Khallouki F, Haubner R, Hull W E, Erben G, Spiegelhalder B, Bartsch H, Owen R W
Division of Toxicology and Cancer Risk Factors, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany.
Food Chem Toxicol. 2007 Mar;45(3):472-85. doi: 10.1016/j.fct.2006.09.011. Epub 2006 Oct 1.
The root bark of Anisophyllea dichostyla R. Br. is traditionally used in the Democratic Republic Congo for the treatment of several conditions such as anorexia, fatigue and intestinal infections. We have identified and quantitated several polyphenol antioxidants in the methanol extract of the root bark (120g). The polyphenol content (3.32g/kg) was predominantly ellagitannins (25%) and polyhydroxyflavan-3-ols (catechins and procyanidins, 75%) with 3'-O-methyl-3,4-methylenedioxo ellagic acid 4'-O-beta-d-glucopyranoside and (-)-epicatechin as the major species in each class. These two compounds and the following species were identified unequivocally by NMR spectroscopy: (+)-catechin, (-)-epicatechin 3-O-gallate, 3-O-methyl ellagic acid, 3,3'-di-O-methyl ellagic acid, 3'-O-methyl-3,4-methylenedioxo ellagic acid, 3'-O-methyl-3,4-methylenedioxo ellagic acid 4'-O-beta-d-glucopyranoside, and 3'-O-methyl ellagic acid 4-O-beta-d-xylopyranoside. The following additional compounds were purified by semi-preparative HPLC and tentatively identified on the basis of UV spectra, HPLC-ESI-MS and nano-ESI-MS-MS: (+)-catechin-3-O-beta-d-glucopyranoside, epicatechin-(4beta-->8)-catechin (procyanidin B(1)), epicatechin-(4beta-->8)-epicatechin (procyanidin B(2)), an (epi)catechin trimer, 3-O-methyl ellagic acid 4-O-beta-d-glucopyranoside, (-)-epicatechin 3-O-vanillate, 3,4-methylenedioxo ellagic acid 4'-O- beta-d-glucopyranoside, and 3,3'-di-O-methyl ellagic acid 4-O-beta-d-xylopyranoside. Fractionation of the raw extract by column chromatography on silicic acid yielded 10 fractions. In the hypoxanthine/xanthine oxidase antioxidant assay system, CC-9 which contained a range of polyphenols dominated by (-)-epicatechin-O-gallate proved to be the most potent antioxidant fraction (IC(50)=52 micro g/mL) in terms of ROS scavenging. In terms of XO inhibition CC-8, dominated by (epi)catechin trimer and which also contained appreciable amounts of 3'-O-methyl ellagic acid 4'-O-beta-d-xylopyranoside, as well as the catechins (+)-catechin-3-O-beta-d-glucopyranoside, epicatechin-(4beta-->8)-catechin (procyanidin B(1)), and (-)-epicatechin 3-O-gallate, proved to be the most potent (IC(50)=36 micro g/mL).
二歧异叶番荔枝的根皮在刚果民主共和国传统上用于治疗多种病症，如厌食、疲劳和肠道感染。我们已鉴定并定量了根皮（120克）甲醇提取物中的几种多酚抗氧化剂。多酚含量（3.32克/千克）主要为鞣花单宁（25%）和多羟基黄烷-3-醇（儿茶素和原花青素，75%），其中3'-O-甲基-3,4-亚甲二氧基鞣花酸4'-O-β-D-吡喃葡萄糖苷和（-）-表儿茶素是每类中的主要成分。通过核磁共振光谱明确鉴定了这两种化合物以及以下成分：（+）-儿茶素、（-）-表儿茶素3-O-没食子酸酯、3-O-甲基鞣花酸、3,3'-二-O-甲基鞣花酸、3'-O-甲基-3,4-亚甲二氧基鞣花酸、3'-O-甲基-3,4-亚甲二氧基鞣花酸4'-O-β-D-吡喃葡萄糖苷和3'-O-甲基鞣花酸4-O-β-D-木糖吡喃糖苷。通过半制备型高效液相色谱法纯化了以下其他化合物，并根据紫外光谱、高效液相色谱-电喷雾电离质谱和纳升电喷雾电离串联质谱进行了初步鉴定：（+）-儿茶素-3-O-β-D-吡喃葡萄糖苷、表儿茶素-（4β→8）-儿茶素（原花青素B(1)）、表儿茶素-（4β→8）-表儿茶素（原花青素B(2)）、一种（表）儿茶素三聚体、3-O-甲基鞣花酸4-O-β-D-吡喃葡萄糖苷、（-）-表儿茶素3-O-香草酸酯、3,4-亚甲二氧基鞣花酸4'-O-β-D-吡喃葡萄糖苷和3,3'-二-O-甲基鞣花酸4-O-β-D-木糖吡喃糖苷。通过硅胶柱色谱法对粗提取物进行分馏得到10个馏分。在次黄嘌呤/黄嘌呤氧化酶抗氧化测定系统中，以（-）-表儿茶素-O-没食子酸酯为主的含有多种多酚的CC-9馏分在清除活性氧方面被证明是最有效的抗氧化馏分（半数抑制浓度=52微克/毫升）。就抑制黄嘌呤氧化酶而言，以（表）儿茶素三聚体为主且还含有相当量的3'-O-甲基鞣花酸4'-O-β-D-木糖吡喃糖苷以及儿茶素（+）-儿茶素-3-O-β-D-吡喃葡萄糖苷、表儿茶素-（4β→8）-儿茶素（原花青素B(1)）和（-）-表儿茶素3-O-没食子酸酯的CC-8馏分被证明是最有效的（半数抑制浓度=36微克/毫升）。
