中心组合设计效应面法优化桔梗中桔梗皂苷D的提取工艺
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摘要
目的找到综合效率最高的方法,从桔梗中提取出桔梗皂苷D。方法采用超声和回流法,以水为溶剂进行提取,根据桔梗皂苷D的含量测定结果,初步选择适宜的提取方法,之后采用Box-Behken法,考察乙醇比例,提取时间和超声功率三个参数,以桔梗皂苷D的含量为指标,通过绘制效应面(RSM)预测出最优提取条件并进行实验验证。含量测定采用HPLC法,使用Supelco Discovery C18色谱柱(4.6×150mm,5μm),以乙腈-0.1%磷酸水溶液(30∶70,v/v)为流动相,流速1.0m L·min-1,进样量10μL,检测波长203nm,柱温25℃。结果 RSM预测结果表明,当乙醇比例为45%,提取时间45min,超声功率200%时,所得供试品中桔梗皂苷D的提取率为0.2959%,综合效率最优。结论 RSM准确、高效,能够以最少的实验次数全面、科学地筛选出从桔梗中提取桔梗皂苷D的最优工艺。
OBJECTIVE To select method for the extraction of kikyosaponin D in Platycodon grandiflorum with optimum efficiency in the whole. METHODS Ultrasonic and heat reflex were first compared,using water as solvent and kikyosaponin D content as index,the resulting suitable method was chosen for further optimization,in which BoxBehken method was used,with ethanol composition,extraction time and ultrasonic power as parameters,and kikyosaponin D content also as index; through the construction of response surface(RSM),the predicted conditions were validated by real experiment; HPLC was used for content determination,with a Supelco Discovery C18 column(4. 6 ×150 mm,5 μm),a mobile phase containing acetonitrile-0. 1 % phosphoric acid(30 ∶ 70,v/v),the flow rate was 1. 0 m L·min-1,injection volume was 10μL,detection wavelength was 203 nm and the column temperature was 25℃. RESULTS RSM predicted that when using the condition of 45 % ethanol,ultrasonic extraction for 45 min with the power set at 200 %,the obtained solution had a kikyosaponin D extraction rate of 0. 2959 %,which was optimum in the whole. CONCLUSION RSM was accurate and efficient,by which optimum parameters extracting kikyosaponin D from Platycodon grandiflorum were comprehensively and scientifically selected.
OBJECTIVE To select method for the extraction of kikyosaponin D in Platycodon grandiflorum with optimum efficiency in the whole. METHODS Ultrasonic and heat reflex were first compared,using water as solvent and kikyosaponin D content as index,the resulting suitable method was chosen for further optimization,in which BoxBehken method was used,with ethanol composition,extraction time and ultrasonic power as parameters,and kikyosaponin D content also as index; through the construction of response surface(RSM),the predicted conditions were validated by real experiment; HPLC was used for content determination,with a Supelco Discovery C18 column(4. 6 ×150 mm,5 μm),a mobile phase containing acetonitrile-0. 1 % phosphoric acid(30 ∶ 70,v/v),the flow rate was 1. 0 m L·min-1,injection volume was 10μL,detection wavelength was 203 nm and the column temperature was 25℃. RESULTS RSM predicted that when using the condition of 45 % ethanol,ultrasonic extraction for 45 min with the power set at 200 %,the obtained solution had a kikyosaponin D extraction rate of 0. 2959 %,which was optimum in the whole. CONCLUSION RSM was accurate and efficient,by which optimum parameters extracting kikyosaponin D from Platycodon grandiflorum were comprehensively and scientifically selected.
引文
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[8]孙印石,王建华,董其亭,等.桔梗中桔梗总皂苷和桔梗皂苷D的超声波提取工艺[J].食品与发酵工业,2008,34(10):178-181.
[2]李婷,徐文珊,李西文,等.中药桔梗的现代药理研究进展[J].中药药理与临床,2013,29(2):205-208.
[3]韩向晖,叶依依,郭保凤,等.桔梗皂苷D配伍不同中药有效成分对乳腺癌4T1和MDA-MB-231细胞增殖及侵袭的影响[J].中西医结合学报,2012,10(1):67-75.
[4]田雨弘,高鸽,刘颖,等.桔梗茎叶皂苷对H22荷瘤小鼠的抗肿瘤作用及机制研究[J].毒理学杂志,2016,30(1):45-48.
[5]洪玮,罗娟敏,郭磊,等.HPLC法测定桔梗药材中桔梗皂苷D的含量[J].江西中医药,2014,45(374):59-61.
[6]曾金祥,方香香,朱继孝,等.不同产地桔梗皂苷成分HPLC指纹图谱比较研究[J].世界科学技术-中医药现代化·中药研究,2015,17(5):1000-1006.
[7]谭玲玲,侯晓敏,胡正海,等.不同产地桔梗药材中桔梗总皂苷和桔梗皂苷D的测定[J].中草药,2015,46(11):1682-1684.
[8]孙印石,王建华,董其亭,等.桔梗中桔梗总皂苷和桔梗皂苷D的超声波提取工艺[J].食品与发酵工业,2008,34(10):178-181.