Department of Bioprocess and Polymer Engineering, School of Chemical and Energy Engineering, Faculty of Engineering, University Teknologi Malaysia, Skudai 81310, Malaysia.
Institute of Marine Biotechnology, Universiti Malaysia Terengganu, Kuala Terengganu 21030, Malaysia.
Molecules. 2020 Jan 24;25(3):517. doi: 10.3390/molecules25030517.
Herbal plants are traditionally utilized to treat various illnesses. They contain phytochemicals that can be extracted using conventional methods such as maceration, soxhlet, and boiling, as well as non-conventional methods including ultrasonic, microwave, and others. leaves have been used for the treatment of dengue, fungal, and bacterial infections as well as an ingredient in anti-aging products. Phytochemicals analysis detected the presence of kaempferol, myricetin, carpaine, pseudocarpaine, dehydrocarpaine I and II, ferulic acid, caffeic acid, chlorogenic acid, β-carotene, lycopene, and anthraquinones glycoside. Conventional preparation by boiling and simple maceration is practical, simple, and safe; however, only polar phytochemicals are extracted. The present study aims to investigate the effects of three different non-conventional extraction techniques (ultrasonic-assisted extraction, reflux, and agitation) on phytochemical constituents, the antioxidant capacity, and wound-healing activities. Among the three techniques, the reflux technique produced the highest extraction yield (17.86%) with the presence of saponins, flavonoids, coumarins, alkaloids, and phenolic metabolites. The reflux technique also produced the highest 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging with an IC value of 0.236 mg/mL followed by ultrasonic-assisted extraction (UAE) (IC: 0.377 mg/mL) and agitation (IC: 0.404 mg/mL). At tested concentrations (3.125 µg/mL to 500 µg/mL), all extracts do not exhibit a cytotoxicity effect on the human skin fibroblast, HSF1184. Interestingly, reflux and UAE were active fibroblast proliferators that support 85% (12.5 µg/mL) and 41% (6.25 µg/mL) better cell growth, respectively. Additionally, during the early 24 h of the scratch assay, the migration rate at 12.5 µg/mL was faster for all extracts with 51.8% (reflux), 49.3% (agitation), and 42.5% (UAE) as compared to control (21.87%). At 48 h, proliferated cells covered 78.7% of the scratch area for reflux extract, 63.1% for UAE, 61% for agitation, and 42.6% for control. Additionally, the collagen synthesis was enhanced for 31.6% and 65% after 24 and 48 h of treatment for reflux. An HPLC-MS/MS-QTOF (quadruple time-of-flight) analysis of reflux identified nine phytochemicals, including carpaine, kaempferol 3-(2G-glucosylrutinoside), kaempferol 3-(2″-rhamnosylgalactoside), 7-rhamnoside, kaempferol 3-rhamnosyl-(1->2)-galactoside-7-rhamnoside, luteolin 7-galactosyl-(1->6)-galactoside, orientin 7-O-rhamnoside, 11-hydroperoxy-12,13-epoxy-9-octadecenoic acid, palmitic amide, and 2-hexaprenyl-6-methoxyphenol. The results suggested that reflux was the best technique as compared to ultrasonic and agitation.
草药传统上被用于治疗各种疾病。它们含有可以通过常规方法（如浸渍、索氏提取和煮沸）以及非传统方法（如超声、微波等）提取的植物化学物质。该植物已被用于治疗登革热、真菌和细菌感染，以及抗衰老产品的成分。植物化学物质分析检测到山奈酚、杨梅素、卡巴碱、伪卡巴碱 I 和 II、去氢卡巴碱 I 和 II、阿魏酸、咖啡酸、绿原酸、β-胡萝卜素、番茄红素和蒽醌糖苷的存在。煮沸和简单浸渍的常规制备既实用、简单又安全；然而，只有极性植物化学物质被提取出来。本研究旨在探讨三种不同非传统提取技术（超声辅助提取、回流和搅拌）对植物化学成分、抗氧化能力和伤口愈合活性的影响。在这三种技术中，回流技术产生了最高的提取产率（17.86%），并存在皂角苷、类黄酮、香豆素、生物碱和酚类代谢物。回流技术还产生了最高的 2,2-二苯基-1-苦基肼（DPPH）自由基清除能力，IC 值为 0.236mg/mL，其次是超声辅助提取（UAE）（IC：0.377mg/mL）和搅拌（IC：0.404mg/mL）。在测试浓度（3.125μg/mL 至 500μg/mL）下，所有提取物对人皮肤成纤维细胞 HSF1184 均没有细胞毒性作用。有趣的是，回流和 UAE 是有效的成纤维细胞增殖剂，分别支持 12.5μg/mL（85%）和 6.25μg/mL（41%）更好的细胞生长。此外，在划痕实验的前 24 小时内，所有提取物的迁移率在 12.5μg/mL 时都更快，其中回流为 51.8%，搅拌为 49.3%，UAE 为 42.5%，而对照为 21.87%。在 48 小时时，回流提取物的增殖细胞覆盖了划痕区域的 78.7%，UAE 为 63.1%，搅拌为 61%，对照为 42.6%。此外，在 24 和 48 小时的治疗后，胶原合成分别增强了 31.6%和 65%。回流的 HPLC-MS/MS-QTOF（四极飞行时间）分析鉴定出了 9 种植物化学物质，包括卡巴碱、山奈酚 3-（2G-葡萄糖基芦丁苷）、山奈酚 3-（2”-鼠李糖苷半乳糖苷）、7-鼠李糖苷、山奈酚 3-鼠李糖苷-（1->2）-半乳糖苷-7-鼠李糖苷、木樨草素 7-半乳糖基-（1->6）-半乳糖苷、橙皮苷 7-O-鼠李糖苷、11-过氧-12,13-环氧-9-十八烯酸、棕榈酰胺和 2-十六烷基-6-甲氧基苯酚。结果表明，回流是最好的技术，优于超声和搅拌。
