超临界流体色谱分离纯化木香中的木香烃内酯和去氢木香内酯
|
摘要
目的:建立超临界流体色谱(supercritical fluid chromatography,SFC)分离纯化木香中的木香烃内酯和去氢木香内酯的方法。方法:采用超临界二氧化碳(supercritical carbon dioxide,SC-CO2)作流动相,探究各个条件对SFC分离过程的影响,利用半制备型超临界流体色谱对木香粗提物进行分离纯化,采用高效液相色谱法和核磁共振对得到的化合物进行纯度分析和结构鉴定,并研究色谱过程的热力学规律。结果:采用YMC-C_(18)色谱柱(10 mm×250 mm,5μm),流动相SC-CO2,0. 13%甲醇为改性剂,流速12. 0 m L·min~(-1),背压13 MPa,柱温318℃,检测波长225 nm,重复进样20次,粗提物进样量4 mg,根据色谱图收集各个目标成分。经高效液相色谱测定分离纯化后的化合物纯度均> 99%,经核磁共振确定其结构为木香烃内酯和去氢木香内酯。在此色谱条件下SFC分离过程为正相色谱过程。结论:该方法分离纯化得到的成分纯度高,有机溶剂残留少,可用于木香中木香烃内酯和去氢木香内酯的高效快速制备。
Objective: To establish a supercritical fluid chromatography( SFC) method for separating and purifying costunolide and dehydrocostus lactone in Aucklandiae Radix. Method: With supercritical carbon dioxide as the mobile phase,the effect of six factors,such as type of chromatographic columns,modifiers and modifiers ratio,flow rate of mobile phase,pressure and temperature,on the separation process of supercritical fluid chromatography were explored. The target components were separated and prepared by semi-preparative supercritical fluid chromatography. High performance liquid chromatography and nuclear magnetic resonance were used to analyze the components and study the thermodynamic regularity of the chromatographic process. Result:C_(18) column( 10 mm × 250 mm,5 μm) was adopted,with supercritical fluid dioxide as the mobile phase,the ratio of methanol was 0. 13%,the flow rate was 12 mL·min~(-1),column pressure was 13 MPa,column temperature was318 ℃,and detection wavelength was 225 nm. The sample was injected for 20 times,crude extract was 4 mg,and each target component was collected according to the chromatogram. Its purity was determined to be more than 99% by HPLC,and its structure was determined as costunolide and dehydrocostus lactone by NMR. Under this condition,the SFC separation process was normal-phase chromatography. Conclusion: The method can be used to prepare effective components of Aucklandiae Radix with a high purity and low solvent residue.
Objective: To establish a supercritical fluid chromatography( SFC) method for separating and purifying costunolide and dehydrocostus lactone in Aucklandiae Radix. Method: With supercritical carbon dioxide as the mobile phase,the effect of six factors,such as type of chromatographic columns,modifiers and modifiers ratio,flow rate of mobile phase,pressure and temperature,on the separation process of supercritical fluid chromatography were explored. The target components were separated and prepared by semi-preparative supercritical fluid chromatography. High performance liquid chromatography and nuclear magnetic resonance were used to analyze the components and study the thermodynamic regularity of the chromatographic process. Result:C_(18) column( 10 mm × 250 mm,5 μm) was adopted,with supercritical fluid dioxide as the mobile phase,the ratio of methanol was 0. 13%,the flow rate was 12 mL·min~(-1),column pressure was 13 MPa,column temperature was318 ℃,and detection wavelength was 225 nm. The sample was injected for 20 times,crude extract was 4 mg,and each target component was collected according to the chromatogram. Its purity was determined to be more than 99% by HPLC,and its structure was determined as costunolide and dehydrocostus lactone by NMR. Under this condition,the SFC separation process was normal-phase chromatography. Conclusion: The method can be used to prepare effective components of Aucklandiae Radix with a high purity and low solvent residue.
引文
[1]吕选民,姬水英.柴草瓜果篇第三十五讲木香[J].中国乡村医药,2018,25(9):45-46.
[2]徐珍珍,樊旭蕾,王淑美.木香化学成分及挥发油提取的研究进展[J].广东化工,2017,44(3):77-78.
[3]徐珍珍,史星星,樊旭蕾,等.基于色差原理分析木香有效成分含量与颜色值的相关性[J].中国实验方剂学杂志,2018,24(13):17-21.
[4]金清,白晓华,邓亚飞,等.木香降血糖有效部位及有效成分研究[J].中草药,2012,43(7):1371-1375.
[5]李爱峰.高速逆流色谱分离纯化大黄、补骨脂等中药活性成分的研究[D].聊城:聊城大学,2005.
[6] LI A F,SUN A L,LIU R M. Preparative isolation and purification of costunolide and dehydrocostuslactone from Aucklandia lappa Decne by high-speed counter-current chromatography[J]. J Chromatogr A,2005,1076(1/2):193-197.
[7]许卉,杨小玲,刘生生,等.土木香的倍半萜类化学成分研究[J].时珍国医国药,2007,18(11):2738-2740.
[8]尹宏权,齐秀兰,华会明,等.云木香化学成分研究[J].中国药物化学杂志,2005,15(4):217-220.
[9]吴玉琼.香连丸的质量标准及其活性组分的药物代谢动力学研究[D].兰州:兰州大学,2010.
[10]莫绪飞,吕惠生,张敏华,等.超临界流体色谱纯化Z-藁本内酯的研究[J].高校化学工程学报,2013,27(5):737-742.
[11] Mazzei J L,Avila L A. Chromatographic models as tools for scale-up of isolation of natural products by semipreparative HPLC[J]. J Liq Chromatogr Relat Technol,2003,26(2):177-193.
[12]魏华,何春年,彭勇,等.川木香化学成分研究[J].中国中药杂志,2012,37(9):1249-1253.
[13] Sander L C,Field L R. Effect of eluent composition on thermodynamic properties in high-performance liquid chromatography[J]. Anal Chem,1980,52(13):2009-2013.
[14] Van Quach. Evaluation of polar-embedded reversedphase liquid chromatography columns and the temperature dependence of the phase ratio[D].Tallahassee:Florida State University,2009.
[2]徐珍珍,樊旭蕾,王淑美.木香化学成分及挥发油提取的研究进展[J].广东化工,2017,44(3):77-78.
[3]徐珍珍,史星星,樊旭蕾,等.基于色差原理分析木香有效成分含量与颜色值的相关性[J].中国实验方剂学杂志,2018,24(13):17-21.
[4]金清,白晓华,邓亚飞,等.木香降血糖有效部位及有效成分研究[J].中草药,2012,43(7):1371-1375.
[5]李爱峰.高速逆流色谱分离纯化大黄、补骨脂等中药活性成分的研究[D].聊城:聊城大学,2005.
[6] LI A F,SUN A L,LIU R M. Preparative isolation and purification of costunolide and dehydrocostuslactone from Aucklandia lappa Decne by high-speed counter-current chromatography[J]. J Chromatogr A,2005,1076(1/2):193-197.
[7]许卉,杨小玲,刘生生,等.土木香的倍半萜类化学成分研究[J].时珍国医国药,2007,18(11):2738-2740.
[8]尹宏权,齐秀兰,华会明,等.云木香化学成分研究[J].中国药物化学杂志,2005,15(4):217-220.
[9]吴玉琼.香连丸的质量标准及其活性组分的药物代谢动力学研究[D].兰州:兰州大学,2010.
[10]莫绪飞,吕惠生,张敏华,等.超临界流体色谱纯化Z-藁本内酯的研究[J].高校化学工程学报,2013,27(5):737-742.
[11] Mazzei J L,Avila L A. Chromatographic models as tools for scale-up of isolation of natural products by semipreparative HPLC[J]. J Liq Chromatogr Relat Technol,2003,26(2):177-193.
[12]魏华,何春年,彭勇,等.川木香化学成分研究[J].中国中药杂志,2012,37(9):1249-1253.
[13] Sander L C,Field L R. Effect of eluent composition on thermodynamic properties in high-performance liquid chromatography[J]. Anal Chem,1980,52(13):2009-2013.
[14] Van Quach. Evaluation of polar-embedded reversedphase liquid chromatography columns and the temperature dependence of the phase ratio[D].Tallahassee:Florida State University,2009.