Moreira Bruno Oliveira, de Carvalho Anaildes Lago, Alves Clayton Queiroz, Morbeck Lorena Lôbo Brito, Cruz Mariluze Peixoto, Yatsuda Regiane, David Juceni Pereira, David Jorge Mauricio
Instituto Multidisciplinar em Saúde, Universidade Federal da Bahia 45029-094 Vitória da Conquista BA Brazil.
Departamento de Ciências Naturais, Universidade Estadual do Sudoeste da Bahia 45083-900 Vitória da Conquista BA Brazil.
RSC Adv. 2019 Dec 13;9(70):41256-41268. doi: 10.1039/c9ra05985g. eCollection 2019 Dec 9.
The stem barks and leaves of are used in Brazilian folk medicines in the treatment of stomach and intestinal diseases. However, there are no reports of chromatographic methods used to evaluate the bioactives of its standardized extracts and for biological evaluation. An analytical method was developed and validated for simultaneous determination and quantification of the bioactive phenolics gallic acid, methyl gallate, ellagic acid and, the biflavonoids agathisflavone and amentoflavone in the leaves and stem bark of . HPLC operating conditions were optimized and the parameters such as selectivity, linearity, precision, accuracy, LOD, LOQ and, robustness of the method were also evaluated. Robustness was evaluated using a multivariate optimization technique. Linear relationships within the range of investigated concentrations were observed with their correlation coefficients greater than 0.9991. The method was validated for repeatability (RSD ≤ 2.88%), intermediate precision (RSD ≤ 3.38%) with recovery between 84.12 and 106.64% and the RSD less than 3.40% and proved to be robust. Besides, antioxidant, acetylcholinesterase inhibition, anti-inflammatory and antinociceptive activities of the standardized hydromethanolic extracts of leaves and stem bark of this species were evaluated. The method was successfully applied in the quantification of the gallic acid, methyl gallate, ellagic acid, agathisflavone and amentoflavone of standardized extracts. The results showed the present method developed was simple, sensitive, reproducible, accurate and precise. The standardized hydromethanolic extracts of leaves and stem bark of showed antioxidant activity (EC 69.09 and 83.06 μg mL), acetylcholinesterase inhibition (52.23 and 83.36%) and they were able to inhibit the formalin-induced nociception and also reduced the edema formations at 100 mg kg doses. The anti-inflammatory potentials were evaluated by the decrease of the Cg-induced neutrophils migrations at the same doses.
该植物的茎皮和叶在巴西民间医学中用于治疗胃肠疾病。然而，尚无关于用于评估其标准化提取物生物活性物质以及进行生物学评估的色谱方法的报道。开发并验证了一种分析方法，用于同时测定和定量该植物叶和茎皮中的生物活性酚类物质没食子酸、没食子酸甲酯、鞣花酸以及双黄酮阿盖黄酮和穗花杉双黄酮。对高效液相色谱（HPLC）操作条件进行了优化，并评估了该方法的选择性、线性、精密度、准确度、检测限、定量限和稳健性等参数。使用多变量优化技术评估稳健性。在所研究的浓度范围内观察到线性关系，其相关系数大于0.9991。该方法在重复性（相对标准偏差≤2.88%）、中间精密度（相对标准偏差≤3.38%）方面得到验证，回收率在84.12%至106.64%之间，相对标准偏差小于3.40%，且证明具有稳健性。此外，还评估了该物种叶和茎皮的标准化水甲醇提取物的抗氧化、乙酰胆碱酯酶抑制、抗炎和抗伤害感受活性。该方法成功应用于标准化提取物中没食子酸、没食子酸甲酯、鞣花酸、阿盖黄酮和穗花杉双黄酮的定量分析。结果表明，所开发的本方法简单、灵敏、可重现且准确精密。该植物叶和茎皮的标准化水甲醇提取物显示出抗氧化活性（半数有效浓度分别为69.09和83.06μg/mL）、乙酰胆碱酯酶抑制活性（分别为52.23%和83.36%），并且在100mg/kg剂量下能够抑制福尔马林诱导的伤害感受，还能减轻水肿形成。通过相同剂量下对角叉菜胶诱导的中性粒细胞迁移的减少来评估其抗炎潜力。
