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Abstract：
Propolis extracts have been widely studied due to their popularity in traditional medicine, presenting incredible biodiversity. This study aimed to analyze propolis extracts\' phytochemical, physicochemical, and biological activities from four different biogeographic zones of the Huila region (Colombia). The raw material samples were collected by the scraping method and the ethanolic extracts (EEPs) were obtained by cold maceration with ethanol (96%). The physicochemical and sensory characterization was carried out according to the protocols recommended by the Brazilian Ministry of Agriculture and the main components of the EEPs were identified by LC-HRMS analysis. The determination of total phenols and flavonoids was carried out using colorimetric techniques. The antioxidant activity, cytotoxicity, and cell cycle regulation analyses in L929 and HGnF cells were evaluated using DPPH, Alamar Blue, and 7-amino actinomycin D (7-AAD) assays. The propolis samples presented an average yield of 33.1%, humidity between 1.6 and 2.8%, melting point between 54 and 62 °C, ashes between 1.40 and 2.19%, and waxes of 6.6-17.9%, respectively. The sensory characteristics of all samples were heterogeneous, complying with the quality specifications established by international standards. The polyphenolic and total flavonoid content was representative in the samples from Quebradon (255.9 ± 9.2 mg GAE/g, 543.1 ± 8.4 mg QE/g) and Arcadia (543.1 ± 8.4 mg GAE/g, 32.5 ± 1.18 g QE/g) (p < 0.05) that correlated with high antioxidant activity (Quebradon: 37.2 ± 1.2 µmol/g, Arcadia: 38.19 ± 0.7 µmol/g). In the chemical composition analysis, 19 compounds were characterized as phenolic acids and flavonoids, the most representative being chrysoeriol-O-methyl-ether, ellagic acid, and 3,4-O-dimethylcaffeic acid. Regarding biological activity, Quebradon and Arcadia propolis presented low toxicity with IC50 of 2.83 ± 2.3 mg/mL and 4.28 ± 1.4 mg/mL in HGnF cells, respectively, and an arrest of the cell cycle in the G2/M phase of 71.6% and 50.8% compared to the control (11.9%) (p < 0.05). In general, the results of this study contribute to the identification of valid quality criteria to evaluate Colombian propolis, contributing to its study and chemical and biological characterization as a source of raw material for industrial and pharmaceutical use. In addition, Quebradon and Arcadia propolis can be important sources of bioactive molecules for the development of new drugs.
摘要：
蜂胶提取物由于其在传统医学中的流行而被广泛研究，呈现令人难以置信的生物多样性。本研究旨在分析蜂胶提取物的植物化学成分，物理化学,和来自威拉地区（哥伦比亚）四个不同生物地理区域的生物活动。通过刮削法收集原料样品，并通过用乙醇（96％）冷浸渍获得乙醇提取物（EEP）。根据巴西农业部推荐的方案进行物理化学和感官表征，并通过LC-HRMS分析鉴定EEP的主要成分。使用比色技术进行总酚和类黄酮的测定。抗氧化活性，细胞毒性，使用DPPH评估L929和HGnF细胞中的细胞周期调控分析，Alamar蓝色,和7-氨基放线菌素D(7-AAD)测定。蜂胶样品的平均产量为33.1%，湿度在1.6和2.8%之间，熔点在54和62°C之间，灰烬在1.40和2.19%之间，蜡为6.6-17.9%，分别。所有样品的感官特性均不均匀，符合国际标准制定的质量规范。Quebradon样品中的多酚和总黄酮含量具有代表性（255.9±9.2mgGAE/g，543.1±8.4mgQE/g）和阿卡迪亚（543.1±8.4mgGAE/g，与高抗氧化活性相关的32.5±1.18gQE/g）（p<0.05）（Quebradon：37.2±1.2µmol/g，阿卡迪亚：38.19±0.7µmol/g）。在化学成分分析中,19个化合物被表征为酚酸和类黄酮，最具代表性的是β-O-甲基醚，鞣花酸,和3,4-O-二甲基咖啡酸。关于生物活性,Quebradon和Arcadia蜂胶在HGnF细胞中表现出低毒性，IC50为2.83±2.3mg/mL和4.28±1.4mg/mL。分别,与对照(11.9%)相比,细胞周期停滞在G2/M期71.6%和50.8%(p<0.05)。总的来说,这项研究的结果有助于确定有效的质量标准来评估哥伦比亚蜂胶，有助于其研究以及作为工业和制药原料的化学和生物学表征。此外,Quebradon和Arcadia蜂胶可能是开发新药的生物活性分子的重要来源。
