东紫苏总黄酮提取、纯化及活性初探

Study on Extraction,Purification and Activity of Total Flavonoids from Elsholtzia bodinieri Van

目的:对东紫苏总黄酮提取、纯化最佳工艺及其活性进行探究。方法:采用单因素试验得到东紫苏总黄酮纯化工艺,利用β-半乳糖苷酶染色观察衰老细胞阳性率,SOD活性实验和ABTS实验检测东紫苏总黄酮抗氧化活性,MTT法检测其对HCT116、A549两种细胞增殖的抑制作用。结果:东紫苏初提物总黄酮含量为19.375 mg/g,AB-8最适合用于东紫苏总黄酮的分离纯化,其静态吸附-解析最优条件为上样质量浓度1.5 mg/mL、pH 5、吸附6 h,解析采用70%酸化乙醇(pH 5),时间2 h,纯度由46.67%提高至81.62%;动态吸附-解析最适工艺条件为上样液流速2 mL/min、洗脱液流速1 mL/min,80 mL即可洗脱,纯度提高至80.33%。抗氧化活性:0.25 mg/mL总黄酮溶液处理组较对照组β-半乳糖苷酶染色观察衰老细胞阳性率降低(P<0.01),东紫苏总黄酮对ABTS自由基清除IC50为0.4844 mg/mL,维生素C对ABTS自由基清除率IC50为0.1545 mg/mL。0.5 mg/mL、1 mg/mL处理组SOD活性较损伤组高(P<0.001)。抗肿瘤活性:东紫苏总黄酮对HCT116和A549生长有一定的抑制作用,IC50分别是0.8486 mg/mL和0.7708 mg/mL,表明东紫苏总黄酮是一种潜在的天然抗氧化剂并有一定的肿瘤抑制作用。结论:利用最佳提取、纯化得到的东紫苏总黄酮具有一定的抗氧化、抗肿瘤活性。

【Objective】To study the purification conditions and activity of total flavonoids from Elsholtzia bodinieri Van.【Method】The single factor experiment was used to obtain the purification process of total flavonoids from Elsholtzia bodinieri Van.Theβ-galactosidase staining was used to observe the positive rate of aging cells.The SOD activity assay and ABTS assay were used to detect the antioxidant activity of total flavonoids from Elsholtzia bodinieri Van.【Result】The results showed that the total flavonoid content of Elsholtzia bodinieri Van was 19.375 mg/g.The AB-8 was the most suitable for the separation and purification of total flavonoids from Elsholtzia bodinieri Van.The optimal conditions for static adsorption and resolution of AB-8 were loading mass concentration 1.5 mg/mL,pH 5,adsorption for 6 h,and resolution by 70%acidified ethanol(pH 5)for 4 h,the purity increased from 46.67%to 81.62%.The optimal process conditions of dynamic adsorption and resolution were the flow rate of loading liquid was 2 mL/min,the flow rate of eluting liquid was 1 mL/min,and the elution could be achieved at 80 mL,the purity was increased to 80.33%.Antioxidant activity:compared with the control group,the posi⁃tive rate of senile cells was decreased by SA-β-gal(P<0.01).The scavenging IC50 of total flavonoids on ABTS was 0.4844 mg/mL,and the scavenging IC50 of VC on ABTS was 0.1545 mg/mL.The SOD activity in 0.5 mg/mL and 1 mg/mL treatment groups was higher than that in injured group(P<0.001).Antitumor activity:Total flavonoids of Elsholtzia bodinieri Van had certain inhibitory effect on the growth of HCT116 and A549,and the IC50 of HCT116 was 0.8486 mg/mL.The IC50 of A549 is 0.7708 mg/mL,indicating that the total flavonoids of Elsholtzia bodinieri Van is a potential natural antioxidant and has certain tumor inhibition.【Conclusion】The total flavonoids obtained by optimum extraction and purification have certain antioxidant and anti-tumor activities.