密闭微波辅助萃取三七中的皂苷类化合物以及细梗胡枝子中的黄酮类化合物
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摘要
三七是我国名贵的传统中药材,主产于云南文山,其主要有效成分是皂苷类化合物。它具有活血化瘀和消肿止痛的功效,目前大多用于冠心病、心绞痛等心血管系统疾病的防治。
豆科胡枝子属(Lespedeza Michx)植物约60余种,分布于东亚至澳大利亚东北部及北美,具有润肺清热、利水通淋、健脾祛湿的功效。根据报道,其主要有效成分为芦丁等黄酮类化合物,具有很好的清除自由基作用及抗氧化性。
本文针对三七中的皂苷类化合物和细梗胡枝子中的黄酮类化合物的微波辅助提取作了一系列的研究工作。
建立了采用微波辅助间歇提取三七样品中的总皂苷,并以分光光度法测定其含量的实验方法。分别通过单因素实验和正交实验设计,优化了萃取溶剂浓度、溶剂用量、微波功率和微波辐射时间等提取条件,确定了微波辅助萃取法提取三七中皂苷类化合物的最佳条件为:使用浓度为60%的乙醇,在液固比100∶1的条件下,以255W的微波功率,微波间歇萃取10×20s。在上述最优条件下进行了精密度和回收率实验,加入回收率在97.7%～101.8%之间,RSD为1.54%(n=5)。与传统的索氏提取法和超声波提取法比较,微波辅助萃取法具有操作简便,快速,重现性好等特点,实验结果表明此方法可推广应用于各类中草药中总皂苷的含量测定。
建立了采用微波辅助间歇提取三七样品中的皂苷类化合物,并以反相高效液相色谱(RP-HPLC)测定其中三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1含量的实验方法。分别通过单因素实验和正交实验设计,优化了萃取溶剂浓度、溶剂用量、微波功率和微波辐射时间等提取条件。与传统的索氏提取法、冷浸法和超声波提取法比较,微波辅助萃取法具有快速、提取率较高、溶剂消耗少等优点,间歇萃取较连续萃取更是大大节省了微波能耗。
建立了微波辅助提取细梗胡枝子[Lespedeza virgata(Thunb.)DC.]不同药用部位中黄酮类化合物,并以分光光度法测定其含量的实验方法。在微波单因素提取的基础上,采取L_9(3~4)正交优化实验设计,优化了萃取乙醇浓度、液固比、微波功率和微波辐射时间等提取条件,确定了微波辅助萃取法提取细梗胡枝子样品中叶、枝和根等不同药用部位中的黄酮类化合物的最佳工艺参数分别为:叶:微波功率40%(340W),乙醇浓度为50%,液固比为80 mL/g,提取时间6min;枝:微波功率40%(340W),乙醇浓度为50%,液固比为80 mL/g,提取时间5min;根:微波功率50%(425W),乙醇浓度为60%,液固比为90 mL/g,提取时间5min。在最优条件下测得黄酮类化合物含量分别为:10.43%(叶),5.91%(枝),5.29%(根)。在上述最优条件下进行了精密度和加入回收率实验,与索氏提取法、超声波提取法以及乙醇冷浸法比较,微波辅助萃取法具有操作简便,快速,重现性好等特点,实验结果表明此方法可推广应用于各类中草药中黄酮类化合物的含量测定。
本课题还利用高效液相色谱-电喷雾质谱联用技术(HPLC-ESI/MS)依据保留特性等规律信息确认了细梗胡枝子中的黄酮类化合物,在此基础上比较了不同提取方式对不同药用部位提取效果的影响,为其质量控制提供了一定依据。
Radix notoginseng(Sanqi),the main active constituent of which is saponin,is an important herb to stanch a cut and alleviate pain.The herb is often used to treat various kinds of bleeding and pains.It is a well-known traditional Chinese medicine and is mainly cultivated in Wenshan of Yunnan, China.
The genus Lespedeza(Leguminosae)has about 60 species distributing from east Asia to north Australia and north America.The whole plants were used in the treatment of acute and chronic nephritis,azothemia,diuresis.The most important bioactive compounds present within this medicinal species are flavones including rutin and so on.It is reported that flavones have shown significant scavenging of free oxygen radicals and antioxidative activity.
The microwave-assisted extraction of active components of Radix notoginseng and Lespedeza virgata DC.was studied in this thesis.
Determination of total saponins from Radix Notoginseng by microwave-assisted extraction and absorption spectroscopy was reported.The effects of solvent concentration,ratio of solution to solid,microwave power and radiation time were evaluated by single factor and orthogonal experiments. The extraction conditions of total saponins from Radix Notoginseng by microwave-assisted method were optimized as solvent:sample ratio 100:1 (v/w)of 60%(v/v)ethanol and microwave power of 255 W for 10×20s.Under the optimized conditions the relative standard deviation and the recovery were 1.54%(n=5)and 97.7%～101.8%respectively.Compared with Soxhlet extraction and ultrasound-assisted extraction,microwave-assisted extraction was simple and accurate with good reproducibility.It is suitable for the determination of saponins in all kinds of traditional Chinese medicines.
A detail procedure for intermittently extracting Notoginseng Saponin R1, Ginseng Saponin Rg1 and Rb1 from Radix Notoginseng was developed by using an obturated microwave-assisted extraction(MAE)device with pressure and temperature controlling units.The effects of solvent concentration,ratio of solution to solid,microwave power and radiation time were evaluated by single factor and orthogonal experiments.Compared with Soxhlet extraction, cold-maceration and ultrasonic extraction,microwave extraction was a rapid method with a higher yield and less solvent consumption.Less microwave energy was consumed by using intermittent microwave extraction than continuous microwave extraction.
Determination of flavonoids from Lespedeza virgata DC.by pressurized microwave-assisted extraction and absorption spectroscopy was reported.The effects of solvent concentration,ratio of solution to solid,microwave power and radiation time were evaluated by single factor and orthogonal experiments. The extraction conditions of flavonoids from leaf of Lespedeza virgata DC.by microwave-assisted extraction method were optimized as:solvent:sample ratio 80:1(v/w),50%(v/v)ethanol and microwave power of 340 W for 6min. The extraction conditions of flavonoids from branch were optimized as: solvent:sample ratio 80:1(v/w),50%(v/v)ethanol and microwave power of 340 W for 5min;and the extraction conditions of flavonoids from root were optimized as:solvent:sample ratio 90:1(v/w),60%(v/v)ethanol and microwave power of 425 W for 5min.Under the optimized conditions the average contents of flavonoids were 10.43%(leaf),5.91%(branch)and 5.29% (root),respectively.Compared with Soxhlet extraction,ultrasound-assisted extraction and cold-maceration by ethol,microwave-assisted extraction was simple and accurate with good reproducibility.It is suitable for the determination of flavonoids in all kinds of traditional Chinese medicines.
High performance liquid chromatography-mass spectrometry method was developed for the analysis and identification of compounds extracted from Lespedeza virgata DC..Moreover,different methods were compared on the species and contents of flavonoids extracted from different parts of Lespedeza virgata DC..
豆科胡枝子属(Lespedeza Michx)植物约60余种,分布于东亚至澳大利亚东北部及北美,具有润肺清热、利水通淋、健脾祛湿的功效。根据报道,其主要有效成分为芦丁等黄酮类化合物,具有很好的清除自由基作用及抗氧化性。
本文针对三七中的皂苷类化合物和细梗胡枝子中的黄酮类化合物的微波辅助提取作了一系列的研究工作。
建立了采用微波辅助间歇提取三七样品中的总皂苷,并以分光光度法测定其含量的实验方法。分别通过单因素实验和正交实验设计,优化了萃取溶剂浓度、溶剂用量、微波功率和微波辐射时间等提取条件,确定了微波辅助萃取法提取三七中皂苷类化合物的最佳条件为:使用浓度为60%的乙醇,在液固比100∶1的条件下,以255W的微波功率,微波间歇萃取10×20s。在上述最优条件下进行了精密度和回收率实验,加入回收率在97.7%～101.8%之间,RSD为1.54%(n=5)。与传统的索氏提取法和超声波提取法比较,微波辅助萃取法具有操作简便,快速,重现性好等特点,实验结果表明此方法可推广应用于各类中草药中总皂苷的含量测定。
建立了采用微波辅助间歇提取三七样品中的皂苷类化合物,并以反相高效液相色谱(RP-HPLC)测定其中三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1含量的实验方法。分别通过单因素实验和正交实验设计,优化了萃取溶剂浓度、溶剂用量、微波功率和微波辐射时间等提取条件。与传统的索氏提取法、冷浸法和超声波提取法比较,微波辅助萃取法具有快速、提取率较高、溶剂消耗少等优点,间歇萃取较连续萃取更是大大节省了微波能耗。
建立了微波辅助提取细梗胡枝子[Lespedeza virgata(Thunb.)DC.]不同药用部位中黄酮类化合物,并以分光光度法测定其含量的实验方法。在微波单因素提取的基础上,采取L_9(3~4)正交优化实验设计,优化了萃取乙醇浓度、液固比、微波功率和微波辐射时间等提取条件,确定了微波辅助萃取法提取细梗胡枝子样品中叶、枝和根等不同药用部位中的黄酮类化合物的最佳工艺参数分别为:叶:微波功率40%(340W),乙醇浓度为50%,液固比为80 mL/g,提取时间6min;枝:微波功率40%(340W),乙醇浓度为50%,液固比为80 mL/g,提取时间5min;根:微波功率50%(425W),乙醇浓度为60%,液固比为90 mL/g,提取时间5min。在最优条件下测得黄酮类化合物含量分别为:10.43%(叶),5.91%(枝),5.29%(根)。在上述最优条件下进行了精密度和加入回收率实验,与索氏提取法、超声波提取法以及乙醇冷浸法比较,微波辅助萃取法具有操作简便,快速,重现性好等特点,实验结果表明此方法可推广应用于各类中草药中黄酮类化合物的含量测定。
本课题还利用高效液相色谱-电喷雾质谱联用技术(HPLC-ESI/MS)依据保留特性等规律信息确认了细梗胡枝子中的黄酮类化合物,在此基础上比较了不同提取方式对不同药用部位提取效果的影响,为其质量控制提供了一定依据。
Radix notoginseng(Sanqi),the main active constituent of which is saponin,is an important herb to stanch a cut and alleviate pain.The herb is often used to treat various kinds of bleeding and pains.It is a well-known traditional Chinese medicine and is mainly cultivated in Wenshan of Yunnan, China.
The genus Lespedeza(Leguminosae)has about 60 species distributing from east Asia to north Australia and north America.The whole plants were used in the treatment of acute and chronic nephritis,azothemia,diuresis.The most important bioactive compounds present within this medicinal species are flavones including rutin and so on.It is reported that flavones have shown significant scavenging of free oxygen radicals and antioxidative activity.
The microwave-assisted extraction of active components of Radix notoginseng and Lespedeza virgata DC.was studied in this thesis.
Determination of total saponins from Radix Notoginseng by microwave-assisted extraction and absorption spectroscopy was reported.The effects of solvent concentration,ratio of solution to solid,microwave power and radiation time were evaluated by single factor and orthogonal experiments. The extraction conditions of total saponins from Radix Notoginseng by microwave-assisted method were optimized as solvent:sample ratio 100:1 (v/w)of 60%(v/v)ethanol and microwave power of 255 W for 10×20s.Under the optimized conditions the relative standard deviation and the recovery were 1.54%(n=5)and 97.7%～101.8%respectively.Compared with Soxhlet extraction and ultrasound-assisted extraction,microwave-assisted extraction was simple and accurate with good reproducibility.It is suitable for the determination of saponins in all kinds of traditional Chinese medicines.
A detail procedure for intermittently extracting Notoginseng Saponin R1, Ginseng Saponin Rg1 and Rb1 from Radix Notoginseng was developed by using an obturated microwave-assisted extraction(MAE)device with pressure and temperature controlling units.The effects of solvent concentration,ratio of solution to solid,microwave power and radiation time were evaluated by single factor and orthogonal experiments.Compared with Soxhlet extraction, cold-maceration and ultrasonic extraction,microwave extraction was a rapid method with a higher yield and less solvent consumption.Less microwave energy was consumed by using intermittent microwave extraction than continuous microwave extraction.
Determination of flavonoids from Lespedeza virgata DC.by pressurized microwave-assisted extraction and absorption spectroscopy was reported.The effects of solvent concentration,ratio of solution to solid,microwave power and radiation time were evaluated by single factor and orthogonal experiments. The extraction conditions of flavonoids from leaf of Lespedeza virgata DC.by microwave-assisted extraction method were optimized as:solvent:sample ratio 80:1(v/w),50%(v/v)ethanol and microwave power of 340 W for 6min. The extraction conditions of flavonoids from branch were optimized as: solvent:sample ratio 80:1(v/w),50%(v/v)ethanol and microwave power of 340 W for 5min;and the extraction conditions of flavonoids from root were optimized as:solvent:sample ratio 90:1(v/w),60%(v/v)ethanol and microwave power of 425 W for 5min.Under the optimized conditions the average contents of flavonoids were 10.43%(leaf),5.91%(branch)and 5.29% (root),respectively.Compared with Soxhlet extraction,ultrasound-assisted extraction and cold-maceration by ethol,microwave-assisted extraction was simple and accurate with good reproducibility.It is suitable for the determination of flavonoids in all kinds of traditional Chinese medicines.
High performance liquid chromatography-mass spectrometry method was developed for the analysis and identification of compounds extracted from Lespedeza virgata DC..Moreover,different methods were compared on the species and contents of flavonoids extracted from different parts of Lespedeza virgata DC..
引文
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