Optimization of the extracting process for alkaloids from Dicranostigma leptopodum by response surface analysis and study on its antibacterial activity in vitro

Abstract

Abstract: In this paper, the Dicranostigma leptopodum was researched, on the basis of single factor level determination, the optimal extraction conditions were obtained by using the response surface analysis, and the concentrated liquid of extract response surface analysis was measured by antibacterial activity, moreover, the transmission electron microscopy images of Escherichia coli of treated with concentrated liquid of extract from D. leptopodum was discussed. The experiment results demonstrated that the optimum conditions were liquid to solid ratio was 15 mL/g, time of ultrasound was 35 min, refluxing time was 2.5 h, alcohol was 65%, and the content of alkaloid from D. leptopodum was 9.33% in the optimum extraction process condition; the optimal extraction conditions were obtained by using the response surface analysis of E.coli, Candida albicans and Pseudomonas aeruginosa of minimal inhibitory concentration and minimal bactericidal concentration were 0.20, 0.15, 0.35 mg/mL and 0.15, 0.10, 0.30 mg/mL. They were observed under transmission electron microscopy to everest to E. coli antibacterial activity effect significantly remarkable. The results also showed that the response surface analysis to D. leptopodum spend alkaloid extraction process optimization are reasonable.

CLC Number:

Q946.88

ZHAO Qiang, YU Si-jiu, WANG Ting-pu, YUAN Yi-jun. Optimization of the extracting process for alkaloids from Dicranostigma leptopodum by response surface analysis and study on its antibacterial activity in vitro[J]. Acta Prataculturae Sinica, 2012, 21(4): 206-214.

References

[1] 江苏医学院. 中药大辞典[M]. 1版. 上海: 上海人民出版社, 1977: 1135.
[2] 畅行若, 王宏新, 马广恩. 秃疮花化学成分的研究[J]. 中国药学杂志, 1981, 2: 52.
[3] 赵祁, 韩寅, 杜宇平, 等. 秃疮花提取物对红细胞氧化性溶血的抑制机制[J]. 兰州大学学报(医学版), 2006, 32(3): 41.
[4] Manske R H F, Rodrigo R G A, Holmes H L, et al. The Alkaloids Chemistry And Physiology[M]. 1nd ed. New York: Academic Press, 1981: 191.
[5] 张兴旺, 何莉莉, 王勤. 红茂草注射液对免疫低下小鼠免疫功能的影响[J]. 兰州大学学报(自然科学版), 2006, 42(6): 55-57.
[6] 赵强. 红茂草生物碱有效成分分析及抑菌活性研究[D]. 兰州: 甘肃农业大学, 2009: 13-16.
[7] 陈荃, 邹亚丽, 赵兰兰. 红茂草营养器官的显微结构观察[J]. 天水师范学院学报, 2010, 30(5): 39-43.
[8] 赵强, 王廷璞, 廖天录. 红茂草生物碱TLC检测技术的建立及抑菌活性研究[J]. 中兽医医药杂志, 2010, (6): 47-51.
[9] 王廷璞, 安建平, 贾志国, 等. 红茂草生物碱提取方法及指纹检测技术建立的研究[J]. 中兽医医药杂志, 2008, (6): 41-44.
[10] 董晓宁, 赵强, 杨明. 秃疮花生物碱的提取与体外抑菌研究[J]. 中兽医学杂志, 2010, (2): 6-11.
[11] 刘大护, 张天才, 柳军玺, 等. 秃疮花生物碱类化学成分研究[J]. 中草药, 2011, 42(8): 1505-1508.
[12] 巩江, 骆蓉芳, 倪士峰, 等. 珍稀中药秃疮花属植物药学研究概况[J]. 西北药学杂志, 2010, 25(2): 153-154.
[13] 赵强, 王廷璞, 董晓宁, 等. 酸性染料比色法测定红茂草中总生物碱含量[J]. 中兽医医药杂志, 2011, (6): 38-40.
[14] 李岩, 梁雪, 刘斌. 伊犁秃疮花基因条形码及系统发生关系研究[A]. 2011年全国系统与进化植物学暨第十届青年学术研讨会论文集[C]. 昆明: 云南大学出版社, 2011: 139.
[15] 朱荣. 秃疮花在Zn、Cd、Cu和Pb胁迫下的耐性和富集特征研究[D]. 成都: 四川农业大学, 2011: 5-7.
[16] 董晓宁, 赵强, 杨明. 苦荞麦中芦丁的提取及体外抑菌研究[J]. 中兽医医药杂志, 2010, (1): 20-21.
[17] 赵强, 王廷璞, 余四九, 等. 红茂草生物碱抑菌活性测定[J]. 中国兽医科学, 2008, 38(12): 1098-1101.
[18] 张芳, 王舟, 宗俊勤, 等. 农杆菌介导的假俭草遗传转化体系的建立[J]. 草业学报, 2011, 20(2): 186-194.
[19] 李振东, 陈秀蓉, 杨成德. 珠芽蓼内生菌Z17抑菌能力测定及其鉴定[J]. 草业科学, 2011, (12): 32-37.
[20] 海洪, 汪坤, 金文英, 等. Box-Behnken响应面法优化超声波提取蚕沙中叶绿素的工艺研究[J]. 食品工业技, 2009, 30(3): 207-211.
[21] 王新雯, 海洪, 金文英, 等. 微波-超声波联合提取银杏叶黄酮工艺的响应面法分析[J]. 食品科技, 2010, 35(3): 189-193.
[22] 何庆元, 吴萍, 张晓红, 等. 不同秋眠性苜蓿SRAP体系优化及遗传多样性分析[J]. 草业学报, 2011, 20(2): 203-211.
[23] 胡甦, 王永清, 陶炼. 三角紫叶酢浆草ISSR反应体系的建立与优化[J]. 草业学报, 2011, 20(5): 145-153.