Study on Sieving and Separation of Active Components of Chinese Angelica by Biochromatography
摘 要
应用分子生物色谱技术研究了筛选和分离当归中的活性成分。以甲基丙烯酸缩水甘油酯为功能单体,通过原位聚合得到了整体柱基质,用二乙胺活化后,以吸附蛋白的方法得到了以牛血清白蛋白(BSA)为固定相的分子生物色谱柱。当归样品经甲醇提取,0.45 μm膜过滤后,以BSA分子生物色谱柱为分离柱,以含2%(体积分数)甲醇的Tris-盐酸缓冲溶液为流动相进行分离,用二极管阵列检测器检测。结果表明:活性组分在BSA上的保留机理是疏水作用、静电作用和特定的与空间结构相关的多种因素的共同作用结果。将BSA柱上的保留组分进行了气相色谱-质谱分析,鉴定了8种成分,包括当归中的主要可挥发性成分(藁本内酯)。
Abstract
Molecular biochromatography (MBC) was applied to the study on sieving and separation of active components of Chinese angelica. The matrix of monolithic column was prepared with glycidylmethacrylate as functional monomer by in-situ copolymerization. After activating the functional group with diethylamine, bovine serum albumin (BSA) was absorbed onto the matrix of monolithic column and a MBC column with BSA as stationary phase was formed. The sample of Chinese angelica was extracted with methanol, and the extract was filtered on 0.45 μm filtering membrane, the filtrate was separated on the MBC column, with Tris-HCl buffer solution (containing 2% by volume of methanol as mobile phase. DAD was adopted in the determination. It was found that the mechanism of retention of the active components on BSA was the combining action of a variety of factors, including hydrophobic and electrostatic actions as well as the spatial structure. The components retained on BSA column were analyzed by GC-MS, and 8 components were identified, including the main volatile component of ligustilide.
中图分类号 O657.7
所属栏目 试验与研究
基金项目 浙江省自然科学基金资助项目(M203030,Y4110536);浙江省公益性技术应用研究(分析测试)项目(2011C37040)
收稿日期 2013/1/22
修改稿日期
网络出版日期
作者单位点击查看
备注潘再法(1976-),男,浙江温岭人,副教授,主要从事色谱分析和荧光光谱等方面的研究。
引用该论文: PAN Zai-fa,TAN Ying,HU Bao-xiang,ZHANG Xu,MO Wei-min. Study on Sieving and Separation of Active Components of Chinese Angelica by Biochromatography[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2014, 50(2): 145~150
潘再法,谭莹,胡宝祥,张旭,莫卫民. 生物色谱用于当归中活性成分筛选及分离研究[J]. 理化检验-化学分册, 2014, 50(2): 145~150
共有人对该论文发表了看法,其中:
人认为该论文很差
人认为该论文较差
人认为该论文一般
人认为该论文较好
人认为该论文很好
参考文献
【1】汪海林,邹汉法,孔亮,等.分子生物色谱用于中药活性成分筛选及质量控制方法的研究[J].色谱, 1999,17(2):123-127.
【2】孔亮,邹汉法,汪海林,等.以人血清白蛋白为固定相的分子生物色谱分析几种中药活性成分的研究[J].高等学校化学学报, 2000,21(1):36-40.
【3】WANG Hai-lin, ZOU Han-fa, NI Jian-yi, et al. Fractionation and analysis of Artemisia capillaris Thunb. by affinity chromatography with human serum albumin as stationary phase[J]. J Chromatogr A, 2000,870(1/2):501-510.
【4】HUANG Xiao-dong, Kong Liang, LI Xin, et al. Strategy for analysis and screening of bioactive compounds in traditional Chinese medicines[J]. J Chromatogr B, 2004,812(1/2):71-84.
【5】WANG Yun, KONG Liang, HU Liang-hai, et al. Biological fingerprinting analysis of the traditional Chinese prescription Longdan Xiegan Decoction by on/off-line comprehensive two-dimensional biochromatography[J]. J Chromatogr B, 2007,860(2):185-194.
【6】HU Liang-hai, LI Xin, FENG Shun, et al. Comprehensive two-dimensional HPLC to study the interaction of multiple components in Rheum palmatum L. with HSA by coupling a silica-bonded HSA column to a silica monolithic ODS column[J]. J Sep Sci, 2006,29(6):881-888.
【7】曹琰,严辉,段金廒,等.不同产地当归药材核苷类成分的分析[J].药物分析杂志, 2010,30(11):2026-2030.
【8】杜俊蓉,白波,余彦,等.当归挥发油研究新进展[J].中国中药杂志, 2005,30(18):1400-1405.
【9】BEIGI F, LUNDAHL P. Immobilized biomembrane chromatography of highly lipophilic drugs[J]. J Chromatogr A, 1999,852(1):313-317.
【10】AFEYAN N B, FULTON S P, REGNIER F E. Perfusion chromatography packing materials for proteins and peptides[J]. J Chromatogr A, 1991,544:267-279.
【11】PAN Zai-fa, ZOU Han-fa, MO Wei-min, et al. Protein A immobilized monolithic capillary column for affinity chromatography[J]. Anal Chim Acta, 2002,466(1):141-150.
【12】潘再法,莫卫民,吴仁安,等.蛋白A连续床亲和毛细管色谱柱的制备及性能考察[J].高等学校化学学报, 2002,23(11):2070-2072.
【13】SVEC F, FRECHET J M J. Modified poly (glycidyl metharylate-co-ethylene dimethacrylate) continuous rod columns for preparative-scale ion-exchange chromatography of proteins[J]. J Chromatogr A, 1995,702(1/2):89-95.
【14】邹汉发,张玉奎,卢佩章.高效液相色谱法[M].北京:科学出版社, 1998.
【15】KIMA M R, ABD EL-ATY A M, KIMA I S, et al. Determination of volatile flavor components in danggui cultivars by solvent free injection and hydrodistillation followed by gas chromatographic-mass spectrometric analysis[J]. J Chromatogr A, 2006,1116(1/2):259-264.
【2】孔亮,邹汉法,汪海林,等.以人血清白蛋白为固定相的分子生物色谱分析几种中药活性成分的研究[J].高等学校化学学报, 2000,21(1):36-40.
【3】WANG Hai-lin, ZOU Han-fa, NI Jian-yi, et al. Fractionation and analysis of Artemisia capillaris Thunb. by affinity chromatography with human serum albumin as stationary phase[J]. J Chromatogr A, 2000,870(1/2):501-510.
【4】HUANG Xiao-dong, Kong Liang, LI Xin, et al. Strategy for analysis and screening of bioactive compounds in traditional Chinese medicines[J]. J Chromatogr B, 2004,812(1/2):71-84.
【5】WANG Yun, KONG Liang, HU Liang-hai, et al. Biological fingerprinting analysis of the traditional Chinese prescription Longdan Xiegan Decoction by on/off-line comprehensive two-dimensional biochromatography[J]. J Chromatogr B, 2007,860(2):185-194.
【6】HU Liang-hai, LI Xin, FENG Shun, et al. Comprehensive two-dimensional HPLC to study the interaction of multiple components in Rheum palmatum L. with HSA by coupling a silica-bonded HSA column to a silica monolithic ODS column[J]. J Sep Sci, 2006,29(6):881-888.
【7】曹琰,严辉,段金廒,等.不同产地当归药材核苷类成分的分析[J].药物分析杂志, 2010,30(11):2026-2030.
【8】杜俊蓉,白波,余彦,等.当归挥发油研究新进展[J].中国中药杂志, 2005,30(18):1400-1405.
【9】BEIGI F, LUNDAHL P. Immobilized biomembrane chromatography of highly lipophilic drugs[J]. J Chromatogr A, 1999,852(1):313-317.
【10】AFEYAN N B, FULTON S P, REGNIER F E. Perfusion chromatography packing materials for proteins and peptides[J]. J Chromatogr A, 1991,544:267-279.
【11】PAN Zai-fa, ZOU Han-fa, MO Wei-min, et al. Protein A immobilized monolithic capillary column for affinity chromatography[J]. Anal Chim Acta, 2002,466(1):141-150.
【12】潘再法,莫卫民,吴仁安,等.蛋白A连续床亲和毛细管色谱柱的制备及性能考察[J].高等学校化学学报, 2002,23(11):2070-2072.
【13】SVEC F, FRECHET J M J. Modified poly (glycidyl metharylate-co-ethylene dimethacrylate) continuous rod columns for preparative-scale ion-exchange chromatography of proteins[J]. J Chromatogr A, 1995,702(1/2):89-95.
【14】邹汉发,张玉奎,卢佩章.高效液相色谱法[M].北京:科学出版社, 1998.
【15】KIMA M R, ABD EL-ATY A M, KIMA I S, et al. Determination of volatile flavor components in danggui cultivars by solvent free injection and hydrodistillation followed by gas chromatographic-mass spectrometric analysis[J]. J Chromatogr A, 2006,1116(1/2):259-264.
相关信息