正交试验优选美洲大蠊抗肺纤维化活性部位的提取工艺

Extraction Technology of the Anti-pulmonary Fibrosis Active Site from Periplaneta americana Optimized by Orthogonal Experiment

目的:结合体外活性实验,优选美洲大蠊抗肺纤维化活性部位(ML-HB)的最佳提取工艺。方法:采用单因素实验和L_9(3~4)正交实验设计考察提取温度、溶剂浓度、提取时间、提取次数等因素对美洲大蠊抗肺纤维化活性部位(ML-HB)提取工艺的影响,并结合MTT取工艺。结果:料液比为1:5时(g/mL)最佳提取工艺为:提取温度为70℃,以70%的乙醇提取3次,提取时间为2.5 h/次。对此工艺进行验证,活性部位得率均值为0.59%,其24 h72.40、71.03和72.26μg/mL,72 h的IC_(50)值分别为67.24、69.76和67.22μg/mL。结论:建立了ML-HB的最佳提取工艺,为美洲大蠊抗肺纤维化的研发提供了依据。

Objective: Combined with the activity test in vitro, the extraction technology of the anti-pulmonary fibrosis active site(ML-HB)from Periplaneta americana was optimized. Methods: The single factor experiment and L9(34)orthogonal experiment were used to optimize extraction temperature, solvent concentration, extraction time and the number of extraction times of anti-pulmonary fibrosis active site of Periplaneta americana, and combined with MTT method, the anti-pulmonary fibrosis activity fraction from the different extracts was detected, the yield and IC50 value of the active site were used as evaluation indicators to determine the optimal extraction process of the active site of Periplaneta americana. Results: When the ratio of material to liquid was 1:5(g/mL), the optimal extraction process of the anti-pulmonary fibrosis active site of Periplaneta americana was determined as follows: the extraction temperature was 70 ℃, the ethanol concentration was 70%, the number of extraction times was 3, and the extraction time was 2.5 h.Verifying the optimum conditions, the ML-HB average extraction yield was 0.59%. The IC50 values at 24 h were 77.77, 75.17 and76.03 μg/mL, respectively;72.40, 71.03 and 72.26 μg/mL at 48 h, respectively;67.24, 69.76 and 67.22 μg/mL at 72 h, respectively.Conclusion: It stablishes the best extraction technology of ML-HB, and it provides the basis for the research and development of antipulmonary fibrosis of Periplaneta americana.