Isolation and Identification of Phloroglucinol Derivatives from Dryopteris crassirhizoma by HPLC-LTQ-Orbitrap Mass Spectrometry
Isolation and Identification of Phloroglucinol Derivatives from Dryopteris crassirhizoma by HPLC-LTQ-Orbitrap Mass Spectrometry
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DOI：
10.1007/s10600-016-1887-x
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发表时间：
2016-11-01
影响因子：
0.8
通讯作者：
Wang, Hui-yun
中科院分区：
文献类型：
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作者：
Ren, Qiang;Quan, Xian-gao;Wang, Hui-yun
药理学研究表明，D.绵马贯众具有多种药理活性，例如抗氧化活性（通过其体外1，1-二苯基-2-苦肼基（DPPH）自由基清除能力确定[1]）和对人前列腺癌细胞的抗癌活性[2]。D.含有黄烷酸AB和PB的绵马贯众对耐甲氧西林金黄色葡萄球菌具有抗菌活性[3，4]。黄曲霉酸AB能抑制猪繁殖与呼吸综合征病毒的复制[5]。本文报道了从D.绵马贯众具有脂肪酸合成酶抑制活性[6]。此外，D.通过抑制ERK/AP-1和TBK 1/IRF 3通路，绵马贯众显示出强的抗炎活性[7]。以往的植物化学研究表明，D.绵马贯众主要含有间苯三酚衍生物、三萜类化合物和黄酮类化合物[8-12]。鳞毛蕨间苯三酚衍生物由通过亚甲基桥接的单体间苯甲酮环组成。这些具有类似组成和类似极性的同系物在纯态下由于分解和还原性碱裂解而非常难以分离。在过去的几十年中，电子碰撞电离[13]，场解吸电离[14]和快原子轰击电离[15]已被用于阐明间苯三酚衍生物的结构。然而，没有足够的信息，在线表征和鉴定的间苯三酚衍生物。本研究的目的是确定潜在的化合物负责的抗白血病活性D。绵马藤首先，用95%乙醇水溶液对D.发现绵马贯众对人白血病Reh细胞系表现出细胞毒活性。根据生物活性定向分级分离，组分B和D是对Reh最具活性的组分，其进一步分离得到黄烷酸AB（3）、去甲黄烷酸AB（5）和3-甲基丁酰间苯三酚（6）以及三种已知化合物二十五烷醇（1）、3-羟基二十七烷醇（2）和E-谷甾醇（4）。化合物6为首次从该植物中分离得到。此外，还报道了D.采用HPLC-LTQ-Orbitrap质谱法进行了定性鉴定。共鉴定了10个间苯三酚衍生物的基础上，准确的质量，多阶段的质量数据，和特征的碎片模式。
Pharmacological investigations have demonstrated that D. crassirhizoma has a variety of pharmacological activities, such as antioxidant activity, determined by its 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability in vitro [1], and anticancer activity against human prostate cancer cells [2]. The hexane fraction of D. crassirhizoma, which contains flavaspidic acids AB and PB has antibacterial activity against methicillin-resistant Staphylococcus aureus [3, 4]. Flavaspidic acid AB can inhibit porcine reproductive and respiratory syndrome virus replication [5]. The acylphloroglucinol derivatives from the rhizomes of D. crassirhizoma have fatty acid synthase inhibitory activity [6]. In addition, the ethanol extract of D. crassirhizoma displays strong anti-inflammatory activity by suppressing ERK/AP-1 and TBK1/IRF3 pathways [7]. Previous phytochemical investigations revealed that D. crassirhizoma contains mainly phloroglucinol derivatives, triterpenoid, and flavonoids [8–12].The Dryopteris phloroglucinol derivatives consist of monomeric phlorophenone rings bridged by a methylene group. These homologues with an analogous composition and similar polarity were very difficult to separate in a pure state due to decomposition and reductive alkaline cleavage. Over the past decades, electron impact ionization [13], field-desorption ionization [14], and fast atom bombardment ionization [15] have been employed to elucidate the structures of phloroglucinol derivatives. However, there is not enough information about on-line characterization and identification of the phloroglucinol derivatives. The purpose of the present study was to identify the potential compounds responsible for the antileukemic activity of D. crassirhizoma Nakai. Firstly, a crude 95% aqueous ethanol extract of D. crassirhizoma Nakai was found to exhibit cytotoxic activity against human leukemic Reh cell lines. According to bioactivity-directed fractionation and isolation, fractions B and D were the most active fractions against Reh, which were further separated to yield flavaspidic acid AB (3), norflavaspidic acid AB (5), and 3-methylbutyrylphloroglucinol (6) together with three known compounds pentacosanol (1), 3-hydroxyheptacosanol (2), and E-sitosterol (4). Compound 6 was isolated from this plant for the first time. In addition, the phloroglucinol derivatives of D. crassirhizoma Nakai were characterized and identified by HPLC-LTQ-Orbitrap mass spectrometry. A total of 10 phloroglucinol derivatives was identified on the basis of accurate mass, multiple-stage mass data, and characteristic fragmentation patterns.