Determination of cytotoxic, antioxidant activities and LC/MS-MS profiles of three endemic Verbascum L. species

Sevim Küçük, M. Has, Fatih Göger, Filiz Özdemir, Z. Incesu
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Abstract

The Verbascum genus includes many species used in folk medicine or the treatment of various diseases. In this study, the cytotoxic, antioxidant activity, and LC/MS-MS profiles of three Verbascum species, which are endemic in Eskişehir and its surroundings, were investigated. The cytotoxic effects of methanol extract of V.detersile, V. eskisehirensis, and V.gypsicola species on the cervical (HeLa) and ovarian cancer (SKOV-3) cells were investigated using a colorimetric assay. The results indicated that cytotoxic effect was not observed after treatment of SKOV-3 cells with Verbascum. On the other hand, the cytotoxic activity of V. detersile was found to be 0.1910 mg/dL and 1.057 mg/dL for HeLa cells after 24 or 48 hours incubation with V.detersile, respectively. The antioxidant activity was determined as 1,1-diphenyl-2-picrylhydrazyl (DPPH•) radical scavenging activity, trolox equivalent antioxidant capacity (TEAC assay), and also the total phenolic content of the samples was found. Total phenols were estimated as Gallic acid equivalents (GAE), expressed as mg Gallic acid/in 1g extract previously described by Singletton. LC/MS profiles separation and detection of phytochemicals of the extract were performed on a Shimadzu UPLC system consisting of a vacuum degasser, an autosampler (SIL20A Shimadzu Autosampler), a binary pump (LC20AD Shimadzu), an oven (CTO20A Shimadzu Column Oven) and DAD dedector (SPD M20A Shimadzu DAD Detector). In terms of antioxidant activity, V. gypsicola was found to have the least antioxidant activity among the three extracts, which is also correlated with the total amount of phenolic content in its content. In this way, it differs from other species. V. detersile exhibits a different chemical profile from the other two species with the iridoid catalpol derivatives it contains. Apigenin pentoside is the only flavonoid molecule detected in V. detersille.
测定三种地方性马鞭草的细胞毒性、抗氧化活性和 LC/MS-MS 图谱
马鞭草属包括许多用于民间医药或治疗各种疾病的物种。本研究调查了埃斯基谢希尔及其周边地区特有的三种马鞭草的细胞毒性、抗氧化活性和 LC/MS-MS 图谱。 采用比色法研究了 V.detersile、V.eskisehirensis 和 V.gypsicola 的甲醇提取物对宫颈癌(HeLa)和卵巢癌(SKOV-3)细胞的细胞毒性作用。结果表明,用马鞭草处理 SKOV-3 细胞后,未观察到细胞毒性作用。另一方面,V.detersile 对 HeLa 细胞的细胞毒性活性分别为 0.1910 mg/dL 和 1.057 mg/dL。抗氧化活性以 1,1-二苯基-2-苦基肼(DPPH-)自由基清除活性、trolox 当量抗氧化能力(TEAC 法)来测定,同时还测定了样品中的总酚含量。根据 Singletton 的描述,总酚以没食子酸当量(GAE)估算,以没食子酸毫克/1 克提取物表示。采用岛津 UPLC 系统对提取物中的植物化学物质进行 LC/MS 图谱分离和检测,该系统由真空脱气机、自动进样器(SIL20A 岛津自动进样器)、二元泵(LC20AD 岛津)、烤箱(CTO20A 岛津柱烤箱)和 DAD 检测器(SPD M20A 岛津 DAD 检测器)组成。在抗氧化活性方面,发现 V. gypsicola 的抗氧化活性在三种提取物中最低,这也与它所含的酚类物质的总量有关。因此,它与其他物种有所不同。V. detersile 所含的鸢尾梓醇衍生物显示出与其他两个物种不同的化学特征。芹菜素戊苷是在 V. detersille 中检测到的唯一黄酮类分子。
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