正交实验法优化发酵蜈蚣粉中总黄酮提取工艺
Optimization of total flavonoids extraction process in centipede fermentation by orthogonal test
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摘要: 目的:采用正交实验法优化发酵蜈蚣粉总黄酮回流提取工艺。方法:在对发酵蜈蚣粉中总黄酮提取条件进行单因素实验的基础上,以提取温度(A)、料液比(B)和提取时间(C)为自变量,发酵蜈蚣粉总黄酮得率为因变量,利用正交实验法探究最佳提取工艺条件。同时采用MTT体外抗肿瘤实验方法,研究发酵蜈蚣粉提取物对肺腺癌A549细胞的抑制作用。结果:影响发酵蜈蚣粉总黄酮得率提取因素的大小次序为:料液比>提取温度>提取时间,在此次实验中,发酵蜈蚣粉最佳提取工艺为:提取温度80℃,料液比1∶40(g∶m L),提取时间2.5 h,在此条件下发酵蜈蚣粉中总黄酮得率为1.970%;提取获得的不同浓度的发酵蜈蚣醇提液对肺腺癌A549细胞的抑制率均大于顺铂,具有较优良的抗肿瘤活性。Abstract: Objective: Orthogonal experiment is used to optimize the extraction process of total flavonoids from fermented centipede powder.Method: Regarding the yield of total flavonoids from fermented centipede as the research index.On the basic of single-factor test, the ultrasonic-assisted extraction process parameters of total flavonoids from fermented centipede including the ratio of extractive temperature ( A) , the ratio of material ( B) and extractive time ( C) were optimized by orthogonal test and the yield of total flavonoids was used as the responsive values. The antineoplastic activity of extraction to lung adenocarcinoma cell A549 were measured by using MTT assay in vitro.Result: The order of the effect of these factors on the yield of total flavonoids from fermented centipede powder was the ratio of material > extractive temperature > extractive time.In this experiment, the best extractive process was under 80 ℃ of the extractive temperature, 1 ∶ 40 ( g ∶ m L) of the ratio of material, 2.5 h of the extraction time.Under these conditions, the yield of total flavonoids was 1.970%. The inhibition ratio of different concentration of alcohol extraction of fermented centipede powder to lung adenocarcinoma cell A549 was all greater than cis-platinum, and it had good antineoplasmic activity.