橘叶化学成分的分离鉴定
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摘要
目的:橘叶为常见中药,具有疏肝解郁、理气止痛、化痰散结的功效,主要用于治疗乳痈肿痛、乳房结块、疝气疼痛等症。现代药理学研究表明,橘叶水煎液对小鼠乳腺炎有抑制作用,该文旨在研究橘叶的化学成分,揭示其发挥药效的物质基础。方法:分别采用95%乙醇和70%乙醇对橘叶粗粉进行浸渍和回流提取,利用小孔树脂(MCI)柱色谱对乙醇提取物进行初步分离,综合运用硅胶,MCI,ODS,聚酰胺,LH-20羟丙基葡聚糖凝胶(Sephadex LH-20)等多种柱色谱方法分离和纯化化合物,根据理化性质,结合NMR及MS等波谱技术鉴定化合物结构。结果:从橘叶中分离得到12个化合物,分别鉴定为川陈皮素(nobiletin,1),5-去甲基川陈皮素(5-demethylnobile,2),橘红素(tangeretin,3),5,7,8,4'-四甲氧基黄酮(5,7,8,4'-tetramethoxyflavone,4),5-羟基-6,7,3',4'-四甲氧基黄酮(5-hydroxy-6,7,3',4'-tetramethoxyflavone,5),β-香树脂醇(β-amyrin,6),羽扇豆醇(lupeol,7),植醇(phytol,8),豆甾-5,22-二烯-3β-醇(stigmastane-5,22-dien-3β-ol,9),β-谷甾醇(β-sitosterol,10),对羟基苯甲醛(p-hydroxybenzaldehyde,11),香草醛(vanillin,12)。结论:化合物4~12为首次从橘叶中分离得到。
Objective: The leaves of Citrus reticulata,a common traditional Chinese medicine,are mainly used for the treatment of various diseases,including acute mastitis,caked breast and hernia symptoms,owing to its effects of relieving the depressed liver,soothing the liver,regulating the circulation of Qi,alleviating pain,dissipating phlegm and resolving masses. Modern pharmacological studies have shown that the hot water extracts from the leaves of C. reticulata have inhibitory effects on mastitis in mice. This paper aims to study their chemical constituents and reveal the effective substances. Method: The coarse powder of C. reticulata leaves was extracted by methods of soaking and refluxing with 95% and 70% ethanol,respectively. Then the ethanol extracts were separated preliminarily by MCI column chromatography( CC). The compounds were isolated and purified by using various chromatographic techniques,including silica gel,MCI,ODS,polyamide and Sephadex LH-20 CC.Their structures were identified on the basis of their physical-chemical properties and spectroscopic techniques,including NMR and MS. Result: Twelve compounds were obtained and identified as nobiletin( 1),5-demethylnobile( 2), tangeretin( 3), 5, 7, 8, 4'-tetramethoxyflavone( 4), 5-hydroxy-6, 7, 3',4'-tetramethoxyflavone( 5), β-amyrin( 6), lupeol( 7), phytol( 8), stigmastane-5,22-dien-3β-ol( 9),β-sitosterol( 10), p-hy-droxybenzaldehyde( 11) and vanillin( 12), respectively. Conclusion: Compounds4-12 were isolated from the leaves of Citrus reticulata for the first time.
Objective: The leaves of Citrus reticulata,a common traditional Chinese medicine,are mainly used for the treatment of various diseases,including acute mastitis,caked breast and hernia symptoms,owing to its effects of relieving the depressed liver,soothing the liver,regulating the circulation of Qi,alleviating pain,dissipating phlegm and resolving masses. Modern pharmacological studies have shown that the hot water extracts from the leaves of C. reticulata have inhibitory effects on mastitis in mice. This paper aims to study their chemical constituents and reveal the effective substances. Method: The coarse powder of C. reticulata leaves was extracted by methods of soaking and refluxing with 95% and 70% ethanol,respectively. Then the ethanol extracts were separated preliminarily by MCI column chromatography( CC). The compounds were isolated and purified by using various chromatographic techniques,including silica gel,MCI,ODS,polyamide and Sephadex LH-20 CC.Their structures were identified on the basis of their physical-chemical properties and spectroscopic techniques,including NMR and MS. Result: Twelve compounds were obtained and identified as nobiletin( 1),5-demethylnobile( 2), tangeretin( 3), 5, 7, 8, 4'-tetramethoxyflavone( 4), 5-hydroxy-6, 7, 3',4'-tetramethoxyflavone( 5), β-amyrin( 6), lupeol( 7), phytol( 8), stigmastane-5,22-dien-3β-ol( 9),β-sitosterol( 10), p-hy-droxybenzaldehyde( 11) and vanillin( 12), respectively. Conclusion: Compounds4-12 were isolated from the leaves of Citrus reticulata for the first time.
引文
[1]中国科学院《中国植物志》编辑委员会.中国植物志.43卷.2分册[M].北京:科学出版社,1997:201-209.
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[5]范玉田,杨玉华.乳癖康胶囊的质量控制[J].中国医院药学杂志,2009,29(16):1407-1409.
[6]展晓日,史新元,张培,等.乳块消片高效液相指纹图谱研究[J].中国实验方剂学杂志,2008,14(4):1-3.
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[8]郝自新,汪玉萍,王和协,等.橘叶药材质量标准研究[J].安徽科技学院学报,2017,31(4):56-61.
[9]刘传夫,石俊英.橘叶药材质量评价研究[J].药学研究,2014,33(1):21-23.
[10]汪金玉,帅欧,林励,等.橘叶HPLC指纹图谱研究[J].中药新药与临床药理,2011,22(1):82-85.
[11]汪金玉,帅欧,林励,等.RP-HPLC法同时测定橘叶中3个黄酮类成分的含量[J].药物分析杂志,2011,31(11):2161-2163.
[12]吴今人,耿平,胡田叶,等.柑橘不同部位中柠檬苦素的分布及含量分析[J].湖北农业科学,2015,54(4):882-885.
[13]王桂红,胡俊杰,肖伊,等.橘叶挥发油化学成分的气相色谱-质谱分析[J].中国医院药学杂志,2011,31(4):272-274.
[14]CAO G H,FU Q R,ZHANG C M,et al.Rapid characterization of 96 chemical constituents in Citri Reticulatae Folium(leaves of‘Fuju’)using HPLCDAD-ESI-MS[J].J Chin Pharmaceut Sci,2016,25(2):91-110.
[15]汪金玉,帅欧,林励,等.橘叶黄酮类成分的研究与薄层色谱鉴别[J].广州中医药大学学报,2011,28(2):191-193.
[16]游元元,祝捷,李建春,等.红橘陈皮与橘叶对小鼠实验性乳腺炎的影响[J].时珍国医国药,2012,23(4):909-910.
[17]李林福,肖海,胡海波,等.九里香叶中的化学成分[J].中国实验方剂学杂志,2016,22(7):50-53.
[18]Hamdan D,Elreadi Z M,Tahrani A,et al.Chemical composition and biological activity of Citrus jambhiri Lush.[J].Food Chem,2011,127(2):394-403.
[19]WANG D D,WANG J,HUANG X H,et al.Identification of polymethoxylated flavones from green tangerine peel(Pericarpium Citri Reticulatae Viride)by chromatographic and spectroscopic techniques[J].J Pharmaceut Biomed Anal,2007,44(1):63-69.
[20]Tatum J H,Berry R E.Flavonoids of the Citrus cultivar calamondin and synthetic 2'β-dihydroxychalcones[J].Phytochemistry,1978,17(3):447-449.
[21]彭爱一,曲学伟,李慧,等.高速逆流色谱分离纯化九里香中的黄酮类化合物[J].色谱,2010,28(4):383-387.
[22]夏新中,周思祥,屠鹏飞.细梗胡枝子三萜类化学成分的研究[J].中国实验方剂学杂志,2010,16(6):62-64.
[23]Reynolds W F,Mclean S,Poplawski J,et al.Total assignment of13C and1H spectra of three isomeric triterpenol derivatives by 2D NMR:an investigation of the potential utility of1H chemical shifts in structural investigations of complex natural products[J].Tetrahedron,1986,42(13):3419-3428.
[24]Sims J J,Pettus J A.Isolation of free cis and trans-phytol from the red alga Gracilaria andersoniana[J].Phytochemistry,1976,15(6):1076-1077.
[25]Leit2o S G,Kaplan M A C,Monache F D,et al.Sterols and sterol glucosides from two Aegiphila species[J].Phytochemistry,1992,31(8):2813-2817.
[26]殷启蒙,张雨昇,季楠,等.见血清化学成分分析[J].中国实验方剂学杂志,2017,23(22):65-69.
[27]余邦良,刘寿柏,黄内利,等.眼树莲的化学成分分析[J].中国实验方剂学杂志,2017,23(16):96-100.
[2]国家中医药管理局《中华本草》编委会.中华本草.4册[M].上海:上海科学技术出版社,1999:885-900.
[3]宋立人,洪恂,丁绪亮,等.现代中药学大辞典.下册[M].北京:人民卫生出版社,2001:2404-2409.
[4]国家药典委员会.中华人民共和国药典.一部[M].北京:中国医药科技出版社,2015:191,378-379.
[5]范玉田,杨玉华.乳癖康胶囊的质量控制[J].中国医院药学杂志,2009,29(16):1407-1409.
[6]展晓日,史新元,张培,等.乳块消片高效液相指纹图谱研究[J].中国实验方剂学杂志,2008,14(4):1-3.
[7]肖培根.新编中药志[M].北京:化学工业出版社,2002:349.
[8]郝自新,汪玉萍,王和协,等.橘叶药材质量标准研究[J].安徽科技学院学报,2017,31(4):56-61.
[9]刘传夫,石俊英.橘叶药材质量评价研究[J].药学研究,2014,33(1):21-23.
[10]汪金玉,帅欧,林励,等.橘叶HPLC指纹图谱研究[J].中药新药与临床药理,2011,22(1):82-85.
[11]汪金玉,帅欧,林励,等.RP-HPLC法同时测定橘叶中3个黄酮类成分的含量[J].药物分析杂志,2011,31(11):2161-2163.
[12]吴今人,耿平,胡田叶,等.柑橘不同部位中柠檬苦素的分布及含量分析[J].湖北农业科学,2015,54(4):882-885.
[13]王桂红,胡俊杰,肖伊,等.橘叶挥发油化学成分的气相色谱-质谱分析[J].中国医院药学杂志,2011,31(4):272-274.
[14]CAO G H,FU Q R,ZHANG C M,et al.Rapid characterization of 96 chemical constituents in Citri Reticulatae Folium(leaves of‘Fuju’)using HPLCDAD-ESI-MS[J].J Chin Pharmaceut Sci,2016,25(2):91-110.
[15]汪金玉,帅欧,林励,等.橘叶黄酮类成分的研究与薄层色谱鉴别[J].广州中医药大学学报,2011,28(2):191-193.
[16]游元元,祝捷,李建春,等.红橘陈皮与橘叶对小鼠实验性乳腺炎的影响[J].时珍国医国药,2012,23(4):909-910.
[17]李林福,肖海,胡海波,等.九里香叶中的化学成分[J].中国实验方剂学杂志,2016,22(7):50-53.
[18]Hamdan D,Elreadi Z M,Tahrani A,et al.Chemical composition and biological activity of Citrus jambhiri Lush.[J].Food Chem,2011,127(2):394-403.
[19]WANG D D,WANG J,HUANG X H,et al.Identification of polymethoxylated flavones from green tangerine peel(Pericarpium Citri Reticulatae Viride)by chromatographic and spectroscopic techniques[J].J Pharmaceut Biomed Anal,2007,44(1):63-69.
[20]Tatum J H,Berry R E.Flavonoids of the Citrus cultivar calamondin and synthetic 2'β-dihydroxychalcones[J].Phytochemistry,1978,17(3):447-449.
[21]彭爱一,曲学伟,李慧,等.高速逆流色谱分离纯化九里香中的黄酮类化合物[J].色谱,2010,28(4):383-387.
[22]夏新中,周思祥,屠鹏飞.细梗胡枝子三萜类化学成分的研究[J].中国实验方剂学杂志,2010,16(6):62-64.
[23]Reynolds W F,Mclean S,Poplawski J,et al.Total assignment of13C and1H spectra of three isomeric triterpenol derivatives by 2D NMR:an investigation of the potential utility of1H chemical shifts in structural investigations of complex natural products[J].Tetrahedron,1986,42(13):3419-3428.
[24]Sims J J,Pettus J A.Isolation of free cis and trans-phytol from the red alga Gracilaria andersoniana[J].Phytochemistry,1976,15(6):1076-1077.
[25]Leit2o S G,Kaplan M A C,Monache F D,et al.Sterols and sterol glucosides from two Aegiphila species[J].Phytochemistry,1992,31(8):2813-2817.
[26]殷启蒙,张雨昇,季楠,等.见血清化学成分分析[J].中国实验方剂学杂志,2017,23(22):65-69.
[27]余邦良,刘寿柏,黄内利,等.眼树莲的化学成分分析[J].中国实验方剂学杂志,2017,23(16):96-100.