| 摘要: | 硫酸鹽泉、碳酸鹽泉及碳酸氫鹽泉相傳浸泡後具有養顏及美白功效，但至今仍缺乏直接科學研究證據。本研究的目的在於探討溫泉對於黑色素生成及聚合影響，並透過體外酪胺酸酶之活性測試，作為本研究評估溫泉影響黑色素形成的之重要依據。實驗組有硫酸鹽泉、碳酸鹽泉及碳酸氫鹽作為本研究之溫泉材料來源，並以磷酸鹽緩衝溶液作為研究之對照組。結果發現，相對於對照組而言，硫酸鹽泉及碳酸氫鹽泉在含酪胺酸(2 mM)之實驗溶液中添加酪胺酸酶(27 units/ml)後的第10, 30 和 60 分鐘，皆具有抑制酪胺酸酶活化黑色素生成的作用p<0.001)，但碳酸鹽泉則無明顯之統計意義。進一步在實驗組及對照組溶液中添加金屬離子螯合劑 (EDTA, 25mg/mL)充分螯合反應後，再經微膜濾紙 (0.22 μm)移除溶液中的兩價以上陽離子，結果發現對黑色素生成作用之抑制率，對照組大約有10%抑制效果，但實驗組之碳酸鹽泉及碳酸氫鹽泉有50%的抑制效果(p<0.01)，而硫酸鹽泉更高達100 %的抑制率(p<0.01)。在探討酸氫鹽成分對黑色素形成之影響評估研究實驗，實驗組及對照組溶液中添加氯化鈣(CaCl2, 25mg/mL)，再經煮沸冷却和微膜濾紙 (0.22 μm)移除溶液中的碳酸氫鹽成分。結果顯示，除了硫酸鹽泉有促進黑色素生成外(p<0.01)，碳酸氫鹽泉和碳酸氫鹽泉相對於對照組並無統計意義。最後本研究針對黑色素形成後之聚合反應進行評估研究，評估時間點為添加酪胺酸酶(27 units/ml)後的第48 小時，並以氫氧化鈉(NaOH,
2M)沸騰方式來溶解聚合反應之黑色素沈澱物。結果顯示，若併合聚合之黑色素計算，實驗組三種溫泉溶液相對於對照組，皆呈現有40-60% 的黑色素形成抑制率， 並能有效防止黑色素聚合反應(p<0.01)，此一防止黑色素聚合反應結果，除了硫酸鹽泉因受添加金屬離子螯合劑而完全阻斷黑色素形成，使統計量呈現100%之大幅偏移結果外，去除兩價金屬離子或碳酸離子，對防止黑色素聚合反應，無論實驗組和對照組並無明顯影響。綜合本研究結果發現，硫酸鹽泉及碳酸氫鹽泉能抑制早期黑色素形成及防止後期之黑色素聚合反應，碳酸鹽泉雖然對早期黑色素形成沒有抑制作用，但對後期黑色素形成及黑色素聚合反應也呈現抑制作用。
The carbonated and sulfated spring water had shown the effects of skin beauty and whiteness, but still lacks the scientific evidences. This research will discuss the possible mechanisms of spring water on melanin pigmentation, melanin polymerization, and focus on the tyrosinase activity in vitro. Study was divided into experimental group and control group. The experimental group was divided into 3 subgroups, as well as the sulfated, carbonated and bicarbonate hot spring water as the source material, and with phosphate buffer solution as the study control group. The results showed that, compared with the control group, the sulfated and bicarbonated spring water, which containing tyrosine (2 mM) and adding the test solution of tyrosinase (27 units / ml) after melamin formation reaction time 10, 30 and 60 minutes, all inhibit tyrosinase activity in the role of melanin production (p <0.001), but carbonated spring water was no clear statistical significance. Further in the experimental group and control group solution, add the chelator (EDTA, 25mg/mL) to fully chelate reaction, and then through the micro-membrane filter (0.22 μm) to remove the solution for more than divalent metal cations, was found on the role of melanin formation inhibition rate of about 10% of the control group, but inhibition of the experimental group of carbonate and bicarbonate salt spring, 50% of the inhibitory effect (p <0.01), while the sulfate spring as high as 100% inhibition rate (p <0.01).Of carbonated and bicarbonated spring water on impact assessment study of melanin formation, before the melanin formation experiment, thecalcium chloride (CaCl2, 25mg/mL) was adding all study groups then boiling, cooling, and through the micro-membrane filter (0.22 μm) to remove the calcium carbonate in solution. The results showed that, in addition to sulfated spring water promoted the formation of melanin (p <0.01), but the bicarbonate and bicarbonated spring water compared to the control group was not statistically significant. Finally, the study focuses on melanin polymerization after 48 hours adding tyrosinase (27 units / ml), then a depolymer reaction was adding the sodium hydroxide (NaOH, 2M) and boiling. The findings indicated that the total melanin formation (monomer and polymer) in all experimental groups appeared 40-60% inhibition rate of melanin formation, compared with control group, and all experimental groups can effectively prevent melanin polymerization (p <0.01). Furthermore, in addition to the sulfated spring water which removal of divalent metal ions was shown completely inhibition of melanin formation, the removal of divalent metal ions or carbonate ions in all study groups to prevent the melanin polymerization, both experimental and control groups did not significant. Basic on this study, that compared with the PBS solution (control group), the sulfated and bicarbonated spring water can inhibit melanin formation in early phase, but the carbonated spring water does not effect. To prevent the later phase of melanin polymerization, all kinds of spring
water had the effect of inhibition compared with control group. |
