| Summary: | 博士 === 國立屏東科技大學 === 食品科學系所 === 106 === Part 1: Antrodia cinnamomea (AC) named “Niu-chang-chih” in Chinese, is a medicinal mushroom being widely used as food dietary supplement for cancer prevention and hepatoprotection. It is an endemic species of macro-fungi in Taiwan. In recent years intensive studies has revealed that the fruiting body of AC possesses significant biological activity such as antioxidant, hepatoprotection, anticancer and anti-inflammatory. AC have been identified more than hundred secondary metabolites. But particular attention has been directed to lanostane and ergostane type triterpenoids which are reputed as the most potent that contribute to it’s pharmacological efficacy. Response surface methodology (RSM) was employed to optimize the extraction parameters of total triterpenoids. The maximum value of triterpenoids was obtained at solvent to solid ratio (v/w) of 11.5, extraction temperature of 63 °C, and extraction time of 51 min. Under this condition, the yield of triterpenoids was 240 mg/g of AC extract. An accurate and rapid LC-ESI-MS/MS analytical method was developed and validated for the simultaneous determination of antcin A, antcin B, antcin C, antcin H, antcin K, in the fruiting body and extract of AC.
Part 2: Hypericum formosanum is a Taiwan's endemic plant. The genus Hypericum are known as a rich source of bioactive secondary metabolites such as xanthones, flavonoids and triterpenes. In particular, Hypericin and hyperforin from H. formosanum have been shown to exhibit significant biological effects such as anti-depressant activities. However, there is no report about its bio-activity and chemistry of H. formosanum. The aims of this study were the isolation of chemical constituents from H. formosana, and the evaluation of their bioactivities by bioassay-guided fractionation technology. In this study, the dried of H. formosanum were extracted by methanol. The extracts were partitioned by ethyl acetate (EA) and n-butanol (n-BuOH) to obtain EA, n-BuOH, and water layers. The EA layer was separated fractions by column chromatography. Furthermore, the subfractions were purified by column chromatography, high performance liquid chromatography to yield compounds including three flavonoids. In the present study, ultrahigh performance liquid chromatography equipped with photodiode array detection-mass (UPLC–DAD-MS) method was developed for the separation, identification, and quantification of bioactive flavonoids. The developed method was also validated for accuracy, precision, limit of detection and quantification (LOD and LOQ). In this method, four flavonoids, viz., hyperoside, astilbin, quercitrin, and quercetin, were quantified in linearity range of 10–200 (μg/mL) with correlation coefficient of greater than 0.997. A high recovery (87.55–92.11%) and good reproducibility was obtained for four flavonoids with the relative standard deviation (RSD) ranging from 1.75-2.26 % (intra-day) and 1.99-2.27% (inter-day). Hence, the proposed method for simultaneous quantification of four bioactive flavonoids in the extract and fractions of H. formosanum using UPLC–DAD-MS detection under the specified conditions is accurate and validated. Besides that, subfraction of ethyl acetate fraction (F8) showed the best anti-glycation and its inhibition was observed at 85.9% and significantly reduced matrix metalloproteinase–1 protein expression in human skin keratinocyte cell induced by advanced glycative end product.
Keywords：Antrodiacinnamomea、Response surface methodology (RSM)、Triterpenoids、Hypericum formosanum, bioassay-guided、antiglycation、matrix metalloprotinases
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