藤黄酸提取分离及其抗肿瘤活性
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摘要
[目的]优化中药藤黄中总藤黄酸的提取分离方法,对藤黄酸抗肿瘤活性进行初步研究。[方法]利用回流法进行提取,通过单因素考察实验,研究不同因素对总藤黄酸提取率的影响,并通过正交实验对提取工艺进行优化。再将总藤黄酸粗品进一步分离纯化得到藤黄酸,并进行表征;采用四氮唑蓝(MTT)比色法考察藤黄酸对两种癌细胞的体外抗肿瘤活性。[结果]总藤黄酸最佳的提取工艺为:提取溶剂为乙酸乙酯,料液比为1∶6(g/m L),提取次数为2次,提取时间为3 h,提取温度为80℃,提取率约为57.0%。通过表征确定纯化后所得是藤黄酸。且藤黄酸对癌细胞具有一定的抑制作用。[结论]正交试验优选的总藤黄酸提取工艺简便、稳定、可行,藤黄酸有较强的抗癌作用。
[Objective]To optimize the extraction and separation method of total gambogic acid from Chinese medicine Garcinia Cambogia, and to study the antitumor activity of gambogic acid.[Methods]The extraction process was carried out by reflux method. The effects of different factors on the extraction rate of total gambogic acid were studied by single factor investigation. The extraction process was optimized by orthogonal experiment. The crude total gambogic acid is further separated and purified to obtain gambogic acid, and characterized. The in vitro antitumor activity of gambogic acid against two cancer cells was investigated by MTT colorimetry. [Results]The best extraction process of total gambogic acid was as follows: the extraction solvent is ethyl acetate, the ratio of material to liquid is 1 ∶ 6( g/m L), the extraction times are 2, the extraction period is 3 hours, the extraction temperature is 80 ℃, and the extraction rate is 57. 0 %. After purification, the obtained product was confirmed by characterization to be gambogic acid, which demonstrated a certain inhibitory effect on cancer cells. [Conclusion]The optimal extraction process of total gambogic acid from orthogonal test is simple, stable and feasible, and gambogic acid has strong anticancer effect.
[Objective]To optimize the extraction and separation method of total gambogic acid from Chinese medicine Garcinia Cambogia, and to study the antitumor activity of gambogic acid.[Methods]The extraction process was carried out by reflux method. The effects of different factors on the extraction rate of total gambogic acid were studied by single factor investigation. The extraction process was optimized by orthogonal experiment. The crude total gambogic acid is further separated and purified to obtain gambogic acid, and characterized. The in vitro antitumor activity of gambogic acid against two cancer cells was investigated by MTT colorimetry. [Results]The best extraction process of total gambogic acid was as follows: the extraction solvent is ethyl acetate, the ratio of material to liquid is 1 ∶ 6( g/m L), the extraction times are 2, the extraction period is 3 hours, the extraction temperature is 80 ℃, and the extraction rate is 57. 0 %. After purification, the obtained product was confirmed by characterization to be gambogic acid, which demonstrated a certain inhibitory effect on cancer cells. [Conclusion]The optimal extraction process of total gambogic acid from orthogonal test is simple, stable and feasible, and gambogic acid has strong anticancer effect.
引文
[1]王丽莉.藤黄和双籽藤黄化学成分及抗肿瘤活性研究[D].沈阳:沈阳药科大学,2008.
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[11]钱婧,王科明.藤黄酸抗肿瘤作用靶点的研究进展[J].中国肿瘤,2011,20(6):441-446.
[12]纪娟,裴来良,王斌,等.紫外分光光度法测定藤黄药材中总藤黄酸含量[J].安徽中医药大学学报,2014,33(2):94-96.
[13]陈优生,冯锋,尤启冬. Zr OCl2比色法测定藤黄中总藤黄酸的含量[J].中国药师,2002,5(12):731-732.
[14]刘举,王洋,周云鹏,等.藤黄酸的提取工艺改进[J].辽宁大学学报(自然科学版),2011,38(2):97-99.
[15]季宇彬,聂凡茹,周欣欣,等.藤黄酸纳米混悬剂的制备及抗肿瘤作用[J].药学学报,2018,53(3):414-416.
[2]殷华芳,钱晓萍.中药藤黄抗肿瘤研究现状[J].现代中西医结合杂志,2008,17(14):2264-2267.
[3]郑赵情,张莉,沈凯凯,等.藤黄的研究进展[J].世界中医药,2016,11(7):1180-1188.
[4]陶斯佳.中药藤黄的化学成分及体内代谢研究[D].北京:中国科学院研究生院,2010.
[5]黎平.藤黄属植物的化学成分及其代谢组学研究[D].北京:中央民族大学,2016.
[6]窦娟.中药藤黄炮制解毒的研究[D].南京:南京中医药大学,2013.
[7]徐文龙,杨柏霖.中药藤黄药理作用的研究进展[J].现代中西医结合杂志,2013,22(11):1239-1241.
[8]张海元.藤黄酸引发结直肠癌细胞脂代谢紊乱所产生的ROS诱导细胞内保护性自噬发生的机制研究[D].武汉:华中科技大学,2014.
[9]姜守刚,张星垚,袁颖洁,等.藤黄酸的提取分离[J].中国卫生标准管理,2017(3):106-108.
[10]吴孟强.新型抗肿瘤药物姜黄素衍生物和藤黄酸衍生物的合成与抗肿瘤活性的研究[D].南京:东华大学,2013.
[11]钱婧,王科明.藤黄酸抗肿瘤作用靶点的研究进展[J].中国肿瘤,2011,20(6):441-446.
[12]纪娟,裴来良,王斌,等.紫外分光光度法测定藤黄药材中总藤黄酸含量[J].安徽中医药大学学报,2014,33(2):94-96.
[13]陈优生,冯锋,尤启冬. Zr OCl2比色法测定藤黄中总藤黄酸的含量[J].中国药师,2002,5(12):731-732.
[14]刘举,王洋,周云鹏,等.藤黄酸的提取工艺改进[J].辽宁大学学报(自然科学版),2011,38(2):97-99.
[15]季宇彬,聂凡茹,周欣欣,等.藤黄酸纳米混悬剂的制备及抗肿瘤作用[J].药学学报,2018,53(3):414-416.