the cells showed North Fraxinus formosana leaves crude extract parts of MeOH and EtOH extracts inhibit tyrosinase activity better. Screening aspects of melanoma cell survival , and Indian rosewood tree and Fraxinus formosana crude extract not significantly completely inhibited the growth of melanoma , the different soluble portion of melanoma cells in high concentrations of toxic survival rate slightly about 50-70 %, survival rate in lower concentration oof about 70-95%.
Direct and indirect inhibition of intracellular tyrosinase , the different soluble portion of Fraxinus formosana and Indian rosewood trees
compared to commercial kojic acid with the control group , there are inhibition of intracellular activity of tyrosinase inhibitory activity, the strength of the inhibitory activity sequence for Water soluble portion ＞ But soluble portion ＞ EA soluble portion ＞ Hex soluble portion , only in different parts of India Rosewood leaf soluble portion soluble portion Hex and EA does not have to directly inhibit tyrosinase activity. Melanin synthesis inhibition , the different portions of the soluble portion of the leaf were effective in suppressing the synthesis of melanin , GC-MS analysis of components in the presence of phenylethyl alcohol , the combined result. Fraxinus formosana can be applied to application development whitening products .

2015年
本研究以欖仁(Terminalia catappa)及楓香(Liguidambar formosana)枝葉部位為材料，利用MeOH、EtOH、熱水溶劑萃取得到粗萃物及液相-液相層析之不同可溶部抽出物，進行抗氧化、抗發炎、細胞內美白活性試驗之試驗，抗氧化活性結果發現，欖仁葉部位中部區域熱水萃取葉粗萃物最有較佳的清除能力及抑制率，IC50為6.75 ± 0.90 μg/mL，楓香葉部位以嫩葉EtOH萃取之粗萃物具較佳的自由基清除能力，IC50為9.98 ± 1.15 μg/mL，粗萃物以欖仁在自由基清除能力方面整體比楓香的自由基清除能力佳，螯合亞鐵離子能力測定，在高濃度100及50 ppm皆有較佳之螯合亞鐵離子能力，濃度愈高，螯合能力愈強，濃度愈低，則反之。抗發炎活性方面發現，欖仁及楓香葉部位粗萃物及不同可溶部Ethyl acetate、Butenol及Water層抽出物之細胞存活率皆大於85 %，且發現有促進細胞增生之功能，欖仁中部地區葉部EtOH粗萃物之Hexane層抽出物及楓香嫩葉MeOH粗萃物之Hexane層抽出物，在濃度100ppm具有毒殺效果，欖仁紅葉及綠葉EtOH之粗萃物之修復發炎活性比欖仁及楓香葉部位MeOH、EtOH及熱水萃取之粗萃物較佳。可溶部Hexane層抽出物最具有較佳的修復發炎反應，其次是Ethyl acetate層抽出物。細胞內美白活性試驗方面，針對黑色素細胞瘤細胞存活率，粗萃物及不同可溶部在高濃度對黑色素細胞存活有些許的毒殺作用，低濃度毒性不強，只有可溶部Water層抽出物無明顯的細胞毒性，細胞內直接及間接抑制酪胺酸酶方面，欖仁及楓香粗萃物及不同可溶部與對照組市售麴酸及熊果素相比較，在濃度100及50 ppm，皆有抑制細胞內酪胺酸酶的活性，低濃度較不明顯；直接抑制細胞內酪胺酸酶活性試驗，結果顯示抑制活性大小分別為Hexane層抽出物＞ Butenol層抽出物＞Water層抽出物＞Ethyl acetate層抽出物，對黑色素含量之影響，四個可溶部皆具有對黑色素生成具抑制的效果，當濃度愈高，黑色素細胞受到抑制後顏色愈淡，抑制黑色素生成愈佳。

2015
In this study, Terminalia (Terminalia catappa) and sweetgum (Liguidambar formosana) leaves for the material, the use of MeOH, EtOH,water crude extracts obtained by solvent extraction and liquid matter -different soluble portion of the extract liquid chromatography carried out anti-oxidation, anti-inflammatory, test the intracellular activity of the test of whitening.The results showed that the antioxidant activity,the central region of Terminalia leaves crude extracts of hot water extraction was the most preferred scavenging and inhibition rate, IC50 of 6.75 ± 0.90μg/L, with sweetgum leaves crude extracts wasextracted with leaves of EtOH preferred free radical scavenging ability, IC50 of 9.98 ± 1.15 μg /mL, crude extracts was to Terminalia radical scavenging ability in overall good than Liquidambar free radicalscavenging ability. Determination of ferrous ion chelating ability, the high concentration of 100 and 50 ppm have preferred the ferrous ion chelating ability, the higher the concentration, the stronger chelating ability, the concentration of the lower, and vice versa.From the analysis of the antiinflammatory activity was found, Terminalia and sweetgum leaves crude extracts thereof and different soluble portion Ethyl acetate, Butenol and Water layer extracted cell viability was greater than 85% of all, and found to promote the proliferation of cell functions.Hexane layer of Terminalia leaves in the central region of EtOH extraction of crude extracts thereof and sweetgum leaves crude methanol extracts of Hexane layer t, at a concentration of 100 ppm have toxic effects. Antiinflammatory activity of crude extracts of Terminalia leaves and green leafy substance EtOH the preferred ratio Terminalia and sweetgum leaves MeOH, EtOH and crude extracts hot water extraction thing. Hexane layer extract soluble portion most preferred repair inflammation, followed Ethyl acetate layer extrac Whitening active test aspects of the cells,the survival rate for melanoma cells, crude extracts of different things and a high concentration of soluble portion in melanoma cell survival a slight toxic effect, low concentration is not strong toxicity, only the soluble extract of Water layer no significant cytotoxicity. Direct and indirect inhibition of intracellular tyrosinase aspect, Terminalia and
sweetgum crude extracts of different soluble portion thereof and the control group commercial Kojic acid and Arbutin compared to the concentration of 100 and 50 ppm, there is inhibition of intracellular tyramine acid activity, low concentration less obvious. Direct inhibition of intracellular tyrosinase activity test was shown to inhibit the activity of sizes of Hexane layer extract＞ Butenol layer extract＞ Water extraction layer＞ Ethyl acetate layer extract effect on melanin content, the four-soluble portion melanin are having a dampening effect, when the concentration is higher, the melanocytes by suppressing color after the more light, the more good inhibit melanin production. Direct inhibition of intracellular tyrosinase activity test results showed that crude extracts inhibits the activity of capacity respectively Hexane layer extract＞ Butenol layer extract＞ Water extraction layer＞ Ethyl acetate layer extract effect on melanin content, the four soluble portion are having a dampening effect melanin, when the concentration is higher, the melanocytes by suppressing color after the more light, the more good inhibit melanin production.