半枝莲总黄酮提取工艺优化及抗氧化、抗肿瘤活性评价

Optimization of Extraction Process and Evaluation of Antioxidant and Antitumor Activities of Total Flavonoids from Scutellaria barbata

本实验旨在优化半枝莲总黄酮提取工艺,评价半枝莲总黄酮纯化物抗氧化、抗肿瘤活性。采用单因素实验结合响应面Box-Behnken设计对半枝莲总黄酮提取工艺进行研究,主要考察了提取时间、料液比、提取温度和乙醇体积分数对黄酮得率的影响,从而得出半枝莲总黄酮提取的最佳工艺;采用DPPH、ABTS法检测半枝莲总黄酮纯化物的抗氧化活性;采用MTT法检测纯化物对NCI-H1299、HepG2、MHCC-97H、HuH-7细胞增殖的影响。结果表明,半枝莲总黄酮提取最佳工艺为提取时间93 min、料液比1:41(g/mL)、提取温度为68℃、乙醇体积分数为75%,该条件下半枝莲总黄酮得率为26.46 mg/g。抗氧化实验结果表明,半枝莲总黄酮纯化物对DPPH、ABTS+自由基有较好的清除作用,IC_(50)值分别为25.41、70.41μg/mL;抗肿瘤实验表明,在一定浓度范围内不同质量浓度的半枝莲总黄酮纯化物对肿瘤细胞增殖均有一定抑制作用,其NCI-H1299、HepG2、MHCC-97H、HuH-7细胞的IC_(50)值分别为168.6、330.5、269.2、335.8μg/mL。综上可知,该方法稳定可行、重复性好,能有效提取出半枝莲中总黄酮成分,且纯化后的半枝莲总黄酮具有良好的抗氧化及抗肿瘤活性,可以进行功能性食品的研发。

The purpose of this experiment was to optimize the extraction process of total flavonoids from Scutellaria barbata and to evaluate the antioxidant and antitumor activity of total flavonoids.A single factor experiment combined with a response surface Box-Behnken design was used to study the extraction process of total flavonoids from Scutellaria barbata.The effects of extraction time,material-to-liquid ratio,extraction temperature and volume fraction of ethanol on the yield of flavonoids were investigated to arrive at the optimal extraction process of total flavonoids from Scutellaria barbata.The antioxidant activity of total flavonoids from Scutellaria barbata was detected by DPPH and ABTS methods,and the effect of purified products on the proliferation of NCI-H1299,HepG2,MHCC-97H and HuH-7 cells were analyzed using the MTT assay.The results showed that the optimum extraction conditions of total flavonoids from Scutellaria barbata were as follows:Extraction time 93 min,ratio of material to liquid 1:41(g/mL),extraction temperature 68℃,volume fraction of ethanol 75%.Under these conditions,the yield of total flavonoids was 26.46 mg/g.The results of antioxidant experiments showed that the purified total flavonoids of Scutellaria barbata had good scavenging effects on DPPH and ABTS+radicals,with IC_(50)values of 25.41 and 70.41μg/mL,respectively.The results of anti-tumor experiments showed that the purified total flavonoids of Scutellaria barbata at different mass concentrations within a certain concentration range had certain inhibitory effects on the proliferation of tumor cells,with the IC_(50)values of NCI-H1299,HepG2,MHCC-97H and HuH-7 cells were 168.6,330.5,269.2 and 335.8μg/mL,respectively.In summary,it could be concluded that the method was stable and feasible,reproducible,and could effectively extract the total flavonoid components from Scutellaria barbata,and the purified Scutellaria barbata total flavonoids had good antioxidant and antitumor activities,which could be used for the development of functional foods or pharmaceuticals.