牡丹叶提取物的含量测定及其协同羧甲基壳聚糖的抗菌活性研究

Determination of the Content of Peony Leaf Extract and its Antibacterial Activity in Synergy with Carboxymethyl Chitosan

目的牡丹(Paeonia suffruticosa Andr.)是毛茛科芍药属的多年生落叶灌木,其根皮被称为“牡丹皮”,药用历史悠久,其传统非药用部位“叶”在民族地区应用广泛,研究牡丹叶的药用价值是提高中药资源综合利用的有益尝试。牡丹叶含有的多酚、黄酮类活性成分,在抑菌、抗氧化和抑制酪氨酸酶等方面作用显著。方法采用甲醇提取法获取牡丹叶提取物,建立同时测定牡丹叶提取物中5种活性成分(没食子酸、芍药苷、山奈素、槲皮素和异鼠李素)含量的高效液相色谱(HPLC)法。同时,采用Folin-Ciocalteu法和硝酸铝-亚硝酸钠比色法测定了牡丹叶提取物中总黄酮和总多酚的含量。立足羧甲基壳聚糖(CMCS)抗菌活性和良好的载药特性,研究其与牡丹叶提取叶的协同机制。结果含量测定结果显示,牡丹叶中没食子酸、芍药苷、山奈素、槲皮素和异鼠李素的含量分别为17.780%、0.470%、1.150%、1.920%和3.160%,其线性关系、稳定性、重复性和精密度等均良好。体外抗菌活性结果显示,牡丹叶提取物(PLE)与CMCS联用的FICI系数均小于0.500,其对金黄色葡萄球菌和大肠埃希菌的抑菌效果均表现为协同作用。结论牡丹叶提取物中含有较高的多酚类和黄酮类成分,并且与CMCS具有较强的协同作用。为进一步开发牡丹叶与羧甲基壳聚糖相关抗菌制剂的研制提供联用数据支撑,为牡丹药用资源的综合利用奠定基础。

Objective Paeony(Paeonia suffruticosa Andr.)is a perennial deciduous shrub belonging to the genus Paeonia in the goldencup family.Its root bark is called"peony bark"and its traditional non-medicinal part"leaf"is widely used in ethnic areas.Tyrosinase inhibition,antioxidant properties,and antibacterial properties are all significantly impacted by the active flavonoids and polyphenols found in peony leaves.Methods A high performance liquid chromatography(HPLC)method was established for the simultaneous determination of five active components(gallic acid,paeoniflorin,kaempferol,quercetin and isorhamnetin)in the extract of peony leaves by methanol extraction.Meanwhile,the contents of total flavonoids and total polyphenols in the extract of peony leaves were determined by the Folin-Ciocalteu method and aluminum nitrate-sodium nitrite colorimetric method.The study examined the synergistic mechanism between carboxymethyl chitosan(CMCS)and peony leaf extract,taking into account CMCS's antibacterial activity and superior drug loading capabilities.Results The results showed that the contents of gallic acid,paeoniflorin,kaempferol,quercetin and isorhamnetin were 17.780%,0.470%,1.150%,1.920%and 3.160%,and the linear relationship,stability,repeatability and precision of the leaves were good.Peonies leaf extract(PLE)in combination with CMCS had a FICI coefficient of less than 0.500 and a synergistic antibacterial action against Staphylococcus aureus and Escherichia coli,according to the results of its antibacterial activity in vitro.Conclusion The extract of peony leaves contains high polyphenols and flavonoids,and has strong synergistic effect with CMCS.This study provided data support for the further development of antibacterial preparations related to carboxymethyl chitosan from peony leaves,and laid a foundation for the comprehensive utilization of peony medicinal resources.